guinea pig anti kir4 1  (Alomone Labs)


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    Name:
    Guinea pig Anti Kir4 1 KCNJ10 Antibody
    Description:
    Guinea pig Anti Kir4 1 KCNJ10 Antibody AGP 012 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize Kir4 1 potassium channel from human rat and mouse samples The antigen used to immunize guinea pigs is the same as Anti Kir4 1 KCNJ10 Antibody APC 035 raised in rabbit Our line of guinea pig antibodies enables more flexibility with our products such as multiplex staining studies immunoprecipitation etc
    Catalog Number:
    AGP-012
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Guinea pig
    Isotype:
    Guinea pig total IgG
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    Structured Review

    Alomone Labs guinea pig anti kir4 1
    Guinea pig Anti Kir4 1 KCNJ10 Antibody
    Guinea pig Anti Kir4 1 KCNJ10 Antibody AGP 012 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunohistochemistry and immunocytochemistry applications It has been designed to recognize Kir4 1 potassium channel from human rat and mouse samples The antigen used to immunize guinea pigs is the same as Anti Kir4 1 KCNJ10 Antibody APC 035 raised in rabbit Our line of guinea pig antibodies enables more flexibility with our products such as multiplex staining studies immunoprecipitation etc
    https://www.bioz.com/result/guinea pig anti kir4 1/product/Alomone Labs
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    guinea pig anti kir4 1 - by Bioz Stars, 2021-09
    88/100 stars

    Images

    1) Product Images from "Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS"

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    Journal: Brain Structure & Function

    doi: 10.1007/s00429-016-1199-8

    Glial Kir5.1 expression is reduced in the absence of Kir4.1 subunit. Immunolabelling for Kir5.1 was determined in optic nerve explants cultures, comparing wild-type mice ( A , Kir4.1 +/+ ) with Kir4.1 knock-out mice ( B , Kir4.1 −/− ), and following transfection with scrambled shRNA ( C ) or Kir4.1 shRNA ( D ); transfected cells were identified by the expression of GFP (appears green ) and insets demonstrate Kir4.1 expression in controls ( Ai , Ci ) and complete ablation in Kir4.1 −/− mice ( Bi ) and Kir4.1 shRNA ( Di ). Scale bars 10 μm. Quantification of expression of Kir4.1 ( E ) and Kir5.1 ( F ) in Kir4.1 +/+ , Kir4.1 −/− , scrambled control and Kir4.1shRNA glia; analysis was performed on 10–12 cells in each group, and data are expressed as mean ± SEM number of voxels per µm 3 , *** p
    Figure Legend Snippet: Glial Kir5.1 expression is reduced in the absence of Kir4.1 subunit. Immunolabelling for Kir5.1 was determined in optic nerve explants cultures, comparing wild-type mice ( A , Kir4.1 +/+ ) with Kir4.1 knock-out mice ( B , Kir4.1 −/− ), and following transfection with scrambled shRNA ( C ) or Kir4.1 shRNA ( D ); transfected cells were identified by the expression of GFP (appears green ) and insets demonstrate Kir4.1 expression in controls ( Ai , Ci ) and complete ablation in Kir4.1 −/− mice ( Bi ) and Kir4.1 shRNA ( Di ). Scale bars 10 μm. Quantification of expression of Kir4.1 ( E ) and Kir5.1 ( F ) in Kir4.1 +/+ , Kir4.1 −/− , scrambled control and Kir4.1shRNA glia; analysis was performed on 10–12 cells in each group, and data are expressed as mean ± SEM number of voxels per µm 3 , *** p

    Techniques Used: Expressing, Mouse Assay, Knock-Out, Transfection, shRNA

    Functional implications of homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels in oligodendrocytes. Oligodendroglial expression of Kir4.1 channels indicates they may be important in uptake of excess K + released during axonal action potential propagation, a function largely attribiuted to astrocytes. Due to their wrapping of axons, oligodendrocytes are exposed to large ionic and pH shifts during axonal electrical activity, and it is likely weakly rectifying homomeric Kir4.1 and strongly rectifying Kir4.1/Kir5.1 heteromeric channels are important in maintaining the negative resting membrane potential, which is essential for oligodendroglial and myelin integrity. Weakly rectifying homomeric Kir4.1 channels may preferentially extrude K + and supply extracellular K + for the Na + –K + -pumps, as described in transporting epithelia. In contrast, the pH sensitivity of heteromeric Kir4.1/Kir5.1 channels is likely to have a role in the CO 2 /pH chemosensation in glia, involving carbonic anhydrase that is enriched in astrocytes and oligodendrocytes. Furthermore, intracellular acidification and inhibition of Kir4.1/Kir5.1 channels has been shown to trigger release of ATP from astrocytes, which would act on oligodendroglial P2X and P2Y receptors to provide a mechanism of astrocyte–oligodendrocyte signaling in response to metabolic challenges, which has important implications for white matter physiology and pathology
    Figure Legend Snippet: Functional implications of homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels in oligodendrocytes. Oligodendroglial expression of Kir4.1 channels indicates they may be important in uptake of excess K + released during axonal action potential propagation, a function largely attribiuted to astrocytes. Due to their wrapping of axons, oligodendrocytes are exposed to large ionic and pH shifts during axonal electrical activity, and it is likely weakly rectifying homomeric Kir4.1 and strongly rectifying Kir4.1/Kir5.1 heteromeric channels are important in maintaining the negative resting membrane potential, which is essential for oligodendroglial and myelin integrity. Weakly rectifying homomeric Kir4.1 channels may preferentially extrude K + and supply extracellular K + for the Na + –K + -pumps, as described in transporting epithelia. In contrast, the pH sensitivity of heteromeric Kir4.1/Kir5.1 channels is likely to have a role in the CO 2 /pH chemosensation in glia, involving carbonic anhydrase that is enriched in astrocytes and oligodendrocytes. Furthermore, intracellular acidification and inhibition of Kir4.1/Kir5.1 channels has been shown to trigger release of ATP from astrocytes, which would act on oligodendroglial P2X and P2Y receptors to provide a mechanism of astrocyte–oligodendrocyte signaling in response to metabolic challenges, which has important implications for white matter physiology and pathology

    Techniques Used: Functional Assay, Expressing, Activity Assay, Inhibition

    Specific reduction in plasmalemmal Kir5.1 in the absence of Kir4.1. Immunolocalization of Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explant astrocytes identified by expression of GFAP, following transfection with scrambled shRNA ( A ) or Kir4.1 shRNA ( B ); transfected cells were identified by co-transfection with GFP (appears green ) and the co-localization channel indicates voxels in which Kir5.1 and Na–K-ATPase immunolabelling was at the same intensity ( Avi , Bvi ). Scale bars 20 μm. C Quantification of plasmalemmal Kir5.1 expressed as percentage of total Kir5.1 + voxels (data are mean ± SEM, n = 11–13 per group; * p
    Figure Legend Snippet: Specific reduction in plasmalemmal Kir5.1 in the absence of Kir4.1. Immunolocalization of Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explant astrocytes identified by expression of GFAP, following transfection with scrambled shRNA ( A ) or Kir4.1 shRNA ( B ); transfected cells were identified by co-transfection with GFP (appears green ) and the co-localization channel indicates voxels in which Kir5.1 and Na–K-ATPase immunolabelling was at the same intensity ( Avi , Bvi ). Scale bars 20 μm. C Quantification of plasmalemmal Kir5.1 expressed as percentage of total Kir5.1 + voxels (data are mean ± SEM, n = 11–13 per group; * p

    Techniques Used: Expressing, Transfection, shRNA, Cotransfection

    Reduction of Kir5.1 in oligodendrocytes and myelin in the absence of Kir4.1. Immunolocalization of Kir5.1 with myelin basic protein, MBP ( A , B ) and the oligodenrocyte marker APC/CC1 ( C – F ), in brain tissue from wild-type Kir4.1 +/+ mice ( A , C , E ) compared to Kir4.1 −/− knock-out mice ( B , D , F ). Scale bars 20 μm. Western blot analysis of Kir5.1 from total lysates of optic nerve ( G ) and brain ( H ) from wild-type Kir4.1 +/+ and Kir4.1 −/− knock-out mice, and mean (±SEM) integrated density normalised against β-actin ( I , n = 3, ** p
    Figure Legend Snippet: Reduction of Kir5.1 in oligodendrocytes and myelin in the absence of Kir4.1. Immunolocalization of Kir5.1 with myelin basic protein, MBP ( A , B ) and the oligodenrocyte marker APC/CC1 ( C – F ), in brain tissue from wild-type Kir4.1 +/+ mice ( A , C , E ) compared to Kir4.1 −/− knock-out mice ( B , D , F ). Scale bars 20 μm. Western blot analysis of Kir5.1 from total lysates of optic nerve ( G ) and brain ( H ) from wild-type Kir4.1 +/+ and Kir4.1 −/− knock-out mice, and mean (±SEM) integrated density normalised against β-actin ( I , n = 3, ** p

    Techniques Used: Marker, Mouse Assay, Knock-Out, Western Blot

    Expression of Kir4.1 and Kir5.1 in oligodendrocytes and astrocytes in the cerebellum. Immunolabelling for Kir4.1 and Kir5.1, in combination with GFAP for astrocytes ( A , C ), and APC/CC1 for oligodendrocytes ( B , D ). Immunolabelling for Kir4.1 ( E ) and Kir5.1 ( F ) in mice in which EGFP is under the control of the oligodendrocyte-specific Sox10 promoter. G Double immunolabelling for Kir4.1 ( red ) and the oligodenrocyte-specific marker Olig2 ( green ). Insets in Aiv and Civ illustrate negative controls, in the Kir4.1 KO mouse ( Aiv ) and following preincubation with the Kir5.1 blocking peptide ( Civ ). Scale bars 20 μm. Western blot analysis of the brain and optic and nerve for Kir4.1 ( I ) and Kir5.1 ( J ); bands were absent in the negative controls, in the Kir4.1 knock-out mouse ( I ) following preincubation in the Kir5.1 blocking peptide ( J )
    Figure Legend Snippet: Expression of Kir4.1 and Kir5.1 in oligodendrocytes and astrocytes in the cerebellum. Immunolabelling for Kir4.1 and Kir5.1, in combination with GFAP for astrocytes ( A , C ), and APC/CC1 for oligodendrocytes ( B , D ). Immunolabelling for Kir4.1 ( E ) and Kir5.1 ( F ) in mice in which EGFP is under the control of the oligodendrocyte-specific Sox10 promoter. G Double immunolabelling for Kir4.1 ( red ) and the oligodenrocyte-specific marker Olig2 ( green ). Insets in Aiv and Civ illustrate negative controls, in the Kir4.1 KO mouse ( Aiv ) and following preincubation with the Kir5.1 blocking peptide ( Civ ). Scale bars 20 μm. Western blot analysis of the brain and optic and nerve for Kir4.1 ( I ) and Kir5.1 ( J ); bands were absent in the negative controls, in the Kir4.1 knock-out mouse ( I ) following preincubation in the Kir5.1 blocking peptide ( J )

    Techniques Used: Expressing, Mouse Assay, Marker, Blocking Assay, Western Blot, Knock-Out

    Expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Immunolabelling for Kir4.1 ( A , C ) and Kir5.1 ( B , D ), in GFAP-GFP mice to identify astrocytes ( A , B ) and PLP-DsRED mice to identify oligodendrocytes ( C , D ). Cellular expression of Kir4.1 and Kir5.1 is demonstrated by the generation of colocalisation channels ( Av , Bv , Cv , Dv ) from confocal z -stacks ( Aiv , Biv , Civ , Div ), and green and red channels of equal intensity appear yellow . Scale bars 20 μm
    Figure Legend Snippet: Expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Immunolabelling for Kir4.1 ( A , C ) and Kir5.1 ( B , D ), in GFAP-GFP mice to identify astrocytes ( A , B ) and PLP-DsRED mice to identify oligodendrocytes ( C , D ). Cellular expression of Kir4.1 and Kir5.1 is demonstrated by the generation of colocalisation channels ( Av , Bv , Cv , Dv ) from confocal z -stacks ( Aiv , Biv , Civ , Div ), and green and red channels of equal intensity appear yellow . Scale bars 20 μm

    Techniques Used: Expressing, Mouse Assay, Plasmid Purification

    Co-expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Co-immunolocalization of Kir4.1 and Kir5.1 in optic nerve explant cultures, in astrocytes identified by GFAP immunolabelling ( A ) and oligodendrocytes identified by PLP-DsRED ( B ). The overlay and individual channels are illustrated, together with the co-localisation channel for Kir4.1/Kir5.1 ( Aii, Bii ). Boxed areas on overlay images ( Ai , Bi ) are enlarged in Avi – Aviii and Bvi – Bviii , to illustrate punctate colocalization of Kir4.1 and Kir5.1 along processes (some indicated by arrows ). Scale bars 20 μm. Quantification of the number of voxels that were positive for Kir4.1 and Kir5.1 alone and of Kir4.1/Kir5.1 together, in astrocytes ( C , n = 15) and oligodendrocytes ( D , n = 13); data are mean ± SEM. Co-immunoprecipitation of Kir4.1 with Kir5.1 ( E ) and of Kir5.1 with Kir4.1 ( F ) from total brain and optic nerve (ON) lysates; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1, and using the blocking peptide for Kir5.1
    Figure Legend Snippet: Co-expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Co-immunolocalization of Kir4.1 and Kir5.1 in optic nerve explant cultures, in astrocytes identified by GFAP immunolabelling ( A ) and oligodendrocytes identified by PLP-DsRED ( B ). The overlay and individual channels are illustrated, together with the co-localisation channel for Kir4.1/Kir5.1 ( Aii, Bii ). Boxed areas on overlay images ( Ai , Bi ) are enlarged in Avi – Aviii and Bvi – Bviii , to illustrate punctate colocalization of Kir4.1 and Kir5.1 along processes (some indicated by arrows ). Scale bars 20 μm. Quantification of the number of voxels that were positive for Kir4.1 and Kir5.1 alone and of Kir4.1/Kir5.1 together, in astrocytes ( C , n = 15) and oligodendrocytes ( D , n = 13); data are mean ± SEM. Co-immunoprecipitation of Kir4.1 with Kir5.1 ( E ) and of Kir5.1 with Kir4.1 ( F ) from total brain and optic nerve (ON) lysates; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1, and using the blocking peptide for Kir5.1

    Techniques Used: Expressing, Plasmid Purification, Immunoprecipitation, Knock-Out, Mouse Assay, Blocking Assay

    Plasmalemmal expression of Kir4.1 and Kir5.1 subunit in optic nerve glia. Immunolocalization of Kir4.1 and Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explants of astrocytes identified by GFAP ( A , B ) and oligodendrocytes identified by PLP-DsRed ( C , D ). Scale bars 20 μm. Quantification in astrocytes and oligodendrocytes of total number of voxels immunopositive for Kir4.1 and Kir5.1, compared to voxels that were identified as colocalized for Kir4.1/Na–K-ATPase ( E ) and Kir5.1/Na–K-ATPase ( F ); data are mean ± SEM, n = 13 cells for each analysis. Western blot analysis of Kir5.1 ( G ) and Kir4.1 ( H ) in total optic nerve lysate and plasma membrane fraction. Co-immunoprecipitation of Kir4.1 ( I ) and Kir5.1 ( J ) with PSD95, in total brain and optic nerve (ON) lysate; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1 and preincubation with the blocking peptide for Kir5.1
    Figure Legend Snippet: Plasmalemmal expression of Kir4.1 and Kir5.1 subunit in optic nerve glia. Immunolocalization of Kir4.1 and Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explants of astrocytes identified by GFAP ( A , B ) and oligodendrocytes identified by PLP-DsRed ( C , D ). Scale bars 20 μm. Quantification in astrocytes and oligodendrocytes of total number of voxels immunopositive for Kir4.1 and Kir5.1, compared to voxels that were identified as colocalized for Kir4.1/Na–K-ATPase ( E ) and Kir5.1/Na–K-ATPase ( F ); data are mean ± SEM, n = 13 cells for each analysis. Western blot analysis of Kir5.1 ( G ) and Kir4.1 ( H ) in total optic nerve lysate and plasma membrane fraction. Co-immunoprecipitation of Kir4.1 ( I ) and Kir5.1 ( J ) with PSD95, in total brain and optic nerve (ON) lysate; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1 and preincubation with the blocking peptide for Kir5.1

    Techniques Used: Expressing, Plasmid Purification, Western Blot, Immunoprecipitation, Knock-Out, Mouse Assay, Blocking Assay

    Related Articles

    Blocking Assay:

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS
    Article Snippet: .. After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo). ..

    Incubation:

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS
    Article Snippet: .. After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo). ..

    Next-Generation Sequencing:

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS
    Article Snippet: .. After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo). ..

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    Alomone Labs guinea pig anti kir4 1
    Glial Kir5.1 expression is reduced in the absence of <t>Kir4.1</t> subunit. Immunolabelling for Kir5.1 was determined in optic nerve explants cultures, comparing wild-type mice ( A , Kir4.1 +/+ ) with Kir4.1 knock-out mice ( B , Kir4.1 −/− ), and following transfection with scrambled shRNA ( C ) or Kir4.1 shRNA ( D ); transfected cells were identified by the expression of GFP (appears green ) and insets demonstrate Kir4.1 expression in controls ( Ai , Ci ) and complete ablation in Kir4.1 −/− mice ( Bi ) and Kir4.1 shRNA ( Di ). Scale bars 10 μm. Quantification of expression of Kir4.1 ( E ) and Kir5.1 ( F ) in Kir4.1 +/+ , Kir4.1 −/− , scrambled control and Kir4.1shRNA glia; analysis was performed on 10–12 cells in each group, and data are expressed as mean ± SEM number of voxels per µm 3 , *** p
    Guinea Pig Anti Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/guinea pig anti kir4 1/product/Alomone Labs
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    guinea pig anti kir4 1 - by Bioz Stars, 2021-09
    88/100 stars
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    Glial Kir5.1 expression is reduced in the absence of Kir4.1 subunit. Immunolabelling for Kir5.1 was determined in optic nerve explants cultures, comparing wild-type mice ( A , Kir4.1 +/+ ) with Kir4.1 knock-out mice ( B , Kir4.1 −/− ), and following transfection with scrambled shRNA ( C ) or Kir4.1 shRNA ( D ); transfected cells were identified by the expression of GFP (appears green ) and insets demonstrate Kir4.1 expression in controls ( Ai , Ci ) and complete ablation in Kir4.1 −/− mice ( Bi ) and Kir4.1 shRNA ( Di ). Scale bars 10 μm. Quantification of expression of Kir4.1 ( E ) and Kir5.1 ( F ) in Kir4.1 +/+ , Kir4.1 −/− , scrambled control and Kir4.1shRNA glia; analysis was performed on 10–12 cells in each group, and data are expressed as mean ± SEM number of voxels per µm 3 , *** p

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Glial Kir5.1 expression is reduced in the absence of Kir4.1 subunit. Immunolabelling for Kir5.1 was determined in optic nerve explants cultures, comparing wild-type mice ( A , Kir4.1 +/+ ) with Kir4.1 knock-out mice ( B , Kir4.1 −/− ), and following transfection with scrambled shRNA ( C ) or Kir4.1 shRNA ( D ); transfected cells were identified by the expression of GFP (appears green ) and insets demonstrate Kir4.1 expression in controls ( Ai , Ci ) and complete ablation in Kir4.1 −/− mice ( Bi ) and Kir4.1 shRNA ( Di ). Scale bars 10 μm. Quantification of expression of Kir4.1 ( E ) and Kir5.1 ( F ) in Kir4.1 +/+ , Kir4.1 −/− , scrambled control and Kir4.1shRNA glia; analysis was performed on 10–12 cells in each group, and data are expressed as mean ± SEM number of voxels per µm 3 , *** p

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Expressing, Mouse Assay, Knock-Out, Transfection, shRNA

    Functional implications of homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels in oligodendrocytes. Oligodendroglial expression of Kir4.1 channels indicates they may be important in uptake of excess K + released during axonal action potential propagation, a function largely attribiuted to astrocytes. Due to their wrapping of axons, oligodendrocytes are exposed to large ionic and pH shifts during axonal electrical activity, and it is likely weakly rectifying homomeric Kir4.1 and strongly rectifying Kir4.1/Kir5.1 heteromeric channels are important in maintaining the negative resting membrane potential, which is essential for oligodendroglial and myelin integrity. Weakly rectifying homomeric Kir4.1 channels may preferentially extrude K + and supply extracellular K + for the Na + –K + -pumps, as described in transporting epithelia. In contrast, the pH sensitivity of heteromeric Kir4.1/Kir5.1 channels is likely to have a role in the CO 2 /pH chemosensation in glia, involving carbonic anhydrase that is enriched in astrocytes and oligodendrocytes. Furthermore, intracellular acidification and inhibition of Kir4.1/Kir5.1 channels has been shown to trigger release of ATP from astrocytes, which would act on oligodendroglial P2X and P2Y receptors to provide a mechanism of astrocyte–oligodendrocyte signaling in response to metabolic challenges, which has important implications for white matter physiology and pathology

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Functional implications of homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels in oligodendrocytes. Oligodendroglial expression of Kir4.1 channels indicates they may be important in uptake of excess K + released during axonal action potential propagation, a function largely attribiuted to astrocytes. Due to their wrapping of axons, oligodendrocytes are exposed to large ionic and pH shifts during axonal electrical activity, and it is likely weakly rectifying homomeric Kir4.1 and strongly rectifying Kir4.1/Kir5.1 heteromeric channels are important in maintaining the negative resting membrane potential, which is essential for oligodendroglial and myelin integrity. Weakly rectifying homomeric Kir4.1 channels may preferentially extrude K + and supply extracellular K + for the Na + –K + -pumps, as described in transporting epithelia. In contrast, the pH sensitivity of heteromeric Kir4.1/Kir5.1 channels is likely to have a role in the CO 2 /pH chemosensation in glia, involving carbonic anhydrase that is enriched in astrocytes and oligodendrocytes. Furthermore, intracellular acidification and inhibition of Kir4.1/Kir5.1 channels has been shown to trigger release of ATP from astrocytes, which would act on oligodendroglial P2X and P2Y receptors to provide a mechanism of astrocyte–oligodendrocyte signaling in response to metabolic challenges, which has important implications for white matter physiology and pathology

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Functional Assay, Expressing, Activity Assay, Inhibition

    Specific reduction in plasmalemmal Kir5.1 in the absence of Kir4.1. Immunolocalization of Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explant astrocytes identified by expression of GFAP, following transfection with scrambled shRNA ( A ) or Kir4.1 shRNA ( B ); transfected cells were identified by co-transfection with GFP (appears green ) and the co-localization channel indicates voxels in which Kir5.1 and Na–K-ATPase immunolabelling was at the same intensity ( Avi , Bvi ). Scale bars 20 μm. C Quantification of plasmalemmal Kir5.1 expressed as percentage of total Kir5.1 + voxels (data are mean ± SEM, n = 11–13 per group; * p

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Specific reduction in plasmalemmal Kir5.1 in the absence of Kir4.1. Immunolocalization of Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explant astrocytes identified by expression of GFAP, following transfection with scrambled shRNA ( A ) or Kir4.1 shRNA ( B ); transfected cells were identified by co-transfection with GFP (appears green ) and the co-localization channel indicates voxels in which Kir5.1 and Na–K-ATPase immunolabelling was at the same intensity ( Avi , Bvi ). Scale bars 20 μm. C Quantification of plasmalemmal Kir5.1 expressed as percentage of total Kir5.1 + voxels (data are mean ± SEM, n = 11–13 per group; * p

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Expressing, Transfection, shRNA, Cotransfection

    Reduction of Kir5.1 in oligodendrocytes and myelin in the absence of Kir4.1. Immunolocalization of Kir5.1 with myelin basic protein, MBP ( A , B ) and the oligodenrocyte marker APC/CC1 ( C – F ), in brain tissue from wild-type Kir4.1 +/+ mice ( A , C , E ) compared to Kir4.1 −/− knock-out mice ( B , D , F ). Scale bars 20 μm. Western blot analysis of Kir5.1 from total lysates of optic nerve ( G ) and brain ( H ) from wild-type Kir4.1 +/+ and Kir4.1 −/− knock-out mice, and mean (±SEM) integrated density normalised against β-actin ( I , n = 3, ** p

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Reduction of Kir5.1 in oligodendrocytes and myelin in the absence of Kir4.1. Immunolocalization of Kir5.1 with myelin basic protein, MBP ( A , B ) and the oligodenrocyte marker APC/CC1 ( C – F ), in brain tissue from wild-type Kir4.1 +/+ mice ( A , C , E ) compared to Kir4.1 −/− knock-out mice ( B , D , F ). Scale bars 20 μm. Western blot analysis of Kir5.1 from total lysates of optic nerve ( G ) and brain ( H ) from wild-type Kir4.1 +/+ and Kir4.1 −/− knock-out mice, and mean (±SEM) integrated density normalised against β-actin ( I , n = 3, ** p

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Marker, Mouse Assay, Knock-Out, Western Blot

    Expression of Kir4.1 and Kir5.1 in oligodendrocytes and astrocytes in the cerebellum. Immunolabelling for Kir4.1 and Kir5.1, in combination with GFAP for astrocytes ( A , C ), and APC/CC1 for oligodendrocytes ( B , D ). Immunolabelling for Kir4.1 ( E ) and Kir5.1 ( F ) in mice in which EGFP is under the control of the oligodendrocyte-specific Sox10 promoter. G Double immunolabelling for Kir4.1 ( red ) and the oligodenrocyte-specific marker Olig2 ( green ). Insets in Aiv and Civ illustrate negative controls, in the Kir4.1 KO mouse ( Aiv ) and following preincubation with the Kir5.1 blocking peptide ( Civ ). Scale bars 20 μm. Western blot analysis of the brain and optic and nerve for Kir4.1 ( I ) and Kir5.1 ( J ); bands were absent in the negative controls, in the Kir4.1 knock-out mouse ( I ) following preincubation in the Kir5.1 blocking peptide ( J )

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Expression of Kir4.1 and Kir5.1 in oligodendrocytes and astrocytes in the cerebellum. Immunolabelling for Kir4.1 and Kir5.1, in combination with GFAP for astrocytes ( A , C ), and APC/CC1 for oligodendrocytes ( B , D ). Immunolabelling for Kir4.1 ( E ) and Kir5.1 ( F ) in mice in which EGFP is under the control of the oligodendrocyte-specific Sox10 promoter. G Double immunolabelling for Kir4.1 ( red ) and the oligodenrocyte-specific marker Olig2 ( green ). Insets in Aiv and Civ illustrate negative controls, in the Kir4.1 KO mouse ( Aiv ) and following preincubation with the Kir5.1 blocking peptide ( Civ ). Scale bars 20 μm. Western blot analysis of the brain and optic and nerve for Kir4.1 ( I ) and Kir5.1 ( J ); bands were absent in the negative controls, in the Kir4.1 knock-out mouse ( I ) following preincubation in the Kir5.1 blocking peptide ( J )

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Expressing, Mouse Assay, Marker, Blocking Assay, Western Blot, Knock-Out

    Expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Immunolabelling for Kir4.1 ( A , C ) and Kir5.1 ( B , D ), in GFAP-GFP mice to identify astrocytes ( A , B ) and PLP-DsRED mice to identify oligodendrocytes ( C , D ). Cellular expression of Kir4.1 and Kir5.1 is demonstrated by the generation of colocalisation channels ( Av , Bv , Cv , Dv ) from confocal z -stacks ( Aiv , Biv , Civ , Div ), and green and red channels of equal intensity appear yellow . Scale bars 20 μm

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Immunolabelling for Kir4.1 ( A , C ) and Kir5.1 ( B , D ), in GFAP-GFP mice to identify astrocytes ( A , B ) and PLP-DsRED mice to identify oligodendrocytes ( C , D ). Cellular expression of Kir4.1 and Kir5.1 is demonstrated by the generation of colocalisation channels ( Av , Bv , Cv , Dv ) from confocal z -stacks ( Aiv , Biv , Civ , Div ), and green and red channels of equal intensity appear yellow . Scale bars 20 μm

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Expressing, Mouse Assay, Plasmid Purification

    Co-expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Co-immunolocalization of Kir4.1 and Kir5.1 in optic nerve explant cultures, in astrocytes identified by GFAP immunolabelling ( A ) and oligodendrocytes identified by PLP-DsRED ( B ). The overlay and individual channels are illustrated, together with the co-localisation channel for Kir4.1/Kir5.1 ( Aii, Bii ). Boxed areas on overlay images ( Ai , Bi ) are enlarged in Avi – Aviii and Bvi – Bviii , to illustrate punctate colocalization of Kir4.1 and Kir5.1 along processes (some indicated by arrows ). Scale bars 20 μm. Quantification of the number of voxels that were positive for Kir4.1 and Kir5.1 alone and of Kir4.1/Kir5.1 together, in astrocytes ( C , n = 15) and oligodendrocytes ( D , n = 13); data are mean ± SEM. Co-immunoprecipitation of Kir4.1 with Kir5.1 ( E ) and of Kir5.1 with Kir4.1 ( F ) from total brain and optic nerve (ON) lysates; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1, and using the blocking peptide for Kir5.1

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Co-expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Co-immunolocalization of Kir4.1 and Kir5.1 in optic nerve explant cultures, in astrocytes identified by GFAP immunolabelling ( A ) and oligodendrocytes identified by PLP-DsRED ( B ). The overlay and individual channels are illustrated, together with the co-localisation channel for Kir4.1/Kir5.1 ( Aii, Bii ). Boxed areas on overlay images ( Ai , Bi ) are enlarged in Avi – Aviii and Bvi – Bviii , to illustrate punctate colocalization of Kir4.1 and Kir5.1 along processes (some indicated by arrows ). Scale bars 20 μm. Quantification of the number of voxels that were positive for Kir4.1 and Kir5.1 alone and of Kir4.1/Kir5.1 together, in astrocytes ( C , n = 15) and oligodendrocytes ( D , n = 13); data are mean ± SEM. Co-immunoprecipitation of Kir4.1 with Kir5.1 ( E ) and of Kir5.1 with Kir4.1 ( F ) from total brain and optic nerve (ON) lysates; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1, and using the blocking peptide for Kir5.1

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Expressing, Plasmid Purification, Immunoprecipitation, Knock-Out, Mouse Assay, Blocking Assay

    Plasmalemmal expression of Kir4.1 and Kir5.1 subunit in optic nerve glia. Immunolocalization of Kir4.1 and Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explants of astrocytes identified by GFAP ( A , B ) and oligodendrocytes identified by PLP-DsRed ( C , D ). Scale bars 20 μm. Quantification in astrocytes and oligodendrocytes of total number of voxels immunopositive for Kir4.1 and Kir5.1, compared to voxels that were identified as colocalized for Kir4.1/Na–K-ATPase ( E ) and Kir5.1/Na–K-ATPase ( F ); data are mean ± SEM, n = 13 cells for each analysis. Western blot analysis of Kir5.1 ( G ) and Kir4.1 ( H ) in total optic nerve lysate and plasma membrane fraction. Co-immunoprecipitation of Kir4.1 ( I ) and Kir5.1 ( J ) with PSD95, in total brain and optic nerve (ON) lysate; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1 and preincubation with the blocking peptide for Kir5.1

    Journal: Brain Structure & Function

    Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS

    doi: 10.1007/s00429-016-1199-8

    Figure Lengend Snippet: Plasmalemmal expression of Kir4.1 and Kir5.1 subunit in optic nerve glia. Immunolocalization of Kir4.1 and Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explants of astrocytes identified by GFAP ( A , B ) and oligodendrocytes identified by PLP-DsRed ( C , D ). Scale bars 20 μm. Quantification in astrocytes and oligodendrocytes of total number of voxels immunopositive for Kir4.1 and Kir5.1, compared to voxels that were identified as colocalized for Kir4.1/Na–K-ATPase ( E ) and Kir5.1/Na–K-ATPase ( F ); data are mean ± SEM, n = 13 cells for each analysis. Western blot analysis of Kir5.1 ( G ) and Kir4.1 ( H ) in total optic nerve lysate and plasma membrane fraction. Co-immunoprecipitation of Kir4.1 ( I ) and Kir5.1 ( J ) with PSD95, in total brain and optic nerve (ON) lysate; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1 and preincubation with the blocking peptide for Kir5.1

    Article Snippet: After blocking, samples were incubated overnight at 4 °C with primary antibodies diluted in NGS-PBS: chicken anti-GFAP, 1:500 (Chemicon); mouse anti-APC (adenomatous polyposis coli) antibody [CC-1], 1:700 (Calbiochem); rabbit anti-Olig2, 1:700 (Millipore); rabbit anti-Kir4.1, 1:400 (Kalsi et al. ); guinea-pig anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir4.1, 1:300 (Alomone); rabbit anti-Kir5.1, 1:300 (Alomone); goat anti-Kir5.1, 1:100 (Santa Cruz); mouse anti-Na/K ATPase α1, 1:300 (Abcam); mouse anti-PSD-95, 1:300 (Thermo).

    Techniques: Expressing, Plasmid Purification, Western Blot, Immunoprecipitation, Knock-Out, Mouse Assay, Blocking Assay