rabbit anti twik 1  (Alomone Labs)


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    Alomone Labs rabbit anti twik 1
    Generation of <t>TWIK-1</t> BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik 1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik 1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression"

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    Journal: Cells

    doi: 10.3390/cells10102751

    Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
    Figure Legend Snippet: Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.

    Techniques Used: Modification, Plasmid Preparation, Sequencing, Slice Preparation, Expressing, Immunofluorescence

    Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in . Data are presented as the Mean ± SEM.
    Figure Legend Snippet: Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in . Data are presented as the Mean ± SEM.

    Techniques Used: Expressing, Immunofluorescence

    High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in <xref ref-type=Supplementary Materials Table S1 . Data are presented as the Mean ± SEM. " title="High TWIK-1 expression in immature neurons of the DG in ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Techniques Used: Expressing, Immunofluorescence, Labeling

    Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.
    Figure Legend Snippet: Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.

    Techniques Used: Expressing, Immunofluorescence

    TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in <xref ref-type=Supplementary Materials Table S1 . **** p < 0.0001; ** p < 0.01, two-tailed t tests. Data are presented as the Mean ± SEM. " title="TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in Supplementary Materials Table S1 . **** p < 0.0001; ** p < 0.01, two-tailed t tests. Data are presented as the Mean ± SEM.

    Techniques Used: Expressing, Slice Preparation, Fluorescence, In Situ Hybridization, Immunofluorescence, Two Tailed Test

    rabbit anti twik 1  (Alomone Labs)


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    Alomone Labs rabbit anti twik 1
    Generation of <t>TWIK-1</t> BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik 1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik 1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression"

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    Journal: Cells

    doi: 10.3390/cells10102751

    Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
    Figure Legend Snippet: Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.

    Techniques Used: Modification, Plasmid Preparation, Sequencing, Slice Preparation, Expressing, Immunofluorescence

    Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in . Data are presented as the Mean ± SEM.
    Figure Legend Snippet: Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in . Data are presented as the Mean ± SEM.

    Techniques Used: Expressing, Immunofluorescence

    High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in <xref ref-type=Supplementary Materials Table S1 . Data are presented as the Mean ± SEM. " title="High TWIK-1 expression in immature neurons of the DG in ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Techniques Used: Expressing, Immunofluorescence, Labeling

    Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.
    Figure Legend Snippet: Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.

    Techniques Used: Expressing, Immunofluorescence

    TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in <xref ref-type=Supplementary Materials Table S1 . **** p < 0.0001; ** p < 0.01, two-tailed t tests. Data are presented as the Mean ± SEM. " title="TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in Supplementary Materials Table S1 . **** p < 0.0001; ** p < 0.01, two-tailed t tests. Data are presented as the Mean ± SEM.

    Techniques Used: Expressing, Slice Preparation, Fluorescence, In Situ Hybridization, Immunofluorescence, Two Tailed Test

    rabbit anti twik1  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
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    Alomone Labs rabbit anti twik1
    Rabbit Anti Twik1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik1 - by Bioz Stars, 2023-02
    93/100 stars

    Images

    rabbit anti twik 1  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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    Structured Review

    Alomone Labs rabbit anti twik 1
    Activation of mGluR3 translocates intracellular <t>TWIK-1</t> to astrocyte membrane. a Three panels representing three identified astrocytes in situ from three experimental groups as indicated. The cells in slices were subjected to GFAP (green)/TWIK-1 (red)/GLT-1 (magenta) triple immunostaining. LY354740: mGluR3 agonist (2 μM, 20 min). LY341495: mGluR3 antagonist (10 μM, 10 min). Scale bar, 10 μm. The somatic cytoplasm in all three cells is encircled by GFAP staining in the areas marked by squares. Topographic image illustrated the distribution and relative density of GFAP, TWIK-1, and GLT-1 positive signal before and after the agonist/antagonist application, obtained from soma area indicated in the column of merged image. A large amount of TWIK-1 punctate staining appeared in the cytoplasm in the control cell, but that was largely reduced after LY354740 treatment. The mGluR3 antagonist prevented agonist induced TWIK-1 translocation. b Fluorescence intensity profiles of TWIK-1 and GLT-1 in soma along the “a-b” lines indicated in the merged images in a, and only in the cell pretreated with mGlu3R agonist (middle panel), TWIK-1 fluorescence intensity shifted from cytoplasm to cell membrane. c The change in relative amount of membrane TWIK-1 is determined by the colocalization of TWIK-1 with astrocyte membrane marker GLT-1. Up panel, three subcellular regions along the primary processes (rectangles) were selected based on GFAP staining that extended from soma for TWIK-1/GLT-1 colocalization analysis. The somatic areas were also used (squares) for TWIK-1/GLT-1 colocalization analysis. Bottom panel, TWIK-1/GLT-1 colocalization in soma and process areas was analyzed by Pearson's correlation coefficient (PCC). To achieve accurate PCC value for each astrocyte, three to four selected process areas, shown in the upper panel, were used and the obtained PCC values were averaged. Data are presented as mean±SEM from indicated number of experiments. a.u. arbitrary unit, NS no significance; *P<0.05; **P<0.01 (color figure online)
    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik 1/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik 1 - by Bioz Stars, 2023-02
    86/100 stars

    Images

    1) Product Images from "mGluR3 Activation Recruits Cytoplasmic TWIK-1 Channels to Membrane that Enhances Ammonium Uptake in Hippocampal Astrocytes"

    Article Title: mGluR3 Activation Recruits Cytoplasmic TWIK-1 Channels to Membrane that Enhances Ammonium Uptake in Hippocampal Astrocytes

    Journal: Molecular neurobiology

    doi: 10.1007/s12035-015-9496-4

    Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane. a Three panels representing three identified astrocytes in situ from three experimental groups as indicated. The cells in slices were subjected to GFAP (green)/TWIK-1 (red)/GLT-1 (magenta) triple immunostaining. LY354740: mGluR3 agonist (2 μM, 20 min). LY341495: mGluR3 antagonist (10 μM, 10 min). Scale bar, 10 μm. The somatic cytoplasm in all three cells is encircled by GFAP staining in the areas marked by squares. Topographic image illustrated the distribution and relative density of GFAP, TWIK-1, and GLT-1 positive signal before and after the agonist/antagonist application, obtained from soma area indicated in the column of merged image. A large amount of TWIK-1 punctate staining appeared in the cytoplasm in the control cell, but that was largely reduced after LY354740 treatment. The mGluR3 antagonist prevented agonist induced TWIK-1 translocation. b Fluorescence intensity profiles of TWIK-1 and GLT-1 in soma along the “a-b” lines indicated in the merged images in a, and only in the cell pretreated with mGlu3R agonist (middle panel), TWIK-1 fluorescence intensity shifted from cytoplasm to cell membrane. c The change in relative amount of membrane TWIK-1 is determined by the colocalization of TWIK-1 with astrocyte membrane marker GLT-1. Up panel, three subcellular regions along the primary processes (rectangles) were selected based on GFAP staining that extended from soma for TWIK-1/GLT-1 colocalization analysis. The somatic areas were also used (squares) for TWIK-1/GLT-1 colocalization analysis. Bottom panel, TWIK-1/GLT-1 colocalization in soma and process areas was analyzed by Pearson's correlation coefficient (PCC). To achieve accurate PCC value for each astrocyte, three to four selected process areas, shown in the upper panel, were used and the obtained PCC values were averaged. Data are presented as mean±SEM from indicated number of experiments. a.u. arbitrary unit, NS no significance; *P<0.05; **P<0.01 (color figure online)
    Figure Legend Snippet: Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane. a Three panels representing three identified astrocytes in situ from three experimental groups as indicated. The cells in slices were subjected to GFAP (green)/TWIK-1 (red)/GLT-1 (magenta) triple immunostaining. LY354740: mGluR3 agonist (2 μM, 20 min). LY341495: mGluR3 antagonist (10 μM, 10 min). Scale bar, 10 μm. The somatic cytoplasm in all three cells is encircled by GFAP staining in the areas marked by squares. Topographic image illustrated the distribution and relative density of GFAP, TWIK-1, and GLT-1 positive signal before and after the agonist/antagonist application, obtained from soma area indicated in the column of merged image. A large amount of TWIK-1 punctate staining appeared in the cytoplasm in the control cell, but that was largely reduced after LY354740 treatment. The mGluR3 antagonist prevented agonist induced TWIK-1 translocation. b Fluorescence intensity profiles of TWIK-1 and GLT-1 in soma along the “a-b” lines indicated in the merged images in a, and only in the cell pretreated with mGlu3R agonist (middle panel), TWIK-1 fluorescence intensity shifted from cytoplasm to cell membrane. c The change in relative amount of membrane TWIK-1 is determined by the colocalization of TWIK-1 with astrocyte membrane marker GLT-1. Up panel, three subcellular regions along the primary processes (rectangles) were selected based on GFAP staining that extended from soma for TWIK-1/GLT-1 colocalization analysis. The somatic areas were also used (squares) for TWIK-1/GLT-1 colocalization analysis. Bottom panel, TWIK-1/GLT-1 colocalization in soma and process areas was analyzed by Pearson's correlation coefficient (PCC). To achieve accurate PCC value for each astrocyte, three to four selected process areas, shown in the upper panel, were used and the obtained PCC values were averaged. Data are presented as mean±SEM from indicated number of experiments. a.u. arbitrary unit, NS no significance; *P<0.05; **P<0.01 (color figure online)

    Techniques Used: Activation Assay, In Situ, Triple Immunostaining, Staining, Translocation Assay, Fluorescence, Marker

    Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane in hippocampal slice. a Western blots show the relative TWIK-1 distribution in cytoplasm (Cyto) versus transmembrane (Mem) from proteins isolated from control hippocampal slices and slices preincubated with mGluR3 agonist LY354740 (2 μM for 20 min). TWIK-1 proteins existed in both monomer and dimer forms in the plasma membrane and cytoplasmic fraction. GFAP (50 kDa) and ATP1α2 (112 kDa) are markers for cytoplasmic and membrane fractions, respectively. b Quantitative densitometric analysis of the relative Mem versus total TWIK-1 proteins, i.e., Cyto + Mem. mGluR3 agonist increases the membrane presence of TWIK-1 significantly compared to the control group. Data are presented as mean±SEM from four independent western blot experiments. *P<0.05
    Figure Legend Snippet: Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane in hippocampal slice. a Western blots show the relative TWIK-1 distribution in cytoplasm (Cyto) versus transmembrane (Mem) from proteins isolated from control hippocampal slices and slices preincubated with mGluR3 agonist LY354740 (2 μM for 20 min). TWIK-1 proteins existed in both monomer and dimer forms in the plasma membrane and cytoplasmic fraction. GFAP (50 kDa) and ATP1α2 (112 kDa) are markers for cytoplasmic and membrane fractions, respectively. b Quantitative densitometric analysis of the relative Mem versus total TWIK-1 proteins, i.e., Cyto + Mem. mGluR3 agonist increases the membrane presence of TWIK-1 significantly compared to the control group. Data are presented as mean±SEM from four independent western blot experiments. *P<0.05

    Techniques Used: Activation Assay, Western Blot, Isolation

    Elevated membrane TWIK-1 expression is associated with astrocyte VM depolarization. a Whole-cell VM recording from WT and TWIK-1−/− astrocytes. The black trace shows a stable VM recording with negligible fluctuation over a 40-min duration. In the absence of 0.5 μM TTX, LY354740 induced an initial hyperpolarization and a following secondary depolarization in WT, but the second phase depolarization was significantly smaller in TWIK-1−/− astrocyte (upper panel). The initial hyperpolarization could be totally blocked by TTX treatment applied 5–10 min before a 20-min LY354740 (2 μM) incubation (lower panel). The amplitude of mGluR3-induced depolarization (ΔV) in the end of 20-min agonist treatment is summarized in b. Data are presented as mean±SE from indicated number of recordings. *P<0.05; **P<0.01
    Figure Legend Snippet: Elevated membrane TWIK-1 expression is associated with astrocyte VM depolarization. a Whole-cell VM recording from WT and TWIK-1−/− astrocytes. The black trace shows a stable VM recording with negligible fluctuation over a 40-min duration. In the absence of 0.5 μM TTX, LY354740 induced an initial hyperpolarization and a following secondary depolarization in WT, but the second phase depolarization was significantly smaller in TWIK-1−/− astrocyte (upper panel). The initial hyperpolarization could be totally blocked by TTX treatment applied 5–10 min before a 20-min LY354740 (2 μM) incubation (lower panel). The amplitude of mGluR3-induced depolarization (ΔV) in the end of 20-min agonist treatment is summarized in b. Data are presented as mean±SE from indicated number of recordings. *P<0.05; **P<0.01

    Techniques Used: Expressing, Incubation

    Enhanced potentiation of mGluR3-mediated NH4+ uptake in the absence of Na+-K+ ATPase activity. a To eliminate Na+-K+ ATPase (NKA) activity, NaCl in the electrode solution (14 mM) and aCSF (125 mM) was substituted by equimolar LiCl, and the perfusate was switched from normal to high LiCl-containing aCSF (Li-aCSF) after whole-cell recording was established. These conditions depolarized VM by ~10 mV resulting from inhibition of electrogenic NKA activity. NH4Cl (5 mM, 10 min) was applied 9~10 min after NKA inhibition. Right panel, the NH4Cl response outlined in the dashed rectangle in left panel is displayed in an enlarged scale (green) and superimposed with a recording with intact NKA activity (black). The differences in NH4Cl response under these two conditions are summarized in b; elimination of NKA activity resulted in a (1) progressive increase in NH4+-induced VM depolarization (left, quantified as “time to peak”), (2) an average of 22.0 % increase in peak amplitude (middle), and (3) a total abolishment of VM undershot upon withdrawal of NH4Cl (right). c Representative recording showing that mGluR3 activation induced potentiation was more pronounced in the absence of NKA activity. d mGluR3-induced potentiation of NH4+ response remained when NKA is inactive in WT but not in TWIK-1 KO astrocytes. Data are presented as mean±SEM from the indicated number of experiments. **P<0.01; NS no significance
    Figure Legend Snippet: Enhanced potentiation of mGluR3-mediated NH4+ uptake in the absence of Na+-K+ ATPase activity. a To eliminate Na+-K+ ATPase (NKA) activity, NaCl in the electrode solution (14 mM) and aCSF (125 mM) was substituted by equimolar LiCl, and the perfusate was switched from normal to high LiCl-containing aCSF (Li-aCSF) after whole-cell recording was established. These conditions depolarized VM by ~10 mV resulting from inhibition of electrogenic NKA activity. NH4Cl (5 mM, 10 min) was applied 9~10 min after NKA inhibition. Right panel, the NH4Cl response outlined in the dashed rectangle in left panel is displayed in an enlarged scale (green) and superimposed with a recording with intact NKA activity (black). The differences in NH4Cl response under these two conditions are summarized in b; elimination of NKA activity resulted in a (1) progressive increase in NH4+-induced VM depolarization (left, quantified as “time to peak”), (2) an average of 22.0 % increase in peak amplitude (middle), and (3) a total abolishment of VM undershot upon withdrawal of NH4Cl (right). c Representative recording showing that mGluR3 activation induced potentiation was more pronounced in the absence of NKA activity. d mGluR3-induced potentiation of NH4+ response remained when NKA is inactive in WT but not in TWIK-1 KO astrocytes. Data are presented as mean±SEM from the indicated number of experiments. **P<0.01; NS no significance

    Techniques Used: Activity Assay, Inhibition, Activation Assay

    mGluR3-mediated potentiation of NH4+ uptake depends on astrocyte TWIK-1 channels. a NH4+-induced VM depolarization was comparable in amplitudes between WT and TWIK-1 KO astrocytes with or without BaCl2 application. Five millimolars of NH4Cl was applied for 10 min and 100 μM BaCl2 was added in bath 5 min before and during NH4Cl treatment. b The peak amplitudes of NH4+-induced VM depolarization were summarized between the two genotypes. Ba2+ significantly reduced VM depolarization by 20 % for WT and 27 % for TWIK-1 KO astrocytes. c mGluR3-mediated potentiation of NH4+-induced VM response reduced in a TWIK-1 gene dose-dependent manner. mGluR3 agonist LY354740 (2 μM) was used in these recordings, and a similar effect could be replicated by another mGluR3 agonist APDC (10 μM) shown in d; in both cases, agonists were added to bath 10 min prior to and during NH4+ application. Note that mGluR3-mediated potentiation was nearly absent in TWIK-1 KO astrocytes. e Representative western blot shows an age-dependent upregulation in TWIK-1 protein expression. The total hippocampal proteins were isolated from animals of different postnatal ages as indicated. GAPDH (36 kDa) and GFAP (50 kDa) were used as loading control. TWIK-1 protein is almost absent in P1 and expressed at very low level at P7 compared to those after P23. Kir4.1 is also expressed in an age-dependent manner, and the expression levels become evident from P13 on. f mGluR3-mediated potentiation of NH4+ uptake was absent in immature astrocytes (P7), which is summarized in g. h qRT-PCR results from freshly dissociated astrocytes show a comparable expression level of mGluR3 mRNA in P7 immature astrocytes to that of P21. The mRNAs were obtained and averaged from three independent cell harvests from three mice; each harvest contained 30 astrocytes. Data are presented as mean±SEM from the indicated number of experiments. *P<0.05; **P<0.01; NS no significance
    Figure Legend Snippet: mGluR3-mediated potentiation of NH4+ uptake depends on astrocyte TWIK-1 channels. a NH4+-induced VM depolarization was comparable in amplitudes between WT and TWIK-1 KO astrocytes with or without BaCl2 application. Five millimolars of NH4Cl was applied for 10 min and 100 μM BaCl2 was added in bath 5 min before and during NH4Cl treatment. b The peak amplitudes of NH4+-induced VM depolarization were summarized between the two genotypes. Ba2+ significantly reduced VM depolarization by 20 % for WT and 27 % for TWIK-1 KO astrocytes. c mGluR3-mediated potentiation of NH4+-induced VM response reduced in a TWIK-1 gene dose-dependent manner. mGluR3 agonist LY354740 (2 μM) was used in these recordings, and a similar effect could be replicated by another mGluR3 agonist APDC (10 μM) shown in d; in both cases, agonists were added to bath 10 min prior to and during NH4+ application. Note that mGluR3-mediated potentiation was nearly absent in TWIK-1 KO astrocytes. e Representative western blot shows an age-dependent upregulation in TWIK-1 protein expression. The total hippocampal proteins were isolated from animals of different postnatal ages as indicated. GAPDH (36 kDa) and GFAP (50 kDa) were used as loading control. TWIK-1 protein is almost absent in P1 and expressed at very low level at P7 compared to those after P23. Kir4.1 is also expressed in an age-dependent manner, and the expression levels become evident from P13 on. f mGluR3-mediated potentiation of NH4+ uptake was absent in immature astrocytes (P7), which is summarized in g. h qRT-PCR results from freshly dissociated astrocytes show a comparable expression level of mGluR3 mRNA in P7 immature astrocytes to that of P21. The mRNAs were obtained and averaged from three independent cell harvests from three mice; each harvest contained 30 astrocytes. Data are presented as mean±SEM from the indicated number of experiments. *P<0.05; **P<0.01; NS no significance

    Techniques Used: Western Blot, Expressing, Isolation, Quantitative RT-PCR

    Exocytotic pathways underlying mGluR3-induced TWIK-1 membrane surface expression. a Inhibition of SNARE complex by N-ethylmaleimide (NEM, 250 μM) eliminated mGluR3-mediated potentiation of NH4+ response. NEM was applied intracellularly for 10 min prior to mGluR3 activation. The schematic illustrations of trafficking pathway and steps are shown in the left panel and recording traces are shown in the right panel. b Inhibition of RabGGTase by 5 μM psoromic acid (PA) abolished mGluR3-mediated potentiation of NH4+ uptake. RabGGTase is the enzyme that prenylates newly synthesized Rabs and delivers them to the membranes of donor compartments. Brian slices were incubated in 5 μM PA for 4 h before patch-clamp recording. c Comparison of the resting membrane potential of astrocyte under various conditions. Note that electrode dialysis of NEM and pretreatment of astrocytes with PA all hyperpolarized the VM to levels comparable to that of the TWIK-1−/− astrocytes. d Summary of the effect of NEM and PA on mGluR3-mediated potentiation of NH4+. Data are presented as mean±SEM from the indicated number of experiments. e Immunostaining showed that TWIK-1 and Rab11 were distributed coordinately in astrocyte soma in hippocampal slices. Both TWIK-1 and Rab11 signals were distributed evenly in astrocyte soma in resting state (control) and shifted to a polarized distribution toward to membrane after mGluR3 agonist treatment (LY354740). Scale bar, 5 μm. *P<0.05; **P<0.01; NS no significance. PM plasma membrane, NSF N-ethylmaleimide sensitive factor, EE early endosome, RE recycling endosome, LE late endosome, REP Rab escort protein, RabGGTase Rab geranylgeranyl transferase
    Figure Legend Snippet: Exocytotic pathways underlying mGluR3-induced TWIK-1 membrane surface expression. a Inhibition of SNARE complex by N-ethylmaleimide (NEM, 250 μM) eliminated mGluR3-mediated potentiation of NH4+ response. NEM was applied intracellularly for 10 min prior to mGluR3 activation. The schematic illustrations of trafficking pathway and steps are shown in the left panel and recording traces are shown in the right panel. b Inhibition of RabGGTase by 5 μM psoromic acid (PA) abolished mGluR3-mediated potentiation of NH4+ uptake. RabGGTase is the enzyme that prenylates newly synthesized Rabs and delivers them to the membranes of donor compartments. Brian slices were incubated in 5 μM PA for 4 h before patch-clamp recording. c Comparison of the resting membrane potential of astrocyte under various conditions. Note that electrode dialysis of NEM and pretreatment of astrocytes with PA all hyperpolarized the VM to levels comparable to that of the TWIK-1−/− astrocytes. d Summary of the effect of NEM and PA on mGluR3-mediated potentiation of NH4+. Data are presented as mean±SEM from the indicated number of experiments. e Immunostaining showed that TWIK-1 and Rab11 were distributed coordinately in astrocyte soma in hippocampal slices. Both TWIK-1 and Rab11 signals were distributed evenly in astrocyte soma in resting state (control) and shifted to a polarized distribution toward to membrane after mGluR3 agonist treatment (LY354740). Scale bar, 5 μm. *P<0.05; **P<0.01; NS no significance. PM plasma membrane, NSF N-ethylmaleimide sensitive factor, EE early endosome, RE recycling endosome, LE late endosome, REP Rab escort protein, RabGGTase Rab geranylgeranyl transferase

    Techniques Used: Expressing, Inhibition, Activation Assay, Synthesized, Incubation, Patch Clamp, Immunostaining

    rabbit anti twik 1  (Alomone Labs)


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    Alomone Labs rabbit anti twik 1
    Activation of mGluR3 translocates intracellular <t>TWIK-1</t> to astrocyte membrane. a Three panels representing three identified astrocytes in situ from three experimental groups as indicated. The cells in slices were subjected to GFAP (green)/TWIK-1 (red)/GLT-1 (magenta) triple immunostaining. LY354740: mGluR3 agonist (2 μM, 20 min). LY341495: mGluR3 antagonist (10 μM, 10 min). Scale bar, 10 μm. The somatic cytoplasm in all three cells is encircled by GFAP staining in the areas marked by squares. Topographic image illustrated the distribution and relative density of GFAP, TWIK-1, and GLT-1 positive signal before and after the agonist/antagonist application, obtained from soma area indicated in the column of merged image. A large amount of TWIK-1 punctate staining appeared in the cytoplasm in the control cell, but that was largely reduced after LY354740 treatment. The mGluR3 antagonist prevented agonist induced TWIK-1 translocation. b Fluorescence intensity profiles of TWIK-1 and GLT-1 in soma along the “a-b” lines indicated in the merged images in a, and only in the cell pretreated with mGlu3R agonist (middle panel), TWIK-1 fluorescence intensity shifted from cytoplasm to cell membrane. c The change in relative amount of membrane TWIK-1 is determined by the colocalization of TWIK-1 with astrocyte membrane marker GLT-1. Up panel, three subcellular regions along the primary processes (rectangles) were selected based on GFAP staining that extended from soma for TWIK-1/GLT-1 colocalization analysis. The somatic areas were also used (squares) for TWIK-1/GLT-1 colocalization analysis. Bottom panel, TWIK-1/GLT-1 colocalization in soma and process areas was analyzed by Pearson's correlation coefficient (PCC). To achieve accurate PCC value for each astrocyte, three to four selected process areas, shown in the upper panel, were used and the obtained PCC values were averaged. Data are presented as mean±SEM from indicated number of experiments. a.u. arbitrary unit, NS no significance; *P<0.05; **P<0.01 (color figure online)
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    1) Product Images from "mGluR3 Activation Recruits Cytoplasmic TWIK-1 Channels to Membrane that Enhances Ammonium Uptake in Hippocampal Astrocytes"

    Article Title: mGluR3 Activation Recruits Cytoplasmic TWIK-1 Channels to Membrane that Enhances Ammonium Uptake in Hippocampal Astrocytes

    Journal: Molecular neurobiology

    doi: 10.1007/s12035-015-9496-4

    Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane. a Three panels representing three identified astrocytes in situ from three experimental groups as indicated. The cells in slices were subjected to GFAP (green)/TWIK-1 (red)/GLT-1 (magenta) triple immunostaining. LY354740: mGluR3 agonist (2 μM, 20 min). LY341495: mGluR3 antagonist (10 μM, 10 min). Scale bar, 10 μm. The somatic cytoplasm in all three cells is encircled by GFAP staining in the areas marked by squares. Topographic image illustrated the distribution and relative density of GFAP, TWIK-1, and GLT-1 positive signal before and after the agonist/antagonist application, obtained from soma area indicated in the column of merged image. A large amount of TWIK-1 punctate staining appeared in the cytoplasm in the control cell, but that was largely reduced after LY354740 treatment. The mGluR3 antagonist prevented agonist induced TWIK-1 translocation. b Fluorescence intensity profiles of TWIK-1 and GLT-1 in soma along the “a-b” lines indicated in the merged images in a, and only in the cell pretreated with mGlu3R agonist (middle panel), TWIK-1 fluorescence intensity shifted from cytoplasm to cell membrane. c The change in relative amount of membrane TWIK-1 is determined by the colocalization of TWIK-1 with astrocyte membrane marker GLT-1. Up panel, three subcellular regions along the primary processes (rectangles) were selected based on GFAP staining that extended from soma for TWIK-1/GLT-1 colocalization analysis. The somatic areas were also used (squares) for TWIK-1/GLT-1 colocalization analysis. Bottom panel, TWIK-1/GLT-1 colocalization in soma and process areas was analyzed by Pearson's correlation coefficient (PCC). To achieve accurate PCC value for each astrocyte, three to four selected process areas, shown in the upper panel, were used and the obtained PCC values were averaged. Data are presented as mean±SEM from indicated number of experiments. a.u. arbitrary unit, NS no significance; *P<0.05; **P<0.01 (color figure online)
    Figure Legend Snippet: Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane. a Three panels representing three identified astrocytes in situ from three experimental groups as indicated. The cells in slices were subjected to GFAP (green)/TWIK-1 (red)/GLT-1 (magenta) triple immunostaining. LY354740: mGluR3 agonist (2 μM, 20 min). LY341495: mGluR3 antagonist (10 μM, 10 min). Scale bar, 10 μm. The somatic cytoplasm in all three cells is encircled by GFAP staining in the areas marked by squares. Topographic image illustrated the distribution and relative density of GFAP, TWIK-1, and GLT-1 positive signal before and after the agonist/antagonist application, obtained from soma area indicated in the column of merged image. A large amount of TWIK-1 punctate staining appeared in the cytoplasm in the control cell, but that was largely reduced after LY354740 treatment. The mGluR3 antagonist prevented agonist induced TWIK-1 translocation. b Fluorescence intensity profiles of TWIK-1 and GLT-1 in soma along the “a-b” lines indicated in the merged images in a, and only in the cell pretreated with mGlu3R agonist (middle panel), TWIK-1 fluorescence intensity shifted from cytoplasm to cell membrane. c The change in relative amount of membrane TWIK-1 is determined by the colocalization of TWIK-1 with astrocyte membrane marker GLT-1. Up panel, three subcellular regions along the primary processes (rectangles) were selected based on GFAP staining that extended from soma for TWIK-1/GLT-1 colocalization analysis. The somatic areas were also used (squares) for TWIK-1/GLT-1 colocalization analysis. Bottom panel, TWIK-1/GLT-1 colocalization in soma and process areas was analyzed by Pearson's correlation coefficient (PCC). To achieve accurate PCC value for each astrocyte, three to four selected process areas, shown in the upper panel, were used and the obtained PCC values were averaged. Data are presented as mean±SEM from indicated number of experiments. a.u. arbitrary unit, NS no significance; *P<0.05; **P<0.01 (color figure online)

    Techniques Used: Activation Assay, In Situ, Triple Immunostaining, Staining, Translocation Assay, Fluorescence, Marker

    Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane in hippocampal slice. a Western blots show the relative TWIK-1 distribution in cytoplasm (Cyto) versus transmembrane (Mem) from proteins isolated from control hippocampal slices and slices preincubated with mGluR3 agonist LY354740 (2 μM for 20 min). TWIK-1 proteins existed in both monomer and dimer forms in the plasma membrane and cytoplasmic fraction. GFAP (50 kDa) and ATP1α2 (112 kDa) are markers for cytoplasmic and membrane fractions, respectively. b Quantitative densitometric analysis of the relative Mem versus total TWIK-1 proteins, i.e., Cyto + Mem. mGluR3 agonist increases the membrane presence of TWIK-1 significantly compared to the control group. Data are presented as mean±SEM from four independent western blot experiments. *P<0.05
    Figure Legend Snippet: Activation of mGluR3 translocates intracellular TWIK-1 to astrocyte membrane in hippocampal slice. a Western blots show the relative TWIK-1 distribution in cytoplasm (Cyto) versus transmembrane (Mem) from proteins isolated from control hippocampal slices and slices preincubated with mGluR3 agonist LY354740 (2 μM for 20 min). TWIK-1 proteins existed in both monomer and dimer forms in the plasma membrane and cytoplasmic fraction. GFAP (50 kDa) and ATP1α2 (112 kDa) are markers for cytoplasmic and membrane fractions, respectively. b Quantitative densitometric analysis of the relative Mem versus total TWIK-1 proteins, i.e., Cyto + Mem. mGluR3 agonist increases the membrane presence of TWIK-1 significantly compared to the control group. Data are presented as mean±SEM from four independent western blot experiments. *P<0.05

    Techniques Used: Activation Assay, Western Blot, Isolation

    Elevated membrane TWIK-1 expression is associated with astrocyte VM depolarization. a Whole-cell VM recording from WT and TWIK-1−/− astrocytes. The black trace shows a stable VM recording with negligible fluctuation over a 40-min duration. In the absence of 0.5 μM TTX, LY354740 induced an initial hyperpolarization and a following secondary depolarization in WT, but the second phase depolarization was significantly smaller in TWIK-1−/− astrocyte (upper panel). The initial hyperpolarization could be totally blocked by TTX treatment applied 5–10 min before a 20-min LY354740 (2 μM) incubation (lower panel). The amplitude of mGluR3-induced depolarization (ΔV) in the end of 20-min agonist treatment is summarized in b. Data are presented as mean±SE from indicated number of recordings. *P<0.05; **P<0.01
    Figure Legend Snippet: Elevated membrane TWIK-1 expression is associated with astrocyte VM depolarization. a Whole-cell VM recording from WT and TWIK-1−/− astrocytes. The black trace shows a stable VM recording with negligible fluctuation over a 40-min duration. In the absence of 0.5 μM TTX, LY354740 induced an initial hyperpolarization and a following secondary depolarization in WT, but the second phase depolarization was significantly smaller in TWIK-1−/− astrocyte (upper panel). The initial hyperpolarization could be totally blocked by TTX treatment applied 5–10 min before a 20-min LY354740 (2 μM) incubation (lower panel). The amplitude of mGluR3-induced depolarization (ΔV) in the end of 20-min agonist treatment is summarized in b. Data are presented as mean±SE from indicated number of recordings. *P<0.05; **P<0.01

    Techniques Used: Expressing, Incubation

    Enhanced potentiation of mGluR3-mediated NH4+ uptake in the absence of Na+-K+ ATPase activity. a To eliminate Na+-K+ ATPase (NKA) activity, NaCl in the electrode solution (14 mM) and aCSF (125 mM) was substituted by equimolar LiCl, and the perfusate was switched from normal to high LiCl-containing aCSF (Li-aCSF) after whole-cell recording was established. These conditions depolarized VM by ~10 mV resulting from inhibition of electrogenic NKA activity. NH4Cl (5 mM, 10 min) was applied 9~10 min after NKA inhibition. Right panel, the NH4Cl response outlined in the dashed rectangle in left panel is displayed in an enlarged scale (green) and superimposed with a recording with intact NKA activity (black). The differences in NH4Cl response under these two conditions are summarized in b; elimination of NKA activity resulted in a (1) progressive increase in NH4+-induced VM depolarization (left, quantified as “time to peak”), (2) an average of 22.0 % increase in peak amplitude (middle), and (3) a total abolishment of VM undershot upon withdrawal of NH4Cl (right). c Representative recording showing that mGluR3 activation induced potentiation was more pronounced in the absence of NKA activity. d mGluR3-induced potentiation of NH4+ response remained when NKA is inactive in WT but not in TWIK-1 KO astrocytes. Data are presented as mean±SEM from the indicated number of experiments. **P<0.01; NS no significance
    Figure Legend Snippet: Enhanced potentiation of mGluR3-mediated NH4+ uptake in the absence of Na+-K+ ATPase activity. a To eliminate Na+-K+ ATPase (NKA) activity, NaCl in the electrode solution (14 mM) and aCSF (125 mM) was substituted by equimolar LiCl, and the perfusate was switched from normal to high LiCl-containing aCSF (Li-aCSF) after whole-cell recording was established. These conditions depolarized VM by ~10 mV resulting from inhibition of electrogenic NKA activity. NH4Cl (5 mM, 10 min) was applied 9~10 min after NKA inhibition. Right panel, the NH4Cl response outlined in the dashed rectangle in left panel is displayed in an enlarged scale (green) and superimposed with a recording with intact NKA activity (black). The differences in NH4Cl response under these two conditions are summarized in b; elimination of NKA activity resulted in a (1) progressive increase in NH4+-induced VM depolarization (left, quantified as “time to peak”), (2) an average of 22.0 % increase in peak amplitude (middle), and (3) a total abolishment of VM undershot upon withdrawal of NH4Cl (right). c Representative recording showing that mGluR3 activation induced potentiation was more pronounced in the absence of NKA activity. d mGluR3-induced potentiation of NH4+ response remained when NKA is inactive in WT but not in TWIK-1 KO astrocytes. Data are presented as mean±SEM from the indicated number of experiments. **P<0.01; NS no significance

    Techniques Used: Activity Assay, Inhibition, Activation Assay

    mGluR3-mediated potentiation of NH4+ uptake depends on astrocyte TWIK-1 channels. a NH4+-induced VM depolarization was comparable in amplitudes between WT and TWIK-1 KO astrocytes with or without BaCl2 application. Five millimolars of NH4Cl was applied for 10 min and 100 μM BaCl2 was added in bath 5 min before and during NH4Cl treatment. b The peak amplitudes of NH4+-induced VM depolarization were summarized between the two genotypes. Ba2+ significantly reduced VM depolarization by 20 % for WT and 27 % for TWIK-1 KO astrocytes. c mGluR3-mediated potentiation of NH4+-induced VM response reduced in a TWIK-1 gene dose-dependent manner. mGluR3 agonist LY354740 (2 μM) was used in these recordings, and a similar effect could be replicated by another mGluR3 agonist APDC (10 μM) shown in d; in both cases, agonists were added to bath 10 min prior to and during NH4+ application. Note that mGluR3-mediated potentiation was nearly absent in TWIK-1 KO astrocytes. e Representative western blot shows an age-dependent upregulation in TWIK-1 protein expression. The total hippocampal proteins were isolated from animals of different postnatal ages as indicated. GAPDH (36 kDa) and GFAP (50 kDa) were used as loading control. TWIK-1 protein is almost absent in P1 and expressed at very low level at P7 compared to those after P23. Kir4.1 is also expressed in an age-dependent manner, and the expression levels become evident from P13 on. f mGluR3-mediated potentiation of NH4+ uptake was absent in immature astrocytes (P7), which is summarized in g. h qRT-PCR results from freshly dissociated astrocytes show a comparable expression level of mGluR3 mRNA in P7 immature astrocytes to that of P21. The mRNAs were obtained and averaged from three independent cell harvests from three mice; each harvest contained 30 astrocytes. Data are presented as mean±SEM from the indicated number of experiments. *P<0.05; **P<0.01; NS no significance
    Figure Legend Snippet: mGluR3-mediated potentiation of NH4+ uptake depends on astrocyte TWIK-1 channels. a NH4+-induced VM depolarization was comparable in amplitudes between WT and TWIK-1 KO astrocytes with or without BaCl2 application. Five millimolars of NH4Cl was applied for 10 min and 100 μM BaCl2 was added in bath 5 min before and during NH4Cl treatment. b The peak amplitudes of NH4+-induced VM depolarization were summarized between the two genotypes. Ba2+ significantly reduced VM depolarization by 20 % for WT and 27 % for TWIK-1 KO astrocytes. c mGluR3-mediated potentiation of NH4+-induced VM response reduced in a TWIK-1 gene dose-dependent manner. mGluR3 agonist LY354740 (2 μM) was used in these recordings, and a similar effect could be replicated by another mGluR3 agonist APDC (10 μM) shown in d; in both cases, agonists were added to bath 10 min prior to and during NH4+ application. Note that mGluR3-mediated potentiation was nearly absent in TWIK-1 KO astrocytes. e Representative western blot shows an age-dependent upregulation in TWIK-1 protein expression. The total hippocampal proteins were isolated from animals of different postnatal ages as indicated. GAPDH (36 kDa) and GFAP (50 kDa) were used as loading control. TWIK-1 protein is almost absent in P1 and expressed at very low level at P7 compared to those after P23. Kir4.1 is also expressed in an age-dependent manner, and the expression levels become evident from P13 on. f mGluR3-mediated potentiation of NH4+ uptake was absent in immature astrocytes (P7), which is summarized in g. h qRT-PCR results from freshly dissociated astrocytes show a comparable expression level of mGluR3 mRNA in P7 immature astrocytes to that of P21. The mRNAs were obtained and averaged from three independent cell harvests from three mice; each harvest contained 30 astrocytes. Data are presented as mean±SEM from the indicated number of experiments. *P<0.05; **P<0.01; NS no significance

    Techniques Used: Western Blot, Expressing, Isolation, Quantitative RT-PCR

    Exocytotic pathways underlying mGluR3-induced TWIK-1 membrane surface expression. a Inhibition of SNARE complex by N-ethylmaleimide (NEM, 250 μM) eliminated mGluR3-mediated potentiation of NH4+ response. NEM was applied intracellularly for 10 min prior to mGluR3 activation. The schematic illustrations of trafficking pathway and steps are shown in the left panel and recording traces are shown in the right panel. b Inhibition of RabGGTase by 5 μM psoromic acid (PA) abolished mGluR3-mediated potentiation of NH4+ uptake. RabGGTase is the enzyme that prenylates newly synthesized Rabs and delivers them to the membranes of donor compartments. Brian slices were incubated in 5 μM PA for 4 h before patch-clamp recording. c Comparison of the resting membrane potential of astrocyte under various conditions. Note that electrode dialysis of NEM and pretreatment of astrocytes with PA all hyperpolarized the VM to levels comparable to that of the TWIK-1−/− astrocytes. d Summary of the effect of NEM and PA on mGluR3-mediated potentiation of NH4+. Data are presented as mean±SEM from the indicated number of experiments. e Immunostaining showed that TWIK-1 and Rab11 were distributed coordinately in astrocyte soma in hippocampal slices. Both TWIK-1 and Rab11 signals were distributed evenly in astrocyte soma in resting state (control) and shifted to a polarized distribution toward to membrane after mGluR3 agonist treatment (LY354740). Scale bar, 5 μm. *P<0.05; **P<0.01; NS no significance. PM plasma membrane, NSF N-ethylmaleimide sensitive factor, EE early endosome, RE recycling endosome, LE late endosome, REP Rab escort protein, RabGGTase Rab geranylgeranyl transferase
    Figure Legend Snippet: Exocytotic pathways underlying mGluR3-induced TWIK-1 membrane surface expression. a Inhibition of SNARE complex by N-ethylmaleimide (NEM, 250 μM) eliminated mGluR3-mediated potentiation of NH4+ response. NEM was applied intracellularly for 10 min prior to mGluR3 activation. The schematic illustrations of trafficking pathway and steps are shown in the left panel and recording traces are shown in the right panel. b Inhibition of RabGGTase by 5 μM psoromic acid (PA) abolished mGluR3-mediated potentiation of NH4+ uptake. RabGGTase is the enzyme that prenylates newly synthesized Rabs and delivers them to the membranes of donor compartments. Brian slices were incubated in 5 μM PA for 4 h before patch-clamp recording. c Comparison of the resting membrane potential of astrocyte under various conditions. Note that electrode dialysis of NEM and pretreatment of astrocytes with PA all hyperpolarized the VM to levels comparable to that of the TWIK-1−/− astrocytes. d Summary of the effect of NEM and PA on mGluR3-mediated potentiation of NH4+. Data are presented as mean±SEM from the indicated number of experiments. e Immunostaining showed that TWIK-1 and Rab11 were distributed coordinately in astrocyte soma in hippocampal slices. Both TWIK-1 and Rab11 signals were distributed evenly in astrocyte soma in resting state (control) and shifted to a polarized distribution toward to membrane after mGluR3 agonist treatment (LY354740). Scale bar, 5 μm. *P<0.05; **P<0.01; NS no significance. PM plasma membrane, NSF N-ethylmaleimide sensitive factor, EE early endosome, RE recycling endosome, LE late endosome, REP Rab escort protein, RabGGTase Rab geranylgeranyl transferase

    Techniques Used: Expressing, Inhibition, Activation Assay, Synthesized, Incubation, Patch Clamp, Immunostaining

    rabbit anti twik 1 polyclonal antibody  (Alomone Labs)


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    Alomone Labs rabbit anti twik 1 polyclonal antibody
    <t>TWIK-1</t> is expressed in mouse hippocampal dentate granule cells. (A) Representative fluorescence immunostaining images show that TWIK-1 ( a ) is highly expressed in dentate granular layer and CA1-3 regions. DAPI staining ( b ) indicates the overall hippocampal sub-regions including dentate gyrus and CA1-3. Merged image ( c ) demonstrates co-localization of TWIK-1 with principal cells in all hippocampal sub-regions. (d) Magnified image of dentate gyrus, showing co-localization of TWIK-1 with dentate granule cells. (e) Magnified image of the dotted area indicated in (d) . (B) Representative Western Blot data for the expression of TWIK-1 in dentate gyrus and CA1-3 region of the hippocampus (N = 3 mice, P < 0.01, Student’s unpaired t -test). (C) Representative immunostaining images with TWIK-1 (a) , MAP2 (b) , and DAPI (c) . ML, molecular layer; GL, granule layer; H. hilus. Merged TWIK-1 and MAP2 staining image (d) showing that TWIK-1 is co-localized with MAP2 in dendrites of dentate granule cells. High magnification image (e) of dotted rectangle in ( d ) shows that MAP2-positive proximal dendrites of granule layer cells are co-localized with TWIK-1. Note the presence of TWIK-1 positive cells in the molecular layer (ML) of the dentate gyrus and the hilus (H). (D) Double immunostaining with TWIK-1 (green) and calbindin D28k (red) demonstrates that TWIK-1 is only co-localized with calbindin D28k in the granule layer (GL) but not in the hilus (H) or CA3. DAPI stains neuronal cells in the granule cell layer and CA3 layer. Scale bar, 50 μm. DG: dentate gyrus, GL: granule layer, CA1: cornu ammonis 1, CA3: cornu ammonis 3, ML: dentate molecular layer, H: dentate hilus, MF: mossy fibers.
    Rabbit Anti Twik 1 Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus"

    Article Title: TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus

    Journal: Molecular Brain

    doi: 10.1186/s13041-014-0080-z

    TWIK-1 is expressed in mouse hippocampal dentate granule cells. (A) Representative fluorescence immunostaining images show that TWIK-1 ( a ) is highly expressed in dentate granular layer and CA1-3 regions. DAPI staining ( b ) indicates the overall hippocampal sub-regions including dentate gyrus and CA1-3. Merged image ( c ) demonstrates co-localization of TWIK-1 with principal cells in all hippocampal sub-regions. (d) Magnified image of dentate gyrus, showing co-localization of TWIK-1 with dentate granule cells. (e) Magnified image of the dotted area indicated in (d) . (B) Representative Western Blot data for the expression of TWIK-1 in dentate gyrus and CA1-3 region of the hippocampus (N = 3 mice, P < 0.01, Student’s unpaired t -test). (C) Representative immunostaining images with TWIK-1 (a) , MAP2 (b) , and DAPI (c) . ML, molecular layer; GL, granule layer; H. hilus. Merged TWIK-1 and MAP2 staining image (d) showing that TWIK-1 is co-localized with MAP2 in dendrites of dentate granule cells. High magnification image (e) of dotted rectangle in ( d ) shows that MAP2-positive proximal dendrites of granule layer cells are co-localized with TWIK-1. Note the presence of TWIK-1 positive cells in the molecular layer (ML) of the dentate gyrus and the hilus (H). (D) Double immunostaining with TWIK-1 (green) and calbindin D28k (red) demonstrates that TWIK-1 is only co-localized with calbindin D28k in the granule layer (GL) but not in the hilus (H) or CA3. DAPI stains neuronal cells in the granule cell layer and CA3 layer. Scale bar, 50 μm. DG: dentate gyrus, GL: granule layer, CA1: cornu ammonis 1, CA3: cornu ammonis 3, ML: dentate molecular layer, H: dentate hilus, MF: mossy fibers.
    Figure Legend Snippet: TWIK-1 is expressed in mouse hippocampal dentate granule cells. (A) Representative fluorescence immunostaining images show that TWIK-1 ( a ) is highly expressed in dentate granular layer and CA1-3 regions. DAPI staining ( b ) indicates the overall hippocampal sub-regions including dentate gyrus and CA1-3. Merged image ( c ) demonstrates co-localization of TWIK-1 with principal cells in all hippocampal sub-regions. (d) Magnified image of dentate gyrus, showing co-localization of TWIK-1 with dentate granule cells. (e) Magnified image of the dotted area indicated in (d) . (B) Representative Western Blot data for the expression of TWIK-1 in dentate gyrus and CA1-3 region of the hippocampus (N = 3 mice, P < 0.01, Student’s unpaired t -test). (C) Representative immunostaining images with TWIK-1 (a) , MAP2 (b) , and DAPI (c) . ML, molecular layer; GL, granule layer; H. hilus. Merged TWIK-1 and MAP2 staining image (d) showing that TWIK-1 is co-localized with MAP2 in dendrites of dentate granule cells. High magnification image (e) of dotted rectangle in ( d ) shows that MAP2-positive proximal dendrites of granule layer cells are co-localized with TWIK-1. Note the presence of TWIK-1 positive cells in the molecular layer (ML) of the dentate gyrus and the hilus (H). (D) Double immunostaining with TWIK-1 (green) and calbindin D28k (red) demonstrates that TWIK-1 is only co-localized with calbindin D28k in the granule layer (GL) but not in the hilus (H) or CA3. DAPI stains neuronal cells in the granule cell layer and CA3 layer. Scale bar, 50 μm. DG: dentate gyrus, GL: granule layer, CA1: cornu ammonis 1, CA3: cornu ammonis 3, ML: dentate molecular layer, H: dentate hilus, MF: mossy fibers.

    Techniques Used: Fluorescence, Immunostaining, Staining, Western Blot, Expressing, Double Immunostaining

    TWIK-1 contributes to outwardly rectifying currents in dentate granule cells. (A) Averaged current-voltage ( I-V ) relationships of whole-cell currents in dentate granule cells measured in standard ACSF and after subsequent application of Cs + /TEA. (B) Current-voltage relationship of the whole-cell currents in naïve (left panel; n = 34 cells, N = 3 mice), Sc shRNA (middle panel; n = 26 cells, N = 3 mice) or TWIK-1 shRNA (right panel; n = 31 cells, N = 3 mice) expressing dentate granule cells in the presence of Cs + /TEA. Whole-cell currents were elicited by 1 s duration ramp pulses descending from 40 mV to -150 mV from a holding potential of -70 mV. (C) Summary bar charts for (B) . Shown are mean values of whole-cell current density in naïve, Sc shRNA or TWIK-1 shRNA expressing granule cells measured after subsequent application of the blockers; current density values are depicted at -150 mV (shadowed bars below baseline) and 40 mV (open bars) . (D) Reversal potential values of the whole-cell currents in naïve (n = 34 cells, N = 3 mice), Sc shRNA (n = 26 cells, N = 3 mice) or TWIK-1 shRNA (n = 31 cells, N = 3 mice) expressing dentate granule cells. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.
    Figure Legend Snippet: TWIK-1 contributes to outwardly rectifying currents in dentate granule cells. (A) Averaged current-voltage ( I-V ) relationships of whole-cell currents in dentate granule cells measured in standard ACSF and after subsequent application of Cs + /TEA. (B) Current-voltage relationship of the whole-cell currents in naïve (left panel; n = 34 cells, N = 3 mice), Sc shRNA (middle panel; n = 26 cells, N = 3 mice) or TWIK-1 shRNA (right panel; n = 31 cells, N = 3 mice) expressing dentate granule cells in the presence of Cs + /TEA. Whole-cell currents were elicited by 1 s duration ramp pulses descending from 40 mV to -150 mV from a holding potential of -70 mV. (C) Summary bar charts for (B) . Shown are mean values of whole-cell current density in naïve, Sc shRNA or TWIK-1 shRNA expressing granule cells measured after subsequent application of the blockers; current density values are depicted at -150 mV (shadowed bars below baseline) and 40 mV (open bars) . (D) Reversal potential values of the whole-cell currents in naïve (n = 34 cells, N = 3 mice), Sc shRNA (n = 26 cells, N = 3 mice) or TWIK-1 shRNA (n = 31 cells, N = 3 mice) expressing dentate granule cells. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.

    Techniques Used: shRNA, Expressing

    TWIK-1 deficiency causes enhanced intrinsic excitability of dentate granule cells. (A) Representative response of membrane potential to stepwise current injections (left panel). The current was injected into cells in 25 pA steps starting from -120 pA and up to 55 pA (1.2 sec step duration). Input-output properties of naïve (n = 36 cells), Sc shRNA (n = 32 cells) or TWIK-1 shRNA (n = 30 cells) expressing granule cells measured as the number of spikes vs. injected current intensity (right panel). (B) Distribution of cells according to excitability patterns. Plotted are percentage of cells with binned number of spikes fired during a 30 pA injected current step. (C) Representative response of membrane potential to stepwise current injections (left panel). Averaged response of membrane potential to stepwise current injection in naïve (n = 27 cells), Sc shRNA (n = 20 cells) or TWIK-1 shRNA (n = 21 cells) expressing cells (right panel). The RMP of cells was maintained at -70 mV. Current injection into the cell body was performed stepwise from -30 pA to 90 pA, in 5 pA steps. The solid lines are an exponential fit of the data plots. Dotted line indicates the spiking threshold level. (D) Representative traces of rheobase current measurements (left panel). The RMP of cells was kept at -70 mV and then depolarizing current was injected stepwise, in 2 pA steps until the membrane potential reached the firing threshold (red traces) . Averaged values of rheobase currents in naïve (n = 12 cells), Sc shRNA (n = 13 cells) or TWIK-1 shRNA (n = 12 cells) expressing cells (right panel). All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.
    Figure Legend Snippet: TWIK-1 deficiency causes enhanced intrinsic excitability of dentate granule cells. (A) Representative response of membrane potential to stepwise current injections (left panel). The current was injected into cells in 25 pA steps starting from -120 pA and up to 55 pA (1.2 sec step duration). Input-output properties of naïve (n = 36 cells), Sc shRNA (n = 32 cells) or TWIK-1 shRNA (n = 30 cells) expressing granule cells measured as the number of spikes vs. injected current intensity (right panel). (B) Distribution of cells according to excitability patterns. Plotted are percentage of cells with binned number of spikes fired during a 30 pA injected current step. (C) Representative response of membrane potential to stepwise current injections (left panel). Averaged response of membrane potential to stepwise current injection in naïve (n = 27 cells), Sc shRNA (n = 20 cells) or TWIK-1 shRNA (n = 21 cells) expressing cells (right panel). The RMP of cells was maintained at -70 mV. Current injection into the cell body was performed stepwise from -30 pA to 90 pA, in 5 pA steps. The solid lines are an exponential fit of the data plots. Dotted line indicates the spiking threshold level. (D) Representative traces of rheobase current measurements (left panel). The RMP of cells was kept at -70 mV and then depolarizing current was injected stepwise, in 2 pA steps until the membrane potential reached the firing threshold (red traces) . Averaged values of rheobase currents in naïve (n = 12 cells), Sc shRNA (n = 13 cells) or TWIK-1 shRNA (n = 12 cells) expressing cells (right panel). All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.

    Techniques Used: Injection, shRNA, Expressing

    TWIK-1 deficiency in dentate granule cells causes enhanced amplitude of evoked EPSPs in response to perforant path stimulation. (A) ( a ) Schematic diagram for synaptically evoked responsiveness of dentate granule cells. PP: perforant path, Stim: stimulation, Rec: recording. ( b ) Representative traces of evoked EPSPs in naïve, Sc shRNA or TWIK-1 shRNA expressing dentate granule cells. Synaptic responses evoked by 150 μA stimulation of perforant path are shown. ( c ) Input-output properties of dentate granule cells. Plotted values are of evoked EPSP amplitudes in naïve (n = 13cells, N = 3 mice), Sc shRNA (n = 14 cells, N = 3 mice) or TWIK-1 shRNA (n = 12 cells, N = 3 mice) expressing granule cells. The RMP was maintained at -80 mV by steady current injection. For each stimulation intensity, ten perforant path stimuli were applied and the evoked EPSP responses were averaged. The solid line is a logistic fit of the data plots. ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. ( B ) Representative traces for paired-pulse ratio measurements (left panel). Paired-pulse ratios in perforant path to granule cell synaptic transmission as a function of interstimulus interval (ISI) in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing cells (right panel). ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.
    Figure Legend Snippet: TWIK-1 deficiency in dentate granule cells causes enhanced amplitude of evoked EPSPs in response to perforant path stimulation. (A) ( a ) Schematic diagram for synaptically evoked responsiveness of dentate granule cells. PP: perforant path, Stim: stimulation, Rec: recording. ( b ) Representative traces of evoked EPSPs in naïve, Sc shRNA or TWIK-1 shRNA expressing dentate granule cells. Synaptic responses evoked by 150 μA stimulation of perforant path are shown. ( c ) Input-output properties of dentate granule cells. Plotted values are of evoked EPSP amplitudes in naïve (n = 13cells, N = 3 mice), Sc shRNA (n = 14 cells, N = 3 mice) or TWIK-1 shRNA (n = 12 cells, N = 3 mice) expressing granule cells. The RMP was maintained at -80 mV by steady current injection. For each stimulation intensity, ten perforant path stimuli were applied and the evoked EPSP responses were averaged. The solid line is a logistic fit of the data plots. ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. ( B ) Representative traces for paired-pulse ratio measurements (left panel). Paired-pulse ratios in perforant path to granule cell synaptic transmission as a function of interstimulus interval (ISI) in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing cells (right panel). ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.

    Techniques Used: shRNA, Expressing, Injection, Transferring, Transmission Assay

    TWIK-1 knockdown in dentate granule cells affects signal input-output properties of the cells. (A) Representative traces from spiking probability measurements (left panel). Synaptic responses evoked by 200 μA stimulation of perforant path are shown. Input-output properties of dentate granule cells (right panel). Plotted are the values of spiking probability in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing dentate granule cells. The solid line is a logistic fit of the data plots. (B) Representative traces from spiking probability measurements (left panel). Traces with multiple spikes are shown in red. Averaged input-output properties of the subpopulation of the burst discharging cells (right panel). Plotted are the averaged values of probability of firing more than one spike per eEPSP in TWIK-1 shRNA or Sc shRNA expressing cells. (C) Representative traces of ten evoked EPSP responses in Sc shRNA and TWIK-1 shRNA expressing dentate granule cells (stimulation intensity was 350 μA) (left panel). The dotted lines indicate RMP, the voltage threshold for the first AP (V th1 ) and the voltage threshold for the second AP (V th2 ) in the burst response. Traces containing the second spike are shown in red. Summary bar graph for averaged depolarization level after the termination of the first AP in Sc shRNA and a population of TWIK-1 shRNA expressing cells with multiple spikes (right panel). The dotted lines show the threshold levels for the first (V th1 ) and the second (V th2 ) AP during a single EPSP. Number of cells are shown within the bars. All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.
    Figure Legend Snippet: TWIK-1 knockdown in dentate granule cells affects signal input-output properties of the cells. (A) Representative traces from spiking probability measurements (left panel). Synaptic responses evoked by 200 μA stimulation of perforant path are shown. Input-output properties of dentate granule cells (right panel). Plotted are the values of spiking probability in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing dentate granule cells. The solid line is a logistic fit of the data plots. (B) Representative traces from spiking probability measurements (left panel). Traces with multiple spikes are shown in red. Averaged input-output properties of the subpopulation of the burst discharging cells (right panel). Plotted are the averaged values of probability of firing more than one spike per eEPSP in TWIK-1 shRNA or Sc shRNA expressing cells. (C) Representative traces of ten evoked EPSP responses in Sc shRNA and TWIK-1 shRNA expressing dentate granule cells (stimulation intensity was 350 μA) (left panel). The dotted lines indicate RMP, the voltage threshold for the first AP (V th1 ) and the voltage threshold for the second AP (V th2 ) in the burst response. Traces containing the second spike are shown in red. Summary bar graph for averaged depolarization level after the termination of the first AP in Sc shRNA and a population of TWIK-1 shRNA expressing cells with multiple spikes (right panel). The dotted lines show the threshold levels for the first (V th1 ) and the second (V th2 ) AP during a single EPSP. Number of cells are shown within the bars. All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.

    Techniques Used: shRNA, Expressing

    rabbit anti twik1  (Alomone Labs)


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    Alomone Labs rabbit anti twik1
    Rabbit Anti Twik1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti twik1  (Alomone Labs)


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    Alomone Labs rabbit anti twik1
    Rabbit Anti Twik1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
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    rabbit anti twik 1  (Alomone Labs)


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    Alomone Labs rabbit anti twik 1
    The relative Cs+ permeability of astrocyte passive conductance and cloned <t>TWIK-1</t> and TREK-1 K+ channels. A, Astrocyte reversal potential VM (at 0 currents) shifted when bath solutions were switched from normal aCSF containing 3.5 mm K+ to the modified aCSF solutions containing, in sequence, 70 mm K+, Rb+, and Cs+. Astrocyte VM (at 0 current) at the steady-state level was recorded for 2 s for each ionic condition by continuous current-clamp recording. B, E, The voltage ramp-induced whole-cell current traces recorded from two CHO cells, one transfected with TWIK-1 (B) and another with TREK-1 (E) K+ channels. In each case, the current traces recorded in 135 mm Na+ (black traces), 135 mm K+ (dashed black traces), and 135 mm Cs+ (gray traces) bath solutions are shown. C, F, Representative traces of TWIK-1 and TREK-1 K+ channel whole-cell currents recorded with Cs+- and K+-based solutions as indicated. For the TWIK-1 K+ channels, the Cs+-conducted whole-cell currents showed a strong outward rectification and the overall whole-cell current at +50 mV is significantly larger than that of the K+-conducted whole-cell currents (56.6 ± 5.9 pA/pF vs 31.6 ± 2.5 pA/pF, n = 6). The Cs+-conducted TREK-1 channel currents remained outwardly rectifying and were smaller compared with K+-mediated conductance (at +50 mV, 49.5 ± 4.8 pA/pF vs 1118.4 ± 137.7 pA/pF, n = 5). Calibration: 100 ms and 1 nA. D, The relative permeability of astrocyte passive conductance (gray bars), TREK-1 (black bars), and TWIK-1 (open bars) K+ channels to different monovalent ions. The relative permeability (PX/PK) values in each test were calculated from Equation 2.
    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "TWIK-1 and TREK-1 Are Potassium Channels Contributing Significantly to Astrocyte Passive Conductance in Rat Hippocampal Slices"

    Article Title: TWIK-1 and TREK-1 Are Potassium Channels Contributing Significantly to Astrocyte Passive Conductance in Rat Hippocampal Slices

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5784-08.2009

    The relative Cs+ permeability of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, Astrocyte reversal potential VM (at 0 currents) shifted when bath solutions were switched from normal aCSF containing 3.5 mm K+ to the modified aCSF solutions containing, in sequence, 70 mm K+, Rb+, and Cs+. Astrocyte VM (at 0 current) at the steady-state level was recorded for 2 s for each ionic condition by continuous current-clamp recording. B, E, The voltage ramp-induced whole-cell current traces recorded from two CHO cells, one transfected with TWIK-1 (B) and another with TREK-1 (E) K+ channels. In each case, the current traces recorded in 135 mm Na+ (black traces), 135 mm K+ (dashed black traces), and 135 mm Cs+ (gray traces) bath solutions are shown. C, F, Representative traces of TWIK-1 and TREK-1 K+ channel whole-cell currents recorded with Cs+- and K+-based solutions as indicated. For the TWIK-1 K+ channels, the Cs+-conducted whole-cell currents showed a strong outward rectification and the overall whole-cell current at +50 mV is significantly larger than that of the K+-conducted whole-cell currents (56.6 ± 5.9 pA/pF vs 31.6 ± 2.5 pA/pF, n = 6). The Cs+-conducted TREK-1 channel currents remained outwardly rectifying and were smaller compared with K+-mediated conductance (at +50 mV, 49.5 ± 4.8 pA/pF vs 1118.4 ± 137.7 pA/pF, n = 5). Calibration: 100 ms and 1 nA. D, The relative permeability of astrocyte passive conductance (gray bars), TREK-1 (black bars), and TWIK-1 (open bars) K+ channels to different monovalent ions. The relative permeability (PX/PK) values in each test were calculated from Equation 2.
    Figure Legend Snippet: The relative Cs+ permeability of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, Astrocyte reversal potential VM (at 0 currents) shifted when bath solutions were switched from normal aCSF containing 3.5 mm K+ to the modified aCSF solutions containing, in sequence, 70 mm K+, Rb+, and Cs+. Astrocyte VM (at 0 current) at the steady-state level was recorded for 2 s for each ionic condition by continuous current-clamp recording. B, E, The voltage ramp-induced whole-cell current traces recorded from two CHO cells, one transfected with TWIK-1 (B) and another with TREK-1 (E) K+ channels. In each case, the current traces recorded in 135 mm Na+ (black traces), 135 mm K+ (dashed black traces), and 135 mm Cs+ (gray traces) bath solutions are shown. C, F, Representative traces of TWIK-1 and TREK-1 K+ channel whole-cell currents recorded with Cs+- and K+-based solutions as indicated. For the TWIK-1 K+ channels, the Cs+-conducted whole-cell currents showed a strong outward rectification and the overall whole-cell current at +50 mV is significantly larger than that of the K+-conducted whole-cell currents (56.6 ± 5.9 pA/pF vs 31.6 ± 2.5 pA/pF, n = 6). The Cs+-conducted TREK-1 channel currents remained outwardly rectifying and were smaller compared with K+-mediated conductance (at +50 mV, 49.5 ± 4.8 pA/pF vs 1118.4 ± 137.7 pA/pF, n = 5). Calibration: 100 ms and 1 nA. D, The relative permeability of astrocyte passive conductance (gray bars), TREK-1 (black bars), and TWIK-1 (open bars) K+ channels to different monovalent ions. The relative permeability (PX/PK) values in each test were calculated from Equation 2.

    Techniques Used: Permeability, Clone Assay, Modification, Sequencing, Transfection

    The sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to quinine. A, B, Representative whole-cell current traces recorded from CHO cells expressing TWIK-1 (A) or TREK-1 (B) K+ channels induced by voltage ramp pulses before (black traces) and after (gray traces) application of 100 μm quinine. After washout of quinine, the traces were the same as control traces. C, The dose–response curves of the quinine blockage on TWIK-1 (open triangles) and TREK-1 (filled triangles) channels. The continuous lines were fitted according to Equation 3 that yielded IC50 values of 41.4 ± 4.8 μm and 85.4 ± 9.7 μm and h values of 0.98 and 0.77 for TREK-1 and TWIK-1, respectively (n = 5–6 for each data point). D, Representative astrocyte whole-cell passive currents recorded first in aCSF and then after addition of 200 μm quinine. The voltage pulse protocol was the same as in Figure 2. Calibration: 5 ms, 2 nA. E, The mean current–voltage relationship shows that 200 μm quinine (filled circles) inhibited 58% of passive conductance at the command voltages of +20 mV and −160 mV (also presented as filled circle in C, n = 5). Quinine (200 μm) similarly inhibited Cs+-mediated TWIK-1, TREK-1, and astrocyte passive conductances by 72.0% (filled square, n = 3), 88.0% (filled triangle, n = 4), and 42.5% (gray circle, n = 4), respectively (C).
    Figure Legend Snippet: The sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to quinine. A, B, Representative whole-cell current traces recorded from CHO cells expressing TWIK-1 (A) or TREK-1 (B) K+ channels induced by voltage ramp pulses before (black traces) and after (gray traces) application of 100 μm quinine. After washout of quinine, the traces were the same as control traces. C, The dose–response curves of the quinine blockage on TWIK-1 (open triangles) and TREK-1 (filled triangles) channels. The continuous lines were fitted according to Equation 3 that yielded IC50 values of 41.4 ± 4.8 μm and 85.4 ± 9.7 μm and h values of 0.98 and 0.77 for TREK-1 and TWIK-1, respectively (n = 5–6 for each data point). D, Representative astrocyte whole-cell passive currents recorded first in aCSF and then after addition of 200 μm quinine. The voltage pulse protocol was the same as in Figure 2. Calibration: 5 ms, 2 nA. E, The mean current–voltage relationship shows that 200 μm quinine (filled circles) inhibited 58% of passive conductance at the command voltages of +20 mV and −160 mV (also presented as filled circle in C, n = 5). Quinine (200 μm) similarly inhibited Cs+-mediated TWIK-1, TREK-1, and astrocyte passive conductances by 72.0% (filled square, n = 3), 88.0% (filled triangle, n = 4), and 42.5% (gray circle, n = 4), respectively (C).

    Techniques Used: Clone Assay, Expressing

    Sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to extracellular pH. A, B, Representative voltage ramp-induced whole-cell current traces of TWIK-1 and TREK-1 in transfected CHO cells at pH 7.4 (black traces) and pH 6.0 (gray traces). Washout traces are shown as the dashed lines. C, Comparison of effects of extracellular low pH, pH 6.0, on astrocyte passive conductance and expressed TWIK-1 and TREK-1 channels. Whole-cell currents at test voltage +20 mV in the pH 6.0 bath solution (gray bars) are compared with the normalized corresponding values in the pH 7.4 bath solution (open bars) (n = 5–7 for each data point; *p < 0.005). D, Representative whole-cell current traces recorded from an astrocyte in pH 7.4 and pH 6.0 bath solutions. Calibration: 10 ms, 1 nA. E, The mean current–voltage relationships for astrocyte recordings obtained in pH 7.4 (open square) and pH 6.0 (gray square) bath solutions (n = 6). The pooled data show that pH 6.0 significantly inhibited 27% of the outward passive currents at +20 mV of test voltage.
    Figure Legend Snippet: Sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to extracellular pH. A, B, Representative voltage ramp-induced whole-cell current traces of TWIK-1 and TREK-1 in transfected CHO cells at pH 7.4 (black traces) and pH 6.0 (gray traces). Washout traces are shown as the dashed lines. C, Comparison of effects of extracellular low pH, pH 6.0, on astrocyte passive conductance and expressed TWIK-1 and TREK-1 channels. Whole-cell currents at test voltage +20 mV in the pH 6.0 bath solution (gray bars) are compared with the normalized corresponding values in the pH 7.4 bath solution (open bars) (n = 5–7 for each data point; *p < 0.005). D, Representative whole-cell current traces recorded from an astrocyte in pH 7.4 and pH 6.0 bath solutions. Calibration: 10 ms, 1 nA. E, The mean current–voltage relationships for astrocyte recordings obtained in pH 7.4 (open square) and pH 6.0 (gray square) bath solutions (n = 6). The pooled data show that pH 6.0 significantly inhibited 27% of the outward passive currents at +20 mV of test voltage.

    Techniques Used: Clone Assay, Transfection

    Effects of Ba2+ on astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, B, Representative whole-cell current traces recorded from TWIK-1 (A) and TREK-1 (B). K+ channels expressed in CHO cells induced by voltage ramp pulses before (black lines) and after (gray lines) bath application of 800 μm Ba2+. The washout recording traces (dashed lines) are superimposable with the control traces. C, Dose–response curves of Ba2+ blockade of TWIK-1 (open squares) K+ channels. The continuous line is a fit of Equation 3, yielding an IC50 of 960 ± 9.8 μm with h = 0.84. Blockade of astrocyte passive conductance (filled circle) and TREK-1 (filled square) K+ channels at test voltage 20 mV by 1.2 mm Ba2+ and 800 μm Ba2+, respectively, are also shown (n = 5–7 for each data point). D, Representative traces of whole-cell currents recorded from an astrocyte before and after application of 1.2 mm Ba2+ in bath solution. E, The mean current–voltage relationships for the representative recordings shown in D (n = 6). Calibration: 10 ms and 2 nA.
    Figure Legend Snippet: Effects of Ba2+ on astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, B, Representative whole-cell current traces recorded from TWIK-1 (A) and TREK-1 (B). K+ channels expressed in CHO cells induced by voltage ramp pulses before (black lines) and after (gray lines) bath application of 800 μm Ba2+. The washout recording traces (dashed lines) are superimposable with the control traces. C, Dose–response curves of Ba2+ blockade of TWIK-1 (open squares) K+ channels. The continuous line is a fit of Equation 3, yielding an IC50 of 960 ± 9.8 μm with h = 0.84. Blockade of astrocyte passive conductance (filled circle) and TREK-1 (filled square) K+ channels at test voltage 20 mV by 1.2 mm Ba2+ and 800 μm Ba2+, respectively, are also shown (n = 5–7 for each data point). D, Representative traces of whole-cell currents recorded from an astrocyte before and after application of 1.2 mm Ba2+ in bath solution. E, The mean current–voltage relationships for the representative recordings shown in D (n = 6). Calibration: 10 ms and 2 nA.

    Techniques Used: Clone Assay

    Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in CA1 pyramidal cells and in the stratum radiatum. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST (A, E, I, M), and NeuN (C) or GFAP (G, K, O) and imaged with confocal microscopy. Each row of images shows the 3 channels and the merged image from a single field in the CA1 pyramidal cell layer (upper left quadrant of each panel) and the stratum radiatum. TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that, compared with the TASK-1 immunoreactivity, anti-TWIK-1 and TREK-1 were more intense in the stratum radiatum neuropil, similar to anti-GLAST. The insets in B, J, and N are from images taken from sections treated with the respective K2P antibody preincubated with the blocking peptide. The scale bar in A represents 50 μm and applies to all images in this figure.
    Figure Legend Snippet: Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in CA1 pyramidal cells and in the stratum radiatum. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST (A, E, I, M), and NeuN (C) or GFAP (G, K, O) and imaged with confocal microscopy. Each row of images shows the 3 channels and the merged image from a single field in the CA1 pyramidal cell layer (upper left quadrant of each panel) and the stratum radiatum. TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that, compared with the TASK-1 immunoreactivity, anti-TWIK-1 and TREK-1 were more intense in the stratum radiatum neuropil, similar to anti-GLAST. The insets in B, J, and N are from images taken from sections treated with the respective K2P antibody preincubated with the blocking peptide. The scale bar in A represents 50 μm and applies to all images in this figure.

    Techniques Used: Immunocytochemistry, Labeling, Confocal Microscopy, Blocking Assay

    Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in hippocampal sections. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST, (A, E, I, M), and NeuN (C) or GFAP (G, K, O). Shown are single confocal planes taken in the CA1 pyramidal cell layer (A–D), or the stratum radiatum (E–P). TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that the TASK-1 immunoreactivity overlaps with that for GFAP and is within the GLAST outline of the astrocyte soma (E–H). Compared with anti-TASK-1, anti-TWIK-1 and TREK-1 bound more in the neuropil, in a similar manner to anti-GLAST. These antibodies also labeled the GFAP(+) somata (see merged images L and P). The scale bar in D represents 10 μm and applies to all images in this figure.
    Figure Legend Snippet: Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in hippocampal sections. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST, (A, E, I, M), and NeuN (C) or GFAP (G, K, O). Shown are single confocal planes taken in the CA1 pyramidal cell layer (A–D), or the stratum radiatum (E–P). TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that the TASK-1 immunoreactivity overlaps with that for GFAP and is within the GLAST outline of the astrocyte soma (E–H). Compared with anti-TASK-1, anti-TWIK-1 and TREK-1 bound more in the neuropil, in a similar manner to anti-GLAST. These antibodies also labeled the GFAP(+) somata (see merged images L and P). The scale bar in D represents 10 μm and applies to all images in this figure.

    Techniques Used: Immunocytochemistry, Labeling

    rabbit anti twik 1  (Alomone Labs)


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    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit anti twik 1  (Alomone Labs)


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    Alomone Labs rabbit anti twik 1
    The relative Cs+ permeability of astrocyte passive conductance and cloned <t>TWIK-1</t> and TREK-1 K+ channels. A, Astrocyte reversal potential VM (at 0 currents) shifted when bath solutions were switched from normal aCSF containing 3.5 mm K+ to the modified aCSF solutions containing, in sequence, 70 mm K+, Rb+, and Cs+. Astrocyte VM (at 0 current) at the steady-state level was recorded for 2 s for each ionic condition by continuous current-clamp recording. B, E, The voltage ramp-induced whole-cell current traces recorded from two CHO cells, one transfected with TWIK-1 (B) and another with TREK-1 (E) K+ channels. In each case, the current traces recorded in 135 mm Na+ (black traces), 135 mm K+ (dashed black traces), and 135 mm Cs+ (gray traces) bath solutions are shown. C, F, Representative traces of TWIK-1 and TREK-1 K+ channel whole-cell currents recorded with Cs+- and K+-based solutions as indicated. For the TWIK-1 K+ channels, the Cs+-conducted whole-cell currents showed a strong outward rectification and the overall whole-cell current at +50 mV is significantly larger than that of the K+-conducted whole-cell currents (56.6 ± 5.9 pA/pF vs 31.6 ± 2.5 pA/pF, n = 6). The Cs+-conducted TREK-1 channel currents remained outwardly rectifying and were smaller compared with K+-mediated conductance (at +50 mV, 49.5 ± 4.8 pA/pF vs 1118.4 ± 137.7 pA/pF, n = 5). Calibration: 100 ms and 1 nA. D, The relative permeability of astrocyte passive conductance (gray bars), TREK-1 (black bars), and TWIK-1 (open bars) K+ channels to different monovalent ions. The relative permeability (PX/PK) values in each test were calculated from Equation 2.
    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik 1/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik 1 - by Bioz Stars, 2023-02
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    1) Product Images from "TWIK-1 and TREK-1 Are Potassium Channels Contributing Significantly to Astrocyte Passive Conductance in Rat Hippocampal Slices"

    Article Title: TWIK-1 and TREK-1 Are Potassium Channels Contributing Significantly to Astrocyte Passive Conductance in Rat Hippocampal Slices

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.5784-08.2009

    The relative Cs+ permeability of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, Astrocyte reversal potential VM (at 0 currents) shifted when bath solutions were switched from normal aCSF containing 3.5 mm K+ to the modified aCSF solutions containing, in sequence, 70 mm K+, Rb+, and Cs+. Astrocyte VM (at 0 current) at the steady-state level was recorded for 2 s for each ionic condition by continuous current-clamp recording. B, E, The voltage ramp-induced whole-cell current traces recorded from two CHO cells, one transfected with TWIK-1 (B) and another with TREK-1 (E) K+ channels. In each case, the current traces recorded in 135 mm Na+ (black traces), 135 mm K+ (dashed black traces), and 135 mm Cs+ (gray traces) bath solutions are shown. C, F, Representative traces of TWIK-1 and TREK-1 K+ channel whole-cell currents recorded with Cs+- and K+-based solutions as indicated. For the TWIK-1 K+ channels, the Cs+-conducted whole-cell currents showed a strong outward rectification and the overall whole-cell current at +50 mV is significantly larger than that of the K+-conducted whole-cell currents (56.6 ± 5.9 pA/pF vs 31.6 ± 2.5 pA/pF, n = 6). The Cs+-conducted TREK-1 channel currents remained outwardly rectifying and were smaller compared with K+-mediated conductance (at +50 mV, 49.5 ± 4.8 pA/pF vs 1118.4 ± 137.7 pA/pF, n = 5). Calibration: 100 ms and 1 nA. D, The relative permeability of astrocyte passive conductance (gray bars), TREK-1 (black bars), and TWIK-1 (open bars) K+ channels to different monovalent ions. The relative permeability (PX/PK) values in each test were calculated from Equation 2.
    Figure Legend Snippet: The relative Cs+ permeability of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, Astrocyte reversal potential VM (at 0 currents) shifted when bath solutions were switched from normal aCSF containing 3.5 mm K+ to the modified aCSF solutions containing, in sequence, 70 mm K+, Rb+, and Cs+. Astrocyte VM (at 0 current) at the steady-state level was recorded for 2 s for each ionic condition by continuous current-clamp recording. B, E, The voltage ramp-induced whole-cell current traces recorded from two CHO cells, one transfected with TWIK-1 (B) and another with TREK-1 (E) K+ channels. In each case, the current traces recorded in 135 mm Na+ (black traces), 135 mm K+ (dashed black traces), and 135 mm Cs+ (gray traces) bath solutions are shown. C, F, Representative traces of TWIK-1 and TREK-1 K+ channel whole-cell currents recorded with Cs+- and K+-based solutions as indicated. For the TWIK-1 K+ channels, the Cs+-conducted whole-cell currents showed a strong outward rectification and the overall whole-cell current at +50 mV is significantly larger than that of the K+-conducted whole-cell currents (56.6 ± 5.9 pA/pF vs 31.6 ± 2.5 pA/pF, n = 6). The Cs+-conducted TREK-1 channel currents remained outwardly rectifying and were smaller compared with K+-mediated conductance (at +50 mV, 49.5 ± 4.8 pA/pF vs 1118.4 ± 137.7 pA/pF, n = 5). Calibration: 100 ms and 1 nA. D, The relative permeability of astrocyte passive conductance (gray bars), TREK-1 (black bars), and TWIK-1 (open bars) K+ channels to different monovalent ions. The relative permeability (PX/PK) values in each test were calculated from Equation 2.

    Techniques Used: Permeability, Clone Assay, Modification, Sequencing, Transfection

    The sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to quinine. A, B, Representative whole-cell current traces recorded from CHO cells expressing TWIK-1 (A) or TREK-1 (B) K+ channels induced by voltage ramp pulses before (black traces) and after (gray traces) application of 100 μm quinine. After washout of quinine, the traces were the same as control traces. C, The dose–response curves of the quinine blockage on TWIK-1 (open triangles) and TREK-1 (filled triangles) channels. The continuous lines were fitted according to Equation 3 that yielded IC50 values of 41.4 ± 4.8 μm and 85.4 ± 9.7 μm and h values of 0.98 and 0.77 for TREK-1 and TWIK-1, respectively (n = 5–6 for each data point). D, Representative astrocyte whole-cell passive currents recorded first in aCSF and then after addition of 200 μm quinine. The voltage pulse protocol was the same as in Figure 2. Calibration: 5 ms, 2 nA. E, The mean current–voltage relationship shows that 200 μm quinine (filled circles) inhibited 58% of passive conductance at the command voltages of +20 mV and −160 mV (also presented as filled circle in C, n = 5). Quinine (200 μm) similarly inhibited Cs+-mediated TWIK-1, TREK-1, and astrocyte passive conductances by 72.0% (filled square, n = 3), 88.0% (filled triangle, n = 4), and 42.5% (gray circle, n = 4), respectively (C).
    Figure Legend Snippet: The sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to quinine. A, B, Representative whole-cell current traces recorded from CHO cells expressing TWIK-1 (A) or TREK-1 (B) K+ channels induced by voltage ramp pulses before (black traces) and after (gray traces) application of 100 μm quinine. After washout of quinine, the traces were the same as control traces. C, The dose–response curves of the quinine blockage on TWIK-1 (open triangles) and TREK-1 (filled triangles) channels. The continuous lines were fitted according to Equation 3 that yielded IC50 values of 41.4 ± 4.8 μm and 85.4 ± 9.7 μm and h values of 0.98 and 0.77 for TREK-1 and TWIK-1, respectively (n = 5–6 for each data point). D, Representative astrocyte whole-cell passive currents recorded first in aCSF and then after addition of 200 μm quinine. The voltage pulse protocol was the same as in Figure 2. Calibration: 5 ms, 2 nA. E, The mean current–voltage relationship shows that 200 μm quinine (filled circles) inhibited 58% of passive conductance at the command voltages of +20 mV and −160 mV (also presented as filled circle in C, n = 5). Quinine (200 μm) similarly inhibited Cs+-mediated TWIK-1, TREK-1, and astrocyte passive conductances by 72.0% (filled square, n = 3), 88.0% (filled triangle, n = 4), and 42.5% (gray circle, n = 4), respectively (C).

    Techniques Used: Clone Assay, Expressing

    Sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to extracellular pH. A, B, Representative voltage ramp-induced whole-cell current traces of TWIK-1 and TREK-1 in transfected CHO cells at pH 7.4 (black traces) and pH 6.0 (gray traces). Washout traces are shown as the dashed lines. C, Comparison of effects of extracellular low pH, pH 6.0, on astrocyte passive conductance and expressed TWIK-1 and TREK-1 channels. Whole-cell currents at test voltage +20 mV in the pH 6.0 bath solution (gray bars) are compared with the normalized corresponding values in the pH 7.4 bath solution (open bars) (n = 5–7 for each data point; *p < 0.005). D, Representative whole-cell current traces recorded from an astrocyte in pH 7.4 and pH 6.0 bath solutions. Calibration: 10 ms, 1 nA. E, The mean current–voltage relationships for astrocyte recordings obtained in pH 7.4 (open square) and pH 6.0 (gray square) bath solutions (n = 6). The pooled data show that pH 6.0 significantly inhibited 27% of the outward passive currents at +20 mV of test voltage.
    Figure Legend Snippet: Sensitivity of astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels to extracellular pH. A, B, Representative voltage ramp-induced whole-cell current traces of TWIK-1 and TREK-1 in transfected CHO cells at pH 7.4 (black traces) and pH 6.0 (gray traces). Washout traces are shown as the dashed lines. C, Comparison of effects of extracellular low pH, pH 6.0, on astrocyte passive conductance and expressed TWIK-1 and TREK-1 channels. Whole-cell currents at test voltage +20 mV in the pH 6.0 bath solution (gray bars) are compared with the normalized corresponding values in the pH 7.4 bath solution (open bars) (n = 5–7 for each data point; *p < 0.005). D, Representative whole-cell current traces recorded from an astrocyte in pH 7.4 and pH 6.0 bath solutions. Calibration: 10 ms, 1 nA. E, The mean current–voltage relationships for astrocyte recordings obtained in pH 7.4 (open square) and pH 6.0 (gray square) bath solutions (n = 6). The pooled data show that pH 6.0 significantly inhibited 27% of the outward passive currents at +20 mV of test voltage.

    Techniques Used: Clone Assay, Transfection

    Effects of Ba2+ on astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, B, Representative whole-cell current traces recorded from TWIK-1 (A) and TREK-1 (B). K+ channels expressed in CHO cells induced by voltage ramp pulses before (black lines) and after (gray lines) bath application of 800 μm Ba2+. The washout recording traces (dashed lines) are superimposable with the control traces. C, Dose–response curves of Ba2+ blockade of TWIK-1 (open squares) K+ channels. The continuous line is a fit of Equation 3, yielding an IC50 of 960 ± 9.8 μm with h = 0.84. Blockade of astrocyte passive conductance (filled circle) and TREK-1 (filled square) K+ channels at test voltage 20 mV by 1.2 mm Ba2+ and 800 μm Ba2+, respectively, are also shown (n = 5–7 for each data point). D, Representative traces of whole-cell currents recorded from an astrocyte before and after application of 1.2 mm Ba2+ in bath solution. E, The mean current–voltage relationships for the representative recordings shown in D (n = 6). Calibration: 10 ms and 2 nA.
    Figure Legend Snippet: Effects of Ba2+ on astrocyte passive conductance and cloned TWIK-1 and TREK-1 K+ channels. A, B, Representative whole-cell current traces recorded from TWIK-1 (A) and TREK-1 (B). K+ channels expressed in CHO cells induced by voltage ramp pulses before (black lines) and after (gray lines) bath application of 800 μm Ba2+. The washout recording traces (dashed lines) are superimposable with the control traces. C, Dose–response curves of Ba2+ blockade of TWIK-1 (open squares) K+ channels. The continuous line is a fit of Equation 3, yielding an IC50 of 960 ± 9.8 μm with h = 0.84. Blockade of astrocyte passive conductance (filled circle) and TREK-1 (filled square) K+ channels at test voltage 20 mV by 1.2 mm Ba2+ and 800 μm Ba2+, respectively, are also shown (n = 5–7 for each data point). D, Representative traces of whole-cell currents recorded from an astrocyte before and after application of 1.2 mm Ba2+ in bath solution. E, The mean current–voltage relationships for the representative recordings shown in D (n = 6). Calibration: 10 ms and 2 nA.

    Techniques Used: Clone Assay

    Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in CA1 pyramidal cells and in the stratum radiatum. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST (A, E, I, M), and NeuN (C) or GFAP (G, K, O) and imaged with confocal microscopy. Each row of images shows the 3 channels and the merged image from a single field in the CA1 pyramidal cell layer (upper left quadrant of each panel) and the stratum radiatum. TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that, compared with the TASK-1 immunoreactivity, anti-TWIK-1 and TREK-1 were more intense in the stratum radiatum neuropil, similar to anti-GLAST. The insets in B, J, and N are from images taken from sections treated with the respective K2P antibody preincubated with the blocking peptide. The scale bar in A represents 50 μm and applies to all images in this figure.
    Figure Legend Snippet: Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in CA1 pyramidal cells and in the stratum radiatum. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST (A, E, I, M), and NeuN (C) or GFAP (G, K, O) and imaged with confocal microscopy. Each row of images shows the 3 channels and the merged image from a single field in the CA1 pyramidal cell layer (upper left quadrant of each panel) and the stratum radiatum. TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that, compared with the TASK-1 immunoreactivity, anti-TWIK-1 and TREK-1 were more intense in the stratum radiatum neuropil, similar to anti-GLAST. The insets in B, J, and N are from images taken from sections treated with the respective K2P antibody preincubated with the blocking peptide. The scale bar in A represents 50 μm and applies to all images in this figure.

    Techniques Used: Immunocytochemistry, Labeling, Confocal Microscopy, Blocking Assay

    Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in hippocampal sections. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST, (A, E, I, M), and NeuN (C) or GFAP (G, K, O). Shown are single confocal planes taken in the CA1 pyramidal cell layer (A–D), or the stratum radiatum (E–P). TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that the TASK-1 immunoreactivity overlaps with that for GFAP and is within the GLAST outline of the astrocyte soma (E–H). Compared with anti-TASK-1, anti-TWIK-1 and TREK-1 bound more in the neuropil, in a similar manner to anti-GLAST. These antibodies also labeled the GFAP(+) somata (see merged images L and P). The scale bar in D represents 10 μm and applies to all images in this figure.
    Figure Legend Snippet: Immunocytochemistry of TASK-1, TWIK-1, and TREK-1 channel proteins in hippocampal sections. Coronal hippocampal sections were immunofluorescently labeled for one of the three K2P channel proteins, namely, TASK-1 (B, F), TWIK-1 (J), and TREK-1 (N), GLAST, (A, E, I, M), and NeuN (C) or GFAP (G, K, O). Shown are single confocal planes taken in the CA1 pyramidal cell layer (A–D), or the stratum radiatum (E–P). TASK-1 immunoreactivity was present in NeuN-immunoreactive pyramidal neurons (A–D) and to a lesser extent in GFAP- and GLAST-immunoreactive astrocytes (E–H). Note that the TASK-1 immunoreactivity overlaps with that for GFAP and is within the GLAST outline of the astrocyte soma (E–H). Compared with anti-TASK-1, anti-TWIK-1 and TREK-1 bound more in the neuropil, in a similar manner to anti-GLAST. These antibodies also labeled the GFAP(+) somata (see merged images L and P). The scale bar in D represents 10 μm and applies to all images in this figure.

    Techniques Used: Immunocytochemistry, Labeling

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    Alomone Labs rabbit anti twik 1
    Generation of <t>TWIK-1</t> BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
    Rabbit Anti Twik 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik 1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik 1 - by Bioz Stars, 2023-02
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    93
    Alomone Labs rabbit anti twik1
    Generation of <t>TWIK-1</t> BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.
    Rabbit Anti Twik1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik1 - by Bioz Stars, 2023-02
    93/100 stars
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    93
    Alomone Labs rabbit anti twik 1 polyclonal antibody
    <t>TWIK-1</t> is expressed in mouse hippocampal dentate granule cells. (A) Representative fluorescence immunostaining images show that TWIK-1 ( a ) is highly expressed in dentate granular layer and CA1-3 regions. DAPI staining ( b ) indicates the overall hippocampal sub-regions including dentate gyrus and CA1-3. Merged image ( c ) demonstrates co-localization of TWIK-1 with principal cells in all hippocampal sub-regions. (d) Magnified image of dentate gyrus, showing co-localization of TWIK-1 with dentate granule cells. (e) Magnified image of the dotted area indicated in (d) . (B) Representative Western Blot data for the expression of TWIK-1 in dentate gyrus and CA1-3 region of the hippocampus (N = 3 mice, P < 0.01, Student’s unpaired t -test). (C) Representative immunostaining images with TWIK-1 (a) , MAP2 (b) , and DAPI (c) . ML, molecular layer; GL, granule layer; H. hilus. Merged TWIK-1 and MAP2 staining image (d) showing that TWIK-1 is co-localized with MAP2 in dendrites of dentate granule cells. High magnification image (e) of dotted rectangle in ( d ) shows that MAP2-positive proximal dendrites of granule layer cells are co-localized with TWIK-1. Note the presence of TWIK-1 positive cells in the molecular layer (ML) of the dentate gyrus and the hilus (H). (D) Double immunostaining with TWIK-1 (green) and calbindin D28k (red) demonstrates that TWIK-1 is only co-localized with calbindin D28k in the granule layer (GL) but not in the hilus (H) or CA3. DAPI stains neuronal cells in the granule cell layer and CA3 layer. Scale bar, 50 μm. DG: dentate gyrus, GL: granule layer, CA1: cornu ammonis 1, CA3: cornu ammonis 3, ML: dentate molecular layer, H: dentate hilus, MF: mossy fibers.
    Rabbit Anti Twik 1 Polyclonal Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti twik 1 polyclonal antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti twik 1 polyclonal antibody - by Bioz Stars, 2023-02
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    Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: Generation of TWIK-1 BAC-GFP Tg mice. ( A ) Schematic design of the modified Kcnk1 BAC vector. GFP coding sequence was inserted just before the start codon of Kcnk1 . ( B ) Genotype PCR results for the designed primer pair in A. ( C ) Representative macroscopic GFP fluorescent images in the adult brain (P56 age) from TWIK-1 BAC-GFP Tg mice. Upper left numbers indicate the relative position of the brain slice from bregma. Strong GFP expression was identified in DG, LEC, and Cb (yellow dashed arrow), Scale bar, 1000 μm. ( D ) Representative co-immunofluorescence images with GFP and TWIK-1 in the DG. Scale bar, 200 μm. ( E ) Enlarged inset from D. Scale bar, 50 μm. ( F ) Representative co-immunofluorescence images with GFP and TWIK-1 in the LEC. Scale bar, 200 μm. ( G ) Enlarged inset from F. Scale bar, 50 μm. ( H ) Representative co-immunofluorescence images with GFP and TWIK-1 in the Cb. Scale bar, 200 μm. ( I ) Enlarged inset from H. Scale bar, 50 μm.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Modification, Plasmid Preparation, Sequencing, Slice Preparation, Expressing, Immunofluorescence

    Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in . Data are presented as the Mean ± SEM.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: Cellular identification of GFP-expressing cells of the DG, LEC, and Cb in P56 of TWIK-1 BAC-GFP Tg mice. ( A ) Overview of GFP expression in DG, LEC, and CB of TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. ( B ) Representative co-immunofluorescence images with GFP, NeuN, and GAD67 antibodies. Scale bar, 200 μm. ( C ) Quantification bar graph of the cell type of GFP-positive cells in each brain area from B. Quantification was analyzed by the percentage of each cell type from all GFP-positive cells. Raw data are listed in . Data are presented as the Mean ± SEM.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, Immunofluorescence

    High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in <xref ref-type=Supplementary Materials Table S1 . Data are presented as the Mean ± SEM. " width="100%" height="100%">

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: High TWIK-1 expression in immature neurons of the DG in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with GFP, DCX, CB, and Ki67 in P56 of DG. Scale bar, 200 μm. ( B ) Enlarged inset from A. Most of strong GFP-expressing cells co-labeled with DCX (yellow arrow), but not with CB and Ki67 (white arrow). Scale bar, 10 μm. ( C , D ) Quantification of the cell type of strong GFP-expressing cells. Raw data are listed in Supplementary Materials Table S1 . Data are presented as the Mean ± SEM.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, Immunofluorescence, Labeling

    Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: Glial expression of TWIK-1 in TWIK-1 BAC-GFP Tg mice. ( A ) Representative co-immunofluorescence images with TWIK-1 and GFAP. Scale bar, 200 μm. ( B ) Enlarged inset from A. Yellow arrow indicates double immunoreactive cells with TWIK-1 and GFAP. Scale bar, 50 μm. ( C ) Representative co-immunofluorescence images with GFP and GFAP. Scale bar, 200 μm. ( D ) Enlarged inset from C. Yellow arrow indicates double immunoreactive cells with GFP and GFAP. Scale bar, 50 μm. ( E ) Representative co-immunofluorescence images with GFP and Iba1. Scale bar, 200 μm. ( F ) Enlarged inset from E. There are no Iba1-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm. ( G ) Representative co-immunofluorescence images with GFP and NG2. Scale bar, 200 μm. ( H ) Enlarged inset from G. There are no NG2-positive glial-like GFP-expressing cells (White arrow). Scale bar, 50 μm.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, Immunofluorescence

    TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in <xref ref-type=Supplementary Materials Table S1 . **** p < 0.0001; ** p < 0.01, two-tailed t tests. Data are presented as the Mean ± SEM. " width="100%" height="100%">

    Journal: Cells

    Article Title: TWIK-1 BAC-GFP Transgenic Mice, an Animal Model for TWIK-1 Expression

    doi: 10.3390/cells10102751

    Figure Lengend Snippet: TWIK-1 BAC-GFP Tg mice represent kainic acid (KA)-induced increase of TWIK-1 expression. ( A ) Representative GFP expression in a brain slice from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 1000 μm. ( B ) Representative GFP expression in the DG from saline or KA-treated TWIK-1 BAC-GFP Tg mice. Scale bar, 200 μm. Enlarged inset from the experiment. Scale bar, 50 μm. ( C ) Quantification of relative mean GFP intensity of the granule cell layer. ( D ) Representative Kcnk1 mRNA fluorescence in situ hybridization (FISH) images of DG from saline- or KA-treated mice. Scale bar, 100 μm. ( E ) Quantification bar graph of Kcnk1 mRNA spot density in the granule cell layer from saline- or KA-treated mice. ( F ) Representative TWIK-1 immunofluorescence images of DG from saline- or KA-treated mice. Scale bar, 200 μm. ( G ) Quantification bar graph of the relative mean TWIK-1 immunofluorescence in the granule cell layer of saline- or KA-treated mice. Raw data are listed in Supplementary Materials Table S1 . **** p < 0.0001; ** p < 0.01, two-tailed t tests. Data are presented as the Mean ± SEM.

    Article Snippet: The following antibodies were used: chicken anti-GFP (Abcam, Cat#; ab136970, 1:300); rabbit anti-TWIK-1 (Alomone labs, Cat#; APC-10, 1:200); mouse anti-NeuN (Abcam, Cat#; ab104224, 1:100); rabbit anti-GAD67 (GeneTex, Cat#; GTX113190, 1:300); guinea pig anti-doublecortin (Millipore, Cat#; AB2253, 1:100); rabbit anti-calbindin (Swant, Cat#; CB38, 1:2000); rat anti-Ki67 (Invitrogen, Cat#; 14-5698-85, 1:200); rat anti-GFAP (Invitrogen, Cat#; 13-0300, 1:500); rabbit anti-NG2 (Millipore, Cat#; AB5320, 1:300); and Alexa Fluor 488-, 594-, and 647-conjugated secondary antibodies (Jackson ImmunoResearch, 1:300).

    Techniques: Expressing, Slice Preparation, Fluorescence, In Situ Hybridization, Immunofluorescence, Two Tailed Test

    TWIK-1 is expressed in mouse hippocampal dentate granule cells. (A) Representative fluorescence immunostaining images show that TWIK-1 ( a ) is highly expressed in dentate granular layer and CA1-3 regions. DAPI staining ( b ) indicates the overall hippocampal sub-regions including dentate gyrus and CA1-3. Merged image ( c ) demonstrates co-localization of TWIK-1 with principal cells in all hippocampal sub-regions. (d) Magnified image of dentate gyrus, showing co-localization of TWIK-1 with dentate granule cells. (e) Magnified image of the dotted area indicated in (d) . (B) Representative Western Blot data for the expression of TWIK-1 in dentate gyrus and CA1-3 region of the hippocampus (N = 3 mice, P < 0.01, Student’s unpaired t -test). (C) Representative immunostaining images with TWIK-1 (a) , MAP2 (b) , and DAPI (c) . ML, molecular layer; GL, granule layer; H. hilus. Merged TWIK-1 and MAP2 staining image (d) showing that TWIK-1 is co-localized with MAP2 in dendrites of dentate granule cells. High magnification image (e) of dotted rectangle in ( d ) shows that MAP2-positive proximal dendrites of granule layer cells are co-localized with TWIK-1. Note the presence of TWIK-1 positive cells in the molecular layer (ML) of the dentate gyrus and the hilus (H). (D) Double immunostaining with TWIK-1 (green) and calbindin D28k (red) demonstrates that TWIK-1 is only co-localized with calbindin D28k in the granule layer (GL) but not in the hilus (H) or CA3. DAPI stains neuronal cells in the granule cell layer and CA3 layer. Scale bar, 50 μm. DG: dentate gyrus, GL: granule layer, CA1: cornu ammonis 1, CA3: cornu ammonis 3, ML: dentate molecular layer, H: dentate hilus, MF: mossy fibers.

    Journal: Molecular Brain

    Article Title: TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus

    doi: 10.1186/s13041-014-0080-z

    Figure Lengend Snippet: TWIK-1 is expressed in mouse hippocampal dentate granule cells. (A) Representative fluorescence immunostaining images show that TWIK-1 ( a ) is highly expressed in dentate granular layer and CA1-3 regions. DAPI staining ( b ) indicates the overall hippocampal sub-regions including dentate gyrus and CA1-3. Merged image ( c ) demonstrates co-localization of TWIK-1 with principal cells in all hippocampal sub-regions. (d) Magnified image of dentate gyrus, showing co-localization of TWIK-1 with dentate granule cells. (e) Magnified image of the dotted area indicated in (d) . (B) Representative Western Blot data for the expression of TWIK-1 in dentate gyrus and CA1-3 region of the hippocampus (N = 3 mice, P < 0.01, Student’s unpaired t -test). (C) Representative immunostaining images with TWIK-1 (a) , MAP2 (b) , and DAPI (c) . ML, molecular layer; GL, granule layer; H. hilus. Merged TWIK-1 and MAP2 staining image (d) showing that TWIK-1 is co-localized with MAP2 in dendrites of dentate granule cells. High magnification image (e) of dotted rectangle in ( d ) shows that MAP2-positive proximal dendrites of granule layer cells are co-localized with TWIK-1. Note the presence of TWIK-1 positive cells in the molecular layer (ML) of the dentate gyrus and the hilus (H). (D) Double immunostaining with TWIK-1 (green) and calbindin D28k (red) demonstrates that TWIK-1 is only co-localized with calbindin D28k in the granule layer (GL) but not in the hilus (H) or CA3. DAPI stains neuronal cells in the granule cell layer and CA3 layer. Scale bar, 50 μm. DG: dentate gyrus, GL: granule layer, CA1: cornu ammonis 1, CA3: cornu ammonis 3, ML: dentate molecular layer, H: dentate hilus, MF: mossy fibers.

    Article Snippet: Tissues were incubated overnight with primary antibodies, such as rabbit anti-TWIK-1 polyclonal antibody (1:100; Alomone, Jerusalem, Israel) or chicken anti-MAP2 polyclonal antibody (1:1000; Abcam, Cambridge, MA), at 4°C.

    Techniques: Fluorescence, Immunostaining, Staining, Western Blot, Expressing, Double Immunostaining

    TWIK-1 contributes to outwardly rectifying currents in dentate granule cells. (A) Averaged current-voltage ( I-V ) relationships of whole-cell currents in dentate granule cells measured in standard ACSF and after subsequent application of Cs + /TEA. (B) Current-voltage relationship of the whole-cell currents in naïve (left panel; n = 34 cells, N = 3 mice), Sc shRNA (middle panel; n = 26 cells, N = 3 mice) or TWIK-1 shRNA (right panel; n = 31 cells, N = 3 mice) expressing dentate granule cells in the presence of Cs + /TEA. Whole-cell currents were elicited by 1 s duration ramp pulses descending from 40 mV to -150 mV from a holding potential of -70 mV. (C) Summary bar charts for (B) . Shown are mean values of whole-cell current density in naïve, Sc shRNA or TWIK-1 shRNA expressing granule cells measured after subsequent application of the blockers; current density values are depicted at -150 mV (shadowed bars below baseline) and 40 mV (open bars) . (D) Reversal potential values of the whole-cell currents in naïve (n = 34 cells, N = 3 mice), Sc shRNA (n = 26 cells, N = 3 mice) or TWIK-1 shRNA (n = 31 cells, N = 3 mice) expressing dentate granule cells. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.

    Journal: Molecular Brain

    Article Title: TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus

    doi: 10.1186/s13041-014-0080-z

    Figure Lengend Snippet: TWIK-1 contributes to outwardly rectifying currents in dentate granule cells. (A) Averaged current-voltage ( I-V ) relationships of whole-cell currents in dentate granule cells measured in standard ACSF and after subsequent application of Cs + /TEA. (B) Current-voltage relationship of the whole-cell currents in naïve (left panel; n = 34 cells, N = 3 mice), Sc shRNA (middle panel; n = 26 cells, N = 3 mice) or TWIK-1 shRNA (right panel; n = 31 cells, N = 3 mice) expressing dentate granule cells in the presence of Cs + /TEA. Whole-cell currents were elicited by 1 s duration ramp pulses descending from 40 mV to -150 mV from a holding potential of -70 mV. (C) Summary bar charts for (B) . Shown are mean values of whole-cell current density in naïve, Sc shRNA or TWIK-1 shRNA expressing granule cells measured after subsequent application of the blockers; current density values are depicted at -150 mV (shadowed bars below baseline) and 40 mV (open bars) . (D) Reversal potential values of the whole-cell currents in naïve (n = 34 cells, N = 3 mice), Sc shRNA (n = 26 cells, N = 3 mice) or TWIK-1 shRNA (n = 31 cells, N = 3 mice) expressing dentate granule cells. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.

    Article Snippet: Tissues were incubated overnight with primary antibodies, such as rabbit anti-TWIK-1 polyclonal antibody (1:100; Alomone, Jerusalem, Israel) or chicken anti-MAP2 polyclonal antibody (1:1000; Abcam, Cambridge, MA), at 4°C.

    Techniques: shRNA, Expressing

    TWIK-1 deficiency causes enhanced intrinsic excitability of dentate granule cells. (A) Representative response of membrane potential to stepwise current injections (left panel). The current was injected into cells in 25 pA steps starting from -120 pA and up to 55 pA (1.2 sec step duration). Input-output properties of naïve (n = 36 cells), Sc shRNA (n = 32 cells) or TWIK-1 shRNA (n = 30 cells) expressing granule cells measured as the number of spikes vs. injected current intensity (right panel). (B) Distribution of cells according to excitability patterns. Plotted are percentage of cells with binned number of spikes fired during a 30 pA injected current step. (C) Representative response of membrane potential to stepwise current injections (left panel). Averaged response of membrane potential to stepwise current injection in naïve (n = 27 cells), Sc shRNA (n = 20 cells) or TWIK-1 shRNA (n = 21 cells) expressing cells (right panel). The RMP of cells was maintained at -70 mV. Current injection into the cell body was performed stepwise from -30 pA to 90 pA, in 5 pA steps. The solid lines are an exponential fit of the data plots. Dotted line indicates the spiking threshold level. (D) Representative traces of rheobase current measurements (left panel). The RMP of cells was kept at -70 mV and then depolarizing current was injected stepwise, in 2 pA steps until the membrane potential reached the firing threshold (red traces) . Averaged values of rheobase currents in naïve (n = 12 cells), Sc shRNA (n = 13 cells) or TWIK-1 shRNA (n = 12 cells) expressing cells (right panel). All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.

    Journal: Molecular Brain

    Article Title: TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus

    doi: 10.1186/s13041-014-0080-z

    Figure Lengend Snippet: TWIK-1 deficiency causes enhanced intrinsic excitability of dentate granule cells. (A) Representative response of membrane potential to stepwise current injections (left panel). The current was injected into cells in 25 pA steps starting from -120 pA and up to 55 pA (1.2 sec step duration). Input-output properties of naïve (n = 36 cells), Sc shRNA (n = 32 cells) or TWIK-1 shRNA (n = 30 cells) expressing granule cells measured as the number of spikes vs. injected current intensity (right panel). (B) Distribution of cells according to excitability patterns. Plotted are percentage of cells with binned number of spikes fired during a 30 pA injected current step. (C) Representative response of membrane potential to stepwise current injections (left panel). Averaged response of membrane potential to stepwise current injection in naïve (n = 27 cells), Sc shRNA (n = 20 cells) or TWIK-1 shRNA (n = 21 cells) expressing cells (right panel). The RMP of cells was maintained at -70 mV. Current injection into the cell body was performed stepwise from -30 pA to 90 pA, in 5 pA steps. The solid lines are an exponential fit of the data plots. Dotted line indicates the spiking threshold level. (D) Representative traces of rheobase current measurements (left panel). The RMP of cells was kept at -70 mV and then depolarizing current was injected stepwise, in 2 pA steps until the membrane potential reached the firing threshold (red traces) . Averaged values of rheobase currents in naïve (n = 12 cells), Sc shRNA (n = 13 cells) or TWIK-1 shRNA (n = 12 cells) expressing cells (right panel). All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.

    Article Snippet: Tissues were incubated overnight with primary antibodies, such as rabbit anti-TWIK-1 polyclonal antibody (1:100; Alomone, Jerusalem, Israel) or chicken anti-MAP2 polyclonal antibody (1:1000; Abcam, Cambridge, MA), at 4°C.

    Techniques: Injection, shRNA, Expressing

    TWIK-1 deficiency in dentate granule cells causes enhanced amplitude of evoked EPSPs in response to perforant path stimulation. (A) ( a ) Schematic diagram for synaptically evoked responsiveness of dentate granule cells. PP: perforant path, Stim: stimulation, Rec: recording. ( b ) Representative traces of evoked EPSPs in naïve, Sc shRNA or TWIK-1 shRNA expressing dentate granule cells. Synaptic responses evoked by 150 μA stimulation of perforant path are shown. ( c ) Input-output properties of dentate granule cells. Plotted values are of evoked EPSP amplitudes in naïve (n = 13cells, N = 3 mice), Sc shRNA (n = 14 cells, N = 3 mice) or TWIK-1 shRNA (n = 12 cells, N = 3 mice) expressing granule cells. The RMP was maintained at -80 mV by steady current injection. For each stimulation intensity, ten perforant path stimuli were applied and the evoked EPSP responses were averaged. The solid line is a logistic fit of the data plots. ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. ( B ) Representative traces for paired-pulse ratio measurements (left panel). Paired-pulse ratios in perforant path to granule cell synaptic transmission as a function of interstimulus interval (ISI) in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing cells (right panel). ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.

    Journal: Molecular Brain

    Article Title: TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus

    doi: 10.1186/s13041-014-0080-z

    Figure Lengend Snippet: TWIK-1 deficiency in dentate granule cells causes enhanced amplitude of evoked EPSPs in response to perforant path stimulation. (A) ( a ) Schematic diagram for synaptically evoked responsiveness of dentate granule cells. PP: perforant path, Stim: stimulation, Rec: recording. ( b ) Representative traces of evoked EPSPs in naïve, Sc shRNA or TWIK-1 shRNA expressing dentate granule cells. Synaptic responses evoked by 150 μA stimulation of perforant path are shown. ( c ) Input-output properties of dentate granule cells. Plotted values are of evoked EPSP amplitudes in naïve (n = 13cells, N = 3 mice), Sc shRNA (n = 14 cells, N = 3 mice) or TWIK-1 shRNA (n = 12 cells, N = 3 mice) expressing granule cells. The RMP was maintained at -80 mV by steady current injection. For each stimulation intensity, ten perforant path stimuli were applied and the evoked EPSP responses were averaged. The solid line is a logistic fit of the data plots. ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. ( B ) Representative traces for paired-pulse ratio measurements (left panel). Paired-pulse ratios in perforant path to granule cell synaptic transmission as a function of interstimulus interval (ISI) in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing cells (right panel). ACSF contained 10 μM bicuculline and 10 μM CGP 55845; pipette solution contained 5 mM QX-314. All data are represented as mean ± S.E.M; *P < 0.05, Student’s unpaired t -test.

    Article Snippet: Tissues were incubated overnight with primary antibodies, such as rabbit anti-TWIK-1 polyclonal antibody (1:100; Alomone, Jerusalem, Israel) or chicken anti-MAP2 polyclonal antibody (1:1000; Abcam, Cambridge, MA), at 4°C.

    Techniques: shRNA, Expressing, Injection, Transferring, Transmission Assay

    TWIK-1 knockdown in dentate granule cells affects signal input-output properties of the cells. (A) Representative traces from spiking probability measurements (left panel). Synaptic responses evoked by 200 μA stimulation of perforant path are shown. Input-output properties of dentate granule cells (right panel). Plotted are the values of spiking probability in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing dentate granule cells. The solid line is a logistic fit of the data plots. (B) Representative traces from spiking probability measurements (left panel). Traces with multiple spikes are shown in red. Averaged input-output properties of the subpopulation of the burst discharging cells (right panel). Plotted are the averaged values of probability of firing more than one spike per eEPSP in TWIK-1 shRNA or Sc shRNA expressing cells. (C) Representative traces of ten evoked EPSP responses in Sc shRNA and TWIK-1 shRNA expressing dentate granule cells (stimulation intensity was 350 μA) (left panel). The dotted lines indicate RMP, the voltage threshold for the first AP (V th1 ) and the voltage threshold for the second AP (V th2 ) in the burst response. Traces containing the second spike are shown in red. Summary bar graph for averaged depolarization level after the termination of the first AP in Sc shRNA and a population of TWIK-1 shRNA expressing cells with multiple spikes (right panel). The dotted lines show the threshold levels for the first (V th1 ) and the second (V th2 ) AP during a single EPSP. Number of cells are shown within the bars. All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.

    Journal: Molecular Brain

    Article Title: TWIK-1 contributes to the intrinsic excitability of dentate granule cells in mouse hippocampus

    doi: 10.1186/s13041-014-0080-z

    Figure Lengend Snippet: TWIK-1 knockdown in dentate granule cells affects signal input-output properties of the cells. (A) Representative traces from spiking probability measurements (left panel). Synaptic responses evoked by 200 μA stimulation of perforant path are shown. Input-output properties of dentate granule cells (right panel). Plotted are the values of spiking probability in naïve (n = 16 cells, N = 3 mice), Sc shRNA (n = 16 cells, N = 3 mice) or TWIK-1 shRNA (n = 16 cells, N = 3 mice) expressing dentate granule cells. The solid line is a logistic fit of the data plots. (B) Representative traces from spiking probability measurements (left panel). Traces with multiple spikes are shown in red. Averaged input-output properties of the subpopulation of the burst discharging cells (right panel). Plotted are the averaged values of probability of firing more than one spike per eEPSP in TWIK-1 shRNA or Sc shRNA expressing cells. (C) Representative traces of ten evoked EPSP responses in Sc shRNA and TWIK-1 shRNA expressing dentate granule cells (stimulation intensity was 350 μA) (left panel). The dotted lines indicate RMP, the voltage threshold for the first AP (V th1 ) and the voltage threshold for the second AP (V th2 ) in the burst response. Traces containing the second spike are shown in red. Summary bar graph for averaged depolarization level after the termination of the first AP in Sc shRNA and a population of TWIK-1 shRNA expressing cells with multiple spikes (right panel). The dotted lines show the threshold levels for the first (V th1 ) and the second (V th2 ) AP during a single EPSP. Number of cells are shown within the bars. All data are represented as mean ± S.E.M; *P < 0.05, **P < 0.01 Student’s unpaired t -test.

    Article Snippet: Tissues were incubated overnight with primary antibodies, such as rabbit anti-TWIK-1 polyclonal antibody (1:100; Alomone, Jerusalem, Israel) or chicken anti-MAP2 polyclonal antibody (1:1000; Abcam, Cambridge, MA), at 4°C.

    Techniques: shRNA, Expressing