girk2  (Alomone Labs)


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    Structured Review

    Alomone Labs girk2
    <t>GIRK2</t> is not regulated by Gα i3 in whole oocytes
    Girk2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 94 stars, based on 2 article reviews
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    Images

    1) Product Images from "Divergent regulation of GIRK1 and GIRK2 subunits of the neuronal G protein gated K+ channel by G?iGDP and G??"

    Article Title: Divergent regulation of GIRK1 and GIRK2 subunits of the neuronal G protein gated K+ channel by G?iGDP and G??

    Journal:

    doi: 10.1113/jphysiol.2009.173229

    GIRK2 is not regulated by Gα i3 in whole oocytes
    Figure Legend Snippet: GIRK2 is not regulated by Gα i3 in whole oocytes

    Techniques Used:

    Functional differences between homomeric GIRK1 and GIRK2
    Figure Legend Snippet: Functional differences between homomeric GIRK1 and GIRK2

    Techniques Used: Functional Assay

    Gα i3 GA does not regulate GIRK2 in excised plasma membrane patches
    Figure Legend Snippet: Gα i3 GA does not regulate GIRK2 in excised plasma membrane patches

    Techniques Used:

    Biochemical and functional differences between GIRK1 and GIRK2
    Figure Legend Snippet: Biochemical and functional differences between GIRK1 and GIRK2

    Techniques Used: Functional Assay

    The basal activity is Gβγ dependent in GIRK1*, but Gβγ independent in GIRK2
    Figure Legend Snippet: The basal activity is Gβγ dependent in GIRK1*, but Gβγ independent in GIRK2

    Techniques Used: Activity Assay

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    Alomone Labs anti trpv4 antibody
    Acute activation of the PKA signaling cascade promotes apical <t>TRPV4</t> translocation. A , distribution of averaged relative fluorescent signals representing TRPV4 localization along a line on z -axis in individual cells from distal nephrons similar to that
    Anti Trpv4 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Alomone Labs dopamine d2r for co ip
    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with <t>D2R</t> from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p
    Dopamine D2r For Co Ip, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Alomone Labs anti kcnn3 kca2 3 sk3 n term antibody
    Summary of nucleus accumbens <t>KCNN3</t> transcript expression in ethanol drinking rhesus macaques and C57BL/6J mice. a-c The relative expression of brain KCNN3 transcripts ( <t>SK3_ex7/8</t> , 1 SK3_ex1B , and SK3_ex4 ) among the three drinking macaque groups ( SK3_ex1B : * p = 0.0381 vs CTRL; SK3_ex4 : ** p = 0.0084 vs CTRL, *** p = 0.0009 vs CTRL). d-f The relative expression of the KCNN3 transcripts in drinking monkeys collapsed by age at onset of ethanol drinking ( SK3_ex4 : ** p
    Anti Kcnn3 Kca2 3 Sk3 N Term Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Alomone Labs shank3 antibody
    Confocal microphotographs showing the distribution of <t>SHANK3</t> in the outer retina. Double-labeled elements (green for SHANK3; red for CB and ChX10) appear yellowish and some double-labeled cells are indicated by arrows. (a–a”) Colocalization of SHANK3 with CB in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the CB (a’) , the merged image of CB and SHANK3 (a”) . CB-labeled horizontal cells (a’) . The CB-positive somata (arrows) are colabeled by SHANK3 (a,a”) . (b–b”) Colocalization of SHANK3 with ChX10 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the ChX10 (b’b”) is the merged image of ChX10 and SHANK3. ChX10 is a marker for bipolar cells. Note that labeling for SHANK3 is detected in ChX10-positive BCs (arrows). Scale bar = 20 μm.
    Shank3 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
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    Image Search Results


    Acute activation of the PKA signaling cascade promotes apical TRPV4 translocation. A , distribution of averaged relative fluorescent signals representing TRPV4 localization along a line on z -axis in individual cells from distal nephrons similar to that

    Journal: The Journal of Biological Chemistry

    Article Title: Discrete Control of TRPV4 Channel Function in the Distal Nephron by Protein Kinases A and C *

    doi: 10.1074/jbc.M113.466797

    Figure Lengend Snippet: Acute activation of the PKA signaling cascade promotes apical TRPV4 translocation. A , distribution of averaged relative fluorescent signals representing TRPV4 localization along a line on z -axis in individual cells from distal nephrons similar to that

    Article Snippet: After washing three times with PBS for 5 min, the samples were incubated for 3 h at room temperature in the dark with anti-TRPV4 antibody (1:1000 dilution; Alomone Labs) in 1% normal goat serum and 0.1% Triton X-100 in PBS.

    Techniques: Activation Assay, Translocation Assay

    Regulation of mechanosensitive [Ca 2+ ] i responses by PKA and PKC cascades occurs in a TRPV4-dependent manner. A , average time course of changes in [Ca 2+ ] i levels in response to a 10-fold elevation in flow over the apical surface ( gray bars ) for individual

    Journal: The Journal of Biological Chemistry

    Article Title: Discrete Control of TRPV4 Channel Function in the Distal Nephron by Protein Kinases A and C *

    doi: 10.1074/jbc.M113.466797

    Figure Lengend Snippet: Regulation of mechanosensitive [Ca 2+ ] i responses by PKA and PKC cascades occurs in a TRPV4-dependent manner. A , average time course of changes in [Ca 2+ ] i levels in response to a 10-fold elevation in flow over the apical surface ( gray bars ) for individual

    Article Snippet: After washing three times with PBS for 5 min, the samples were incubated for 3 h at room temperature in the dark with anti-TRPV4 antibody (1:1000 dilution; Alomone Labs) in 1% normal goat serum and 0.1% Triton X-100 in PBS.

    Techniques: Flow Cytometry

    Distinct effects of PKC- and PKA-dependent signaling cascades on subcellular TRPV4 localization in distal nephron cells. Shown are representative confocal plane micrographs (axes are shown) and corresponding cross-sections (indicated by arrows ) demonstrating

    Journal: The Journal of Biological Chemistry

    Article Title: Discrete Control of TRPV4 Channel Function in the Distal Nephron by Protein Kinases A and C *

    doi: 10.1074/jbc.M113.466797

    Figure Lengend Snippet: Distinct effects of PKC- and PKA-dependent signaling cascades on subcellular TRPV4 localization in distal nephron cells. Shown are representative confocal plane micrographs (axes are shown) and corresponding cross-sections (indicated by arrows ) demonstrating

    Article Snippet: After washing three times with PBS for 5 min, the samples were incubated for 3 h at room temperature in the dark with anti-TRPV4 antibody (1:1000 dilution; Alomone Labs) in 1% normal goat serum and 0.1% Triton X-100 in PBS.

    Techniques:

    The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p

    Journal: Frontiers in Pharmacology

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB

    doi: 10.3389/fphar.2017.00924

    Figure Lengend Snippet: The effects of D1-D2 heteromer stimulation and inactivation on basal conditioned place preference. (A) Vehicle-conditioned rats did not exhibit a preference toward a particular chamber. D1-D2 heteromer stimulation by SKF 83959 (1.5 mg/kg, s.c.) induced conditioned place aversion (CPA) as the animals spent significantly less time in the drug paired chamber. (B) SKF 83959-induced CPA was abolished by pre-treatment by the D1-D2 heteromer selective disrupting peptide, TAT-D1, but not the control TAT-Sc peptide. (C) Inactivation of D1-D2 heteromer by TAT-D1 resulted in conditioned place preference (CPP) as the rats spent significantly more time in the drug paired chamber, not observed with the control TAT-Sc. (D) Representative western blots (inset) and histogram showing the amount of D1R co-immunoprecipitated with D2R from the NAc of rats treated with saline or SKF 83959. Pretreatment with TAT-D1 led to decreased co-immunoprecipitated receptors. An aliquot of each sample was used as a control for WB (input control). (E,F) The CPA induced by D1-D2 heteromer stimulation was abolished by Cdk5 inhibitor roscovitine pre-treatment (200 nmol, i.c.v, E ) or intra-accumbal injection (30 nmol, F ). (G) Representative western blot and histogram showing the density of Thr75-DARPP-32 phosphorylation (pT75) relative to GAPDH (as loading control). Data represent means ± SEM of n = 8–10 rats/group. ( * p

    Article Snippet: AntibodiesThe primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit).

    Techniques: Conditioned Place Preference, Western Blot, Immunoprecipitation, Injection

    Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.

    Journal: Frontiers in Pharmacology

    Article Title: Activation of Dopamine D1-D2 Receptor Complex Attenuates Cocaine Reward and Reinstatement of Cocaine-Seeking through Inhibition of DARPP-32, ERK, and ΔFosB

    doi: 10.3389/fphar.2017.00924

    Figure Lengend Snippet: Evidence for the existence of dopamine D1-D2 receptor heteromer in rat NAc. (A) Proximity ligation assay (PLA) was used to visualize and detect D1R and D2R close proximity. (A1) A scheme depicts the PLA probes used in the present study. (A2–A4) Representative images of PLA signals (red dots) in neurons (nuclei stained by DAPI) in rat caudate putamen (CPu), nucleus accumbens core (NAc-core) and shell (NAc-shell) subregions. (A5) Graph representing the percent of neurons with a positive PLA signal. (A6) Representative image of PLA signals in neurons (nuclei stained by DAPI) in rat NAc-core using the second set of antibodies. (B) Representative images of immunohistochemistry using D1R antibody (D1R-Ab) or D2R antibody (D2R-Ab) directly conjugated to Alexa-488 or Alexa-568, respectively, in the NAc-shell. Direct confocal FRET analysis was performed, reflected by FRET efficiency (FRET E) and the distance between the dipoles, less than 10 nm (100 Å). (C) A representative close-up of a single MSN cell body from NAc showing D1R-D2R colocalization (left), D1-D2 heteromer FRET efficiency (center) and relative distance between receptors (right). (D,E) Histograms showing FRET E ratios (D) and distance (E) obtained from MSN cell bodies from NAc ( n = 24). Bars are 10 μm.

    Article Snippet: AntibodiesThe primary antibodies used in the present study were raised against: Dopamine D1R (Sigma, D2944, rat); dopamine D2R for IHC and PLA (1st set) (Millipore; AB5084P; rabbit), dopamine D2R for co-IP (Alomone; Rabbit), dopamine D2R for second set PLA (Millipore, ABN 462), phospho-Thr75-DARPP-32 (Cell signaling, 2301s; rabbit), phospho-Thr34-DARPP-32 (Cell Signaling, 2304; rabbit), enkephalin (Millipore, MAB350, mouse), phospho-ERK1/2 (Sigma, E7028; rabbit), ΔFosB (Cell signaling, 14695S, rabbit), GAPDH (Millipore, rabbit).

    Techniques: Proximity Ligation Assay, Staining, Immunohistochemistry

    Summary of nucleus accumbens KCNN3 transcript expression in ethanol drinking rhesus macaques and C57BL/6J mice. a-c The relative expression of brain KCNN3 transcripts ( SK3_ex7/8 , 1 SK3_ex1B , and SK3_ex4 ) among the three drinking macaque groups ( SK3_ex1B : * p = 0.0381 vs CTRL; SK3_ex4 : ** p = 0.0084 vs CTRL, *** p = 0.0009 vs CTRL). d-f The relative expression of the KCNN3 transcripts in drinking monkeys collapsed by age at onset of ethanol drinking ( SK3_ex4 : ** p

    Journal: bioRxiv

    Article Title: Cross-species epigenetic regulation of nucleus accumbens KCNN3 transcript variants by excessive ethanol drinking and dependence

    doi: 10.1101/713826

    Figure Lengend Snippet: Summary of nucleus accumbens KCNN3 transcript expression in ethanol drinking rhesus macaques and C57BL/6J mice. a-c The relative expression of brain KCNN3 transcripts ( SK3_ex7/8 , 1 SK3_ex1B , and SK3_ex4 ) among the three drinking macaque groups ( SK3_ex1B : * p = 0.0381 vs CTRL; SK3_ex4 : ** p = 0.0084 vs CTRL, *** p = 0.0009 vs CTRL). d-f The relative expression of the KCNN3 transcripts in drinking monkeys collapsed by age at onset of ethanol drinking ( SK3_ex4 : ** p

    Article Snippet: We first performed a series of western blots using different titrations of sample and antibody to establish the linear range for KCa 2.3 (Alomone Labs, Jerusalem, Israel; Catalog #: APC-025; epitope AA 2-21 of human KCa 2.3) in primate tissue samples.

    Techniques: Expressing, Mouse Assay

    Adaptations in nucleus accumbens KCNN3 transcript and protein expression in ethanol drinking female rhesus macaques. a-c The relative expression of KCNN3 transcripts ( SK3_ex7/8 , 1 SK3_ex1B , and SK3_ex4 ) among control and very heavy drinking macaque groups ( SK3_ex1B : * p = 0.037 vs CTRL; SK3_ex4 : * p = 0.024 vs CTRL). d Characterization of anti-K Ca 2.3 channel western blot in macaque accumbens tissue (protein loading range, 1.25 – 40 µg). e Positive correlation between the amount of protein loaded and anti-K Ca 2.3 channel optical density values. f,g The full K Ca 2.3 channel blot and quantitation of normalized K Ca 2.3 channel protein expression in controls and drinkers (* p = 0.0324 vs CTRL).

    Journal: bioRxiv

    Article Title: Cross-species epigenetic regulation of nucleus accumbens KCNN3 transcript variants by excessive ethanol drinking and dependence

    doi: 10.1101/713826

    Figure Lengend Snippet: Adaptations in nucleus accumbens KCNN3 transcript and protein expression in ethanol drinking female rhesus macaques. a-c The relative expression of KCNN3 transcripts ( SK3_ex7/8 , 1 SK3_ex1B , and SK3_ex4 ) among control and very heavy drinking macaque groups ( SK3_ex1B : * p = 0.037 vs CTRL; SK3_ex4 : * p = 0.024 vs CTRL). d Characterization of anti-K Ca 2.3 channel western blot in macaque accumbens tissue (protein loading range, 1.25 – 40 µg). e Positive correlation between the amount of protein loaded and anti-K Ca 2.3 channel optical density values. f,g The full K Ca 2.3 channel blot and quantitation of normalized K Ca 2.3 channel protein expression in controls and drinkers (* p = 0.0324 vs CTRL).

    Article Snippet: We first performed a series of western blots using different titrations of sample and antibody to establish the linear range for KCa 2.3 (Alomone Labs, Jerusalem, Israel; Catalog #: APC-025; epitope AA 2-21 of human KCa 2.3) in primate tissue samples.

    Techniques: Expressing, Western Blot, Quantitation Assay

    KCNN3 methylation levels within MR-ex1-200 of ethanol drinking monkeys and dependent mice. The average methylation rates of individual CpGs included in the methylation region under study are shown. a Exon organization of the KCNN3 locus showing the location of exons and the dual CAG trinucleotide repeat arrays and methylated region in exon 1 (MR-ex1). b In rhesus macaque, the following CpGs showed elevated rates of methylation in heavy/very heavy drinking macaques vs controls: CpG 129130376 : F(2, 11.846) = 7.9, * p = 0.04; CpG 129130680 : F(2, 15.955) = 3.661, * p = 0.036; CpG 129130739 : F(2, 15.468) = 3.817, * p = 0.04; CpG 129130770 : F(2, 13.57) = 5.047, * p = 0.041; CpG 129130792 : F(2, 13.945) = 7.047, * p = 0.01; CpG 129130816 : F(2, 15.63) = 5.836, * p = 0.015; CpG 129130832 : F(2, 15.473) = 3.95, * p = 0.033; CpG 129130931 : F(2, 14.393) = 4.016, * p = 0.038; CpG 129130964 : F(2, 15.708) = 3.985, * p = 0.031. c In female macaques, elevated rates of methylation were observed at the following CpGs in very heavy drinking macaques vs controls: CpG 129130612 : F(1, 7) = 6.122, * p = 0.048; CpG 129130632 : F(1, 7) = 7.64, * p = 0.033; CpG 129130685 : F(1, 7) = 13.370, * p = 0.011; CpG 129130699 : F(1, 7) = 7.799, * p = 0.031. d In mouse accumbens, the following CpGs showed elevated rates of methylation between non-drinking mice and drinking dependent mice: Independent t-tests: CpG 89555945, t (16)= −2.946, *** p = 0.0009; CpG 89555974, t (17) = −2.860, * p = 0.011; CpG 89556220, t (17) = −2.206, * p = 0.042.

    Journal: bioRxiv

    Article Title: Cross-species epigenetic regulation of nucleus accumbens KCNN3 transcript variants by excessive ethanol drinking and dependence

    doi: 10.1101/713826

    Figure Lengend Snippet: KCNN3 methylation levels within MR-ex1-200 of ethanol drinking monkeys and dependent mice. The average methylation rates of individual CpGs included in the methylation region under study are shown. a Exon organization of the KCNN3 locus showing the location of exons and the dual CAG trinucleotide repeat arrays and methylated region in exon 1 (MR-ex1). b In rhesus macaque, the following CpGs showed elevated rates of methylation in heavy/very heavy drinking macaques vs controls: CpG 129130376 : F(2, 11.846) = 7.9, * p = 0.04; CpG 129130680 : F(2, 15.955) = 3.661, * p = 0.036; CpG 129130739 : F(2, 15.468) = 3.817, * p = 0.04; CpG 129130770 : F(2, 13.57) = 5.047, * p = 0.041; CpG 129130792 : F(2, 13.945) = 7.047, * p = 0.01; CpG 129130816 : F(2, 15.63) = 5.836, * p = 0.015; CpG 129130832 : F(2, 15.473) = 3.95, * p = 0.033; CpG 129130931 : F(2, 14.393) = 4.016, * p = 0.038; CpG 129130964 : F(2, 15.708) = 3.985, * p = 0.031. c In female macaques, elevated rates of methylation were observed at the following CpGs in very heavy drinking macaques vs controls: CpG 129130612 : F(1, 7) = 6.122, * p = 0.048; CpG 129130632 : F(1, 7) = 7.64, * p = 0.033; CpG 129130685 : F(1, 7) = 13.370, * p = 0.011; CpG 129130699 : F(1, 7) = 7.799, * p = 0.031. d In mouse accumbens, the following CpGs showed elevated rates of methylation between non-drinking mice and drinking dependent mice: Independent t-tests: CpG 89555945, t (16)= −2.946, *** p = 0.0009; CpG 89555974, t (17) = −2.860, * p = 0.011; CpG 89556220, t (17) = −2.206, * p = 0.042.

    Article Snippet: We first performed a series of western blots using different titrations of sample and antibody to establish the linear range for KCa 2.3 (Alomone Labs, Jerusalem, Israel; Catalog #: APC-025; epitope AA 2-21 of human KCa 2.3) in primate tissue samples.

    Techniques: Methylation, Mouse Assay

    KCNN3 -CAG n allele frequency distribution among male and female rhesus macaques. a The frequency distribution of (CAG) n alleles is shown for low drinkers (LD), binge drinkers (BD), heavy drinkers (HD), and very heavy drinkers (VHD). b Correlation between CAG repeat sum and average ethanol intake. c-e Correlations between CAG repeat sum and KCNN3 transcript expression in long-term drinking rhesus macaques.

    Journal: bioRxiv

    Article Title: Cross-species epigenetic regulation of nucleus accumbens KCNN3 transcript variants by excessive ethanol drinking and dependence

    doi: 10.1101/713826

    Figure Lengend Snippet: KCNN3 -CAG n allele frequency distribution among male and female rhesus macaques. a The frequency distribution of (CAG) n alleles is shown for low drinkers (LD), binge drinkers (BD), heavy drinkers (HD), and very heavy drinkers (VHD). b Correlation between CAG repeat sum and average ethanol intake. c-e Correlations between CAG repeat sum and KCNN3 transcript expression in long-term drinking rhesus macaques.

    Article Snippet: We first performed a series of western blots using different titrations of sample and antibody to establish the linear range for KCa 2.3 (Alomone Labs, Jerusalem, Israel; Catalog #: APC-025; epitope AA 2-21 of human KCa 2.3) in primate tissue samples.

    Techniques: Expressing

    Confocal microphotographs showing the distribution of SHANK3 in the outer retina. Double-labeled elements (green for SHANK3; red for CB and ChX10) appear yellowish and some double-labeled cells are indicated by arrows. (a–a”) Colocalization of SHANK3 with CB in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the CB (a’) , the merged image of CB and SHANK3 (a”) . CB-labeled horizontal cells (a’) . The CB-positive somata (arrows) are colabeled by SHANK3 (a,a”) . (b–b”) Colocalization of SHANK3 with ChX10 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the ChX10 (b’b”) is the merged image of ChX10 and SHANK3. ChX10 is a marker for bipolar cells. Note that labeling for SHANK3 is detected in ChX10-positive BCs (arrows). Scale bar = 20 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Expression of SH3 and Multiple Ankyrin Repeat Domains Protein 3 in Mouse Retina

    doi: 10.3389/fncel.2022.795668

    Figure Lengend Snippet: Confocal microphotographs showing the distribution of SHANK3 in the outer retina. Double-labeled elements (green for SHANK3; red for CB and ChX10) appear yellowish and some double-labeled cells are indicated by arrows. (a–a”) Colocalization of SHANK3 with CB in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the CB (a’) , the merged image of CB and SHANK3 (a”) . CB-labeled horizontal cells (a’) . The CB-positive somata (arrows) are colabeled by SHANK3 (a,a”) . (b–b”) Colocalization of SHANK3 with ChX10 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the ChX10 (b’b”) is the merged image of ChX10 and SHANK3. ChX10 is a marker for bipolar cells. Note that labeling for SHANK3 is detected in ChX10-positive BCs (arrows). Scale bar = 20 μm.

    Article Snippet: No band was detected when the SHANK3 antibody was preabsorbed with the immunizing peptide for 3 h at room temperature.

    Techniques: Labeling, Fluorescence, Marker

    Confocal microphotographs showing the distribution of SHANK3 in the photoreceptors. Double-labeled elements (green for SHANK3; red for L/M-opsin, S-opsin and Rho4D2) appear yellowish and some double-labeled cells are indicated by arrows. (a–a”) Colocalization of SHANK3 with L/M- opsin in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the L/M-opsin (a’) , the merged image of L/M-opsin and SHANK3 (a”) . L/M-opsin-labeled the outer segment (OS) of red/green cones (a’) . Part of the L/M-opsin-positive IS (arrows) are co-labeled by SHANK3 (a,a”) . (b–b”) Colocalization of SHANK3 with S-opsin in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the S-opsin (b’) , and b” is the merged image of S-opsin and SHANK3. S-opsin is a marker for blue cone. Note that labeling for SHANK3 is detected in S-opsin-positive blue cones (arrows). (c–c”) Colocalization of SHANK3 with Rho4D2 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (c) and the Rho4D2 (c’) , the merged image of Rho4D2 and SHANK3 (c”) . Rho4D2-labeled the outer segment (OS) of rods (c’) . Rho4D2-positive OS are partially labeled by SHANK3 (c,c”) . OS, outer segment; IS, inner segment. Scale bar = 20 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Expression of SH3 and Multiple Ankyrin Repeat Domains Protein 3 in Mouse Retina

    doi: 10.3389/fncel.2022.795668

    Figure Lengend Snippet: Confocal microphotographs showing the distribution of SHANK3 in the photoreceptors. Double-labeled elements (green for SHANK3; red for L/M-opsin, S-opsin and Rho4D2) appear yellowish and some double-labeled cells are indicated by arrows. (a–a”) Colocalization of SHANK3 with L/M- opsin in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the L/M-opsin (a’) , the merged image of L/M-opsin and SHANK3 (a”) . L/M-opsin-labeled the outer segment (OS) of red/green cones (a’) . Part of the L/M-opsin-positive IS (arrows) are co-labeled by SHANK3 (a,a”) . (b–b”) Colocalization of SHANK3 with S-opsin in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the S-opsin (b’) , and b” is the merged image of S-opsin and SHANK3. S-opsin is a marker for blue cone. Note that labeling for SHANK3 is detected in S-opsin-positive blue cones (arrows). (c–c”) Colocalization of SHANK3 with Rho4D2 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (c) and the Rho4D2 (c’) , the merged image of Rho4D2 and SHANK3 (c”) . Rho4D2-labeled the outer segment (OS) of rods (c’) . Rho4D2-positive OS are partially labeled by SHANK3 (c,c”) . OS, outer segment; IS, inner segment. Scale bar = 20 μm.

    Article Snippet: No band was detected when the SHANK3 antibody was preabsorbed with the immunizing peptide for 3 h at room temperature.

    Techniques: Labeling, Fluorescence, Marker

    Confocal microphotographs showing the distribution of SHANK3 in synaptic. Double staining (green for SHANK3; red for Synaptophysin, PSD95 and VGluT1) appear yellowish. (a–a”) Colocalization of SHANK3 with Synaptophysin in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the Synaptophysin (a’,a”) is the merged image of Synaptophysin and SHANK3. (b–b”) are the locally magnified images for (a–a”) . The merged image shows that the presynaptic membrane is not labeled by SHANK3. (c–c”) Colocalization of SHANK3 with PSD95 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (c) and the PSD95 (c’,c”) is the merged image of PSD95 and SHANK3. (d–d”) are the locally magnified images for (c–c”) . The merged image shows that the postsynaptic membrane is SHANK3-positive. (e–e”) Colocalization of SHANK3 with VGluT1 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (e) and the VGluT1 (e’,e”) is the merged image of VGluT1 and SHANK3. Scale bar = 20 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Expression of SH3 and Multiple Ankyrin Repeat Domains Protein 3 in Mouse Retina

    doi: 10.3389/fncel.2022.795668

    Figure Lengend Snippet: Confocal microphotographs showing the distribution of SHANK3 in synaptic. Double staining (green for SHANK3; red for Synaptophysin, PSD95 and VGluT1) appear yellowish. (a–a”) Colocalization of SHANK3 with Synaptophysin in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the Synaptophysin (a’,a”) is the merged image of Synaptophysin and SHANK3. (b–b”) are the locally magnified images for (a–a”) . The merged image shows that the presynaptic membrane is not labeled by SHANK3. (c–c”) Colocalization of SHANK3 with PSD95 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (c) and the PSD95 (c’,c”) is the merged image of PSD95 and SHANK3. (d–d”) are the locally magnified images for (c–c”) . The merged image shows that the postsynaptic membrane is SHANK3-positive. (e–e”) Colocalization of SHANK3 with VGluT1 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (e) and the VGluT1 (e’,e”) is the merged image of VGluT1 and SHANK3. Scale bar = 20 μm.

    Article Snippet: No band was detected when the SHANK3 antibody was preabsorbed with the immunizing peptide for 3 h at room temperature.

    Techniques: Double Staining, Fluorescence, Labeling

    Confocal microphotographs showing the distribution of SHANK3 in different AC subtypes. Double staining (green for SHANK3; red for GAD 65, TH, and ChAT) appear yellowish and arrows indicate some double-labeled cells. (a–a”) Colocalization of SHANK3 with GAD 65 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the GAD 65 (a’) . Note that the GABAergic ACs in the innermost part of the INL are GAD 65-labeled, and also strongly labeled numerous neuronal processes of these cells in the IPL (a’) . All the GAD 65-positive GABAergic ACs somata are labeled by SHANK3 (a”) . (b–b”) Colocalization of SHANK3 with TH in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the TH (b’) . The soma and processes of the dopaminergic ACs are strongly labeled by TH (b’,b”) , and the soma is clearly labeled by SHANK3 (b”) . (c–c”) Colocalization of SHANK3 with ChAT in a retinal vertical section. Fluorescence double stainings show that SHANK3 (c) and the ChAT (c’) . ChAT-labeled mirror-symmetric cholinergic ACs and their processes, forming two bands in the IPL. The merged image shows that the somata of cholinergic ACs in INL and GCL were both SHANK3-positive (c”) . Scale bar = 20 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Expression of SH3 and Multiple Ankyrin Repeat Domains Protein 3 in Mouse Retina

    doi: 10.3389/fncel.2022.795668

    Figure Lengend Snippet: Confocal microphotographs showing the distribution of SHANK3 in different AC subtypes. Double staining (green for SHANK3; red for GAD 65, TH, and ChAT) appear yellowish and arrows indicate some double-labeled cells. (a–a”) Colocalization of SHANK3 with GAD 65 in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the GAD 65 (a’) . Note that the GABAergic ACs in the innermost part of the INL are GAD 65-labeled, and also strongly labeled numerous neuronal processes of these cells in the IPL (a’) . All the GAD 65-positive GABAergic ACs somata are labeled by SHANK3 (a”) . (b–b”) Colocalization of SHANK3 with TH in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the TH (b’) . The soma and processes of the dopaminergic ACs are strongly labeled by TH (b’,b”) , and the soma is clearly labeled by SHANK3 (b”) . (c–c”) Colocalization of SHANK3 with ChAT in a retinal vertical section. Fluorescence double stainings show that SHANK3 (c) and the ChAT (c’) . ChAT-labeled mirror-symmetric cholinergic ACs and their processes, forming two bands in the IPL. The merged image shows that the somata of cholinergic ACs in INL and GCL were both SHANK3-positive (c”) . Scale bar = 20 μm.

    Article Snippet: No band was detected when the SHANK3 antibody was preabsorbed with the immunizing peptide for 3 h at room temperature.

    Techniques: Double Staining, Labeling, Fluorescence

    Confocal microphotographs showing the distribution of SHANK3 in GCL and Müller cells. Double staining (green for SHANK3; red for Brn3a and CRALBP) appear yellowish, and arrows indicate some of the double-labeled cells. (a–a”) Colocalization of SHANK3 with Brn3a in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the Brn3a (a’) , (a”) are the merged image of Brn3a and SHANK3. Almost all the ganglion cells are clearly labeled by Brn3a (a’) . SHANK3 immunoreactivity is observed in all Brn3a (arrows). Meanwhile, some SHANK3-positive cells in the GCL are not immunoreactive for Brn3a (arrowheads in a–a” ), which could be displaced ACs or Brn3a-negative GCs. (b–b”) Colocalization of SHANK3 with CRALBP in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the CRALBP (b’,b”) is the merged image of CRALBP and SHANK3. CRALBP is a specific Müller cell marker. Note that SHANK3 immunostaining is observed in somata of CRALBP-positive Müller cells. Scale bar = 20 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Expression of SH3 and Multiple Ankyrin Repeat Domains Protein 3 in Mouse Retina

    doi: 10.3389/fncel.2022.795668

    Figure Lengend Snippet: Confocal microphotographs showing the distribution of SHANK3 in GCL and Müller cells. Double staining (green for SHANK3; red for Brn3a and CRALBP) appear yellowish, and arrows indicate some of the double-labeled cells. (a–a”) Colocalization of SHANK3 with Brn3a in a retinal vertical section. Fluorescence double stainings show that SHANK3 (a) and the Brn3a (a’) , (a”) are the merged image of Brn3a and SHANK3. Almost all the ganglion cells are clearly labeled by Brn3a (a’) . SHANK3 immunoreactivity is observed in all Brn3a (arrows). Meanwhile, some SHANK3-positive cells in the GCL are not immunoreactive for Brn3a (arrowheads in a–a” ), which could be displaced ACs or Brn3a-negative GCs. (b–b”) Colocalization of SHANK3 with CRALBP in a retinal vertical section. Fluorescence double stainings show that SHANK3 (b) and the CRALBP (b’,b”) is the merged image of CRALBP and SHANK3. CRALBP is a specific Müller cell marker. Note that SHANK3 immunostaining is observed in somata of CRALBP-positive Müller cells. Scale bar = 20 μm.

    Article Snippet: No band was detected when the SHANK3 antibody was preabsorbed with the immunizing peptide for 3 h at room temperature.

    Techniques: Double Staining, Labeling, Fluorescence, Marker, Immunostaining

    Expression of SH3 and multiple ankyrin repeat domains protein 3 (SHANK3) in mice retina. (A) Western blot analysis confirmed that SHANK3 was specifically expressed in the retinal homogenates, with three SHANK3 isoforms at 150–250 kDa. No band was detected when the SHANK3 antibody was pre-absorbed with the immunizing antigen. The similar was find when the primary antibody was omitted. And when the shank3 was mutant, the bands of SHANK3 were reduced. (B) Confocal fluorescence microphotographs of a vertical section of the mouse retina, labeled by SHANK3. Fluorescence staining shows that the SHANK3 is widely distributed through the whole retina. (C) Micrographs of a retinal vertical section, showing that no signal was detectable when the primary antibody for SHANK3 was pre-absorbed with the immunizing antigen. (D) Micrographs of a retinal vertical section, showing that no signal was detectable when the primary antibody was omitted. (E) Micrographs of a retinal vertical section, showing that the shank3 was mutant, the signal of SHANK3 was reduced. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale bar = 20 μm.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Expression of SH3 and Multiple Ankyrin Repeat Domains Protein 3 in Mouse Retina

    doi: 10.3389/fncel.2022.795668

    Figure Lengend Snippet: Expression of SH3 and multiple ankyrin repeat domains protein 3 (SHANK3) in mice retina. (A) Western blot analysis confirmed that SHANK3 was specifically expressed in the retinal homogenates, with three SHANK3 isoforms at 150–250 kDa. No band was detected when the SHANK3 antibody was pre-absorbed with the immunizing antigen. The similar was find when the primary antibody was omitted. And when the shank3 was mutant, the bands of SHANK3 were reduced. (B) Confocal fluorescence microphotographs of a vertical section of the mouse retina, labeled by SHANK3. Fluorescence staining shows that the SHANK3 is widely distributed through the whole retina. (C) Micrographs of a retinal vertical section, showing that no signal was detectable when the primary antibody for SHANK3 was pre-absorbed with the immunizing antigen. (D) Micrographs of a retinal vertical section, showing that no signal was detectable when the primary antibody was omitted. (E) Micrographs of a retinal vertical section, showing that the shank3 was mutant, the signal of SHANK3 was reduced. ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale bar = 20 μm.

    Article Snippet: No band was detected when the SHANK3 antibody was preabsorbed with the immunizing peptide for 3 h at room temperature.

    Techniques: Expressing, Mouse Assay, Western Blot, Mutagenesis, Fluorescence, Labeling, Staining