primary rabbit anti glua1 antibody  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs primary rabbit anti glua1 antibody
    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on <t>GluA1</t> currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Primary Rabbit Anti Glua1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary rabbit anti glua1 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary rabbit anti glua1 antibody - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling"

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0033889

    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on GluA1 currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Figure Legend Snippet: ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on GluA1 currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.

    Techniques Used: Purification, Recombinant, Mutagenesis, Western Blot, Incubation, Stripping Membranes, Reverse Transcription Polymerase Chain Reaction, Expressing, Injection

    ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with SGK3, the inactive form of PIKfyve (PIKfyve(S318A)), or wild type PIKfyve. Numbers of oocytes are n = 20–30. Significant changes (p<0.001) are indicated by *** (p = 0.00049; 0.00036; 0.000073, respectively). ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.
    Figure Legend Snippet: ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with SGK3, the inactive form of PIKfyve (PIKfyve(S318A)), or wild type PIKfyve. Numbers of oocytes are n = 20–30. Significant changes (p<0.001) are indicated by *** (p = 0.00049; 0.00036; 0.000073, respectively). ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Techniques Used: Expressing, SDS-Gel, Western Blot, Molecular Weight, Software

    ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with Rab11, the dominant negative form of Rab11 (Rab11DN), PIKfyve, or SGK3. Numbers of oocytes are n = 15–30. Significant changes (p<0.001, p<0.05) are indicated by *** (p = 0.00015) and * (p = 0.036), respectively. ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.
    Figure Legend Snippet: ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with Rab11, the dominant negative form of Rab11 (Rab11DN), PIKfyve, or SGK3. Numbers of oocytes are n = 15–30. Significant changes (p<0.001, p<0.05) are indicated by *** (p = 0.00015) and * (p = 0.036), respectively. ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Techniques Used: Expressing, Dominant Negative Mutation, SDS-Gel, Western Blot, Molecular Weight, Software

    ( A ) GluA1 current amplitudes in oocytes before and after acute injection of a water-soluble analog of PI(3,5)P 2 . Significant change (p<0.05) is indicated by * (p = 0.0015). ( B ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with Rab11 or the dominant negative form of Rab11 (Rab11DN), before and after acute injection of a water-soluble analog of PI (3,5)P 2 . Numbers of oocytes are n = 7–28. Significant change (p<0.05) is indicated by * (p = 0.025). ( C ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with SGK3, myosin Vb, or the mutated form of myosin Vb (myosin del). Number of oocytes are n = 14–28.
    Figure Legend Snippet: ( A ) GluA1 current amplitudes in oocytes before and after acute injection of a water-soluble analog of PI(3,5)P 2 . Significant change (p<0.05) is indicated by * (p = 0.0015). ( B ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with Rab11 or the dominant negative form of Rab11 (Rab11DN), before and after acute injection of a water-soluble analog of PI (3,5)P 2 . Numbers of oocytes are n = 7–28. Significant change (p<0.05) is indicated by * (p = 0.025). ( C ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with SGK3, myosin Vb, or the mutated form of myosin Vb (myosin del). Number of oocytes are n = 14–28.

    Techniques Used: Injection, Expressing, Dominant Negative Mutation

    ( A , B ) Statistical analysis of GluA1 surface expression and colocalization of GluA1 with neuroligin revealed increased surface and synaptic GluA1 expression after treatment with NMDA. This effect is abolished by SGK or PIKfyve inhibition. Control = untreated, NMDA = incubated with NMDA, PIK-In = treated with PIKfyve inhibitor, SGK-In = treated with SGK inhibitor. (C–H) Representative confocal images of dendrites stained with GluA1 (green) and neuroligin (red –Neuro) in controls versus NMDA-treated neurons under different conditions: ( C ) control versus ( D ) NMDA, ( E ) PIKfyve inhibitor (PIK inhibitor), ( F ) PIKfyve inhibitor and NMDA (PIK inhibitor+NMDA), ( G ) SGK inhibitor (SGK inhibitor), ( H ) SGK inhibitor and NMDA (SGK inhibitor+NMDA). The magnification is equal for all images - scale bar indicates 5 µm. Number of images analyzed n = 10–15, obtained from independent neuron culture preparations.
    Figure Legend Snippet: ( A , B ) Statistical analysis of GluA1 surface expression and colocalization of GluA1 with neuroligin revealed increased surface and synaptic GluA1 expression after treatment with NMDA. This effect is abolished by SGK or PIKfyve inhibition. Control = untreated, NMDA = incubated with NMDA, PIK-In = treated with PIKfyve inhibitor, SGK-In = treated with SGK inhibitor. (C–H) Representative confocal images of dendrites stained with GluA1 (green) and neuroligin (red –Neuro) in controls versus NMDA-treated neurons under different conditions: ( C ) control versus ( D ) NMDA, ( E ) PIKfyve inhibitor (PIK inhibitor), ( F ) PIKfyve inhibitor and NMDA (PIK inhibitor+NMDA), ( G ) SGK inhibitor (SGK inhibitor), ( H ) SGK inhibitor and NMDA (SGK inhibitor+NMDA). The magnification is equal for all images - scale bar indicates 5 µm. Number of images analyzed n = 10–15, obtained from independent neuron culture preparations.

    Techniques Used: Expressing, Inhibition, Incubation, Staining

    Upon stimulation of NMDA receptors SGK3 is transcriptionally upregulated. SGK3 in turn phosphorylates/activates PIKfyve, which leads to local production of PI(3,5)P 2 in PIKfyve-containing recycling vesicles. PI(3,5)P 2 in turn stimulates Rab11-dependent plasma membrane-directed trafficking of GluA1-containing vesicles.
    Figure Legend Snippet: Upon stimulation of NMDA receptors SGK3 is transcriptionally upregulated. SGK3 in turn phosphorylates/activates PIKfyve, which leads to local production of PI(3,5)P 2 in PIKfyve-containing recycling vesicles. PI(3,5)P 2 in turn stimulates Rab11-dependent plasma membrane-directed trafficking of GluA1-containing vesicles.

    Techniques Used:

    primary rabbit anti glua1 antibody  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs primary rabbit anti glua1 antibody
    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on <t>GluA1</t> currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Primary Rabbit Anti Glua1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary rabbit anti glua1 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary rabbit anti glua1 antibody - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling"

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0033889

    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on GluA1 currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Figure Legend Snippet: ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on GluA1 currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.

    Techniques Used: Purification, Recombinant, Mutagenesis, Western Blot, Incubation, Stripping Membranes, Reverse Transcription Polymerase Chain Reaction, Expressing, Injection

    ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with SGK3, the inactive form of PIKfyve (PIKfyve(S318A)), or wild type PIKfyve. Numbers of oocytes are n = 20–30. Significant changes (p<0.001) are indicated by *** (p = 0.00049; 0.00036; 0.000073, respectively). ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.
    Figure Legend Snippet: ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with SGK3, the inactive form of PIKfyve (PIKfyve(S318A)), or wild type PIKfyve. Numbers of oocytes are n = 20–30. Significant changes (p<0.001) are indicated by *** (p = 0.00049; 0.00036; 0.000073, respectively). ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Techniques Used: Expressing, SDS-Gel, Western Blot, Molecular Weight, Software

    ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with Rab11, the dominant negative form of Rab11 (Rab11DN), PIKfyve, or SGK3. Numbers of oocytes are n = 15–30. Significant changes (p<0.001, p<0.05) are indicated by *** (p = 0.00015) and * (p = 0.036), respectively. ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.
    Figure Legend Snippet: ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with Rab11, the dominant negative form of Rab11 (Rab11DN), PIKfyve, or SGK3. Numbers of oocytes are n = 15–30. Significant changes (p<0.001, p<0.05) are indicated by *** (p = 0.00015) and * (p = 0.036), respectively. ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Techniques Used: Expressing, Dominant Negative Mutation, SDS-Gel, Western Blot, Molecular Weight, Software

    ( A ) GluA1 current amplitudes in oocytes before and after acute injection of a water-soluble analog of PI(3,5)P 2 . Significant change (p<0.05) is indicated by * (p = 0.0015). ( B ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with Rab11 or the dominant negative form of Rab11 (Rab11DN), before and after acute injection of a water-soluble analog of PI (3,5)P 2 . Numbers of oocytes are n = 7–28. Significant change (p<0.05) is indicated by * (p = 0.025). ( C ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with SGK3, myosin Vb, or the mutated form of myosin Vb (myosin del). Number of oocytes are n = 14–28.
    Figure Legend Snippet: ( A ) GluA1 current amplitudes in oocytes before and after acute injection of a water-soluble analog of PI(3,5)P 2 . Significant change (p<0.05) is indicated by * (p = 0.0015). ( B ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with Rab11 or the dominant negative form of Rab11 (Rab11DN), before and after acute injection of a water-soluble analog of PI (3,5)P 2 . Numbers of oocytes are n = 7–28. Significant change (p<0.05) is indicated by * (p = 0.025). ( C ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with SGK3, myosin Vb, or the mutated form of myosin Vb (myosin del). Number of oocytes are n = 14–28.

    Techniques Used: Injection, Expressing, Dominant Negative Mutation

    ( A , B ) Statistical analysis of GluA1 surface expression and colocalization of GluA1 with neuroligin revealed increased surface and synaptic GluA1 expression after treatment with NMDA. This effect is abolished by SGK or PIKfyve inhibition. Control = untreated, NMDA = incubated with NMDA, PIK-In = treated with PIKfyve inhibitor, SGK-In = treated with SGK inhibitor. (C–H) Representative confocal images of dendrites stained with GluA1 (green) and neuroligin (red –Neuro) in controls versus NMDA-treated neurons under different conditions: ( C ) control versus ( D ) NMDA, ( E ) PIKfyve inhibitor (PIK inhibitor), ( F ) PIKfyve inhibitor and NMDA (PIK inhibitor+NMDA), ( G ) SGK inhibitor (SGK inhibitor), ( H ) SGK inhibitor and NMDA (SGK inhibitor+NMDA). The magnification is equal for all images - scale bar indicates 5 µm. Number of images analyzed n = 10–15, obtained from independent neuron culture preparations.
    Figure Legend Snippet: ( A , B ) Statistical analysis of GluA1 surface expression and colocalization of GluA1 with neuroligin revealed increased surface and synaptic GluA1 expression after treatment with NMDA. This effect is abolished by SGK or PIKfyve inhibition. Control = untreated, NMDA = incubated with NMDA, PIK-In = treated with PIKfyve inhibitor, SGK-In = treated with SGK inhibitor. (C–H) Representative confocal images of dendrites stained with GluA1 (green) and neuroligin (red –Neuro) in controls versus NMDA-treated neurons under different conditions: ( C ) control versus ( D ) NMDA, ( E ) PIKfyve inhibitor (PIK inhibitor), ( F ) PIKfyve inhibitor and NMDA (PIK inhibitor+NMDA), ( G ) SGK inhibitor (SGK inhibitor), ( H ) SGK inhibitor and NMDA (SGK inhibitor+NMDA). The magnification is equal for all images - scale bar indicates 5 µm. Number of images analyzed n = 10–15, obtained from independent neuron culture preparations.

    Techniques Used: Expressing, Inhibition, Incubation, Staining

    Upon stimulation of NMDA receptors SGK3 is transcriptionally upregulated. SGK3 in turn phosphorylates/activates PIKfyve, which leads to local production of PI(3,5)P 2 in PIKfyve-containing recycling vesicles. PI(3,5)P 2 in turn stimulates Rab11-dependent plasma membrane-directed trafficking of GluA1-containing vesicles.
    Figure Legend Snippet: Upon stimulation of NMDA receptors SGK3 is transcriptionally upregulated. SGK3 in turn phosphorylates/activates PIKfyve, which leads to local production of PI(3,5)P 2 in PIKfyve-containing recycling vesicles. PI(3,5)P 2 in turn stimulates Rab11-dependent plasma membrane-directed trafficking of GluA1-containing vesicles.

    Techniques Used:

    agc004  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs agc004
    Agc004, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/agc004/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agc004 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    agc  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs agc
    Agc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/agc/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agc - by Bioz Stars, 2023-02
    94/100 stars

    Images

    rabbit polyclonal anti ampa receptor 1  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs rabbit polyclonal anti ampa receptor 1
    (A and B) Upper: representative images of individual endogenous <t>GluA1</t> (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.
    Rabbit Polyclonal Anti Ampa Receptor 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti ampa receptor 1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti ampa receptor 1 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X"

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.110853

    (A and B) Upper: representative images of individual endogenous GluA1 (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.
    Figure Legend Snippet: (A and B) Upper: representative images of individual endogenous GluA1 (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.

    Techniques Used: Immunofluorescence, Marker

    (A–D) Representative images of endogenous single mRNAs molecules encoding GluA1 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (A) quantified in (B). (C) Higher-magnification images of boxed region in (A) quantified in (D). (E–H) Representative images of endogenous single mRNAs molecules encoding GluA2 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (E) quantified in (F). (G) Higher-magnification images of boxed region in (E) quantified in (H) (GluA1, WT: 101 dendrites, 35 neurons; KO: 134 dendrites, 63 neurons, n = 4 per group; GluA2, WT: 190 dendrites, 82 neurons; KO: 99 dendrites, 82 neurons, n = 4 per group). (I) Representative images of a dual FISH experiment with probes targeting either the GluA2 coding (upper left) or intronic (upper right) sequences. (J) Representative images showing the number of GluA1 and GluA2 mRNA molecules at transcription hotspots in the nucleus of neurons from WT and Fmr1 KO mice. (K) Summary data of images like those illustrated in (J) (GluA1, WT: 35 neurons; KO: 63 neurons; GluA2, WT: 82 neurons, KO: 86 neurons, n = 4). (L) Summary qRT-PCR data from whole hippocampus showing GluA1 (upper) and GluA2 (lower) mRNA expression (n = 4 per group). For all the quantitative graphs: Data are mean ± SEM. *p < 0.05; **p < 0.01.
    Figure Legend Snippet: (A–D) Representative images of endogenous single mRNAs molecules encoding GluA1 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (A) quantified in (B). (C) Higher-magnification images of boxed region in (A) quantified in (D). (E–H) Representative images of endogenous single mRNAs molecules encoding GluA2 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (E) quantified in (F). (G) Higher-magnification images of boxed region in (E) quantified in (H) (GluA1, WT: 101 dendrites, 35 neurons; KO: 134 dendrites, 63 neurons, n = 4 per group; GluA2, WT: 190 dendrites, 82 neurons; KO: 99 dendrites, 82 neurons, n = 4 per group). (I) Representative images of a dual FISH experiment with probes targeting either the GluA2 coding (upper left) or intronic (upper right) sequences. (J) Representative images showing the number of GluA1 and GluA2 mRNA molecules at transcription hotspots in the nucleus of neurons from WT and Fmr1 KO mice. (K) Summary data of images like those illustrated in (J) (GluA1, WT: 35 neurons; KO: 63 neurons; GluA2, WT: 82 neurons, KO: 86 neurons, n = 4). (L) Summary qRT-PCR data from whole hippocampus showing GluA1 (upper) and GluA2 (lower) mRNA expression (n = 4 per group). For all the quantitative graphs: Data are mean ± SEM. *p < 0.05; **p < 0.01.

    Techniques Used: Quantitative RT-PCR, Expressing

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: shRNA, Recombinant, Sequencing, Binding Assay, Plasmid Preparation, Software

    agc004  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs agc004
    Agc004, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/agc004/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agc004 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    anti glua1 antibody produced in guinea pig  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93

    Structured Review

    Alomone Labs anti glua1 antibody produced in guinea pig
    List of primary antibodies used
    Anti Glua1 Antibody Produced In Guinea Pig, 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/anti glua1 antibody produced in guinea pig/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti glua1 antibody produced in guinea pig - by Bioz Stars, 2023-02
    93/100 stars

    Images

    1) Product Images from "Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain"

    Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

    Journal: Journal of Neuroscience Research

    doi: 10.1002/jnr.24967

    List of primary antibodies used
    Figure Legend Snippet: List of primary antibodies used

    Techniques Used: Concentration Assay, Produced, Recombinant, Purification, Derivative Assay, Transduction

    Antibodies used and amino acid conservation between species
    Figure Legend Snippet: Antibodies used and amino acid conservation between species

    Techniques Used:

    Molecular heterogeneity of astrocytes in the marmoset cerebellum. (A) Lower magnification images showing labeling for astrocytic proteins with respect to the molecular layer (ML), Purkinje cell layer (PCL), granule cell layer (GCL), and white matter (WM). Calbindin is used as a Purkinje cell marker. DAPI staining (blue) shows the nuclear staining of interneurons in the ML and granule cells in GCL. Scale bar: 100 μm. (B) Detailed images of ML and GCL stained with specific markers for Bergmann glia (BG) such as GluA1, Kirk4.1, and velate astrocytes (VA) such as AQP4. EAAT2 was expressed in both BGs and VAs with higher expression in VAs. Purkinje cell bodies are indicated with a white asterisk. Scale bar: 20 μm. bv, blood vessel
    Figure Legend Snippet: Molecular heterogeneity of astrocytes in the marmoset cerebellum. (A) Lower magnification images showing labeling for astrocytic proteins with respect to the molecular layer (ML), Purkinje cell layer (PCL), granule cell layer (GCL), and white matter (WM). Calbindin is used as a Purkinje cell marker. DAPI staining (blue) shows the nuclear staining of interneurons in the ML and granule cells in GCL. Scale bar: 100 μm. (B) Detailed images of ML and GCL stained with specific markers for Bergmann glia (BG) such as GluA1, Kirk4.1, and velate astrocytes (VA) such as AQP4. EAAT2 was expressed in both BGs and VAs with higher expression in VAs. Purkinje cell bodies are indicated with a white asterisk. Scale bar: 20 μm. bv, blood vessel

    Techniques Used: Labeling, Marker, Staining, Expressing

    anti ampar 1 glua1  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs anti ampar 1 glua1
    Nogo-A loss-of-function shifts the excitation/inhibition balance toward a higher excitation at a single cell level. ( A – C ) Patch-Clamp recordings in organotypic hippocampal cultures of simultaneously measuring mEPSCs and mIPSCs before (0 min) and after (5–10 min) control antibody, in all experiments mouse IgG1, FG12 (black, n = 14) or Nogo-A mouse IgG1, 11C7 function-blocking antibody (red, n = 14) treatment. ( A ) Example traces of mPSCs recorded by whole-cell patch clamp. Each trace shows representative mEPSCs (magenta) and mIPSCs (orange) distinguished by their decay time according to fast (mEPSCs < 25 ms) and slow (mIPSC > 25 ms) decay kinetics. Scale bars are 20 pA vertical and 200 ms horizontal ( B ) Normalized data for mIPSC and mEPSC ratio. ( C ) Normalized values for mIPSC and mEPSC amplitude plotted individually for each recorded neuron and the mean values visualized by the dotted line. ( D , E ) The percent change in the amplitude of mEPSC and mIPSC of individual neurons over time and their respective mean values (( D ) Ctrl-Ab: black, ( E ) Nogo-A Ab: dark red, mEPSCs: # p , mIPSCs: *** p ). ( F , G ) The percent change of individual recorded neurons in mIPSCs ( F ) and mEPSCs ( G ) exceeding average ± SEM color coded by heat mapping changes between 0–100% changes. ( H – J ) Live-cell labeling of surface GABA A and <t>GluA1</t> receptors in dissociated hippocampal neurons after 10 min Nogo-A loss-of-function (red, n = 40) compared to the control condition (black, n = 40). ( H ) Representative image of surface stained GABA A Rs (green) and AMPARs subunit GluA1 (magenta). All of the images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values. Scale bars, 2 µm. ( I ) Normalized values for fluorescence intensity of GABA A R cluster and GluA1 immuno-positive puncta ratio upon Nogo-A loss-of-function for 10 min. ( J ) Normalized fluorescence intensity of GABA A R and GluA1 positive clusters plotted individually for each recorded neuron and the mean alteration visualized by the dotted line. Values represent means ± SEMs. # p < 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001; n = number of neurons.
    Anti Ampar 1 Glua1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti ampar 1 glua1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti ampar 1 glua1 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Nogo-A Modulates the Synaptic Excitation of Hippocampal Neurons in a Ca 2+ -Dependent Manner"

    Article Title: Nogo-A Modulates the Synaptic Excitation of Hippocampal Neurons in a Ca 2+ -Dependent Manner

    Journal: Cells

    doi: 10.3390/cells10092299

    Nogo-A loss-of-function shifts the excitation/inhibition balance toward a higher excitation at a single cell level. ( A – C ) Patch-Clamp recordings in organotypic hippocampal cultures of simultaneously measuring mEPSCs and mIPSCs before (0 min) and after (5–10 min) control antibody, in all experiments mouse IgG1, FG12 (black, n = 14) or Nogo-A mouse IgG1, 11C7 function-blocking antibody (red, n = 14) treatment. ( A ) Example traces of mPSCs recorded by whole-cell patch clamp. Each trace shows representative mEPSCs (magenta) and mIPSCs (orange) distinguished by their decay time according to fast (mEPSCs < 25 ms) and slow (mIPSC > 25 ms) decay kinetics. Scale bars are 20 pA vertical and 200 ms horizontal ( B ) Normalized data for mIPSC and mEPSC ratio. ( C ) Normalized values for mIPSC and mEPSC amplitude plotted individually for each recorded neuron and the mean values visualized by the dotted line. ( D , E ) The percent change in the amplitude of mEPSC and mIPSC of individual neurons over time and their respective mean values (( D ) Ctrl-Ab: black, ( E ) Nogo-A Ab: dark red, mEPSCs: # p , mIPSCs: *** p ). ( F , G ) The percent change of individual recorded neurons in mIPSCs ( F ) and mEPSCs ( G ) exceeding average ± SEM color coded by heat mapping changes between 0–100% changes. ( H – J ) Live-cell labeling of surface GABA A and GluA1 receptors in dissociated hippocampal neurons after 10 min Nogo-A loss-of-function (red, n = 40) compared to the control condition (black, n = 40). ( H ) Representative image of surface stained GABA A Rs (green) and AMPARs subunit GluA1 (magenta). All of the images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values. Scale bars, 2 µm. ( I ) Normalized values for fluorescence intensity of GABA A R cluster and GluA1 immuno-positive puncta ratio upon Nogo-A loss-of-function for 10 min. ( J ) Normalized fluorescence intensity of GABA A R and GluA1 positive clusters plotted individually for each recorded neuron and the mean alteration visualized by the dotted line. Values represent means ± SEMs. # p < 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001; n = number of neurons.
    Figure Legend Snippet: Nogo-A loss-of-function shifts the excitation/inhibition balance toward a higher excitation at a single cell level. ( A – C ) Patch-Clamp recordings in organotypic hippocampal cultures of simultaneously measuring mEPSCs and mIPSCs before (0 min) and after (5–10 min) control antibody, in all experiments mouse IgG1, FG12 (black, n = 14) or Nogo-A mouse IgG1, 11C7 function-blocking antibody (red, n = 14) treatment. ( A ) Example traces of mPSCs recorded by whole-cell patch clamp. Each trace shows representative mEPSCs (magenta) and mIPSCs (orange) distinguished by their decay time according to fast (mEPSCs < 25 ms) and slow (mIPSC > 25 ms) decay kinetics. Scale bars are 20 pA vertical and 200 ms horizontal ( B ) Normalized data for mIPSC and mEPSC ratio. ( C ) Normalized values for mIPSC and mEPSC amplitude plotted individually for each recorded neuron and the mean values visualized by the dotted line. ( D , E ) The percent change in the amplitude of mEPSC and mIPSC of individual neurons over time and their respective mean values (( D ) Ctrl-Ab: black, ( E ) Nogo-A Ab: dark red, mEPSCs: # p , mIPSCs: *** p ). ( F , G ) The percent change of individual recorded neurons in mIPSCs ( F ) and mEPSCs ( G ) exceeding average ± SEM color coded by heat mapping changes between 0–100% changes. ( H – J ) Live-cell labeling of surface GABA A and GluA1 receptors in dissociated hippocampal neurons after 10 min Nogo-A loss-of-function (red, n = 40) compared to the control condition (black, n = 40). ( H ) Representative image of surface stained GABA A Rs (green) and AMPARs subunit GluA1 (magenta). All of the images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values. Scale bars, 2 µm. ( I ) Normalized values for fluorescence intensity of GABA A R cluster and GluA1 immuno-positive puncta ratio upon Nogo-A loss-of-function for 10 min. ( J ) Normalized fluorescence intensity of GABA A R and GluA1 positive clusters plotted individually for each recorded neuron and the mean alteration visualized by the dotted line. Values represent means ± SEMs. # p < 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001; n = number of neurons.

    Techniques Used: Inhibition, Patch Clamp, Blocking Assay, Labeling, Staining, Fluorescence

    Nogo-A regulates the synaptic insertion of calcium permeable-AMPARs. ( A , B ) Live-cell immunolabeling of surface AMPAR subunit (magenta) GluA1 ( A ) or GluA2 ( B ) followed by immunofluorescence for presynaptic marker synapsin (Syn1/2; green) and their merged images (bottom) in primary hippocampal neurons treated for 10 min either with the control (left) or the Nogo-A function-blocking (right) antibody. For illustration, all images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values for visibility. Scale bar 2 μm. ( C , D ) Normalized data for density ( C ) and fluorescence intensity ( D ) of GluA1 immuno-positive puncta in hippocampal neurons treated with either control antibody (black, n = 40) or Nogo-A function-blocking antibody (red, n = 39) for 10 min. ( E ) Normalized values for the density of GluA1 clusters colocalized with Syn 1/2 immuno-positive puncta. ( F , G ) Normalized GluA2 cluster density ( F ) and fluorescence intensity ( G ) in hippocampal neurons upon 10 min application with control antibody (black, n = 36) or Nogo-A function-blocking antibody (red, n = 35). ( H ) Normalized density of GluA2 immuno-positive puncta colocalized with Syn 1/2. Data are presented as mean ± SEM. * p < 0.05.
    Figure Legend Snippet: Nogo-A regulates the synaptic insertion of calcium permeable-AMPARs. ( A , B ) Live-cell immunolabeling of surface AMPAR subunit (magenta) GluA1 ( A ) or GluA2 ( B ) followed by immunofluorescence for presynaptic marker synapsin (Syn1/2; green) and their merged images (bottom) in primary hippocampal neurons treated for 10 min either with the control (left) or the Nogo-A function-blocking (right) antibody. For illustration, all images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values for visibility. Scale bar 2 μm. ( C , D ) Normalized data for density ( C ) and fluorescence intensity ( D ) of GluA1 immuno-positive puncta in hippocampal neurons treated with either control antibody (black, n = 40) or Nogo-A function-blocking antibody (red, n = 39) for 10 min. ( E ) Normalized values for the density of GluA1 clusters colocalized with Syn 1/2 immuno-positive puncta. ( F , G ) Normalized GluA2 cluster density ( F ) and fluorescence intensity ( G ) in hippocampal neurons upon 10 min application with control antibody (black, n = 36) or Nogo-A function-blocking antibody (red, n = 35). ( H ) Normalized density of GluA2 immuno-positive puncta colocalized with Syn 1/2. Data are presented as mean ± SEM. * p < 0.05.

    Techniques Used: Immunolabeling, Immunofluorescence, Marker, Blocking Assay, Fluorescence

    rabbit anti glun1and rabbit anti glua1  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs rabbit anti glun1and rabbit anti glua1
    Summary of antibodies used for the Western blotting.
    Rabbit Anti Glun1and Rabbit Anti Glua1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti glun1and rabbit anti glua1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti glun1and rabbit anti glua1 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "Endocannabinoids Released in the Ventral Tegmental Area During Copulation to Satiety Modulate Changes in Glutamate Receptors Associated With Synaptic Plasticity Processes"

    Article Title: Endocannabinoids Released in the Ventral Tegmental Area During Copulation to Satiety Modulate Changes in Glutamate Receptors Associated With Synaptic Plasticity Processes

    Journal: Frontiers in Synaptic Neuroscience

    doi: 10.3389/fnsyn.2021.701290

    Summary of antibodies used for the Western blotting.
    Figure Legend Snippet: Summary of antibodies used for the Western blotting.

    Techniques Used: Western Blot

    Changes in AMPAR density and subunit composition in the VTA of sexually satiated rats and involvement of eCBs in their induction. Western blot analysis showing changes in GluA1 (A) and GluA2 AMPAR subunit (B) densities in the VTA of control sexually experienced unmated rats (Control), males that ejaculated once (1 Ejac) or copulated to satiety (Sexually satiated) 24 h earlier, and of males that copulated to satiety in the presence of the CB1R antagonist AM251 (Satiated + AM251). Differences among untreated groups with different sexual conditions were determined by means of a one-way ANOVA followed by Tukey test, *** P < 0.001; * P < 0.05. A comparison between the rats that copulated to satiety in the absence or presence of AM251 was conducted with the unpaired t -test ** P < 0.01. Values are mean ± S.E.M of the protein/β-Actin optical density (O.D.) ratios of 5 rats per group.
    Figure Legend Snippet: Changes in AMPAR density and subunit composition in the VTA of sexually satiated rats and involvement of eCBs in their induction. Western blot analysis showing changes in GluA1 (A) and GluA2 AMPAR subunit (B) densities in the VTA of control sexually experienced unmated rats (Control), males that ejaculated once (1 Ejac) or copulated to satiety (Sexually satiated) 24 h earlier, and of males that copulated to satiety in the presence of the CB1R antagonist AM251 (Satiated + AM251). Differences among untreated groups with different sexual conditions were determined by means of a one-way ANOVA followed by Tukey test, *** P < 0.001; * P < 0.05. A comparison between the rats that copulated to satiety in the absence or presence of AM251 was conducted with the unpaired t -test ** P < 0.01. Values are mean ± S.E.M of the protein/β-Actin optical density (O.D.) ratios of 5 rats per group.

    Techniques Used: Western Blot

    glua1 extra  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs glua1 extra
    Glua1 Extra, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glua1 extra/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    glua1 extra - by Bioz Stars, 2023-02
    94/100 stars

    Images

    glua1  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Alomone Labs glua1
    Primary antibodies. WB , dilution for Western blotting; IF , dilution for immunofluorescence staining
    Glua1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glua1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    glua1 - by Bioz Stars, 2023-02
    94/100 stars

    Images

    1) Product Images from "The Golgi-Associated PDZ Domain Protein Gopc/PIST Is Required for Synaptic Targeting of mGluR5"

    Article Title: The Golgi-Associated PDZ Domain Protein Gopc/PIST Is Required for Synaptic Targeting of mGluR5

    Journal: Molecular Neurobiology

    doi: 10.1007/s12035-021-02504-9

    Primary antibodies. WB , dilution for Western blotting; IF , dilution for immunofluorescence staining
    Figure Legend Snippet: Primary antibodies. WB , dilution for Western blotting; IF , dilution for immunofluorescence staining

    Techniques Used: Western Blot, Immunofluorescence

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Alomone Labs primary rabbit anti glua1 antibody
    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on <t>GluA1</t> currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Primary Rabbit Anti Glua1 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/primary rabbit anti glua1 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    primary rabbit anti glua1 antibody - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs agc004
    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on <t>GluA1</t> currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Agc004, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/agc004/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agc004 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs agc
    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on <t>GluA1</t> currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.
    Agc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/agc/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agc - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs rabbit polyclonal anti ampa receptor 1
    (A and B) Upper: representative images of individual endogenous <t>GluA1</t> (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.
    Rabbit Polyclonal Anti Ampa Receptor 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal anti ampa receptor 1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti ampa receptor 1 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    93
    Alomone Labs anti glua1 antibody produced in guinea pig
    List of primary antibodies used
    Anti Glua1 Antibody Produced In Guinea Pig, 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/anti glua1 antibody produced in guinea pig/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti glua1 antibody produced in guinea pig - by Bioz Stars, 2023-02
    93/100 stars
      Buy from Supplier

    94
    Alomone Labs anti ampar 1 glua1
    Nogo-A loss-of-function shifts the excitation/inhibition balance toward a higher excitation at a single cell level. ( A – C ) Patch-Clamp recordings in organotypic hippocampal cultures of simultaneously measuring mEPSCs and mIPSCs before (0 min) and after (5–10 min) control antibody, in all experiments mouse IgG1, FG12 (black, n = 14) or Nogo-A mouse IgG1, 11C7 function-blocking antibody (red, n = 14) treatment. ( A ) Example traces of mPSCs recorded by whole-cell patch clamp. Each trace shows representative mEPSCs (magenta) and mIPSCs (orange) distinguished by their decay time according to fast (mEPSCs < 25 ms) and slow (mIPSC > 25 ms) decay kinetics. Scale bars are 20 pA vertical and 200 ms horizontal ( B ) Normalized data for mIPSC and mEPSC ratio. ( C ) Normalized values for mIPSC and mEPSC amplitude plotted individually for each recorded neuron and the mean values visualized by the dotted line. ( D , E ) The percent change in the amplitude of mEPSC and mIPSC of individual neurons over time and their respective mean values (( D ) Ctrl-Ab: black, ( E ) Nogo-A Ab: dark red, mEPSCs: # p , mIPSCs: *** p ). ( F , G ) The percent change of individual recorded neurons in mIPSCs ( F ) and mEPSCs ( G ) exceeding average ± SEM color coded by heat mapping changes between 0–100% changes. ( H – J ) Live-cell labeling of surface GABA A and <t>GluA1</t> receptors in dissociated hippocampal neurons after 10 min Nogo-A loss-of-function (red, n = 40) compared to the control condition (black, n = 40). ( H ) Representative image of surface stained GABA A Rs (green) and AMPARs subunit GluA1 (magenta). All of the images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values. Scale bars, 2 µm. ( I ) Normalized values for fluorescence intensity of GABA A R cluster and GluA1 immuno-positive puncta ratio upon Nogo-A loss-of-function for 10 min. ( J ) Normalized fluorescence intensity of GABA A R and GluA1 positive clusters plotted individually for each recorded neuron and the mean alteration visualized by the dotted line. Values represent means ± SEMs. # p < 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001; n = number of neurons.
    Anti Ampar 1 Glua1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti ampar 1 glua1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti ampar 1 glua1 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs rabbit anti glun1and rabbit anti glua1
    Summary of antibodies used for the Western blotting.
    Rabbit Anti Glun1and Rabbit Anti Glua1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti glun1and rabbit anti glua1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti glun1and rabbit anti glua1 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs glua1 extra
    Summary of antibodies used for the Western blotting.
    Glua1 Extra, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glua1 extra/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    glua1 extra - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    94
    Alomone Labs glua1
    Primary antibodies. WB , dilution for Western blotting; IF , dilution for immunofluorescence staining
    Glua1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glua1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    glua1 - by Bioz Stars, 2023-02
    94/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on GluA1 currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.

    Journal: PLoS ONE

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    doi: 10.1371/journal.pone.0033889

    Figure Lengend Snippet: ( A ) PIKfyve is phosphorylated at Ser318 by SGK3 and at Ser318 by PKB. Purified recombinant GST-tagged wild-type (WT) or S318A mutant (SA) of PIKfyve was subjected to Western blotting. Blots were incubated with rabbit anti-GST antibody (PIKfyve; lower panel), followed by stripping and reprobing with a rabbit anti-PIKfyve antibody specific for phosphoserine 318 (apS318; top panel). ( B ) RT-PCR demonstrating that PIKfyve is expressed in hippocampus. Lane 1: cDNA from hippocampus, the two primers bind in exons 19 and 20, respectively, and amplify a 315 bp fragment of PIKfyve; lane 2: control reaction to exclude genomic contamination; lane 3: control reaction without reverse transcriptase; lane 4: control reaction without cDNA. ( C ) The PIKfyve inhibitor YM201636 and SGK inhibitor EMD638683 suppress the upregulating effect of SGK3 on GluA1 currents. GluA1 current amplitudes in oocytes expressing GluA1. Acute injection of purified active SGK3 protein led to an increase in GluA1 currents. The effects of YM201636 and EMD638683 on GluA1 currents were measured in oocytes before and after acute injection of SGK3 protein. Significant change to GluA1 control (p = 0.013) is indicated by *. Numbers of oocytes were n = 7–36.

    Article Snippet: Thereafter, cell cultures were washed three times in PBS and incubated with primary rabbit anti-GluA1 antibody (kind gift of R. Huganir) and mouse anti-PSD95 antibody (Thermo Scientific, Rockford, IL, USA, #MA1-046), or primary rabbit anti-GluA1 antibody (Alomone, Jerusalem, Israel, #AGC-004) and mouse anti- neuroligin-1 antibody (Millipore, Temecula, CA, USA, #MABN38) overnight at 4°C.

    Techniques: Purification, Recombinant, Mutagenesis, Western Blot, Incubation, Stripping Membranes, Reverse Transcription Polymerase Chain Reaction, Expressing, Injection

    ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with SGK3, the inactive form of PIKfyve (PIKfyve(S318A)), or wild type PIKfyve. Numbers of oocytes are n = 20–30. Significant changes (p<0.001) are indicated by *** (p = 0.00049; 0.00036; 0.000073, respectively). ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Journal: PLoS ONE

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    doi: 10.1371/journal.pone.0033889

    Figure Lengend Snippet: ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with SGK3, the inactive form of PIKfyve (PIKfyve(S318A)), or wild type PIKfyve. Numbers of oocytes are n = 20–30. Significant changes (p<0.001) are indicated by *** (p = 0.00049; 0.00036; 0.000073, respectively). ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Article Snippet: Thereafter, cell cultures were washed three times in PBS and incubated with primary rabbit anti-GluA1 antibody (kind gift of R. Huganir) and mouse anti-PSD95 antibody (Thermo Scientific, Rockford, IL, USA, #MA1-046), or primary rabbit anti-GluA1 antibody (Alomone, Jerusalem, Israel, #AGC-004) and mouse anti- neuroligin-1 antibody (Millipore, Temecula, CA, USA, #MABN38) overnight at 4°C.

    Techniques: Expressing, SDS-Gel, Western Blot, Molecular Weight, Software

    ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with Rab11, the dominant negative form of Rab11 (Rab11DN), PIKfyve, or SGK3. Numbers of oocytes are n = 15–30. Significant changes (p<0.001, p<0.05) are indicated by *** (p = 0.00015) and * (p = 0.036), respectively. ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Journal: PLoS ONE

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    doi: 10.1371/journal.pone.0033889

    Figure Lengend Snippet: ( A ) Representative current traces measured in Xenopus oocytes in response to superfusion with 300 µM glutamate plus 100 µM cyclo-thiacide. All currents were measured at −70 mV. Vertical scale-bar, 0,5 µA; horizontal bar, 4 s. ( B ) GluA1 current amplitudes in oocytes expressing GluA1, or combinations of GluA1 with Rab11, the dominant negative form of Rab11 (Rab11DN), PIKfyve, or SGK3. Numbers of oocytes are n = 15–30. Significant changes (p<0.001, p<0.05) are indicated by *** (p = 0.00015) and * (p = 0.036), respectively. ( C ) Representative samples including controls from uninjected oocytes were biotinylated to isolate plasma membrane GluA1, then separated on an SDS gel, Western-blotted and probed with a primary rabbit anti-GluA1 antibody. The GluA1 protein has an apparent molecular weight of ∼105 kDa. ( D ) Bar graph showing relative abundance of GluA1 plasma membrane protein. The band intensity was quantified by densitometric analysis using the software Scion image.

    Article Snippet: Thereafter, cell cultures were washed three times in PBS and incubated with primary rabbit anti-GluA1 antibody (kind gift of R. Huganir) and mouse anti-PSD95 antibody (Thermo Scientific, Rockford, IL, USA, #MA1-046), or primary rabbit anti-GluA1 antibody (Alomone, Jerusalem, Israel, #AGC-004) and mouse anti- neuroligin-1 antibody (Millipore, Temecula, CA, USA, #MABN38) overnight at 4°C.

    Techniques: Expressing, Dominant Negative Mutation, SDS-Gel, Western Blot, Molecular Weight, Software

    ( A ) GluA1 current amplitudes in oocytes before and after acute injection of a water-soluble analog of PI(3,5)P 2 . Significant change (p<0.05) is indicated by * (p = 0.0015). ( B ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with Rab11 or the dominant negative form of Rab11 (Rab11DN), before and after acute injection of a water-soluble analog of PI (3,5)P 2 . Numbers of oocytes are n = 7–28. Significant change (p<0.05) is indicated by * (p = 0.025). ( C ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with SGK3, myosin Vb, or the mutated form of myosin Vb (myosin del). Number of oocytes are n = 14–28.

    Journal: PLoS ONE

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    doi: 10.1371/journal.pone.0033889

    Figure Lengend Snippet: ( A ) GluA1 current amplitudes in oocytes before and after acute injection of a water-soluble analog of PI(3,5)P 2 . Significant change (p<0.05) is indicated by * (p = 0.0015). ( B ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with Rab11 or the dominant negative form of Rab11 (Rab11DN), before and after acute injection of a water-soluble analog of PI (3,5)P 2 . Numbers of oocytes are n = 7–28. Significant change (p<0.05) is indicated by * (p = 0.025). ( C ) GluA1 current amplitudes in oocytes expressing GluA1 or combinations of GluA1 with SGK3, myosin Vb, or the mutated form of myosin Vb (myosin del). Number of oocytes are n = 14–28.

    Article Snippet: Thereafter, cell cultures were washed three times in PBS and incubated with primary rabbit anti-GluA1 antibody (kind gift of R. Huganir) and mouse anti-PSD95 antibody (Thermo Scientific, Rockford, IL, USA, #MA1-046), or primary rabbit anti-GluA1 antibody (Alomone, Jerusalem, Israel, #AGC-004) and mouse anti- neuroligin-1 antibody (Millipore, Temecula, CA, USA, #MABN38) overnight at 4°C.

    Techniques: Injection, Expressing, Dominant Negative Mutation

    ( A , B ) Statistical analysis of GluA1 surface expression and colocalization of GluA1 with neuroligin revealed increased surface and synaptic GluA1 expression after treatment with NMDA. This effect is abolished by SGK or PIKfyve inhibition. Control = untreated, NMDA = incubated with NMDA, PIK-In = treated with PIKfyve inhibitor, SGK-In = treated with SGK inhibitor. (C–H) Representative confocal images of dendrites stained with GluA1 (green) and neuroligin (red –Neuro) in controls versus NMDA-treated neurons under different conditions: ( C ) control versus ( D ) NMDA, ( E ) PIKfyve inhibitor (PIK inhibitor), ( F ) PIKfyve inhibitor and NMDA (PIK inhibitor+NMDA), ( G ) SGK inhibitor (SGK inhibitor), ( H ) SGK inhibitor and NMDA (SGK inhibitor+NMDA). The magnification is equal for all images - scale bar indicates 5 µm. Number of images analyzed n = 10–15, obtained from independent neuron culture preparations.

    Journal: PLoS ONE

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    doi: 10.1371/journal.pone.0033889

    Figure Lengend Snippet: ( A , B ) Statistical analysis of GluA1 surface expression and colocalization of GluA1 with neuroligin revealed increased surface and synaptic GluA1 expression after treatment with NMDA. This effect is abolished by SGK or PIKfyve inhibition. Control = untreated, NMDA = incubated with NMDA, PIK-In = treated with PIKfyve inhibitor, SGK-In = treated with SGK inhibitor. (C–H) Representative confocal images of dendrites stained with GluA1 (green) and neuroligin (red –Neuro) in controls versus NMDA-treated neurons under different conditions: ( C ) control versus ( D ) NMDA, ( E ) PIKfyve inhibitor (PIK inhibitor), ( F ) PIKfyve inhibitor and NMDA (PIK inhibitor+NMDA), ( G ) SGK inhibitor (SGK inhibitor), ( H ) SGK inhibitor and NMDA (SGK inhibitor+NMDA). The magnification is equal for all images - scale bar indicates 5 µm. Number of images analyzed n = 10–15, obtained from independent neuron culture preparations.

    Article Snippet: Thereafter, cell cultures were washed three times in PBS and incubated with primary rabbit anti-GluA1 antibody (kind gift of R. Huganir) and mouse anti-PSD95 antibody (Thermo Scientific, Rockford, IL, USA, #MA1-046), or primary rabbit anti-GluA1 antibody (Alomone, Jerusalem, Israel, #AGC-004) and mouse anti- neuroligin-1 antibody (Millipore, Temecula, CA, USA, #MABN38) overnight at 4°C.

    Techniques: Expressing, Inhibition, Incubation, Staining

    Upon stimulation of NMDA receptors SGK3 is transcriptionally upregulated. SGK3 in turn phosphorylates/activates PIKfyve, which leads to local production of PI(3,5)P 2 in PIKfyve-containing recycling vesicles. PI(3,5)P 2 in turn stimulates Rab11-dependent plasma membrane-directed trafficking of GluA1-containing vesicles.

    Journal: PLoS ONE

    Article Title: Identification of a Novel Signaling Pathway and Its Relevance for GluA1 Recycling

    doi: 10.1371/journal.pone.0033889

    Figure Lengend Snippet: Upon stimulation of NMDA receptors SGK3 is transcriptionally upregulated. SGK3 in turn phosphorylates/activates PIKfyve, which leads to local production of PI(3,5)P 2 in PIKfyve-containing recycling vesicles. PI(3,5)P 2 in turn stimulates Rab11-dependent plasma membrane-directed trafficking of GluA1-containing vesicles.

    Article Snippet: Thereafter, cell cultures were washed three times in PBS and incubated with primary rabbit anti-GluA1 antibody (kind gift of R. Huganir) and mouse anti-PSD95 antibody (Thermo Scientific, Rockford, IL, USA, #MA1-046), or primary rabbit anti-GluA1 antibody (Alomone, Jerusalem, Israel, #AGC-004) and mouse anti- neuroligin-1 antibody (Millipore, Temecula, CA, USA, #MABN38) overnight at 4°C.

    Techniques:

    (A and B) Upper: representative images of individual endogenous GluA1 (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.

    Journal: Cell reports

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    doi: 10.1016/j.celrep.2022.110853

    Figure Lengend Snippet: (A and B) Upper: representative images of individual endogenous GluA1 (red) (A) or GluA2 (red) (B) mRNA molecules within a single hippocampal neuron detected by single-molecule FISH, followed by immunofluorescence with antibodies directed to the presynaptic marker bassoon (green), the dendritic marker MAP2 (blue), and merge. Lower: higher-magnification images of boxed region in the upper panel. (C) Upper: representative images of individual GluA1 (red) and GluA2 (green) mRNA molecules in the same neuron. Lower: higher-magnification images. (D) Summary data of images like those in (A and B) showing the number of AMPAR GluA1 (black) and GluA2 (red) mRNAs that juxtapose to synaptic sites marked by basoon (first two bars) or that colocalize with each other (third bar). GluA1 and GluA2, 30dendrites, 30 neurons, n = 4. (E) Summary data for images like those in (C) showing the number of GluA1 and GluA2 molecules per dendrite. (F) GluA1 and GluA2 mRNA molecules as a function of the distance from the soma (GluA1, 101 dendrites, 35 neurons, n = 4; GluA2, 190 dendrites, 82 neurons, n = 4). The number of individual GluA1 and GluA2 mRNA molecules in all dendrites of all hippocampal neurons that met the criteria for identification as mRNAs were analyzed by an individual blinded to the treatment. Scale bar, 10 μm. For (D–F): Data are mean ± SEM. **p < 0.01. NS, not significant. Here and in , , (with the exception of ), , and , n is defined as number of independent experiments each involving a different batch of neurons. In , n is defined as number of animals.

    Article Snippet: Rabbit polyclonal anti-AMPA Receptor 1 (GluR1)/GluA1 , Alomone Labs , Cat# AGC-004; RRID:AB_2039878.

    Techniques: Immunofluorescence, Marker

    (A–D) Representative images of endogenous single mRNAs molecules encoding GluA1 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (A) quantified in (B). (C) Higher-magnification images of boxed region in (A) quantified in (D). (E–H) Representative images of endogenous single mRNAs molecules encoding GluA2 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (E) quantified in (F). (G) Higher-magnification images of boxed region in (E) quantified in (H) (GluA1, WT: 101 dendrites, 35 neurons; KO: 134 dendrites, 63 neurons, n = 4 per group; GluA2, WT: 190 dendrites, 82 neurons; KO: 99 dendrites, 82 neurons, n = 4 per group). (I) Representative images of a dual FISH experiment with probes targeting either the GluA2 coding (upper left) or intronic (upper right) sequences. (J) Representative images showing the number of GluA1 and GluA2 mRNA molecules at transcription hotspots in the nucleus of neurons from WT and Fmr1 KO mice. (K) Summary data of images like those illustrated in (J) (GluA1, WT: 35 neurons; KO: 63 neurons; GluA2, WT: 82 neurons, KO: 86 neurons, n = 4). (L) Summary qRT-PCR data from whole hippocampus showing GluA1 (upper) and GluA2 (lower) mRNA expression (n = 4 per group). For all the quantitative graphs: Data are mean ± SEM. *p < 0.05; **p < 0.01.

    Journal: Cell reports

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    doi: 10.1016/j.celrep.2022.110853

    Figure Lengend Snippet: (A–D) Representative images of endogenous single mRNAs molecules encoding GluA1 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (A) quantified in (B). (C) Higher-magnification images of boxed region in (A) quantified in (D). (E–H) Representative images of endogenous single mRNAs molecules encoding GluA2 were detected by single-molecule FISH in primary cultures of hippocampal neurons from WT and Fmr1 KO mice (E) quantified in (F). (G) Higher-magnification images of boxed region in (E) quantified in (H) (GluA1, WT: 101 dendrites, 35 neurons; KO: 134 dendrites, 63 neurons, n = 4 per group; GluA2, WT: 190 dendrites, 82 neurons; KO: 99 dendrites, 82 neurons, n = 4 per group). (I) Representative images of a dual FISH experiment with probes targeting either the GluA2 coding (upper left) or intronic (upper right) sequences. (J) Representative images showing the number of GluA1 and GluA2 mRNA molecules at transcription hotspots in the nucleus of neurons from WT and Fmr1 KO mice. (K) Summary data of images like those illustrated in (J) (GluA1, WT: 35 neurons; KO: 63 neurons; GluA2, WT: 82 neurons, KO: 86 neurons, n = 4). (L) Summary qRT-PCR data from whole hippocampus showing GluA1 (upper) and GluA2 (lower) mRNA expression (n = 4 per group). For all the quantitative graphs: Data are mean ± SEM. *p < 0.05; **p < 0.01.

    Article Snippet: Rabbit polyclonal anti-AMPA Receptor 1 (GluR1)/GluA1 , Alomone Labs , Cat# AGC-004; RRID:AB_2039878.

    Techniques: Quantitative RT-PCR, Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: CPEB3-dependent increase in GluA2 subunits impairs excitatory transmission onto inhibitory interneurons in a mouse model of fragile X

    doi: 10.1016/j.celrep.2022.110853

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Rabbit polyclonal anti-AMPA Receptor 1 (GluR1)/GluA1 , Alomone Labs , Cat# AGC-004; RRID:AB_2039878.

    Techniques: shRNA, Recombinant, Sequencing, Binding Assay, Plasmid Preparation, Software

    List of primary antibodies used

    Journal: Journal of Neuroscience Research

    Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

    doi: 10.1002/jnr.24967

    Figure Lengend Snippet: List of primary antibodies used

    Article Snippet: Anti‐GluA1 antibody produced in guinea pig , Peptide corresponding to AA 271‐285 of rat GluR1 , Alomone lab, # AGP‐009, Polyclonal antibody, RRID:AB_2340961 , 3.2 μg/ml/1:250.

    Techniques: Concentration Assay, Produced, Recombinant, Purification, Derivative Assay, Transduction

    Antibodies used and amino acid conservation between species

    Journal: Journal of Neuroscience Research

    Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

    doi: 10.1002/jnr.24967

    Figure Lengend Snippet: Antibodies used and amino acid conservation between species

    Article Snippet: Anti‐GluA1 antibody produced in guinea pig , Peptide corresponding to AA 271‐285 of rat GluR1 , Alomone lab, # AGP‐009, Polyclonal antibody, RRID:AB_2340961 , 3.2 μg/ml/1:250.

    Techniques:

    Molecular heterogeneity of astrocytes in the marmoset cerebellum. (A) Lower magnification images showing labeling for astrocytic proteins with respect to the molecular layer (ML), Purkinje cell layer (PCL), granule cell layer (GCL), and white matter (WM). Calbindin is used as a Purkinje cell marker. DAPI staining (blue) shows the nuclear staining of interneurons in the ML and granule cells in GCL. Scale bar: 100 μm. (B) Detailed images of ML and GCL stained with specific markers for Bergmann glia (BG) such as GluA1, Kirk4.1, and velate astrocytes (VA) such as AQP4. EAAT2 was expressed in both BGs and VAs with higher expression in VAs. Purkinje cell bodies are indicated with a white asterisk. Scale bar: 20 μm. bv, blood vessel

    Journal: Journal of Neuroscience Research

    Article Title: Light microscopic and heterogeneity analysis of astrocytes in the common marmoset brain

    doi: 10.1002/jnr.24967

    Figure Lengend Snippet: Molecular heterogeneity of astrocytes in the marmoset cerebellum. (A) Lower magnification images showing labeling for astrocytic proteins with respect to the molecular layer (ML), Purkinje cell layer (PCL), granule cell layer (GCL), and white matter (WM). Calbindin is used as a Purkinje cell marker. DAPI staining (blue) shows the nuclear staining of interneurons in the ML and granule cells in GCL. Scale bar: 100 μm. (B) Detailed images of ML and GCL stained with specific markers for Bergmann glia (BG) such as GluA1, Kirk4.1, and velate astrocytes (VA) such as AQP4. EAAT2 was expressed in both BGs and VAs with higher expression in VAs. Purkinje cell bodies are indicated with a white asterisk. Scale bar: 20 μm. bv, blood vessel

    Article Snippet: Anti‐GluA1 antibody produced in guinea pig , Peptide corresponding to AA 271‐285 of rat GluR1 , Alomone lab, # AGP‐009, Polyclonal antibody, RRID:AB_2340961 , 3.2 μg/ml/1:250.

    Techniques: Labeling, Marker, Staining, Expressing

    Nogo-A loss-of-function shifts the excitation/inhibition balance toward a higher excitation at a single cell level. ( A – C ) Patch-Clamp recordings in organotypic hippocampal cultures of simultaneously measuring mEPSCs and mIPSCs before (0 min) and after (5–10 min) control antibody, in all experiments mouse IgG1, FG12 (black, n = 14) or Nogo-A mouse IgG1, 11C7 function-blocking antibody (red, n = 14) treatment. ( A ) Example traces of mPSCs recorded by whole-cell patch clamp. Each trace shows representative mEPSCs (magenta) and mIPSCs (orange) distinguished by their decay time according to fast (mEPSCs < 25 ms) and slow (mIPSC > 25 ms) decay kinetics. Scale bars are 20 pA vertical and 200 ms horizontal ( B ) Normalized data for mIPSC and mEPSC ratio. ( C ) Normalized values for mIPSC and mEPSC amplitude plotted individually for each recorded neuron and the mean values visualized by the dotted line. ( D , E ) The percent change in the amplitude of mEPSC and mIPSC of individual neurons over time and their respective mean values (( D ) Ctrl-Ab: black, ( E ) Nogo-A Ab: dark red, mEPSCs: # p , mIPSCs: *** p ). ( F , G ) The percent change of individual recorded neurons in mIPSCs ( F ) and mEPSCs ( G ) exceeding average ± SEM color coded by heat mapping changes between 0–100% changes. ( H – J ) Live-cell labeling of surface GABA A and GluA1 receptors in dissociated hippocampal neurons after 10 min Nogo-A loss-of-function (red, n = 40) compared to the control condition (black, n = 40). ( H ) Representative image of surface stained GABA A Rs (green) and AMPARs subunit GluA1 (magenta). All of the images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values. Scale bars, 2 µm. ( I ) Normalized values for fluorescence intensity of GABA A R cluster and GluA1 immuno-positive puncta ratio upon Nogo-A loss-of-function for 10 min. ( J ) Normalized fluorescence intensity of GABA A R and GluA1 positive clusters plotted individually for each recorded neuron and the mean alteration visualized by the dotted line. Values represent means ± SEMs. # p < 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001; n = number of neurons.

    Journal: Cells

    Article Title: Nogo-A Modulates the Synaptic Excitation of Hippocampal Neurons in a Ca 2+ -Dependent Manner

    doi: 10.3390/cells10092299

    Figure Lengend Snippet: Nogo-A loss-of-function shifts the excitation/inhibition balance toward a higher excitation at a single cell level. ( A – C ) Patch-Clamp recordings in organotypic hippocampal cultures of simultaneously measuring mEPSCs and mIPSCs before (0 min) and after (5–10 min) control antibody, in all experiments mouse IgG1, FG12 (black, n = 14) or Nogo-A mouse IgG1, 11C7 function-blocking antibody (red, n = 14) treatment. ( A ) Example traces of mPSCs recorded by whole-cell patch clamp. Each trace shows representative mEPSCs (magenta) and mIPSCs (orange) distinguished by their decay time according to fast (mEPSCs < 25 ms) and slow (mIPSC > 25 ms) decay kinetics. Scale bars are 20 pA vertical and 200 ms horizontal ( B ) Normalized data for mIPSC and mEPSC ratio. ( C ) Normalized values for mIPSC and mEPSC amplitude plotted individually for each recorded neuron and the mean values visualized by the dotted line. ( D , E ) The percent change in the amplitude of mEPSC and mIPSC of individual neurons over time and their respective mean values (( D ) Ctrl-Ab: black, ( E ) Nogo-A Ab: dark red, mEPSCs: # p , mIPSCs: *** p ). ( F , G ) The percent change of individual recorded neurons in mIPSCs ( F ) and mEPSCs ( G ) exceeding average ± SEM color coded by heat mapping changes between 0–100% changes. ( H – J ) Live-cell labeling of surface GABA A and GluA1 receptors in dissociated hippocampal neurons after 10 min Nogo-A loss-of-function (red, n = 40) compared to the control condition (black, n = 40). ( H ) Representative image of surface stained GABA A Rs (green) and AMPARs subunit GluA1 (magenta). All of the images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values. Scale bars, 2 µm. ( I ) Normalized values for fluorescence intensity of GABA A R cluster and GluA1 immuno-positive puncta ratio upon Nogo-A loss-of-function for 10 min. ( J ) Normalized fluorescence intensity of GABA A R and GluA1 positive clusters plotted individually for each recorded neuron and the mean alteration visualized by the dotted line. Values represent means ± SEMs. # p < 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001; n = number of neurons.

    Article Snippet: In the case of the AMPA receptors, the anti-AMPAR 1 GluA1 (Alomone Labs, Jerusalem, Israel, Cat# AGP-009, 1:50) and anti-AMPAR 2 GluA2 (Alomone Labs, Cat# AGC-005, 1:50) were co-applied with the Nogo-A or control antibodies for 10 min. After completion of the treatment, the coverslips were rinsed with pre-warmed NB - medium and fixed with 4% paraformaldehyde (PFA) in phosphate buffer (PB containing in mM 50 NaH 2 PO 4 *2H 2 O, 85 Na 2 HPO 4 *2H 2 O) for 10 min at room temperature (RT).

    Techniques: Inhibition, Patch Clamp, Blocking Assay, Labeling, Staining, Fluorescence

    Nogo-A regulates the synaptic insertion of calcium permeable-AMPARs. ( A , B ) Live-cell immunolabeling of surface AMPAR subunit (magenta) GluA1 ( A ) or GluA2 ( B ) followed by immunofluorescence for presynaptic marker synapsin (Syn1/2; green) and their merged images (bottom) in primary hippocampal neurons treated for 10 min either with the control (left) or the Nogo-A function-blocking (right) antibody. For illustration, all images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values for visibility. Scale bar 2 μm. ( C , D ) Normalized data for density ( C ) and fluorescence intensity ( D ) of GluA1 immuno-positive puncta in hippocampal neurons treated with either control antibody (black, n = 40) or Nogo-A function-blocking antibody (red, n = 39) for 10 min. ( E ) Normalized values for the density of GluA1 clusters colocalized with Syn 1/2 immuno-positive puncta. ( F , G ) Normalized GluA2 cluster density ( F ) and fluorescence intensity ( G ) in hippocampal neurons upon 10 min application with control antibody (black, n = 36) or Nogo-A function-blocking antibody (red, n = 35). ( H ) Normalized density of GluA2 immuno-positive puncta colocalized with Syn 1/2. Data are presented as mean ± SEM. * p < 0.05.

    Journal: Cells

    Article Title: Nogo-A Modulates the Synaptic Excitation of Hippocampal Neurons in a Ca 2+ -Dependent Manner

    doi: 10.3390/cells10092299

    Figure Lengend Snippet: Nogo-A regulates the synaptic insertion of calcium permeable-AMPARs. ( A , B ) Live-cell immunolabeling of surface AMPAR subunit (magenta) GluA1 ( A ) or GluA2 ( B ) followed by immunofluorescence for presynaptic marker synapsin (Syn1/2; green) and their merged images (bottom) in primary hippocampal neurons treated for 10 min either with the control (left) or the Nogo-A function-blocking (right) antibody. For illustration, all images underwent deconvolution and were equally increased in brightness and contrast by the same absolute values for visibility. Scale bar 2 μm. ( C , D ) Normalized data for density ( C ) and fluorescence intensity ( D ) of GluA1 immuno-positive puncta in hippocampal neurons treated with either control antibody (black, n = 40) or Nogo-A function-blocking antibody (red, n = 39) for 10 min. ( E ) Normalized values for the density of GluA1 clusters colocalized with Syn 1/2 immuno-positive puncta. ( F , G ) Normalized GluA2 cluster density ( F ) and fluorescence intensity ( G ) in hippocampal neurons upon 10 min application with control antibody (black, n = 36) or Nogo-A function-blocking antibody (red, n = 35). ( H ) Normalized density of GluA2 immuno-positive puncta colocalized with Syn 1/2. Data are presented as mean ± SEM. * p < 0.05.

    Article Snippet: In the case of the AMPA receptors, the anti-AMPAR 1 GluA1 (Alomone Labs, Jerusalem, Israel, Cat# AGP-009, 1:50) and anti-AMPAR 2 GluA2 (Alomone Labs, Cat# AGC-005, 1:50) were co-applied with the Nogo-A or control antibodies for 10 min. After completion of the treatment, the coverslips were rinsed with pre-warmed NB - medium and fixed with 4% paraformaldehyde (PFA) in phosphate buffer (PB containing in mM 50 NaH 2 PO 4 *2H 2 O, 85 Na 2 HPO 4 *2H 2 O) for 10 min at room temperature (RT).

    Techniques: Immunolabeling, Immunofluorescence, Marker, Blocking Assay, Fluorescence

    Summary of antibodies used for the Western blotting.

    Journal: Frontiers in Synaptic Neuroscience

    Article Title: Endocannabinoids Released in the Ventral Tegmental Area During Copulation to Satiety Modulate Changes in Glutamate Receptors Associated With Synaptic Plasticity Processes

    doi: 10.3389/fnsyn.2021.701290

    Figure Lengend Snippet: Summary of antibodies used for the Western blotting.

    Article Snippet: The following primary antibodies were used: rabbit anti-CB1R, mouse anti-β-actin, goat anti-β-Arrestin2 (β-A2), mouse anti-GluN2B and mouse anti-GLUA2 (Santa Cruz Biotechnology, Dallas, TX, USA); rabbit anti-GluN2A, rabbit anti-GluN1and rabbit anti-GluA1 (Alomone Labs, Jerusalem, Israel); rabbit anti-CB1R phospho S316 (pCB1R) (Abcam, Cambridge, UK), rabbit anti-ERK 1/2 phospho Thr202/Tyr204 (pERK 1/2) (Cell Signaling Technology, Danvers, MA, USA) and mouse anti-VGlut2 (Merck-Millipore, Burlington, MA, USA).

    Techniques: Western Blot

    Changes in AMPAR density and subunit composition in the VTA of sexually satiated rats and involvement of eCBs in their induction. Western blot analysis showing changes in GluA1 (A) and GluA2 AMPAR subunit (B) densities in the VTA of control sexually experienced unmated rats (Control), males that ejaculated once (1 Ejac) or copulated to satiety (Sexually satiated) 24 h earlier, and of males that copulated to satiety in the presence of the CB1R antagonist AM251 (Satiated + AM251). Differences among untreated groups with different sexual conditions were determined by means of a one-way ANOVA followed by Tukey test, *** P < 0.001; * P < 0.05. A comparison between the rats that copulated to satiety in the absence or presence of AM251 was conducted with the unpaired t -test ** P < 0.01. Values are mean ± S.E.M of the protein/β-Actin optical density (O.D.) ratios of 5 rats per group.

    Journal: Frontiers in Synaptic Neuroscience

    Article Title: Endocannabinoids Released in the Ventral Tegmental Area During Copulation to Satiety Modulate Changes in Glutamate Receptors Associated With Synaptic Plasticity Processes

    doi: 10.3389/fnsyn.2021.701290

    Figure Lengend Snippet: Changes in AMPAR density and subunit composition in the VTA of sexually satiated rats and involvement of eCBs in their induction. Western blot analysis showing changes in GluA1 (A) and GluA2 AMPAR subunit (B) densities in the VTA of control sexually experienced unmated rats (Control), males that ejaculated once (1 Ejac) or copulated to satiety (Sexually satiated) 24 h earlier, and of males that copulated to satiety in the presence of the CB1R antagonist AM251 (Satiated + AM251). Differences among untreated groups with different sexual conditions were determined by means of a one-way ANOVA followed by Tukey test, *** P < 0.001; * P < 0.05. A comparison between the rats that copulated to satiety in the absence or presence of AM251 was conducted with the unpaired t -test ** P < 0.01. Values are mean ± S.E.M of the protein/β-Actin optical density (O.D.) ratios of 5 rats per group.

    Article Snippet: The following primary antibodies were used: rabbit anti-CB1R, mouse anti-β-actin, goat anti-β-Arrestin2 (β-A2), mouse anti-GluN2B and mouse anti-GLUA2 (Santa Cruz Biotechnology, Dallas, TX, USA); rabbit anti-GluN2A, rabbit anti-GluN1and rabbit anti-GluA1 (Alomone Labs, Jerusalem, Israel); rabbit anti-CB1R phospho S316 (pCB1R) (Abcam, Cambridge, UK), rabbit anti-ERK 1/2 phospho Thr202/Tyr204 (pERK 1/2) (Cell Signaling Technology, Danvers, MA, USA) and mouse anti-VGlut2 (Merck-Millipore, Burlington, MA, USA).

    Techniques: Western Blot

    Primary antibodies. WB , dilution for Western blotting; IF , dilution for immunofluorescence staining

    Journal: Molecular Neurobiology

    Article Title: The Golgi-Associated PDZ Domain Protein Gopc/PIST Is Required for Synaptic Targeting of mGluR5

    doi: 10.1007/s12035-021-02504-9

    Figure Lengend Snippet: Primary antibodies. WB , dilution for Western blotting; IF , dilution for immunofluorescence staining

    Article Snippet: GluA1 , Rabbit , WB 1:1000 , 5% milk powder/TBS-T , Alomone (AGC-004).

    Techniques: Western Blot, Immunofluorescence