kir 4 1  (Alomone Labs)


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    Structured Review

    Alomone Labs kir 4 1
    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video   [Link] ,   [Link]  and Video   [Link] ,   [Link]
    Kir 4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kir 4 1/product/Alomone Labs
    Average 93 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    kir 4 1 - by Bioz Stars, 2022-12
    93/100 stars

    Images

    1) Product Images from "Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia. Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia"

    Article Title: Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia. Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.16701

    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video   [Link] ,   [Link]  and Video   [Link] ,   [Link]
    Figure Legend Snippet: 3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video [Link] , [Link] and Video [Link] , [Link]

    Techniques Used: Formalin-fixed Paraffin-Embedded, Marker, Staining

    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded murine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with Bisbenzimide (blue). GS‐ / Kir4.1‐positive satellite glial cells (SGCs) form a tight sheath around the neuronal bodies. The zoomed in images (B, D) clearly illustrate the close contact between SGCs and neurons. For GS/NeuN staining (A‐B), 39 z‐stack frames (6.4 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 39 z‐stack frames (4.7 µm total size; approx. 0.12 µm steps) were obtained. Scale bars: 20 µm (A, B, C); 10 µm (D). A movie of 3D confocal reconstructions is provided in Video   [Link] ,   [Link]  and Video   [Link] ,   [Link]
    Figure Legend Snippet: 3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded murine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with Bisbenzimide (blue). GS‐ / Kir4.1‐positive satellite glial cells (SGCs) form a tight sheath around the neuronal bodies. The zoomed in images (B, D) clearly illustrate the close contact between SGCs and neurons. For GS/NeuN staining (A‐B), 39 z‐stack frames (6.4 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 39 z‐stack frames (4.7 µm total size; approx. 0.12 µm steps) were obtained. Scale bars: 20 µm (A, B, C); 10 µm (D). A movie of 3D confocal reconstructions is provided in Video [Link] , [Link] and Video [Link] , [Link]

    Techniques Used: Formalin-fixed Paraffin-Embedded, Marker, Staining

    2) Product Images from "Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity"

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    Journal: eLife

    doi: 10.7554/eLife.36428

    Validation of OL-encoded Kcnj10 cKO efficiency Kir4.1 channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .
    Figure Legend Snippet: Validation of OL-encoded Kcnj10 cKO efficiency Kir4.1 channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .

    Techniques Used: Mouse Assay, Expressing, Purification, In Vitro, Cell Culture, Activation Assay, Labeling, Antibody Labeling, MANN-WHITNEY

    OL-Kir4.1 controls motor performance and visual function in adult mice. Mice lacking OL-Kir4.1 channels had increased mortality with survival rates of 96% in the control group (n = 29), 54% in cKO-1 (n = 13) and only 33% in cKO-2 (n = 9) mice at P180 ( A ). Log-rank (Mantel-Cox) test was performed and p-value shown in A . Kcnj10 cKO-1 (n = 10) and cKO-2 (n = 9) mice were significantly smaller than control littermates (n = 30) at P140 ( B ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in B ; **p≤0.01, ***p≤0.001. Motor dysfunction with reduced rotarod performance has been observed in both cKO-1 (n = 7) and cKO-2 (n = 7) mice as compared to controls (n = 21) ( C ). Two-way ANOVA with Tukey’s multiple comparisons test was performed in C ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. Visual function was measured by single-flash light VEP recordings from control and Kcnj10 cKO mice ( D–E ). VEPs were delayed in cKO-1 (n = 7) and cKO-2 (n = 4) mice versus controls (n = 14) ( E ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in E ; *p≤0.05. Retina integrity was measured by OCT imaging at P140 and revealed IRL thinning in cKO-1 (n = 5) and cKO-2 (n = 4) mice as compared to controls (n = 15; F ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in F ; *p≤0.05, ***p≤0.001.
    Figure Legend Snippet: OL-Kir4.1 controls motor performance and visual function in adult mice. Mice lacking OL-Kir4.1 channels had increased mortality with survival rates of 96% in the control group (n = 29), 54% in cKO-1 (n = 13) and only 33% in cKO-2 (n = 9) mice at P180 ( A ). Log-rank (Mantel-Cox) test was performed and p-value shown in A . Kcnj10 cKO-1 (n = 10) and cKO-2 (n = 9) mice were significantly smaller than control littermates (n = 30) at P140 ( B ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in B ; **p≤0.01, ***p≤0.001. Motor dysfunction with reduced rotarod performance has been observed in both cKO-1 (n = 7) and cKO-2 (n = 7) mice as compared to controls (n = 21) ( C ). Two-way ANOVA with Tukey’s multiple comparisons test was performed in C ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. Visual function was measured by single-flash light VEP recordings from control and Kcnj10 cKO mice ( D–E ). VEPs were delayed in cKO-1 (n = 7) and cKO-2 (n = 4) mice versus controls (n = 14) ( E ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in E ; *p≤0.05. Retina integrity was measured by OCT imaging at P140 and revealed IRL thinning in cKO-1 (n = 5) and cKO-2 (n = 4) mice as compared to controls (n = 15; F ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in F ; *p≤0.05, ***p≤0.001.

    Techniques Used: Mouse Assay, Imaging

    Long-term white matter pathologies in chronic OL-encoded Kcnj10 loss of function Note Kir4.1 protein levels were significantly reduced in ONs from cKO-1 mice at P180 (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, Kir4.1 protein levels were substantially decreased in spinal cord tissue from cKO-1 mice at P180 ( B–C ); note that Mbp protein levels are slightly reduced, but not significantly different between control and OL- Kcnj10 cKO-1 animals (ctrl: n = 4, cKO-1: n = 4; B–C ); Mann-Whitney tests were performed in A and C ; *p≤0.05, p=0.06 (Mbp, C ). Representative toluidine blue staining of control and cKO-1 spinal WM at P140 ( D ); note disorganized WM tracts in Kcnj10 cKO mice. By electron microscopy, dystrophic myelin and altered axonal integrity was observed in spinal WM tracts of OL-specific cKO-1 but not in control or AS-specific Kcnj10 cKO ( Aldh1l1-cre ) mice ( E ). At P140, g-ratios (146 axons from 4 control mice, 139 axons from 4 cKO-1 mice), densities and counts of intra-axonal mitochondria (73 axons from 4 control mice, 86 axons from 4 cKO-1 mice) were not altered in ONs between control and cKO-1 mice ( F–G ). Mann-Whitney tests were performed in F–G ; p=0.18 (g-ratios, F ), p=0.23 (mitochondrial densities, G ) and p=0.20 (mitochondria/axon, G ).
    Figure Legend Snippet: Long-term white matter pathologies in chronic OL-encoded Kcnj10 loss of function Note Kir4.1 protein levels were significantly reduced in ONs from cKO-1 mice at P180 (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, Kir4.1 protein levels were substantially decreased in spinal cord tissue from cKO-1 mice at P180 ( B–C ); note that Mbp protein levels are slightly reduced, but not significantly different between control and OL- Kcnj10 cKO-1 animals (ctrl: n = 4, cKO-1: n = 4; B–C ); Mann-Whitney tests were performed in A and C ; *p≤0.05, p=0.06 (Mbp, C ). Representative toluidine blue staining of control and cKO-1 spinal WM at P140 ( D ); note disorganized WM tracts in Kcnj10 cKO mice. By electron microscopy, dystrophic myelin and altered axonal integrity was observed in spinal WM tracts of OL-specific cKO-1 but not in control or AS-specific Kcnj10 cKO ( Aldh1l1-cre ) mice ( E ). At P140, g-ratios (146 axons from 4 control mice, 139 axons from 4 cKO-1 mice), densities and counts of intra-axonal mitochondria (73 axons from 4 control mice, 86 axons from 4 cKO-1 mice) were not altered in ONs between control and cKO-1 mice ( F–G ). Mann-Whitney tests were performed in F–G ; p=0.18 (g-ratios, F ), p=0.23 (mitochondrial densities, G ) and p=0.20 (mitochondria/axon, G ).

    Techniques Used: Mouse Assay, MANN-WHITNEY, Staining, Electron Microscopy

    Long-term retinal changes in chronic OL-encoded Kcnj10 loss-of-function Cartoon shows cross-section of inner retinal layers with glial cells expressing Kir4.1 ( A ); note that Kir4.1 immunoreactivity was increased in inner retinal layers of cKO-1 mice at P180 ( B–C ). Conversely, IRs for Gfap ( B–C ), Aqp4 ( D–E ) and Iba1 ( F–G ) were not different in retinal layers between control and cKO-1 mice at P180 (ctrl: n = 3, cKO: n = 3; A–F ). Mann-Whitney tests were performed in C , E and G ; *p≤0.05, p=0.1 (Gfap, C ), p=0.2 (Aqp4, E ), p=0.7 (Iba1, G ). Data are presented as mean ±s.e.m in C , E and G .
    Figure Legend Snippet: Long-term retinal changes in chronic OL-encoded Kcnj10 loss-of-function Cartoon shows cross-section of inner retinal layers with glial cells expressing Kir4.1 ( A ); note that Kir4.1 immunoreactivity was increased in inner retinal layers of cKO-1 mice at P180 ( B–C ). Conversely, IRs for Gfap ( B–C ), Aqp4 ( D–E ) and Iba1 ( F–G ) were not different in retinal layers between control and cKO-1 mice at P180 (ctrl: n = 3, cKO: n = 3; A–F ). Mann-Whitney tests were performed in C , E and G ; *p≤0.05, p=0.1 (Gfap, C ), p=0.2 (Aqp4, E ), p=0.7 (Iba1, G ). Data are presented as mean ±s.e.m in C , E and G .

    Techniques Used: Expressing, Mouse Assay, MANN-WHITNEY

    Early developmental changes in OL-encoded Kcnj10 loss-of-function Kcnj10 -deficient OPCs exhibited less BrdU incorporation in spinal cord tissue of P1 mice suggesting precocious exit from the cell cycle (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, purified and immunopanned Kcnj10 -deficient OPCs exhibited less EdU incorporation but no difference in the mitosis marker phospho-histone H3 (pH3) in-vitro (ctrl: n = 3, cKO-1: n = 3; B ). cKO-1 mice showed enhanced myelination in spinal cord WM at P1 by Mbp IHC (ctrl: n = 4, cKO-1: n = 4; C ), and Kcnj10 -deficient OLs showed more myelination during differentiating culture conditions in-vitro (ctrl: n = 4, cKO-1: n = 4; D ). Mann-Whitney tests were performed in A–D ; *p≤0.05, ***p≤0.001. Transcript levels for Cdk1 and Cdk2 were not different between control and Kcnj10 -deficient OPCs in-vitro, whereas mRNA levels for Uhrf1 and Nkx2-2 were reduced in Kcnj10 -deficient OPCs (ctrl: n = 3, cKO: n = 3; E ). Note transcript levels for Nkx2-2 and Cnp were not different between control and Kcnj10 -deficient OLs in-vitro after switching to differentiating culture conditions, however, Mbp mRNA levels increased in Kir4.1 -deficient OPCs (ctrl: n = 3, cKO-1: n = 3; F ). Multiple t tests were performed in E–F ; **p≤0.01, ***p≤0.001; E : p=0.25 ( Cdk1 ), p=0.88 ( Cdk2 ) and F ): p=0.97 ( Nkx2-2 ), p=0.42 ( Cnp ). Data are presented as mean ±s.e.m in A–F .
    Figure Legend Snippet: Early developmental changes in OL-encoded Kcnj10 loss-of-function Kcnj10 -deficient OPCs exhibited less BrdU incorporation in spinal cord tissue of P1 mice suggesting precocious exit from the cell cycle (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, purified and immunopanned Kcnj10 -deficient OPCs exhibited less EdU incorporation but no difference in the mitosis marker phospho-histone H3 (pH3) in-vitro (ctrl: n = 3, cKO-1: n = 3; B ). cKO-1 mice showed enhanced myelination in spinal cord WM at P1 by Mbp IHC (ctrl: n = 4, cKO-1: n = 4; C ), and Kcnj10 -deficient OLs showed more myelination during differentiating culture conditions in-vitro (ctrl: n = 4, cKO-1: n = 4; D ). Mann-Whitney tests were performed in A–D ; *p≤0.05, ***p≤0.001. Transcript levels for Cdk1 and Cdk2 were not different between control and Kcnj10 -deficient OPCs in-vitro, whereas mRNA levels for Uhrf1 and Nkx2-2 were reduced in Kcnj10 -deficient OPCs (ctrl: n = 3, cKO: n = 3; E ). Note transcript levels for Nkx2-2 and Cnp were not different between control and Kcnj10 -deficient OLs in-vitro after switching to differentiating culture conditions, however, Mbp mRNA levels increased in Kir4.1 -deficient OPCs (ctrl: n = 3, cKO-1: n = 3; F ). Multiple t tests were performed in E–F ; **p≤0.01, ***p≤0.001; E : p=0.25 ( Cdk1 ), p=0.88 ( Cdk2 ) and F ): p=0.97 ( Nkx2-2 ), p=0.42 ( Cnp ). Data are presented as mean ±s.e.m in A–F .

    Techniques Used: BrdU Incorporation Assay, Mouse Assay, Purification, Marker, In Vitro, Immunohistochemistry, MANN-WHITNEY

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    Alomone Labs kir 4 1
    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video   [Link] ,   [Link]  and Video   [Link] ,   [Link]
    Kir 4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kir 4 1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kir 4 1 - by Bioz Stars, 2022-12
    93/100 stars
      Buy from Supplier

    93
    Alomone Labs rabbit anti kir4 1
    Validation of OL-encoded Kcnj10 cKO efficiency <t>Kir4.1</t> channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .
    Rabbit Anti Kir4 1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti kir4 1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti kir4 1 - by Bioz Stars, 2022-12
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    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video   [Link] ,   [Link]  and Video   [Link] ,   [Link]

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia. Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

    doi: 10.1111/jcmm.16701

    Figure Lengend Snippet: 3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded canine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with bisbenzimide (blue). The zoomed in pictures (B, D) show that GS‐, respectively, Kir4.1‐positive satellite glial cells (SGCs) tightly envelop NeuN‐positive neurons. For GS/NeuN staining (A‐B), 32 z‐stack frames (5.2 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 31 z‐stack frames (5.0 µm total size; approx. 0.16 µm steps) were collected. Scale bars: 20 µm. A movie of 3D confocal reconstructions is provided in Video [Link] , [Link] and Video [Link] , [Link]

    Article Snippet: In addition, an antibody specifically targeting an extracellularly located epitope of Kir 4.1 (APC‐165, Alomone laboratories Ltd.) was also found in mSGCs and cSGCs (data not shown).

    Techniques: Formalin-fixed Paraffin-Embedded, Marker, Staining

    3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded murine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with Bisbenzimide (blue). GS‐ / Kir4.1‐positive satellite glial cells (SGCs) form a tight sheath around the neuronal bodies. The zoomed in images (B, D) clearly illustrate the close contact between SGCs and neurons. For GS/NeuN staining (A‐B), 39 z‐stack frames (6.4 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 39 z‐stack frames (4.7 µm total size; approx. 0.12 µm steps) were obtained. Scale bars: 20 µm (A, B, C); 10 µm (D). A movie of 3D confocal reconstructions is provided in Video   [Link] ,   [Link]  and Video   [Link] ,   [Link]

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia. Phenotypical peculiarities and species‐specific differences of canine and murine satellite glial cells of spinal ganglia

    doi: 10.1111/jcmm.16701

    Figure Lengend Snippet: 3D‐reconstructed confocal laser images of formalin‐fixed paraffin‐embedded murine spinal ganglia (A‐D): Double labelling with glutamine synthetase (GS; green; A, B) or inwardly rectifying potassium channel Kir 4.1 (green; C, D), respectively, and the neuronal marker NeuN (magenta). Nuclei are counterstained with Bisbenzimide (blue). GS‐ / Kir4.1‐positive satellite glial cells (SGCs) form a tight sheath around the neuronal bodies. The zoomed in images (B, D) clearly illustrate the close contact between SGCs and neurons. For GS/NeuN staining (A‐B), 39 z‐stack frames (6.4 µm total size; approx. 0.16 µm steps) and for Kir4.1/NeuN staining (C‐D), 39 z‐stack frames (4.7 µm total size; approx. 0.12 µm steps) were obtained. Scale bars: 20 µm (A, B, C); 10 µm (D). A movie of 3D confocal reconstructions is provided in Video [Link] , [Link] and Video [Link] , [Link]

    Article Snippet: In addition, an antibody specifically targeting an extracellularly located epitope of Kir 4.1 (APC‐165, Alomone laboratories Ltd.) was also found in mSGCs and cSGCs (data not shown).

    Techniques: Formalin-fixed Paraffin-Embedded, Marker, Staining

    Validation of OL-encoded Kcnj10 cKO efficiency Kir4.1 channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Validation of OL-encoded Kcnj10 cKO efficiency Kir4.1 channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Mouse Assay, Expressing, Purification, In Vitro, Cell Culture, Activation Assay, Labeling, Antibody Labeling, MANN-WHITNEY

    OL-Kir4.1 controls motor performance and visual function in adult mice. Mice lacking OL-Kir4.1 channels had increased mortality with survival rates of 96% in the control group (n = 29), 54% in cKO-1 (n = 13) and only 33% in cKO-2 (n = 9) mice at P180 ( A ). Log-rank (Mantel-Cox) test was performed and p-value shown in A . Kcnj10 cKO-1 (n = 10) and cKO-2 (n = 9) mice were significantly smaller than control littermates (n = 30) at P140 ( B ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in B ; **p≤0.01, ***p≤0.001. Motor dysfunction with reduced rotarod performance has been observed in both cKO-1 (n = 7) and cKO-2 (n = 7) mice as compared to controls (n = 21) ( C ). Two-way ANOVA with Tukey’s multiple comparisons test was performed in C ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. Visual function was measured by single-flash light VEP recordings from control and Kcnj10 cKO mice ( D–E ). VEPs were delayed in cKO-1 (n = 7) and cKO-2 (n = 4) mice versus controls (n = 14) ( E ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in E ; *p≤0.05. Retina integrity was measured by OCT imaging at P140 and revealed IRL thinning in cKO-1 (n = 5) and cKO-2 (n = 4) mice as compared to controls (n = 15; F ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in F ; *p≤0.05, ***p≤0.001.

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: OL-Kir4.1 controls motor performance and visual function in adult mice. Mice lacking OL-Kir4.1 channels had increased mortality with survival rates of 96% in the control group (n = 29), 54% in cKO-1 (n = 13) and only 33% in cKO-2 (n = 9) mice at P180 ( A ). Log-rank (Mantel-Cox) test was performed and p-value shown in A . Kcnj10 cKO-1 (n = 10) and cKO-2 (n = 9) mice were significantly smaller than control littermates (n = 30) at P140 ( B ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in B ; **p≤0.01, ***p≤0.001. Motor dysfunction with reduced rotarod performance has been observed in both cKO-1 (n = 7) and cKO-2 (n = 7) mice as compared to controls (n = 21) ( C ). Two-way ANOVA with Tukey’s multiple comparisons test was performed in C ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. Visual function was measured by single-flash light VEP recordings from control and Kcnj10 cKO mice ( D–E ). VEPs were delayed in cKO-1 (n = 7) and cKO-2 (n = 4) mice versus controls (n = 14) ( E ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in E ; *p≤0.05. Retina integrity was measured by OCT imaging at P140 and revealed IRL thinning in cKO-1 (n = 5) and cKO-2 (n = 4) mice as compared to controls (n = 15; F ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in F ; *p≤0.05, ***p≤0.001.

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Mouse Assay, Imaging

    Long-term white matter pathologies in chronic OL-encoded Kcnj10 loss of function Note Kir4.1 protein levels were significantly reduced in ONs from cKO-1 mice at P180 (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, Kir4.1 protein levels were substantially decreased in spinal cord tissue from cKO-1 mice at P180 ( B–C ); note that Mbp protein levels are slightly reduced, but not significantly different between control and OL- Kcnj10 cKO-1 animals (ctrl: n = 4, cKO-1: n = 4; B–C ); Mann-Whitney tests were performed in A and C ; *p≤0.05, p=0.06 (Mbp, C ). Representative toluidine blue staining of control and cKO-1 spinal WM at P140 ( D ); note disorganized WM tracts in Kcnj10 cKO mice. By electron microscopy, dystrophic myelin and altered axonal integrity was observed in spinal WM tracts of OL-specific cKO-1 but not in control or AS-specific Kcnj10 cKO ( Aldh1l1-cre ) mice ( E ). At P140, g-ratios (146 axons from 4 control mice, 139 axons from 4 cKO-1 mice), densities and counts of intra-axonal mitochondria (73 axons from 4 control mice, 86 axons from 4 cKO-1 mice) were not altered in ONs between control and cKO-1 mice ( F–G ). Mann-Whitney tests were performed in F–G ; p=0.18 (g-ratios, F ), p=0.23 (mitochondrial densities, G ) and p=0.20 (mitochondria/axon, G ).

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Long-term white matter pathologies in chronic OL-encoded Kcnj10 loss of function Note Kir4.1 protein levels were significantly reduced in ONs from cKO-1 mice at P180 (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, Kir4.1 protein levels were substantially decreased in spinal cord tissue from cKO-1 mice at P180 ( B–C ); note that Mbp protein levels are slightly reduced, but not significantly different between control and OL- Kcnj10 cKO-1 animals (ctrl: n = 4, cKO-1: n = 4; B–C ); Mann-Whitney tests were performed in A and C ; *p≤0.05, p=0.06 (Mbp, C ). Representative toluidine blue staining of control and cKO-1 spinal WM at P140 ( D ); note disorganized WM tracts in Kcnj10 cKO mice. By electron microscopy, dystrophic myelin and altered axonal integrity was observed in spinal WM tracts of OL-specific cKO-1 but not in control or AS-specific Kcnj10 cKO ( Aldh1l1-cre ) mice ( E ). At P140, g-ratios (146 axons from 4 control mice, 139 axons from 4 cKO-1 mice), densities and counts of intra-axonal mitochondria (73 axons from 4 control mice, 86 axons from 4 cKO-1 mice) were not altered in ONs between control and cKO-1 mice ( F–G ). Mann-Whitney tests were performed in F–G ; p=0.18 (g-ratios, F ), p=0.23 (mitochondrial densities, G ) and p=0.20 (mitochondria/axon, G ).

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Mouse Assay, MANN-WHITNEY, Staining, Electron Microscopy

    Long-term retinal changes in chronic OL-encoded Kcnj10 loss-of-function Cartoon shows cross-section of inner retinal layers with glial cells expressing Kir4.1 ( A ); note that Kir4.1 immunoreactivity was increased in inner retinal layers of cKO-1 mice at P180 ( B–C ). Conversely, IRs for Gfap ( B–C ), Aqp4 ( D–E ) and Iba1 ( F–G ) were not different in retinal layers between control and cKO-1 mice at P180 (ctrl: n = 3, cKO: n = 3; A–F ). Mann-Whitney tests were performed in C , E and G ; *p≤0.05, p=0.1 (Gfap, C ), p=0.2 (Aqp4, E ), p=0.7 (Iba1, G ). Data are presented as mean ±s.e.m in C , E and G .

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Long-term retinal changes in chronic OL-encoded Kcnj10 loss-of-function Cartoon shows cross-section of inner retinal layers with glial cells expressing Kir4.1 ( A ); note that Kir4.1 immunoreactivity was increased in inner retinal layers of cKO-1 mice at P180 ( B–C ). Conversely, IRs for Gfap ( B–C ), Aqp4 ( D–E ) and Iba1 ( F–G ) were not different in retinal layers between control and cKO-1 mice at P180 (ctrl: n = 3, cKO: n = 3; A–F ). Mann-Whitney tests were performed in C , E and G ; *p≤0.05, p=0.1 (Gfap, C ), p=0.2 (Aqp4, E ), p=0.7 (Iba1, G ). Data are presented as mean ±s.e.m in C , E and G .

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Expressing, Mouse Assay, MANN-WHITNEY

    Early developmental changes in OL-encoded Kcnj10 loss-of-function Kcnj10 -deficient OPCs exhibited less BrdU incorporation in spinal cord tissue of P1 mice suggesting precocious exit from the cell cycle (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, purified and immunopanned Kcnj10 -deficient OPCs exhibited less EdU incorporation but no difference in the mitosis marker phospho-histone H3 (pH3) in-vitro (ctrl: n = 3, cKO-1: n = 3; B ). cKO-1 mice showed enhanced myelination in spinal cord WM at P1 by Mbp IHC (ctrl: n = 4, cKO-1: n = 4; C ), and Kcnj10 -deficient OLs showed more myelination during differentiating culture conditions in-vitro (ctrl: n = 4, cKO-1: n = 4; D ). Mann-Whitney tests were performed in A–D ; *p≤0.05, ***p≤0.001. Transcript levels for Cdk1 and Cdk2 were not different between control and Kcnj10 -deficient OPCs in-vitro, whereas mRNA levels for Uhrf1 and Nkx2-2 were reduced in Kcnj10 -deficient OPCs (ctrl: n = 3, cKO: n = 3; E ). Note transcript levels for Nkx2-2 and Cnp were not different between control and Kcnj10 -deficient OLs in-vitro after switching to differentiating culture conditions, however, Mbp mRNA levels increased in Kir4.1 -deficient OPCs (ctrl: n = 3, cKO-1: n = 3; F ). Multiple t tests were performed in E–F ; **p≤0.01, ***p≤0.001; E : p=0.25 ( Cdk1 ), p=0.88 ( Cdk2 ) and F ): p=0.97 ( Nkx2-2 ), p=0.42 ( Cnp ). Data are presented as mean ±s.e.m in A–F .

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Early developmental changes in OL-encoded Kcnj10 loss-of-function Kcnj10 -deficient OPCs exhibited less BrdU incorporation in spinal cord tissue of P1 mice suggesting precocious exit from the cell cycle (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, purified and immunopanned Kcnj10 -deficient OPCs exhibited less EdU incorporation but no difference in the mitosis marker phospho-histone H3 (pH3) in-vitro (ctrl: n = 3, cKO-1: n = 3; B ). cKO-1 mice showed enhanced myelination in spinal cord WM at P1 by Mbp IHC (ctrl: n = 4, cKO-1: n = 4; C ), and Kcnj10 -deficient OLs showed more myelination during differentiating culture conditions in-vitro (ctrl: n = 4, cKO-1: n = 4; D ). Mann-Whitney tests were performed in A–D ; *p≤0.05, ***p≤0.001. Transcript levels for Cdk1 and Cdk2 were not different between control and Kcnj10 -deficient OPCs in-vitro, whereas mRNA levels for Uhrf1 and Nkx2-2 were reduced in Kcnj10 -deficient OPCs (ctrl: n = 3, cKO: n = 3; E ). Note transcript levels for Nkx2-2 and Cnp were not different between control and Kcnj10 -deficient OLs in-vitro after switching to differentiating culture conditions, however, Mbp mRNA levels increased in Kir4.1 -deficient OPCs (ctrl: n = 3, cKO-1: n = 3; F ). Multiple t tests were performed in E–F ; **p≤0.01, ***p≤0.001; E : p=0.25 ( Cdk1 ), p=0.88 ( Cdk2 ) and F ): p=0.97 ( Nkx2-2 ), p=0.42 ( Cnp ). Data are presented as mean ±s.e.m in A–F .

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: BrdU Incorporation Assay, Mouse Assay, Purification, Marker, In Vitro, Immunohistochemistry, MANN-WHITNEY

    Validation of OL-encoded Kcnj10 cKO efficiency Kir4.1 channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Validation of OL-encoded Kcnj10 cKO efficiency Kir4.1 channels were efficiently ablated from ON OLs in cKO-1 (n = 5) and cKO-2 (n = 4) mice versus control ONs (n = 5; A ). One-way ANOVA with Tukey’s multiple comparisons test was performed in A ; ****p≤0.0001. Kcnj10 was upregulated during OL differentiation, and expression significantly suppressed in purified and immunopanned OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice ( B ). Conversely, Kcnj16 was not downregulated in OPCs (ctrl: n = 3, cKO-1: n = 3) and OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice in vitro, however, note Kcnj16 downregulation during OPC-OL maturation ( C ). Cacna1c mRNA levels were increased in cultured OPCs (ctrl: n = 3, cKO-1: n = 3) but not OLs (ctrl: n = 3, cKO-1: n = 3) from cKO-1 mice suggesting a partial activation of Cav1.2 channels in OPCs ( D ). One-way ANOVA with Tukey’s multiple comparison tests were performed in B–D ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. No difference in outer tongue Kir4.1 IEM labeling between controls and ON cKO-1 tissue ( E ). Trend towards decreased Kir4.1 IEM labeling in cKO-2 ON tissue as compared to controls with respect to myelin compartments but not AS fibers ( F ). Background IEM labeling without primary antibody confirmed specificity of Kir4.1 IEM antibody labeling of myelin compartments and astrocyte fibers ( G ). Mann-Whitney tests were performed in E–G ; ****p≤0.0001, p=0.8 ( E , outer tongue IEM), p=0.07 ( F , inner tongue IEM), p=0.6 ( F , compact myelin IEM), p=0.3 ( F , outer tongue IEM), p=0.81 ( F , AS fiber IEM), p=0.12 ( G , compact myelin IEM), p=0.08 ( G , outer tongue IEM), p=0.03 ( G , AS fiber IEM). Data are presented as mean ±s.e.m in A–G .

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Mouse Assay, Expressing, Purification, In Vitro, Cell Culture, Activation Assay, Labeling, Antibody Labeling, MANN-WHITNEY

    OL-Kir4.1 controls motor performance and visual function in adult mice. Mice lacking OL-Kir4.1 channels had increased mortality with survival rates of 96% in the control group (n = 29), 54% in cKO-1 (n = 13) and only 33% in cKO-2 (n = 9) mice at P180 ( A ). Log-rank (Mantel-Cox) test was performed and p-value shown in A . Kcnj10 cKO-1 (n = 10) and cKO-2 (n = 9) mice were significantly smaller than control littermates (n = 30) at P140 ( B ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in B ; **p≤0.01, ***p≤0.001. Motor dysfunction with reduced rotarod performance has been observed in both cKO-1 (n = 7) and cKO-2 (n = 7) mice as compared to controls (n = 21) ( C ). Two-way ANOVA with Tukey’s multiple comparisons test was performed in C ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. Visual function was measured by single-flash light VEP recordings from control and Kcnj10 cKO mice ( D–E ). VEPs were delayed in cKO-1 (n = 7) and cKO-2 (n = 4) mice versus controls (n = 14) ( E ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in E ; *p≤0.05. Retina integrity was measured by OCT imaging at P140 and revealed IRL thinning in cKO-1 (n = 5) and cKO-2 (n = 4) mice as compared to controls (n = 15; F ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in F ; *p≤0.05, ***p≤0.001.

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: OL-Kir4.1 controls motor performance and visual function in adult mice. Mice lacking OL-Kir4.1 channels had increased mortality with survival rates of 96% in the control group (n = 29), 54% in cKO-1 (n = 13) and only 33% in cKO-2 (n = 9) mice at P180 ( A ). Log-rank (Mantel-Cox) test was performed and p-value shown in A . Kcnj10 cKO-1 (n = 10) and cKO-2 (n = 9) mice were significantly smaller than control littermates (n = 30) at P140 ( B ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in B ; **p≤0.01, ***p≤0.001. Motor dysfunction with reduced rotarod performance has been observed in both cKO-1 (n = 7) and cKO-2 (n = 7) mice as compared to controls (n = 21) ( C ). Two-way ANOVA with Tukey’s multiple comparisons test was performed in C ; *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. Visual function was measured by single-flash light VEP recordings from control and Kcnj10 cKO mice ( D–E ). VEPs were delayed in cKO-1 (n = 7) and cKO-2 (n = 4) mice versus controls (n = 14) ( E ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in E ; *p≤0.05. Retina integrity was measured by OCT imaging at P140 and revealed IRL thinning in cKO-1 (n = 5) and cKO-2 (n = 4) mice as compared to controls (n = 15; F ). Kruskal-Wallis with Dunn’s multiple comparisons test was performed in F ; *p≤0.05, ***p≤0.001.

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Mouse Assay, Imaging

    Long-term white matter pathologies in chronic OL-encoded Kcnj10 loss of function Note Kir4.1 protein levels were significantly reduced in ONs from cKO-1 mice at P180 (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, Kir4.1 protein levels were substantially decreased in spinal cord tissue from cKO-1 mice at P180 ( B–C ); note that Mbp protein levels are slightly reduced, but not significantly different between control and OL- Kcnj10 cKO-1 animals (ctrl: n = 4, cKO-1: n = 4; B–C ); Mann-Whitney tests were performed in A and C ; *p≤0.05, p=0.06 (Mbp, C ). Representative toluidine blue staining of control and cKO-1 spinal WM at P140 ( D ); note disorganized WM tracts in Kcnj10 cKO mice. By electron microscopy, dystrophic myelin and altered axonal integrity was observed in spinal WM tracts of OL-specific cKO-1 but not in control or AS-specific Kcnj10 cKO ( Aldh1l1-cre ) mice ( E ). At P140, g-ratios (146 axons from 4 control mice, 139 axons from 4 cKO-1 mice), densities and counts of intra-axonal mitochondria (73 axons from 4 control mice, 86 axons from 4 cKO-1 mice) were not altered in ONs between control and cKO-1 mice ( F–G ). Mann-Whitney tests were performed in F–G ; p=0.18 (g-ratios, F ), p=0.23 (mitochondrial densities, G ) and p=0.20 (mitochondria/axon, G ).

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Long-term white matter pathologies in chronic OL-encoded Kcnj10 loss of function Note Kir4.1 protein levels were significantly reduced in ONs from cKO-1 mice at P180 (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, Kir4.1 protein levels were substantially decreased in spinal cord tissue from cKO-1 mice at P180 ( B–C ); note that Mbp protein levels are slightly reduced, but not significantly different between control and OL- Kcnj10 cKO-1 animals (ctrl: n = 4, cKO-1: n = 4; B–C ); Mann-Whitney tests were performed in A and C ; *p≤0.05, p=0.06 (Mbp, C ). Representative toluidine blue staining of control and cKO-1 spinal WM at P140 ( D ); note disorganized WM tracts in Kcnj10 cKO mice. By electron microscopy, dystrophic myelin and altered axonal integrity was observed in spinal WM tracts of OL-specific cKO-1 but not in control or AS-specific Kcnj10 cKO ( Aldh1l1-cre ) mice ( E ). At P140, g-ratios (146 axons from 4 control mice, 139 axons from 4 cKO-1 mice), densities and counts of intra-axonal mitochondria (73 axons from 4 control mice, 86 axons from 4 cKO-1 mice) were not altered in ONs between control and cKO-1 mice ( F–G ). Mann-Whitney tests were performed in F–G ; p=0.18 (g-ratios, F ), p=0.23 (mitochondrial densities, G ) and p=0.20 (mitochondria/axon, G ).

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Mouse Assay, MANN-WHITNEY, Staining, Electron Microscopy

    Long-term retinal changes in chronic OL-encoded Kcnj10 loss-of-function Cartoon shows cross-section of inner retinal layers with glial cells expressing Kir4.1 ( A ); note that Kir4.1 immunoreactivity was increased in inner retinal layers of cKO-1 mice at P180 ( B–C ). Conversely, IRs for Gfap ( B–C ), Aqp4 ( D–E ) and Iba1 ( F–G ) were not different in retinal layers between control and cKO-1 mice at P180 (ctrl: n = 3, cKO: n = 3; A–F ). Mann-Whitney tests were performed in C , E and G ; *p≤0.05, p=0.1 (Gfap, C ), p=0.2 (Aqp4, E ), p=0.7 (Iba1, G ). Data are presented as mean ±s.e.m in C , E and G .

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Long-term retinal changes in chronic OL-encoded Kcnj10 loss-of-function Cartoon shows cross-section of inner retinal layers with glial cells expressing Kir4.1 ( A ); note that Kir4.1 immunoreactivity was increased in inner retinal layers of cKO-1 mice at P180 ( B–C ). Conversely, IRs for Gfap ( B–C ), Aqp4 ( D–E ) and Iba1 ( F–G ) were not different in retinal layers between control and cKO-1 mice at P180 (ctrl: n = 3, cKO: n = 3; A–F ). Mann-Whitney tests were performed in C , E and G ; *p≤0.05, p=0.1 (Gfap, C ), p=0.2 (Aqp4, E ), p=0.7 (Iba1, G ). Data are presented as mean ±s.e.m in C , E and G .

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: Expressing, Mouse Assay, MANN-WHITNEY

    Early developmental changes in OL-encoded Kcnj10 loss-of-function Kcnj10 -deficient OPCs exhibited less BrdU incorporation in spinal cord tissue of P1 mice suggesting precocious exit from the cell cycle (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, purified and immunopanned Kcnj10 -deficient OPCs exhibited less EdU incorporation but no difference in the mitosis marker phospho-histone H3 (pH3) in-vitro (ctrl: n = 3, cKO-1: n = 3; B ). cKO-1 mice showed enhanced myelination in spinal cord WM at P1 by Mbp IHC (ctrl: n = 4, cKO-1: n = 4; C ), and Kcnj10 -deficient OLs showed more myelination during differentiating culture conditions in-vitro (ctrl: n = 4, cKO-1: n = 4; D ). Mann-Whitney tests were performed in A–D ; *p≤0.05, ***p≤0.001. Transcript levels for Cdk1 and Cdk2 were not different between control and Kcnj10 -deficient OPCs in-vitro, whereas mRNA levels for Uhrf1 and Nkx2-2 were reduced in Kcnj10 -deficient OPCs (ctrl: n = 3, cKO: n = 3; E ). Note transcript levels for Nkx2-2 and Cnp were not different between control and Kcnj10 -deficient OLs in-vitro after switching to differentiating culture conditions, however, Mbp mRNA levels increased in Kir4.1 -deficient OPCs (ctrl: n = 3, cKO-1: n = 3; F ). Multiple t tests were performed in E–F ; **p≤0.01, ***p≤0.001; E : p=0.25 ( Cdk1 ), p=0.88 ( Cdk2 ) and F ): p=0.97 ( Nkx2-2 ), p=0.42 ( Cnp ). Data are presented as mean ±s.e.m in A–F .

    Journal: eLife

    Article Title: Oligodendrocyte-encoded Kir4.1 function is required for axonal integrity

    doi: 10.7554/eLife.36428

    Figure Lengend Snippet: Early developmental changes in OL-encoded Kcnj10 loss-of-function Kcnj10 -deficient OPCs exhibited less BrdU incorporation in spinal cord tissue of P1 mice suggesting precocious exit from the cell cycle (ctrl: n = 4, cKO-1: n = 4; A ). Likewise, purified and immunopanned Kcnj10 -deficient OPCs exhibited less EdU incorporation but no difference in the mitosis marker phospho-histone H3 (pH3) in-vitro (ctrl: n = 3, cKO-1: n = 3; B ). cKO-1 mice showed enhanced myelination in spinal cord WM at P1 by Mbp IHC (ctrl: n = 4, cKO-1: n = 4; C ), and Kcnj10 -deficient OLs showed more myelination during differentiating culture conditions in-vitro (ctrl: n = 4, cKO-1: n = 4; D ). Mann-Whitney tests were performed in A–D ; *p≤0.05, ***p≤0.001. Transcript levels for Cdk1 and Cdk2 were not different between control and Kcnj10 -deficient OPCs in-vitro, whereas mRNA levels for Uhrf1 and Nkx2-2 were reduced in Kcnj10 -deficient OPCs (ctrl: n = 3, cKO: n = 3; E ). Note transcript levels for Nkx2-2 and Cnp were not different between control and Kcnj10 -deficient OLs in-vitro after switching to differentiating culture conditions, however, Mbp mRNA levels increased in Kir4.1 -deficient OPCs (ctrl: n = 3, cKO-1: n = 3; F ). Multiple t tests were performed in E–F ; **p≤0.01, ***p≤0.001; E : p=0.25 ( Cdk1 ), p=0.88 ( Cdk2 ) and F ): p=0.97 ( Nkx2-2 ), p=0.42 ( Cnp ). Data are presented as mean ±s.e.m in A–F .

    Article Snippet: Schwab, 1:3,000), rat anti-MBP (ab7349, Abcam, 1:500), mouse anti-MOG (clone 8–18 C5, 1:1,000, Millipore Sigma), rat anti-GFAP (clone 2.2B10, 13–0300, Invitrogen, 1:1,000), rabbit anti-AQP4 (AB3594, 1:500, Millipore Sigma), rabbit anti-KIR4.1 (APC035, Alomone Labs, 1:3,000), rabbit anti-KIR4.1 (APC-165, Alomone Labs, 1:1,000), rabbit anti-KIR5.1 (APC123, Alomone Labs, 1:500), mouse anti-Neurofilament H (NF-H), nonphosphorylated (clone SMI32, 801701, Biolegend, 1:10,000), mouse anti-Neurofilament H (NF-H), phosphorylated (clone SMI312, 837904, Biolegend, 1:1,000), rabbit anti-IBA1 (019–19741, Wako, 1:500), goat anti-BRN3a (sc-31984, Santa Cruz, 1:200), rabbit anti-KCNQ2 (ab22897, Abcam, 1:200) rabbit anti-CASPR (ab34151, Abcam, 1:1,000), mouse anti-BRDU (347580, BD Biosciences, 1:200), rabbit anti-phospho-Histone H3 (pH3, 9701, Cell Signaling, 1:500), rat anti-CD140a (558774, BD Biosciences, 1:500), mouse anti-β-ACTIN (A5316, Sigma, 1:7,000).

    Techniques: BrdU Incorporation Assay, Mouse Assay, Purification, Marker, In Vitro, Immunohistochemistry, MANN-WHITNEY