Guinea Pig Anti Kir4 1 Kcnj10 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Olfactory ensheathing cells from the nasal mucosa and olfactory bulb have distinct membrane properties"
Article Title: Olfactory ensheathing cells from the nasal mucosa and olfactory bulb have distinct membrane properties
Figure Legend Snippet: OECs in the outer olfactory nerve layer of the rat olfactory bulb lack expression of Kir4.1. A Sagittal section of rat OB showing OSN axons (labelled for the neuronal marker βIII-tubulin) running through the outer neuronal layer (ONL), before entering the glomerular layer (GL). Glial cells, including OECs in the ONL, are immuno-labelled for S100. Kir4.1 immunofluorescence was largely undetectable in the ONL, but was evident in cells of the deeper layers. The overlays have cell nuclei counter-stained using DAPI. B A different sagittal section of rat OB, immuno-labelled for the OSN marker OMP and Kir4.1. Kir4.1 immunofluorescence was largely undetectable in the ONL, and was restricted to large multi-polar cells within the GL. The overlay has cell nuclei counter-stained using DAPI. Scale bars: 50 µ m.
Techniques Used: Expressing, Marker, Immunofluorescence, Staining
Figure Legend Snippet: OECs in the rat olfactory mucosa express Kir4.1 channel subunits. A Horizontal section of rat OM, immuno-labelled for the OEC marker S100 ( left ) and the neuronal marker tubulin ( center ). The overlay has cell nuclei counter-stained using DAPI ( right ). The cell bodies of OSNs reside within the olfactory epithelium (OE), and their axons fasciculate within lamina propria (LP). S100+ OECs lie beneath the basement membrane and in close association with SN axons. B and C show detail of a fascicle highlighted in A (*), labelled for S100 and βIII-tubulin ( B ), and for the weakly rectifying K + channel subunit Kir4.1 ( C ). D shows detail of fascicles where OSN axons are labelled using anti-OMP and OECs are labelled using anti-Kir4.1. Scale bars: A 50 µ m, B-D 20 µ m.
Techniques Used: Marker, Staining
Figure Legend Snippet: Membrane properties of OECs from the human olfactory mucosa are comparable to those from the rat olfactory mucosa. A Whole-cell voltage clamp recording from a representative human OEC following three days in vitro (3 DIV), showing membrane currents activated by sequential 200 ms voltage steps between −153 mV and 17 mV, in 10 mV increments from a holding potential of −73 mV. Hyperpolarizing steps activated large inward currents, whereas depolarizing steps typically activated outward currents. B Group (black squares) and individual (gray squares) I-V relationships of 11 human OECs after 1-3 DIV. C Normalized group and individual I-V plots demonstrated the weak voltage-dependence of the whole-cell currents. D Currents in a representative human OEC before (control) and after bath application of 100 µ M Ba 2+ . E Group I-V relationships before (open squares) and after (filled squares) Ba 2+ application in 6 OECs. F Bath applied Ba 2+ depolarized the resting membrane potential (V m ) of all cells reversibly. G Digital subtraction of the currents in the presence of Ba 2+ from control currents revealed a weak inwardly rectifying Ba 2+ -sensitive current. H Group (black squares) and individual (gray squares) I-V relationships of Ba 2+ -sensitive currents in six OECs. Inset , comparison of the Ba 2+ -sensitive component of whole-cell currents recorded in OECs from human OM and rat OM (group data shown in Fig 6H and Fig 1H , respectively). I In a 3 DIV culture, an OEC co-labeled with anti-S100 ( left panel) and anti-Kir4.1 antibodies ( center ). The overlay ( right ) has cell nuclei counter-stained using DAPI (blue), and F-actin using phalloidin (cyan). Scale bar: 50 µ m.
Techniques Used: In Vitro, Labeling, Staining
2) Product Images from "Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS"
Article Title: Expression of Kir4.1 and Kir5.1 inwardly rectifying potassium channels in oligodendrocytes, the myelinating cells of the CNS
Journal: Brain Structure & Function
Figure Legend Snippet: Glial Kir5.1 expression is reduced in the absence of Kir4.1 subunit. Immunolabelling for Kir5.1 was determined in optic nerve explants cultures, comparing wild-type mice ( A , Kir4.1 +/+ ) with Kir4.1 knock-out mice ( B , Kir4.1 −/− ), and following transfection with scrambled shRNA ( C ) or Kir4.1 shRNA ( D ); transfected cells were identified by the expression of GFP (appears green ) and insets demonstrate Kir4.1 expression in controls ( Ai , Ci ) and complete ablation in Kir4.1 −/− mice ( Bi ) and Kir4.1 shRNA ( Di ). Scale bars 10 μm. Quantification of expression of Kir4.1 ( E ) and Kir5.1 ( F ) in Kir4.1 +/+ , Kir4.1 −/− , scrambled control and Kir4.1shRNA glia; analysis was performed on 10–12 cells in each group, and data are expressed as mean ± SEM number of voxels per µm 3 , *** p
Techniques Used: Expressing, Mouse Assay, Knock-Out, Transfection, shRNA
Figure Legend Snippet: Functional implications of homomeric Kir4.1 and heteromeric Kir4.1/Kir5.1 channels in oligodendrocytes. Oligodendroglial expression of Kir4.1 channels indicates they may be important in uptake of excess K + released during axonal action potential propagation, a function largely attribiuted to astrocytes. Due to their wrapping of axons, oligodendrocytes are exposed to large ionic and pH shifts during axonal electrical activity, and it is likely weakly rectifying homomeric Kir4.1 and strongly rectifying Kir4.1/Kir5.1 heteromeric channels are important in maintaining the negative resting membrane potential, which is essential for oligodendroglial and myelin integrity. Weakly rectifying homomeric Kir4.1 channels may preferentially extrude K + and supply extracellular K + for the Na + –K + -pumps, as described in transporting epithelia. In contrast, the pH sensitivity of heteromeric Kir4.1/Kir5.1 channels is likely to have a role in the CO 2 /pH chemosensation in glia, involving carbonic anhydrase that is enriched in astrocytes and oligodendrocytes. Furthermore, intracellular acidification and inhibition of Kir4.1/Kir5.1 channels has been shown to trigger release of ATP from astrocytes, which would act on oligodendroglial P2X and P2Y receptors to provide a mechanism of astrocyte–oligodendrocyte signaling in response to metabolic challenges, which has important implications for white matter physiology and pathology
Techniques Used: Functional Assay, Expressing, Activity Assay, Inhibition
Figure Legend Snippet: Specific reduction in plasmalemmal Kir5.1 in the absence of Kir4.1. Immunolocalization of Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explant astrocytes identified by expression of GFAP, following transfection with scrambled shRNA ( A ) or Kir4.1 shRNA ( B ); transfected cells were identified by co-transfection with GFP (appears green ) and the co-localization channel indicates voxels in which Kir5.1 and Na–K-ATPase immunolabelling was at the same intensity ( Avi , Bvi ). Scale bars 20 μm. C Quantification of plasmalemmal Kir5.1 expressed as percentage of total Kir5.1 + voxels (data are mean ± SEM, n = 11–13 per group; * p
Techniques Used: Expressing, Transfection, shRNA, Cotransfection
Figure Legend Snippet: Reduction of Kir5.1 in oligodendrocytes and myelin in the absence of Kir4.1. Immunolocalization of Kir5.1 with myelin basic protein, MBP ( A , B ) and the oligodenrocyte marker APC/CC1 ( C – F ), in brain tissue from wild-type Kir4.1 +/+ mice ( A , C , E ) compared to Kir4.1 −/− knock-out mice ( B , D , F ). Scale bars 20 μm. Western blot analysis of Kir5.1 from total lysates of optic nerve ( G ) and brain ( H ) from wild-type Kir4.1 +/+ and Kir4.1 −/− knock-out mice, and mean (±SEM) integrated density normalised against β-actin ( I , n = 3, ** p
Techniques Used: Marker, Mouse Assay, Knock-Out, Western Blot
Figure Legend Snippet: Expression of Kir4.1 and Kir5.1 in oligodendrocytes and astrocytes in the cerebellum. Immunolabelling for Kir4.1 and Kir5.1, in combination with GFAP for astrocytes ( A , C ), and APC/CC1 for oligodendrocytes ( B , D ). Immunolabelling for Kir4.1 ( E ) and Kir5.1 ( F ) in mice in which EGFP is under the control of the oligodendrocyte-specific Sox10 promoter. G Double immunolabelling for Kir4.1 ( red ) and the oligodenrocyte-specific marker Olig2 ( green ). Insets in Aiv and Civ illustrate negative controls, in the Kir4.1 KO mouse ( Aiv ) and following preincubation with the Kir5.1 blocking peptide ( Civ ). Scale bars 20 μm. Western blot analysis of the brain and optic and nerve for Kir4.1 ( I ) and Kir5.1 ( J ); bands were absent in the negative controls, in the Kir4.1 knock-out mouse ( I ) following preincubation in the Kir5.1 blocking peptide ( J )
Techniques Used: Expressing, Mouse Assay, Marker, Blocking Assay, Western Blot, Knock-Out
Figure Legend Snippet: Expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Immunolabelling for Kir4.1 ( A , C ) and Kir5.1 ( B , D ), in GFAP-GFP mice to identify astrocytes ( A , B ) and PLP-DsRED mice to identify oligodendrocytes ( C , D ). Cellular expression of Kir4.1 and Kir5.1 is demonstrated by the generation of colocalisation channels ( Av , Bv , Cv , Dv ) from confocal z -stacks ( Aiv , Biv , Civ , Div ), and green and red channels of equal intensity appear yellow . Scale bars 20 μm
Techniques Used: Expressing, Mouse Assay, Plasmid Purification
Figure Legend Snippet: Co-expression of Kir4.1 and Kir5.1 in optic nerve oligodendrocytes and astrocytes. Co-immunolocalization of Kir4.1 and Kir5.1 in optic nerve explant cultures, in astrocytes identified by GFAP immunolabelling ( A ) and oligodendrocytes identified by PLP-DsRED ( B ). The overlay and individual channels are illustrated, together with the co-localisation channel for Kir4.1/Kir5.1 ( Aii, Bii ). Boxed areas on overlay images ( Ai , Bi ) are enlarged in Avi – Aviii and Bvi – Bviii , to illustrate punctate colocalization of Kir4.1 and Kir5.1 along processes (some indicated by arrows ). Scale bars 20 μm. Quantification of the number of voxels that were positive for Kir4.1 and Kir5.1 alone and of Kir4.1/Kir5.1 together, in astrocytes ( C , n = 15) and oligodendrocytes ( D , n = 13); data are mean ± SEM. Co-immunoprecipitation of Kir4.1 with Kir5.1 ( E ) and of Kir5.1 with Kir4.1 ( F ) from total brain and optic nerve (ON) lysates; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1, and using the blocking peptide for Kir5.1
Techniques Used: Expressing, Plasmid Purification, Immunoprecipitation, Knock-Out, Mouse Assay, Blocking Assay
Figure Legend Snippet: Plasmalemmal expression of Kir4.1 and Kir5.1 subunit in optic nerve glia. Immunolocalization of Kir4.1 and Kir5.1 with the membrane bound Na–K-ATPase α1 subunit in optic nerve explants of astrocytes identified by GFAP ( A , B ) and oligodendrocytes identified by PLP-DsRed ( C , D ). Scale bars 20 μm. Quantification in astrocytes and oligodendrocytes of total number of voxels immunopositive for Kir4.1 and Kir5.1, compared to voxels that were identified as colocalized for Kir4.1/Na–K-ATPase ( E ) and Kir5.1/Na–K-ATPase ( F ); data are mean ± SEM, n = 13 cells for each analysis. Western blot analysis of Kir5.1 ( G ) and Kir4.1 ( H ) in total optic nerve lysate and plasma membrane fraction. Co-immunoprecipitation of Kir4.1 ( I ) and Kir5.1 ( J ) with PSD95, in total brain and optic nerve (ON) lysate; negative controls were Kir4.1 knock-out mice (−/−) for Kir4.1 and preincubation with the blocking peptide for Kir5.1
Techniques Used: Expressing, Plasmid Purification, Western Blot, Immunoprecipitation, Knock-Out, Mouse Assay, Blocking Assay