kv4 2  (Alomone Labs)


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    Name:
    Anti Kv4 2 Antibody
    Description:
    Anti KV4 2 Antibody APC 023 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunoprecipitation immunocytochemistry and immunohistochemistry applications It has been designed to recognize KV4 2 from human rat and mouse samples
    Catalog Number:
    APC-023
    Price:
    397.0
    Category:
    Primary Antibody
    Applications:
    Immunocytochemistry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
    Purity:
    Affinity purified on immobilized antigen.
    Immunogen:
    Synthetic peptide
    Size:
    25 mcl
    Antibody Type:
    Polyclonal Primary Antibodies
    Format:
    Lyophilized Powder
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Alomone Labs kv4 2
    Anti Kv4 2 Antibody
    Anti KV4 2 Antibody APC 023 is a highly specific antibody directed against an epitope of the rat protein The antibody can be used in western blot immunoprecipitation immunocytochemistry and immunohistochemistry applications It has been designed to recognize KV4 2 from human rat and mouse samples
    https://www.bioz.com/result/kv4 2/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kv4 2 - by Bioz Stars, 2021-09
    93/100 stars

    Images

    1) Product Images from "G9a Is Essential for Epigenetic Silencing of K+ Channel Genes in Acute-to-Chronic Pain Transition"

    Article Title: G9a Is Essential for Epigenetic Silencing of K+ Channel Genes in Acute-to-Chronic Pain Transition

    Journal: Nature neuroscience

    doi: 10.1038/nn.4165

    Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, Kv4.2, Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P
    Figure Legend Snippet: Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, Kv4.2, Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P

    Techniques Used: Inhibition, Activity Assay, Expressing, Injection, MANN-WHITNEY

    2) Product Images from "Physiological Characterization of Vestibular Efferent Brainstem Neurons Using a Transgenic Mouse Model"

    Article Title: Physiological Characterization of Vestibular Efferent Brainstem Neurons Using a Transgenic Mouse Model

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0098277

    VE and LOC neurons express Kv4.3 and Kv 4.2 subunits in the mouse. Adult mouse immunofluorescently labeled against ChAT (red) and Kv4.2 or Kv4.3 α-subunits (green) using shock-frozen tissue. The ChAT labeling overlaps with the Kv4.3 labeling in the mouse VE neurons (A–C). Likewise, both the ChAT-positive LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) show Kv4.3 expression in the mouse (D–F). In a similar fashion, both the VE neurons (G–I) and the LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) (J–L) are immunolabeled against Kv4.2, which indicate that the Kv4 channels are hetero-meric in these auditory brainstem nuclei. Scale bars: 100 µm.
    Figure Legend Snippet: VE and LOC neurons express Kv4.3 and Kv 4.2 subunits in the mouse. Adult mouse immunofluorescently labeled against ChAT (red) and Kv4.2 or Kv4.3 α-subunits (green) using shock-frozen tissue. The ChAT labeling overlaps with the Kv4.3 labeling in the mouse VE neurons (A–C). Likewise, both the ChAT-positive LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) show Kv4.3 expression in the mouse (D–F). In a similar fashion, both the VE neurons (G–I) and the LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) (J–L) are immunolabeled against Kv4.2, which indicate that the Kv4 channels are hetero-meric in these auditory brainstem nuclei. Scale bars: 100 µm.

    Techniques Used: Labeling, Expressing, Immunolabeling

    3) Product Images from "Kv4.2 is a locus for PKC and ERK/MAPK cross-talk"

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    Journal: The Biochemical journal

    doi: 10.1042/BJ20081213

    Blockade of phosphorylation at the PKC sites increases the surface expression of Kv4.2 channels in COS-7 cells
    Figure Legend Snippet: Blockade of phosphorylation at the PKC sites increases the surface expression of Kv4.2 channels in COS-7 cells

    Techniques Used: Expressing

    PKC phosphorylates the Kv4.2 C-terminal but not the N-terminal cytoplasmic domains in vitro
    Figure Legend Snippet: PKC phosphorylates the Kv4.2 C-terminal but not the N-terminal cytoplasmic domains in vitro

    Techniques Used: In Vitro

    Generation and characterization of the Kv4.2 phospho-specific antibodies for the Ser447 and Ser537 site
    Figure Legend Snippet: Generation and characterization of the Kv4.2 phospho-specific antibodies for the Ser447 and Ser537 site

    Techniques Used:

    PKC phosphorylation of the Kv4.2 C-terminal augments ERK phosphorylation of the Kv4.2 C-terminal in vitro
    Figure Legend Snippet: PKC phosphorylation of the Kv4.2 C-terminal augments ERK phosphorylation of the Kv4.2 C-terminal in vitro

    Techniques Used: In Vitro

    Functional characterization of PKC sites within Kv4.2
    Figure Legend Snippet: Functional characterization of PKC sites within Kv4.2

    Techniques Used: Functional Assay

    Candidate PKC consensus sites within the Kv4.2 channel subunit
    Figure Legend Snippet: Candidate PKC consensus sites within the Kv4.2 channel subunit

    Techniques Used:

    4) Product Images from "Transient Outward K+ Current (Ito) Underlies the Right Ventricular Initiation of Polymorphic Ventricular Tachycardia in a Transgenic Rabbit Model of Long QT Type 1"

    Article Title: Transient Outward K+ Current (Ito) Underlies the Right Ventricular Initiation of Polymorphic Ventricular Tachycardia in a Transgenic Rabbit Model of Long QT Type 1

    Journal: Circulation. Arrhythmia and electrophysiology

    doi: 10.1161/CIRCEP.117.005414

    I to recovery from inactivation. A) The recovery kinetics was tested by a double-pulse protocol with interpulse time varying from 50 ms to 15 sec (n=12 RV and 7 LV cells from n=3 hearts). B) The amplitudes of the slow and fast inactivating components of I to (I to,si and I I to,fi ) as a function of inter-pulse interval were determined by fitting the time course of I to decay during the second pulse to a double exponential function. The x-axis of inter-pulse intervals is in a logarithmic scale. C) The amplitudes of I to,fi and I to,si from RV and LV. Fast and slow-inactivating components (I to,fi and I to,si ) of each I to,f and I to,s were calculated as described in Methods and represented as a stacked column plot. D) Western blots of Kv4.2, Kv1.4, and KChIP2 from LQT1 hearts. E). The accessory unit of I to , KChIP2, known to affect inactivation and recovery kinetics, was twofold higher in RV (ANOVA, p .
    Figure Legend Snippet: I to recovery from inactivation. A) The recovery kinetics was tested by a double-pulse protocol with interpulse time varying from 50 ms to 15 sec (n=12 RV and 7 LV cells from n=3 hearts). B) The amplitudes of the slow and fast inactivating components of I to (I to,si and I I to,fi ) as a function of inter-pulse interval were determined by fitting the time course of I to decay during the second pulse to a double exponential function. The x-axis of inter-pulse intervals is in a logarithmic scale. C) The amplitudes of I to,fi and I to,si from RV and LV. Fast and slow-inactivating components (I to,fi and I to,si ) of each I to,f and I to,s were calculated as described in Methods and represented as a stacked column plot. D) Western blots of Kv4.2, Kv1.4, and KChIP2 from LQT1 hearts. E). The accessory unit of I to , KChIP2, known to affect inactivation and recovery kinetics, was twofold higher in RV (ANOVA, p .

    Techniques Used: Mass Spectrometry, Size-exclusion Chromatography, Western Blot

    5) Product Images from "Characterization of the A-type potassium current in murine gastric antrum"

    Article Title: Characterization of the A-type potassium current in murine gastric antrum

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2002.025171

    Quantification of Kv4 transcripts in antrum A , detection of Kv4 transcripts in isolated antral myocytes. From left to right: 100 bp marker; Kv4.1 (amplicon = 116 bp); Kv4.2 (amplicon = 111 bp); Kv4.3, long isoform (amplicon = 176 bp). Amplicon identity was confirmed by DNA sequencing; see Methods for primer sequences. B , Kv4.1, Kv4.2 and Kv4.3 gene expression relative to β-actin in the antrum as determined by real-time PCR. *Significantly greater expression of Kv4.3 transcripts relative to Kv4.1 and Kv4.2 ( P
    Figure Legend Snippet: Quantification of Kv4 transcripts in antrum A , detection of Kv4 transcripts in isolated antral myocytes. From left to right: 100 bp marker; Kv4.1 (amplicon = 116 bp); Kv4.2 (amplicon = 111 bp); Kv4.3, long isoform (amplicon = 176 bp). Amplicon identity was confirmed by DNA sequencing; see Methods for primer sequences. B , Kv4.1, Kv4.2 and Kv4.3 gene expression relative to β-actin in the antrum as determined by real-time PCR. *Significantly greater expression of Kv4.3 transcripts relative to Kv4.1 and Kv4.2 ( P

    Techniques Used: Isolation, Marker, Amplification, DNA Sequencing, Expressing, Real-time Polymerase Chain Reaction

    Kv4.2- and Kv4.3-like immunoreactivity in the tunica muscularis of the murine antrum Haematoxylin counterstain. A and B , Kv4.2-like ( A ) and Kv4.3-like ( B ) immunoreactivity (in brown) throughout the circular (cm) and longitudinal (lm) muscle layers of the tunica muscularis in murine antrum. Arrowheads indicate Kv4-like immunoreactivity found within myenteric ganglia. Scale bars, 20 μm.
    Figure Legend Snippet: Kv4.2- and Kv4.3-like immunoreactivity in the tunica muscularis of the murine antrum Haematoxylin counterstain. A and B , Kv4.2-like ( A ) and Kv4.3-like ( B ) immunoreactivity (in brown) throughout the circular (cm) and longitudinal (lm) muscle layers of the tunica muscularis in murine antrum. Arrowheads indicate Kv4-like immunoreactivity found within myenteric ganglia. Scale bars, 20 μm.

    Techniques Used:

    6) Product Images from "Glucagon-Like Peptide-1 Cleavage Product GLP-1 (9-36) Amide Enhances Hippocampal Long-term Synaptic Plasticity in Correlation with Suppression of Kv4.2 expression and eEF2 Phosphorylation"

    Article Title: Glucagon-Like Peptide-1 Cleavage Product GLP-1 (9-36) Amide Enhances Hippocampal Long-term Synaptic Plasticity in Correlation with Suppression of Kv4.2 expression and eEF2 Phosphorylation

    Journal: Hippocampus

    doi: 10.1002/hipo.22795

    GLP-1 (9-36) administration in vivo results into decrease of Kv4.2 expression in hippocampus. (a) Western blotting demonstrated significant reduction of Kv4.2 protein levels in hippocampal slices from mice with chronic GLP-1 (9-36) treatment. n=6, * p
    Figure Legend Snippet: GLP-1 (9-36) administration in vivo results into decrease of Kv4.2 expression in hippocampus. (a) Western blotting demonstrated significant reduction of Kv4.2 protein levels in hippocampal slices from mice with chronic GLP-1 (9-36) treatment. n=6, * p

    Techniques Used: In Vivo, Expressing, Western Blot, Mouse Assay

    7) Product Images from "Unique somato-dendritic distribution pattern of Kv4.2 channels on hippocampal CA1 pyramidal cells"

    Article Title: Unique somato-dendritic distribution pattern of Kv4.2 channels on hippocampal CA1 pyramidal cells

    Journal: The European Journal of Neuroscience

    doi: 10.1111/j.1460-9568.2011.07907.x

    Densities of Kv4.2 immunogold particles in different somato-dendritic compartments of rat CA1 PCs. Bar graphs illustrate the background subtracted densities (mean ± SD, in gold/μm 2 ) of immunogold particles in the somato-dendritic compartments. The bars are colour coded to different subcellular compartments as indicated in the schematic drawing of a CA1 PC. Note the moderate increase in the density along the proximo-distal axis of PCs within the stratum radiatum and the subsequent decrease in the stratum lacunosum-moleculare.
    Figure Legend Snippet: Densities of Kv4.2 immunogold particles in different somato-dendritic compartments of rat CA1 PCs. Bar graphs illustrate the background subtracted densities (mean ± SD, in gold/μm 2 ) of immunogold particles in the somato-dendritic compartments. The bars are colour coded to different subcellular compartments as indicated in the schematic drawing of a CA1 PC. Note the moderate increase in the density along the proximo-distal axis of PCs within the stratum radiatum and the subsequent decrease in the stratum lacunosum-moleculare.

    Techniques Used:

    The Kv4.2 subunit is excluded from postsynaptic membrane specializations. (A) Electron micrograph illustrating an excitatory synapse, revealed by the accumulation of the postsynaptic density marker PSD-95 (15-nm gold). Arrowheads point to gold particles labelling the Kv4.2 subunit (10-nm gold) around the PSD. (B) High-magnification image of an inhibitory synapse identified by the enrichment of gold particles (5-nm gold) labelling the γ-aminobutyric acid (GABA) A receptor β3 subunit. Note the clustering of gold particles labelling the Kv4.2 subunit (10-nm gold) in the extrasynaptic membrane, but not within the inhibitory synapse. Scale bars: 100 nm (A); 50 nm (B).
    Figure Legend Snippet: The Kv4.2 subunit is excluded from postsynaptic membrane specializations. (A) Electron micrograph illustrating an excitatory synapse, revealed by the accumulation of the postsynaptic density marker PSD-95 (15-nm gold). Arrowheads point to gold particles labelling the Kv4.2 subunit (10-nm gold) around the PSD. (B) High-magnification image of an inhibitory synapse identified by the enrichment of gold particles (5-nm gold) labelling the γ-aminobutyric acid (GABA) A receptor β3 subunit. Note the clustering of gold particles labelling the Kv4.2 subunit (10-nm gold) in the extrasynaptic membrane, but not within the inhibitory synapse. Scale bars: 100 nm (A); 50 nm (B).

    Techniques Used: Marker

    Specificity test for Kv4.2 subunit labelling on the axo-somato-dendritic surface of CA1 PCs using SDS-FRL. (A) Electron micrograph illustrating the P-face of a spiny CA1 PC dendrite of wild-type mouse immunolabelled for the Kv4.2 subunit. Note that the density and distribution pattern of the immunogold labelling for the Kv4.2 subunit in mouse is similar to that seen in rat. (B) The lack of immunogold labelling for the Kv4.2 subunit in a spiny dendrite of a Kv4.2 −/− mouse demonstrates the specificity of the labelling using SDS-FRL. (C) Axon terminals in rat identified by immunolabelling for SNAP-25 (15-nm gold) contain a low number of immunogold particles (arrows) for the Kv4.2 subunit (10-nm gold). (D and E) Immunogold particles for the Kv4.2 subunit were present in both Kv4.2 +/+ (D) and Kv4.2 −/− mice (E) at approximately the same density. dendr, dendrite; sp, spine. Scale bars: 250 nm (A–E).
    Figure Legend Snippet: Specificity test for Kv4.2 subunit labelling on the axo-somato-dendritic surface of CA1 PCs using SDS-FRL. (A) Electron micrograph illustrating the P-face of a spiny CA1 PC dendrite of wild-type mouse immunolabelled for the Kv4.2 subunit. Note that the density and distribution pattern of the immunogold labelling for the Kv4.2 subunit in mouse is similar to that seen in rat. (B) The lack of immunogold labelling for the Kv4.2 subunit in a spiny dendrite of a Kv4.2 −/− mouse demonstrates the specificity of the labelling using SDS-FRL. (C) Axon terminals in rat identified by immunolabelling for SNAP-25 (15-nm gold) contain a low number of immunogold particles (arrows) for the Kv4.2 subunit (10-nm gold). (D and E) Immunogold particles for the Kv4.2 subunit were present in both Kv4.2 +/+ (D) and Kv4.2 −/− mice (E) at approximately the same density. dendr, dendrite; sp, spine. Scale bars: 250 nm (A–E).

    Techniques Used: Mouse Assay

    Distribution of immunogold particles for the Kv4.2 subunit in oblique dendrites and spines of CA1 PCs in the stratum radiatum. (A) Immunogold particles are homogenously distributed along the P-face of an oblique dendrite (obl.d.) in the proximal stratum radiatum (prox. sr). (B) Slightly higher immunogold particle density for the Kv4.2 subunit can be seen on an oblique dendrite from the middle stratum radiatum (mid. sr). Note that a spine (sp) with high density of immunogold particles emerges from the dendrite and faces an axon terminal (at) E-face. (C) A spiny oblique dendrite in the distal stratum radiatum (dist. sr) contains a large number of immunogold particles for the Kv4.2 subunit. Scale bars: 250 nm (A–C).
    Figure Legend Snippet: Distribution of immunogold particles for the Kv4.2 subunit in oblique dendrites and spines of CA1 PCs in the stratum radiatum. (A) Immunogold particles are homogenously distributed along the P-face of an oblique dendrite (obl.d.) in the proximal stratum radiatum (prox. sr). (B) Slightly higher immunogold particle density for the Kv4.2 subunit can be seen on an oblique dendrite from the middle stratum radiatum (mid. sr). Note that a spine (sp) with high density of immunogold particles emerges from the dendrite and faces an axon terminal (at) E-face. (C) A spiny oblique dendrite in the distal stratum radiatum (dist. sr) contains a large number of immunogold particles for the Kv4.2 subunit. Scale bars: 250 nm (A–C).

    Techniques Used:

    Distribution of the Kv4.2 subunit immunoreactivity in the hippocampus and the specificity of immunoreactions. (A) The immunfluorescent reaction shows strong, homogenous neuropil labelling for the Kv4.2 subunit in strata oriens (so) and radiatum (sr) of rat CA1 region, with a slight reduction of the immunolabelling in stratum lacunosum-moleculare (slm). (B) A very similar labelling pattern was observed in the mouse hippocampus. (C) The labelling was absent in the CA1 area of Kv4.2 −/− mice, demonstrating the specificity of the immunoreaction. sp, stratum pyramidale. All images are single confocal sections. Scale bars: 50 μm (A–C).
    Figure Legend Snippet: Distribution of the Kv4.2 subunit immunoreactivity in the hippocampus and the specificity of immunoreactions. (A) The immunfluorescent reaction shows strong, homogenous neuropil labelling for the Kv4.2 subunit in strata oriens (so) and radiatum (sr) of rat CA1 region, with a slight reduction of the immunolabelling in stratum lacunosum-moleculare (slm). (B) A very similar labelling pattern was observed in the mouse hippocampus. (C) The labelling was absent in the CA1 area of Kv4.2 −/− mice, demonstrating the specificity of the immunoreaction. sp, stratum pyramidale. All images are single confocal sections. Scale bars: 50 μm (A–C).

    Techniques Used: Mouse Assay

    High-resolution immunogold localization of the Kv4.2 subunit in somata and apical dendrites of CA1 PCs. (A–E) Low-magnification images of P-faces of a PC soma (A) and spiny apical dendrites from the proximal (B), middle (C) and distal (D) stratum radiatum and stratum lacunosum-moleculare (E) show a rather uniform immunogold labelling pattern. (A′–E′) High-magnification images of the boxed region in (A–E). (F) Immunogold particles for the Kv4.2 subunit are homogenously distributed around a branch point on a PC dendrite. (F′) High-magnification view of the boxed region in (F). dist. sr, distal stratum radiatum; mid. sr, middle stratum radiatum; prox. sr, proximal stratum radiatum; slm, stratum lacunosum-moleculare; sp, stratum pyramidale. Scale bars: 1 μm (A); 500 nm (B–F); 100 nm (A′–F′).
    Figure Legend Snippet: High-resolution immunogold localization of the Kv4.2 subunit in somata and apical dendrites of CA1 PCs. (A–E) Low-magnification images of P-faces of a PC soma (A) and spiny apical dendrites from the proximal (B), middle (C) and distal (D) stratum radiatum and stratum lacunosum-moleculare (E) show a rather uniform immunogold labelling pattern. (A′–E′) High-magnification images of the boxed region in (A–E). (F) Immunogold particles for the Kv4.2 subunit are homogenously distributed around a branch point on a PC dendrite. (F′) High-magnification view of the boxed region in (F). dist. sr, distal stratum radiatum; mid. sr, middle stratum radiatum; prox. sr, proximal stratum radiatum; slm, stratum lacunosum-moleculare; sp, stratum pyramidale. Scale bars: 1 μm (A); 500 nm (B–F); 100 nm (A′–F′).

    Techniques Used:

    8) Product Images from "Differential Expression of IA Channel Subunits Kv4.2 and Kv4.3 in Mouse Visual Cortical Neurons and Synapses"

    Article Title: Differential Expression of IA Channel Subunits Kv4.2 and Kv4.3 in Mouse Visual Cortical Neurons and Synapses

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2599-06.2006

    Schematic representation of Kv4.2 (red dots) and Kv4.3 (black dots) subunit distribution in neocortical pyramidal cells (top) and GABAergic interneurons (bottom). A close-up at the bottom right applies to both Kv4.2 and Kv4.3 and shows that Kv4 channels are excluded from excitatory synapses (green) but are present at GABAergic inhibitory synapses (purple). PC, Pyramidal cell.
    Figure Legend Snippet: Schematic representation of Kv4.2 (red dots) and Kv4.3 (black dots) subunit distribution in neocortical pyramidal cells (top) and GABAergic interneurons (bottom). A close-up at the bottom right applies to both Kv4.2 and Kv4.3 and shows that Kv4 channels are excluded from excitatory synapses (green) but are present at GABAergic inhibitory synapses (purple). PC, Pyramidal cell.

    Techniques Used:

    Expression levels of Kv4.2 and Kv4.3 in mouse primary visual and primary somatosensory cortex are layer specific. A , B , Immunofluorescence images of Kv4.2 and Kv4.3 expression in coronal sections through mouse primary visual cortex. Staining is densest in layers 1 and 4 and the layer 5/6 border. C , D , Kv4.2 and Kv4.3 immunofluorescence in tangential sections through mouse primary somatosensory cortex. Labeling is most intense in layer 4, in the center of barrels. Expression in septa between barrels is sparse. Scale bars: A , B , 0.2 mm; C , D , 1 mm.
    Figure Legend Snippet: Expression levels of Kv4.2 and Kv4.3 in mouse primary visual and primary somatosensory cortex are layer specific. A , B , Immunofluorescence images of Kv4.2 and Kv4.3 expression in coronal sections through mouse primary visual cortex. Staining is densest in layers 1 and 4 and the layer 5/6 border. C , D , Kv4.2 and Kv4.3 immunofluorescence in tangential sections through mouse primary somatosensory cortex. Labeling is most intense in layer 4, in the center of barrels. Expression in septa between barrels is sparse. Scale bars: A , B , 0.2 mm; C , D , 1 mm.

    Techniques Used: Expressing, Immunofluorescence, Staining, Labeling

    Kv4.2 expression in GABAergic neurons of mouse primary visual cortex. Confocal microscopic images of immunolabeled GABAergic neurons (red) in layer 2/3 ( A – C ) and layer 5 ( D – F ). Kv4.2 (green) is coexpressed in somata and dendrites of a subset of GABAergic neurons. Arrows in A – C mark Kv4.2-expressing GABAergic neuron. Scale bars, 10 μm.
    Figure Legend Snippet: Kv4.2 expression in GABAergic neurons of mouse primary visual cortex. Confocal microscopic images of immunolabeled GABAergic neurons (red) in layer 2/3 ( A – C ) and layer 5 ( D – F ). Kv4.2 (green) is coexpressed in somata and dendrites of a subset of GABAergic neurons. Arrows in A – C mark Kv4.2-expressing GABAergic neuron. Scale bars, 10 μm.

    Techniques Used: Expressing, Immunolabeling

    Kv4.2 clusters are expressed in pyramidal cell somata, dendrites, and spines. A , Confocal microscopic images of punctate Kv4.2 immunolabeling (red) in layers 4 and 5 of mouse primary visual cortex. The distribution of Kv4.2 clusters is non-uniform, showing a higher density over the neuropil than the pyramidal cell soma in the same focal plane (arrows; A , B ). B , C , Kv4.2 clusters (red, yellow) are associated with YFP-labeled (green) pyramidal cell somata and with basal and apical dendrites. D – F , High-magnification images of the boxed apical dendrite of a YFP-labeled layer 5 pyramidal neuron ( B , C ), showing close association of Kv4.2-labeled clusters (red and yellow puncta marked with arrows) with dendritic shaft and spines. The numbers indicate layers. Scale bars: A – C , 10 μm; D – F , 2 μm.
    Figure Legend Snippet: Kv4.2 clusters are expressed in pyramidal cell somata, dendrites, and spines. A , Confocal microscopic images of punctate Kv4.2 immunolabeling (red) in layers 4 and 5 of mouse primary visual cortex. The distribution of Kv4.2 clusters is non-uniform, showing a higher density over the neuropil than the pyramidal cell soma in the same focal plane (arrows; A , B ). B , C , Kv4.2 clusters (red, yellow) are associated with YFP-labeled (green) pyramidal cell somata and with basal and apical dendrites. D – F , High-magnification images of the boxed apical dendrite of a YFP-labeled layer 5 pyramidal neuron ( B , C ), showing close association of Kv4.2-labeled clusters (red and yellow puncta marked with arrows) with dendritic shaft and spines. The numbers indicate layers. Scale bars: A – C , 10 μm; D – F , 2 μm.

    Techniques Used: Immunolabeling, Labeling

    Electron micrographs showing that Kv4.2 is differentially expressed at putative inhibitory and excitatory synapses in mouse primary visual cortex. A , Extrasynaptic localization of Kv4.2 immunoperoxidase labeling (arrows) at an asymmetric synapse onto a thin dendrite (D). The dark immunoperoxidase reaction product is easily distinguished from the much lighter postsynaptic density. B , Kv4.2 immunoperoxidase at the base and neck of spine. Kv4.2 is absent from the asymmetric synapse. C , Extrasynaptic localization of Kv4.2 at an asymmetric synapse onto spine. D , Extrasynaptic localization of Kv4.2 (arrows) at an asymmetric synapse onto GABA immunogold (black dots)-labeled dendrite. E , Synaptic localization of Kv4.2 at a symmetric synapse (AT2) onto a pyramidal cell soma (P). Notice that Kv4.2 is contained in the postsynaptic membrane associated with the synaptic cleft, the margins of which are indicated by arrowheads. Scale bars, 0.5 μm. AT1, Asymmetric synapse; Sp, spine.
    Figure Legend Snippet: Electron micrographs showing that Kv4.2 is differentially expressed at putative inhibitory and excitatory synapses in mouse primary visual cortex. A , Extrasynaptic localization of Kv4.2 immunoperoxidase labeling (arrows) at an asymmetric synapse onto a thin dendrite (D). The dark immunoperoxidase reaction product is easily distinguished from the much lighter postsynaptic density. B , Kv4.2 immunoperoxidase at the base and neck of spine. Kv4.2 is absent from the asymmetric synapse. C , Extrasynaptic localization of Kv4.2 at an asymmetric synapse onto spine. D , Extrasynaptic localization of Kv4.2 (arrows) at an asymmetric synapse onto GABA immunogold (black dots)-labeled dendrite. E , Synaptic localization of Kv4.2 at a symmetric synapse (AT2) onto a pyramidal cell soma (P). Notice that Kv4.2 is contained in the postsynaptic membrane associated with the synaptic cleft, the margins of which are indicated by arrowheads. Scale bars, 0.5 μm. AT1, Asymmetric synapse; Sp, spine.

    Techniques Used: Labeling

    9) Product Images from "Glucagon-Like Peptide-1 Cleavage Product GLP-1 (9-36) Amide Enhances Hippocampal Long-term Synaptic Plasticity in Correlation with Suppression of Kv4.2 expression and eEF2 Phosphorylation"

    Article Title: Glucagon-Like Peptide-1 Cleavage Product GLP-1 (9-36) Amide Enhances Hippocampal Long-term Synaptic Plasticity in Correlation with Suppression of Kv4.2 expression and eEF2 Phosphorylation

    Journal: Hippocampus

    doi: 10.1002/hipo.22795

    GLP-1 (9-36) administration in vivo results into decrease of Kv4.2 expression in hippocampus. (a) Western blotting demonstrated significant reduction of Kv4.2 protein levels in hippocampal slices from mice with chronic GLP-1 (9-36) treatment. n=6, * p
    Figure Legend Snippet: GLP-1 (9-36) administration in vivo results into decrease of Kv4.2 expression in hippocampus. (a) Western blotting demonstrated significant reduction of Kv4.2 protein levels in hippocampal slices from mice with chronic GLP-1 (9-36) treatment. n=6, * p

    Techniques Used: In Vivo, Expressing, Western Blot, Mouse Assay

    10) Product Images from "Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels"

    Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

    Journal: Annals of neurology

    doi: 10.1002/ana.23756

    Immunoprecipitation of DPPX In cultures of dissociated rat hippocampal neurons, patients’ antibodies showed intense reactivity with the neuronal cell surface (A), bar = 10 μm. Immunoprecipitation of the antigen with serum of the index case is shown in B, where the precipitated proteins were run in a gel and subsequently stained with EZblue. Note that patient’s antibodies precipitated a protein (band close to 102 kDa in lane P), which was excised from the gel and analyzed by mass spectrometry, demonstrating sequences of DPPX. Lane N is the precipitate obtained from control serum. Immunoblot of these proteins with a rabbit polyclonal antibody against DPPX (1:1000, developed by BR) confirmed that the band corresponded to DPPX (C).
    Figure Legend Snippet: Immunoprecipitation of DPPX In cultures of dissociated rat hippocampal neurons, patients’ antibodies showed intense reactivity with the neuronal cell surface (A), bar = 10 μm. Immunoprecipitation of the antigen with serum of the index case is shown in B, where the precipitated proteins were run in a gel and subsequently stained with EZblue. Note that patient’s antibodies precipitated a protein (band close to 102 kDa in lane P), which was excised from the gel and analyzed by mass spectrometry, demonstrating sequences of DPPX. Lane N is the precipitate obtained from control serum. Immunoblot of these proteins with a rabbit polyclonal antibody against DPPX (1:1000, developed by BR) confirmed that the band corresponded to DPPX (C).

    Techniques Used: Immunoprecipitation, Staining, Mass Spectrometry

    Expression of DPPX in myenteric plexus Transverse section of small bowel of rat showing the longitudinal muscular layer (LM), circular muscular layer (CM), submucosal layer (SM), and glans (G). The myenteric plexus (Plex) is revealed as clusters of large neurons between the two muscular layers. In the 3 panels (A–C) the nuclei of the neurons (red) was labeled with anti-Hu (a highly specific neuronal marker). Panel A, shows in green the DPPX immunostaining using a rabbit polyclonal antibody (1:1000, developed by BD); panel B shows the DPPX reactivity of serum from one of the patients with encephalitis, and panel C shows the lack of reactivity of serum from a healthy subject. Note that DPPX is predominantly expressed in the cytoplasm-membrane of the large clustered neurons of the myenteric plexus, and is also detected in a fine longitudinal pattern in CM and SM where the submucosal plexus is located. Bar = 20μm.
    Figure Legend Snippet: Expression of DPPX in myenteric plexus Transverse section of small bowel of rat showing the longitudinal muscular layer (LM), circular muscular layer (CM), submucosal layer (SM), and glans (G). The myenteric plexus (Plex) is revealed as clusters of large neurons between the two muscular layers. In the 3 panels (A–C) the nuclei of the neurons (red) was labeled with anti-Hu (a highly specific neuronal marker). Panel A, shows in green the DPPX immunostaining using a rabbit polyclonal antibody (1:1000, developed by BD); panel B shows the DPPX reactivity of serum from one of the patients with encephalitis, and panel C shows the lack of reactivity of serum from a healthy subject. Note that DPPX is predominantly expressed in the cytoplasm-membrane of the large clustered neurons of the myenteric plexus, and is also detected in a fine longitudinal pattern in CM and SM where the submucosal plexus is located. Bar = 20μm.

    Techniques Used: Expressing, Labeling, Marker, Immunostaining

    11) Product Images from "Expression, Cellular and Subcellular Localisation of Kv4.2 and Kv4.3 Channels in the Rodent Hippocampus"

    Article Title: Expression, Cellular and Subcellular Localisation of Kv4.2 and Kv4.3 Channels in the Rodent Hippocampus

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20020246

    Distribution of immunoreactivity for Kv4.2 in the hippocampus. ( A ) At the light microscopic level, immunoreactivity for Kv4.2 was widely distributed in the hippocampus but its intensity varied consistently; ( B ) In the CA1 region, immunolabelling for Kv4.2 was generally moderate-to-strong, with the strata oriens (so) and radiatum (sr) showing the highest and the stratum lacunosum-moleculare (slm) showing weak immunoreactivity. The weakest immunolabelling for Kv4.2 was observed in the stratum pyramidale (sp); ( C ) In the CA3 region, moderate immunolabelling for Kv4.2 was observed in the strata oriens (so), radiatum (sr) and stratum lacunosum-moleculare (slm), weak in the stratum lucidum and very weak in the stratum pyramidale (sp). Kv4.2 immunolabelling was also seen outlining somata and dendrites of scattered interneurons (arrows); ( D ) In the dentate gyrus (DG), immunolabelling was strong in the molecular layer (ml), weak in the hilus (h) and the weakest in the granule cell layer (gc). Scale bars: A , 500 µm; B – D , 150 µm.
    Figure Legend Snippet: Distribution of immunoreactivity for Kv4.2 in the hippocampus. ( A ) At the light microscopic level, immunoreactivity for Kv4.2 was widely distributed in the hippocampus but its intensity varied consistently; ( B ) In the CA1 region, immunolabelling for Kv4.2 was generally moderate-to-strong, with the strata oriens (so) and radiatum (sr) showing the highest and the stratum lacunosum-moleculare (slm) showing weak immunoreactivity. The weakest immunolabelling for Kv4.2 was observed in the stratum pyramidale (sp); ( C ) In the CA3 region, moderate immunolabelling for Kv4.2 was observed in the strata oriens (so), radiatum (sr) and stratum lacunosum-moleculare (slm), weak in the stratum lucidum and very weak in the stratum pyramidale (sp). Kv4.2 immunolabelling was also seen outlining somata and dendrites of scattered interneurons (arrows); ( D ) In the dentate gyrus (DG), immunolabelling was strong in the molecular layer (ml), weak in the hilus (h) and the weakest in the granule cell layer (gc). Scale bars: A , 500 µm; B – D , 150 µm.

    Techniques Used:

    Subcellular localisation of Kv4.2 in the adult hippocampus. Electron micrographs showing immunoparticles for Kv4.2 in the hippocampus, as detected using the pre-embedding immunogold technique at P60. ( A – D ) Immunoreactivity for Kv4.2 was detected in similar neuronal compartments in the CA1 (panels A , B and C ) and CA3 (panels D and E ) regions and dentate gyrus (DG, panels F and G ). Kv4.2 immunoparticles were abundant along the extrasynaptic plasma membrane (arrows) of dendritic spines (s) contacted by axon terminals (at) and dendritic shafts (Den) of pyramidal cells and granule cells. Immunoparticles were also observed at intracellular sites (crossed arrows) in dendritic spines (s) and shafts (Den). Very few immunoparticles for Kv4.2 were also localised to the extrasynaptic plasma membrane (arrowheads) of axon terminals (at) establishing asymmetrical synapses with spines (s). Kv4.2 immunoparticles (double arrowheads) were detected along the main body of the postsynaptic membrane specialisation of GABAergic synapses in the CA1 (panel C ), CA3 (panel E ) and DG (panel G ); ( H ) Compartmentalisation of Kv4.2 in CA1 pyramidal cells, CA3 pyramidal cells and DG granule cells. Bar graphs showing the percentage of immunoparticles for Kv4.2 at post- and presynaptic compartments, and along the plasma membrane and intracellular sites in dendritic spines, dendritic shafts and axon terminals. A total of 1355 immunoparticles in the CA1, 1223 in the CA3 and 1037 in the DG were analysed. Most immunoparticles were postsynaptic, both along the plasma membrane and at cytoplasmic sites. Postsynaptically, immunoparticles were detected in dendritic spines and in dendritic shafts; ( I ) Histogram showing the distribution of immunoreactive Kv4.2 in relation to glutamate release sites in dendritic spines of CA1 and CA3 pyramidal cells, and DG granule cells. Data are expressed as the proportion of immunoparticles at a given distance from the edge of the synaptic specialisation. These data show that Kv4.2 immunoparticles were distributed similarly in spines in CA1, CA3 and DG and in the proximity of asymmetrical synapses on dendritic spines. Scale bars: A , D , F , G , 200 nm; B , C , E , 500 nm.
    Figure Legend Snippet: Subcellular localisation of Kv4.2 in the adult hippocampus. Electron micrographs showing immunoparticles for Kv4.2 in the hippocampus, as detected using the pre-embedding immunogold technique at P60. ( A – D ) Immunoreactivity for Kv4.2 was detected in similar neuronal compartments in the CA1 (panels A , B and C ) and CA3 (panels D and E ) regions and dentate gyrus (DG, panels F and G ). Kv4.2 immunoparticles were abundant along the extrasynaptic plasma membrane (arrows) of dendritic spines (s) contacted by axon terminals (at) and dendritic shafts (Den) of pyramidal cells and granule cells. Immunoparticles were also observed at intracellular sites (crossed arrows) in dendritic spines (s) and shafts (Den). Very few immunoparticles for Kv4.2 were also localised to the extrasynaptic plasma membrane (arrowheads) of axon terminals (at) establishing asymmetrical synapses with spines (s). Kv4.2 immunoparticles (double arrowheads) were detected along the main body of the postsynaptic membrane specialisation of GABAergic synapses in the CA1 (panel C ), CA3 (panel E ) and DG (panel G ); ( H ) Compartmentalisation of Kv4.2 in CA1 pyramidal cells, CA3 pyramidal cells and DG granule cells. Bar graphs showing the percentage of immunoparticles for Kv4.2 at post- and presynaptic compartments, and along the plasma membrane and intracellular sites in dendritic spines, dendritic shafts and axon terminals. A total of 1355 immunoparticles in the CA1, 1223 in the CA3 and 1037 in the DG were analysed. Most immunoparticles were postsynaptic, both along the plasma membrane and at cytoplasmic sites. Postsynaptically, immunoparticles were detected in dendritic spines and in dendritic shafts; ( I ) Histogram showing the distribution of immunoreactive Kv4.2 in relation to glutamate release sites in dendritic spines of CA1 and CA3 pyramidal cells, and DG granule cells. Data are expressed as the proportion of immunoparticles at a given distance from the edge of the synaptic specialisation. These data show that Kv4.2 immunoparticles were distributed similarly in spines in CA1, CA3 and DG and in the proximity of asymmetrical synapses on dendritic spines. Scale bars: A , D , F , G , 200 nm; B , C , E , 500 nm.

    Techniques Used:

    Co-localisation of Kv4.2 and Kv4.3 in granule cells. ( A – C ) Electron micrographs showing co-localisation for Kv4.2 with Kv4.3, as detected using a double-labelling pre-embedding immunogold method at P60. Labelling is shown with immunoperoxidase reaction for Kv4.2, and with silver-intensified immunogold reaction for Kv4.3. Immunoparticles for Kv4.3 were seen in dendritic spines (s) and shafts (Den) of granule cells immunopositive for Kv4.2, both along the plasma membrane (arrows) and at intracellular sites (crossed arrows); ( D ) Change in the density of Kv4.2 and Kv4.3 in DG granule cells as a function of distance from the soma in six somato-dendritic domains. Density of immunoparticles for the two channel subtypes increased significantly from soma to dendritic spines (Sp) in the inner one-third and outer two-thirds of the molecular layer. This analysis demonstrated their similar non-uniform distributions over the neuronal surface of granule cells. Inner SB, spiny branchlets in the inner one-third; Outer SB, spiny branchlets in the outer two-third; Inner Sp, spines in the inner one-third; Outer Sp, spines in the outer two-third. Scale bars: A – C , 500 nm.
    Figure Legend Snippet: Co-localisation of Kv4.2 and Kv4.3 in granule cells. ( A – C ) Electron micrographs showing co-localisation for Kv4.2 with Kv4.3, as detected using a double-labelling pre-embedding immunogold method at P60. Labelling is shown with immunoperoxidase reaction for Kv4.2, and with silver-intensified immunogold reaction for Kv4.3. Immunoparticles for Kv4.3 were seen in dendritic spines (s) and shafts (Den) of granule cells immunopositive for Kv4.2, both along the plasma membrane (arrows) and at intracellular sites (crossed arrows); ( D ) Change in the density of Kv4.2 and Kv4.3 in DG granule cells as a function of distance from the soma in six somato-dendritic domains. Density of immunoparticles for the two channel subtypes increased significantly from soma to dendritic spines (Sp) in the inner one-third and outer two-thirds of the molecular layer. This analysis demonstrated their similar non-uniform distributions over the neuronal surface of granule cells. Inner SB, spiny branchlets in the inner one-third; Outer SB, spiny branchlets in the outer two-third; Inner Sp, spines in the inner one-third; Outer Sp, spines in the outer two-third. Scale bars: A – C , 500 nm.

    Techniques Used:

    Developmental and regional distribution of the Kv4.2 channel in the mouse brain. ( A ) Kv4.2 protein distribution was visualised on histoblots of brain horizontal sections at various stages of postnatal development using an affinity-purified anti-Kv4.2 antibody. Kv4.2 was expressed in the brain since the day of birth (P0), and at all stages the strongest expression was detected in the cerebellum (Cb), caudate putamen (CPu), hippocampus (Hp) and thalamus (Th), with the lowest intensity in the cortex (Cx) and septum (Sp); ( B ) The histoblots were scanned and densitometric measurements from four independent experiments were averaged to compare the protein densities for each developmental time point. The analysis revealed a differential Kv4.2 expression in a developmental stage- and region-specific manner. Kv4.2 expression was detected at P0, increased progressively to reach a peak at P10, P15 or P21 depending on the brain region, and then decreasing at P30. Error bars indicate SEM. Scale bars, 2 mm.
    Figure Legend Snippet: Developmental and regional distribution of the Kv4.2 channel in the mouse brain. ( A ) Kv4.2 protein distribution was visualised on histoblots of brain horizontal sections at various stages of postnatal development using an affinity-purified anti-Kv4.2 antibody. Kv4.2 was expressed in the brain since the day of birth (P0), and at all stages the strongest expression was detected in the cerebellum (Cb), caudate putamen (CPu), hippocampus (Hp) and thalamus (Th), with the lowest intensity in the cortex (Cx) and septum (Sp); ( B ) The histoblots were scanned and densitometric measurements from four independent experiments were averaged to compare the protein densities for each developmental time point. The analysis revealed a differential Kv4.2 expression in a developmental stage- and region-specific manner. Kv4.2 expression was detected at P0, increased progressively to reach a peak at P10, P15 or P21 depending on the brain region, and then decreasing at P30. Error bars indicate SEM. Scale bars, 2 mm.

    Techniques Used: Affinity Purification, Expressing

    Regional distribution of the Kv4.2 channel in the adult mouse brain. ( A , B ) The distribution of the Kv4.2 protein was visualised in histoblots of horizontal brain sections at P60 using an affinity-purified anti-Kv4.2 antibody. The expression of Kv4.2 in different brain regions was determined by densitometric analysis of the scanned histoblots. The strongest expression was detected in the cerebellum (Cb) and hippocampus (Hp), with moderate expression in the caudate putamen (CPu) and thalamus (Th). Weak expression level was detected in the cortex (Cx) and septum (Sp); ( C , D ) In the hippocampus, very strong Kv4.2 expression was detected in the strata oriens (so) and radiatum (sr) of the CA1 region and the molecular layer (ml) of the dentate gyrus (DG); ( C , D ) Moderate staining was observed in all dendritic layers of CA3, in the stratum lacunosum-moleculare (slm) of the CA1 region and the hilus of the dentate gyrus. Very weak Kv4.2 staining was observed in the stratum pyramidale of the CA1 and CA3 regions and in the granule cell layer of the dentate gyrusso, stratum oriens ; sr, stratum radiatum ; DG, dentate gyrus; h, hilus .; ( E , F ) In the cerebellum, the strongest expression level was detected in the granule cell layer (gc), with weak expression in the molecular layer (ml) and very weak in the white matter (wm). Error bars indicate SEM; * p
    Figure Legend Snippet: Regional distribution of the Kv4.2 channel in the adult mouse brain. ( A , B ) The distribution of the Kv4.2 protein was visualised in histoblots of horizontal brain sections at P60 using an affinity-purified anti-Kv4.2 antibody. The expression of Kv4.2 in different brain regions was determined by densitometric analysis of the scanned histoblots. The strongest expression was detected in the cerebellum (Cb) and hippocampus (Hp), with moderate expression in the caudate putamen (CPu) and thalamus (Th). Weak expression level was detected in the cortex (Cx) and septum (Sp); ( C , D ) In the hippocampus, very strong Kv4.2 expression was detected in the strata oriens (so) and radiatum (sr) of the CA1 region and the molecular layer (ml) of the dentate gyrus (DG); ( C , D ) Moderate staining was observed in all dendritic layers of CA3, in the stratum lacunosum-moleculare (slm) of the CA1 region and the hilus of the dentate gyrus. Very weak Kv4.2 staining was observed in the stratum pyramidale of the CA1 and CA3 regions and in the granule cell layer of the dentate gyrusso, stratum oriens ; sr, stratum radiatum ; DG, dentate gyrus; h, hilus .; ( E , F ) In the cerebellum, the strongest expression level was detected in the granule cell layer (gc), with weak expression in the molecular layer (ml) and very weak in the white matter (wm). Error bars indicate SEM; * p

    Techniques Used: Affinity Purification, Expressing, Staining

    12) Product Images from "Overexpression of M3 Muscarinic Receptor Is a Novel Strategy for Preventing Sudden Cardiac Death in Transgenic Mice"

    Article Title: Overexpression of M3 Muscarinic Receptor Is a Novel Strategy for Preventing Sudden Cardiac Death in Transgenic Mice

    Journal: Molecular Medicine

    doi: 10.2119/molmed.2011.00093

    Alterations of protein and mRNA levels of I K1 and I to revealed by Western blot analysis and real-time RT-PCR. (A) Top, examples of Western blot bands of Kir2.1; bottom, quantification of Kir2.1 (B) Top, examples of Western blot bands of Kv4.2; bottom,
    Figure Legend Snippet: Alterations of protein and mRNA levels of I K1 and I to revealed by Western blot analysis and real-time RT-PCR. (A) Top, examples of Western blot bands of Kir2.1; bottom, quantification of Kir2.1 (B) Top, examples of Western blot bands of Kv4.2; bottom,

    Techniques Used: Western Blot, Quantitative RT-PCR

    13) Product Images from "Kv4.2 potassium channels segregate to extrasynaptic domains and influence intrasynaptic NMDA receptor NR2B subunit expression"

    Article Title: Kv4.2 potassium channels segregate to extrasynaptic domains and influence intrasynaptic NMDA receptor NR2B subunit expression

    Journal: Brain Structure & Function

    doi: 10.1007/s00429-012-0450-1

    Modulatory action of Kv4.2 on the synaptic expression of NR2B. a In a double-immunolabeling experiment for NMDA-type glutamate receptor subunit 1 (NR1) and subunit 2B (NR2B), both NR1 (10 nm gold) and NR2B (5 nm gold; arrowheads ) are observed on the P-face of the postsynaptic membrane specialization of an excitatory synapse in an ITC neuron from a Kv4.2 +/+ mouse. b In an ITC neuron from a Kv4.2 −/− mouse, NR1 (10 nm gold) and NR2B immunolabeling (5 nm gold; arrowheads ) are observed in the postsynaptic membrane of an excitatory synapse at a density similar to that observed in Kv4.2 +/+ neurons. Yet, the frequency of such synapses, co-immunolabeled for both the NR1 and NR2B subunits, was increased in ITC neurons of Kv4.2 −/− mice. Scale bar 150 nm ( a , b )
    Figure Legend Snippet: Modulatory action of Kv4.2 on the synaptic expression of NR2B. a In a double-immunolabeling experiment for NMDA-type glutamate receptor subunit 1 (NR1) and subunit 2B (NR2B), both NR1 (10 nm gold) and NR2B (5 nm gold; arrowheads ) are observed on the P-face of the postsynaptic membrane specialization of an excitatory synapse in an ITC neuron from a Kv4.2 +/+ mouse. b In an ITC neuron from a Kv4.2 −/− mouse, NR1 (10 nm gold) and NR2B immunolabeling (5 nm gold; arrowheads ) are observed in the postsynaptic membrane of an excitatory synapse at a density similar to that observed in Kv4.2 +/+ neurons. Yet, the frequency of such synapses, co-immunolabeled for both the NR1 and NR2B subunits, was increased in ITC neurons of Kv4.2 −/− mice. Scale bar 150 nm ( a , b )

    Techniques Used: Expressing, Immunolabeling, Mouse Assay

    SDS-FRL confirms the localization of Kv4.2 in the somato-dendritic plasma membrane of ITC neurons. a Immunoparticles labeling Kv4.2 subunits (10 nm gold) are scattered on the plasma membrane of a large dendritic trunk of a rat ITC neuron. The immunolabeling is restricted to the plasma membrane P-face. b Scattered immunopartices are also observed in the plasma membrane P-face of a large dendritic trunk of a mouse ITC neuron. c In addition to the labeling of the dendritic trunk, Kv4.2 immunoparticles are localized to the head and neck of spines. d Both the plasma membrane E- and P-face of an ITC dendrite from a Kv4.2 −/− mouse are immunonegative when reacted with the Kv4.2 (209–225) antibody (10 nm gold). e An ITC dendritic trunk from a wild-type mouse (Kv4.2 +/+ ) is immunopositive for both Kv4.2 (209–225) (5 nm gold) and MOR (15 nm gold) in a double-immunolabeling experiment. f At higher resolution ( boxed area in e ), both 5 and 15 nm gold particles revealing Kv4.2 subunits and MOR, respectively, are visible on the P-face of the dendritic plasma membrane. MOR μ-opioid receptor. Scale bar 300 nm ( a , c ); 200 nm ( b , d ); 400 nm ( e ); 150 nm ( f )
    Figure Legend Snippet: SDS-FRL confirms the localization of Kv4.2 in the somato-dendritic plasma membrane of ITC neurons. a Immunoparticles labeling Kv4.2 subunits (10 nm gold) are scattered on the plasma membrane of a large dendritic trunk of a rat ITC neuron. The immunolabeling is restricted to the plasma membrane P-face. b Scattered immunopartices are also observed in the plasma membrane P-face of a large dendritic trunk of a mouse ITC neuron. c In addition to the labeling of the dendritic trunk, Kv4.2 immunoparticles are localized to the head and neck of spines. d Both the plasma membrane E- and P-face of an ITC dendrite from a Kv4.2 −/− mouse are immunonegative when reacted with the Kv4.2 (209–225) antibody (10 nm gold). e An ITC dendritic trunk from a wild-type mouse (Kv4.2 +/+ ) is immunopositive for both Kv4.2 (209–225) (5 nm gold) and MOR (15 nm gold) in a double-immunolabeling experiment. f At higher resolution ( boxed area in e ), both 5 and 15 nm gold particles revealing Kv4.2 subunits and MOR, respectively, are visible on the P-face of the dendritic plasma membrane. MOR μ-opioid receptor. Scale bar 300 nm ( a , c ); 200 nm ( b , d ); 400 nm ( e ); 150 nm ( f )

    Techniques Used: Labeling, Immunolabeling

    Kv4.2 does not reside within the postsynaptic membrane specializations of excitatory synapses as revealed by a double-replica approach. a Immunogold particles revealing NR1 subunits (10 nm gold) are localized to the E-face of an ITC dendrite. b On the mirror replica, immunogold particles revealing Kv4.2 subunits (10 nm gold) are observed on the P-face of the same dendrite. c At higher resolution ( boxed area in a ), immunolabeling for NR1 subunits is evident on the E-face of a postsynaptic membrane specialization of a glutamatergic synapse (shown in orange ), which is characterized by the clustering of intramembrane particles. d Immunolabeling for Kv4.2 subunits is present on the P-face of the extrasynaptic plasma membrane at higher resolution ( boxed area in b ). The postsynaptic membrane specialization by itself is free of any Kv4.2 immunolabeling. Sp spine. Scale bar 300 nm ( a , b ); 150 nm ( c , d )
    Figure Legend Snippet: Kv4.2 does not reside within the postsynaptic membrane specializations of excitatory synapses as revealed by a double-replica approach. a Immunogold particles revealing NR1 subunits (10 nm gold) are localized to the E-face of an ITC dendrite. b On the mirror replica, immunogold particles revealing Kv4.2 subunits (10 nm gold) are observed on the P-face of the same dendrite. c At higher resolution ( boxed area in a ), immunolabeling for NR1 subunits is evident on the E-face of a postsynaptic membrane specialization of a glutamatergic synapse (shown in orange ), which is characterized by the clustering of intramembrane particles. d Immunolabeling for Kv4.2 subunits is present on the P-face of the extrasynaptic plasma membrane at higher resolution ( boxed area in b ). The postsynaptic membrane specialization by itself is free of any Kv4.2 immunolabeling. Sp spine. Scale bar 300 nm ( a , b ); 150 nm ( c , d )

    Techniques Used: Immunolabeling

    Quantitative characterization of the distribution of Kv4.2 immunogold particles in ITC neurons and other central principal cells. a A sample SDS-FRL image of a portion of an ITC neuron soma. The area of the relevant profile is colored in blue . Immunogold particles labeling Kv4.2 subunits are marked with a black dot , a 20-nm radius circle around each particle is shown in yellow and the centroid of overlapping circles is marked with a red open circle . A tight cluster of particles is defined as particles residing within the overlapping yellow circles . b Cumulative probability curves for the nearest neighbor distances (NNDs) between individual immunogold particles (in black ) and tight cluster centers along with single particles (in red ), and NNDs of the calculated random distribution (in blue ). c A sample image of a portion of an ITC dendrite and d respective NND analysis. e – h Sample images and NND analyses for CA1 pyramidal neuron soma and dendrite. i – j Sample image and NND analysis for cerebellar granule cell soma. k Histogram showing the immunogold particle density in subcellular domains of ITC neurons (soma: 8 profiles with a mean area of 2.60 μm 2 ; dendrite: 22 profiles with a mean area of 2.05 μm 2 ), CA1 pyramidal neurons (soma: 6 profiles with a mean area of 2.52 μm 2 ; dendrite: 19 profiles with a mean area of 1.52 μm 2 ) and cerebellar granule cells (soma: 25 profiles with a mean area of 2.54 μm 2 ). Cb cerebellar granule cell, NND nearest neighbor distance. Error bars , SEM. * p
    Figure Legend Snippet: Quantitative characterization of the distribution of Kv4.2 immunogold particles in ITC neurons and other central principal cells. a A sample SDS-FRL image of a portion of an ITC neuron soma. The area of the relevant profile is colored in blue . Immunogold particles labeling Kv4.2 subunits are marked with a black dot , a 20-nm radius circle around each particle is shown in yellow and the centroid of overlapping circles is marked with a red open circle . A tight cluster of particles is defined as particles residing within the overlapping yellow circles . b Cumulative probability curves for the nearest neighbor distances (NNDs) between individual immunogold particles (in black ) and tight cluster centers along with single particles (in red ), and NNDs of the calculated random distribution (in blue ). c A sample image of a portion of an ITC dendrite and d respective NND analysis. e – h Sample images and NND analyses for CA1 pyramidal neuron soma and dendrite. i – j Sample image and NND analysis for cerebellar granule cell soma. k Histogram showing the immunogold particle density in subcellular domains of ITC neurons (soma: 8 profiles with a mean area of 2.60 μm 2 ; dendrite: 22 profiles with a mean area of 2.05 μm 2 ), CA1 pyramidal neurons (soma: 6 profiles with a mean area of 2.52 μm 2 ; dendrite: 19 profiles with a mean area of 1.52 μm 2 ) and cerebellar granule cells (soma: 25 profiles with a mean area of 2.54 μm 2 ). Cb cerebellar granule cell, NND nearest neighbor distance. Error bars , SEM. * p

    Techniques Used: Labeling

    Segregation of Kv4.2 to extrasynaptic domains. a In a double-immunolabeling experiment for PSD-95, visualized with 15 nm gold particles, and Kv4.2, visualized with 10 nm gold particles ( arrowheads ), Kv4.2 subunits are observed at extrasynaptic sites of an ITC dendritic spine in a wild-type mouse (Kv4.2 +/+ ). Conversely, PSD-95 labeling can be seen within the postsynaptic membrane specialization of an excitatory synapse. b In a Kv4.2 −/− mouse, labeling for PSD-95 (10 nm gold) can clearly be detected within the postsynaptic membrane specialization of a dendritic spine. No immunoreactivity for Kv4.2 (15 nm gold) is observed in Kv4.2 −/− mouse tissue. Scale bar 200 nm ( a , b )
    Figure Legend Snippet: Segregation of Kv4.2 to extrasynaptic domains. a In a double-immunolabeling experiment for PSD-95, visualized with 15 nm gold particles, and Kv4.2, visualized with 10 nm gold particles ( arrowheads ), Kv4.2 subunits are observed at extrasynaptic sites of an ITC dendritic spine in a wild-type mouse (Kv4.2 +/+ ). Conversely, PSD-95 labeling can be seen within the postsynaptic membrane specialization of an excitatory synapse. b In a Kv4.2 −/− mouse, labeling for PSD-95 (10 nm gold) can clearly be detected within the postsynaptic membrane specialization of a dendritic spine. No immunoreactivity for Kv4.2 (15 nm gold) is observed in Kv4.2 −/− mouse tissue. Scale bar 200 nm ( a , b )

    Techniques Used: Immunolabeling, Labeling

    Distribution of Kv4.2 in the mouse amygdala revealed by immunoperoxidase labeling. a Prominent Kv4.2 immunoreactivity is detected in all ITC clusters and the ITC nucleus in C57Bl/6 wild-type mice (Kv4.2 +/+ ). Only low to moderate levels of immunoreactivity are observed in other amygdaloid areas than the ITC. b Higher magnification of the Imp cluster ( boxed area in a ). Diffuse Kv4.2 immunoreactivity is primarily observed in the neuropil. c A similar immunostaining pattern for Kv4.2 is observed in the ITC nucleus ( boxed area in a ). d Specificity of Kv4.2 immunolabeling is confirmed on respective brain areas from a Kv4.2 −/− mouse. BL basolateral amygala, Ce central nucleus of amygdala, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, Is LA supralateral ITC cluster, La lateral amygdala. Scale bar 200 μm ( a , d ); 40 μm ( b , c )
    Figure Legend Snippet: Distribution of Kv4.2 in the mouse amygdala revealed by immunoperoxidase labeling. a Prominent Kv4.2 immunoreactivity is detected in all ITC clusters and the ITC nucleus in C57Bl/6 wild-type mice (Kv4.2 +/+ ). Only low to moderate levels of immunoreactivity are observed in other amygdaloid areas than the ITC. b Higher magnification of the Imp cluster ( boxed area in a ). Diffuse Kv4.2 immunoreactivity is primarily observed in the neuropil. c A similar immunostaining pattern for Kv4.2 is observed in the ITC nucleus ( boxed area in a ). d Specificity of Kv4.2 immunolabeling is confirmed on respective brain areas from a Kv4.2 −/− mouse. BL basolateral amygala, Ce central nucleus of amygdala, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, Is LA supralateral ITC cluster, La lateral amygdala. Scale bar 200 μm ( a , d ); 40 μm ( b , c )

    Techniques Used: Labeling, Mouse Assay, Immunostaining, Immunolabeling

    Kv4.2 immunolabeling in the rat amygdala using antibodies directed against different regions of the Kv4.2 protein. a – c The Kv4.2 (454–469) antibody reveals prominent immunolabeling of all ITC clusters and the ITC nucleus from rostral ( a ) to medial ( b ) and caudal ( c ) levels of the amygdala using an immunoperoxidase staining technique. Other amygdaloid areas are only faintly labeled. d – f An equivalent immunolabeling pattern is observed with the Kv4.2 (209–225) antibody. BL basolateral amygdala, Ce central nucleus of amygdala, Ii LA intralateral paracapsular ITC cluster, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral nucleus of amygdala. Scale bar 350 μm ( a – f )
    Figure Legend Snippet: Kv4.2 immunolabeling in the rat amygdala using antibodies directed against different regions of the Kv4.2 protein. a – c The Kv4.2 (454–469) antibody reveals prominent immunolabeling of all ITC clusters and the ITC nucleus from rostral ( a ) to medial ( b ) and caudal ( c ) levels of the amygdala using an immunoperoxidase staining technique. Other amygdaloid areas are only faintly labeled. d – f An equivalent immunolabeling pattern is observed with the Kv4.2 (209–225) antibody. BL basolateral amygdala, Ce central nucleus of amygdala, Ii LA intralateral paracapsular ITC cluster, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral nucleus of amygdala. Scale bar 350 μm ( a – f )

    Techniques Used: Immunolabeling, Immunoperoxidase Staining, Labeling

    Subcellular localization of Kv4.2 to somato-dendritic domains of ITC neurons revealed by pre-embedding immunoperoxidase and immunometal electron microscopy. a A small dendritic trunk of a neuron in the rat ITC nucleus displays dense and diffuse immunolabeling for Kv4.2 [Kv4.2 (454–469) antibody]. The immunolabeling appears as an electron-opaque reaction product. A synaptic terminal, contacting this dendrite, is free of any immunolabeling. b A dendritic spine of a neuron in the mouse ITC nucleus shows dense and diffuse labeling for Kv4.2 [Kv4.2 (209–225) antibody]. An axon terminal contacting this spine, as well as other cellular profiles, is free of any immunolabeling. c A dendritic spine of a mouse Imp neuron shows the same dense and diffuse labeling pattern for Kv4.2 [Kv4.2 (454–469) antibody]. d In a double-labeling experiment, a Kv4.2 immunopositive spine of a rat Imp neuron (immunoperoxidase reaction) is also immunoreactive for MOR (immunometal reaction). e A small dendrite of a mouse Imp neuron, immunolabeled for Kv4.2 (immunoperoxidase reaction), is also immunoreactive for MOR (immunometal reaction). An axon terminal, contacting this dendrite, is free of any immunolabeling. f In a sample from a Kv4.2 −/− mouse, a large dendrite of an Imp neuron is labeled for MOR (immunometal reaction), but devoid of Kv4.2 immunolabeling (immunoperoxidase reaction). g Immunometal particles (indicated by arrowheads ) for Kv4.2, when applying the Kv4.2 (454–469) antibody, are observed at the intracellular side of the plasma membrane of a rat Imp neuron. The postsynaptic specialization of an excitatory synapse is free of any immunolabeling. Some immunoparticles appear localized perisynaptically. h Immunometal particles (indicated by arrowheads) for Kv4.2 appear at the intracellular side of an ITC neuron dendrite also when applying the Kv4.2 (209–225) antibody. Both the axon forming a synapse with this dendrite and the postsynaptic specialization are free of any immunolabeling. i The plasma membrane of an Imp dendrite is decorated with immunoparticles at the intracellular side applying the Kv4.2 (209–225) antibody. The postsynaptic specialization of an inhibitory synapse on this dendrite is also free of any immunolabeling. At axon terminal, MOR μ-opioid receptor. Scale bar 200 nm ( a – c , e , h , i ); 300 nm ( d , f , g )
    Figure Legend Snippet: Subcellular localization of Kv4.2 to somato-dendritic domains of ITC neurons revealed by pre-embedding immunoperoxidase and immunometal electron microscopy. a A small dendritic trunk of a neuron in the rat ITC nucleus displays dense and diffuse immunolabeling for Kv4.2 [Kv4.2 (454–469) antibody]. The immunolabeling appears as an electron-opaque reaction product. A synaptic terminal, contacting this dendrite, is free of any immunolabeling. b A dendritic spine of a neuron in the mouse ITC nucleus shows dense and diffuse labeling for Kv4.2 [Kv4.2 (209–225) antibody]. An axon terminal contacting this spine, as well as other cellular profiles, is free of any immunolabeling. c A dendritic spine of a mouse Imp neuron shows the same dense and diffuse labeling pattern for Kv4.2 [Kv4.2 (454–469) antibody]. d In a double-labeling experiment, a Kv4.2 immunopositive spine of a rat Imp neuron (immunoperoxidase reaction) is also immunoreactive for MOR (immunometal reaction). e A small dendrite of a mouse Imp neuron, immunolabeled for Kv4.2 (immunoperoxidase reaction), is also immunoreactive for MOR (immunometal reaction). An axon terminal, contacting this dendrite, is free of any immunolabeling. f In a sample from a Kv4.2 −/− mouse, a large dendrite of an Imp neuron is labeled for MOR (immunometal reaction), but devoid of Kv4.2 immunolabeling (immunoperoxidase reaction). g Immunometal particles (indicated by arrowheads ) for Kv4.2, when applying the Kv4.2 (454–469) antibody, are observed at the intracellular side of the plasma membrane of a rat Imp neuron. The postsynaptic specialization of an excitatory synapse is free of any immunolabeling. Some immunoparticles appear localized perisynaptically. h Immunometal particles (indicated by arrowheads) for Kv4.2 appear at the intracellular side of an ITC neuron dendrite also when applying the Kv4.2 (209–225) antibody. Both the axon forming a synapse with this dendrite and the postsynaptic specialization are free of any immunolabeling. i The plasma membrane of an Imp dendrite is decorated with immunoparticles at the intracellular side applying the Kv4.2 (209–225) antibody. The postsynaptic specialization of an inhibitory synapse on this dendrite is also free of any immunolabeling. At axon terminal, MOR μ-opioid receptor. Scale bar 200 nm ( a – c , e , h , i ); 300 nm ( d , f , g )

    Techniques Used: Electron Microscopy, Immunolabeling, Labeling

    Colocalization of Kv4.2 and μ-opioid receptors (MOR) in the ITCs. a Double-labeling immunofluorescence for Kv4.2 (in green ) and MOR (in red ) reveals a high degree of coexistence in the amygdala, which is evident in the merged images. b At higher resolution, immunoreactivity for Kv4.2 (in green ) as well as for MOR (in red ) appears dense and diffuse in ITC clusters such as the Imp. The distribution profile of these proteins is highly similar ( merge ). BL basolateral amygdala, Ce central nucleus of amygdala, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral amygdala. Scale bar 350 μm ( a ), 50 μm ( b )
    Figure Legend Snippet: Colocalization of Kv4.2 and μ-opioid receptors (MOR) in the ITCs. a Double-labeling immunofluorescence for Kv4.2 (in green ) and MOR (in red ) reveals a high degree of coexistence in the amygdala, which is evident in the merged images. b At higher resolution, immunoreactivity for Kv4.2 (in green ) as well as for MOR (in red ) appears dense and diffuse in ITC clusters such as the Imp. The distribution profile of these proteins is highly similar ( merge ). BL basolateral amygdala, Ce central nucleus of amygdala, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral amygdala. Scale bar 350 μm ( a ), 50 μm ( b )

    Techniques Used: Labeling, Immunofluorescence

    14) Product Images from "Memory Decline and Behavioral Inflexibility in Aged Mice Are Correlated With Dysregulation of Protein Synthesis Capacity"

    Article Title: Memory Decline and Behavioral Inflexibility in Aged Mice Are Correlated With Dysregulation of Protein Synthesis Capacity

    Journal: Frontiers in Aging Neuroscience

    doi: 10.3389/fnagi.2019.00246

    Dysregulations of protein synthesis capacity in aged mice. Western blot performed on hippocampal tissues demonstrated that compared to young mice, old mice exhibited: (A) Increased levels of mTOR phosphorylation (Ser2448); (B) Increased levels of p70S6K phosphorylation (Thr389); (C) Unaltered levels of 4EBP1 phosphorylation; (D) Increased levels of AKT phosphorylation (Ser473); (E) Increased levels of GSK3β (Ser9); (F) Increased levels of eEF2 phosphorylation (Thr56); (G) Increased levels of AMPKα phosphorylation (Thr172); (H) Unaltered levels of Kv4.2 phosphorylation; (I) Decreased levels of GluA1 expression. Except for GluA1, no change on levels of total proteins was observed between young and old mice. n = 7 for young mice and n = 6 for old mice (representative bands from three mice per group). ∗ p
    Figure Legend Snippet: Dysregulations of protein synthesis capacity in aged mice. Western blot performed on hippocampal tissues demonstrated that compared to young mice, old mice exhibited: (A) Increased levels of mTOR phosphorylation (Ser2448); (B) Increased levels of p70S6K phosphorylation (Thr389); (C) Unaltered levels of 4EBP1 phosphorylation; (D) Increased levels of AKT phosphorylation (Ser473); (E) Increased levels of GSK3β (Ser9); (F) Increased levels of eEF2 phosphorylation (Thr56); (G) Increased levels of AMPKα phosphorylation (Thr172); (H) Unaltered levels of Kv4.2 phosphorylation; (I) Decreased levels of GluA1 expression. Except for GluA1, no change on levels of total proteins was observed between young and old mice. n = 7 for young mice and n = 6 for old mice (representative bands from three mice per group). ∗ p

    Techniques Used: Mouse Assay, Western Blot, Expressing

    15) Product Images from "Kv4.2 channels tagged in the S1-S2 loop for alpha-bungarotoxin binding provide a new tool for studies of channel expression and localization"

    Article Title: Kv4.2 channels tagged in the S1-S2 loop for alpha-bungarotoxin binding provide a new tool for studies of channel expression and localization

    Journal: Channels (Austin, Tex.)

    doi:

    Rate of recovery from inactivation. (a) Representative current traces of the recovery from inactivation for Kv4.2, Kv4.2-HAP and Kv4.2-EGFP-HAP respectively are shown. A paired pulse protocol shown as an insert was applied to transfected tsA201 cells.
    Figure Legend Snippet: Rate of recovery from inactivation. (a) Representative current traces of the recovery from inactivation for Kv4.2, Kv4.2-HAP and Kv4.2-EGFP-HAP respectively are shown. A paired pulse protocol shown as an insert was applied to transfected tsA201 cells.

    Techniques Used: Transfection

    Rhodamine-Bgtx binds Kv4.2-HAP-GFP and GluR2-HAP-GFP expressed in primary rat hippocampal neurons. (A) A neuron transfected with Kv4.2-HAP-GFP. Bright Field, bright-field image of the neuron; GFP, fluorescence image of GFP (detecting all Kv4.2-HAP-GFP
    Figure Legend Snippet: Rhodamine-Bgtx binds Kv4.2-HAP-GFP and GluR2-HAP-GFP expressed in primary rat hippocampal neurons. (A) A neuron transfected with Kv4.2-HAP-GFP. Bright Field, bright-field image of the neuron; GFP, fluorescence image of GFP (detecting all Kv4.2-HAP-GFP

    Techniques Used: Transfection, Fluorescence

    Alexa Fluor-Bgtx binds Kv4.2-HAP expressed in tsA201 cells. DIC, image of differential interference contrast; GFP, fluorescent image of GFP (detecting co-transfected GFP, green); Alexa Fluor-Bgtx, fluorescent image of Alexa Fluor-Bgtx (detecting Kv4.2-HAP,
    Figure Legend Snippet: Alexa Fluor-Bgtx binds Kv4.2-HAP expressed in tsA201 cells. DIC, image of differential interference contrast; GFP, fluorescent image of GFP (detecting co-transfected GFP, green); Alexa Fluor-Bgtx, fluorescent image of Alexa Fluor-Bgtx (detecting Kv4.2-HAP,

    Techniques Used: Transfection

    Heterologous expression of wild-type Kv4.2, HAP-tagged Kv4.2 (Kv4.2-HAP) and EGFP plus HAP–tagged Kv4.2 (Kv4.2-HAP-EGFP) in tsA201 cells. (a) K+ currents in tsA201 cells were elicited by voltage clamp steps delivered at 10-mV increments from a
    Figure Legend Snippet: Heterologous expression of wild-type Kv4.2, HAP-tagged Kv4.2 (Kv4.2-HAP) and EGFP plus HAP–tagged Kv4.2 (Kv4.2-HAP-EGFP) in tsA201 cells. (a) K+ currents in tsA201 cells were elicited by voltage clamp steps delivered at 10-mV increments from a

    Techniques Used: Expressing

    Activation kinetics for Kv4.2 (■) and Kv4.2-HAP ( ). (a) Time to peak current analysis. Representative current traces of inactivation for Kv4.2 and Kv4.2-HAP are shown in . The upper panel shows detailed superimposed peak currents for Kv4.2
    Figure Legend Snippet: Activation kinetics for Kv4.2 (■) and Kv4.2-HAP ( ). (a) Time to peak current analysis. Representative current traces of inactivation for Kv4.2 and Kv4.2-HAP are shown in . The upper panel shows detailed superimposed peak currents for Kv4.2

    Techniques Used: Activation Assay

    Inactivation time constants τ Kv4.2 (■) and τ Kv4.2-HAP ( ). The upper panel shows inactivation kinetics of currents in response to step membrane depolarization. The time course of fast inactivation fits mono-exponentially. The
    Figure Legend Snippet: Inactivation time constants τ Kv4.2 (■) and τ Kv4.2-HAP ( ). The upper panel shows inactivation kinetics of currents in response to step membrane depolarization. The time course of fast inactivation fits mono-exponentially. The

    Techniques Used:

    Binding of 125 I -Bgtx to Kv4.2-HAP and effect of Bgtx on Kv4.2-HAP currents. (A) Binding of 125 I -Bgtx to Kv4.2-HAP. tsA201 cells were incubated with 125 I for 2 h. Specific Bgtx binding was observed for Kv4.2-HAP as well as muscle-type nAChR (p
    Figure Legend Snippet: Binding of 125 I -Bgtx to Kv4.2-HAP and effect of Bgtx on Kv4.2-HAP currents. (A) Binding of 125 I -Bgtx to Kv4.2-HAP. tsA201 cells were incubated with 125 I for 2 h. Specific Bgtx binding was observed for Kv4.2-HAP as well as muscle-type nAChR (p

    Techniques Used: Binding Assay, Incubation

    Schematic representation of the insertion of the HAP and Tα1 tag into the S1–S2 linker of Kv4.2. The diagrams depicting the structure of the neuronal nicotinic receptor ( left ) next to the structure of alpha-Bgtx ( right ) and the topology
    Figure Legend Snippet: Schematic representation of the insertion of the HAP and Tα1 tag into the S1–S2 linker of Kv4.2. The diagrams depicting the structure of the neuronal nicotinic receptor ( left ) next to the structure of alpha-Bgtx ( right ) and the topology

    Techniques Used:

    16) Product Images from "Opposite Actions of Brain-Derived Neurotrophic Factor and Neurotrophin-3 on Firing Features and Ion Channel Composition of Murine Spiral Ganglion Neurons"

    Article Title: Opposite Actions of Brain-Derived Neurotrophic Factor and Neurotrophin-3 on Firing Features and Ion Channel Composition of Murine Spiral Ganglion Neurons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.22-04-01385.2002

    Exposure to NT-3 increased Kv4.2 antibody staining, whereas BDNF had relatively little effect. a–l , Representative images of spiral ganglion neurons from the apex and base in control, BDNF, and NT-3 conditions. The top panels ( a, c, e, g, i, k ) show neurons labeled with FITC-conjugated Kv4.2 antibody, and the bottom panels ( b, d, f, h, j, l ) show the overlay of the green FITC-conjugated Kv4.2 antibody and the red TRITC-conjugated NF200 antibody. a–d , Spiral ganglion neurons cultured without exogenously added neurotrophins. Apical neurons showed significantly more anti-Kv4.2 label than basal neurons. a, Anti-Kv4.2 stained a population of neurons removed from the apex of the cochlea. The intensity of Kv4.2 label, however, was not uniform. Neurons from the same cochlear location were either intensely labeled ( arrow ) or weakly labeled ( arrowhead ). b, A double exposure of Kv4.2 ( green ) and NF200 ( red ) staining revealed that not all neurons observed in b labeled with Kv4.2 (compare with neurons in a ). c, Spiral ganglion neurons labeled lightly with anti-Kv4.2. d, The double exposure shows NF200-positive labeling, indicating that neurons were present in the culture, however, the intensity of label with the Kv4.2 antibody ( green ) was lower in basal ( c ) than apical neurons ( a ). e–h , Neurons isolated from the cochlea and grown in media supplemented with 5 ng/ml BDNF. Kv4.2 antibody staining was increased in neurons isolated from the basal cochlea but not the apex, therefore eliminating the apex–base difference observed in control cultures. e, The majority of neurons from the apex stained with anti-Kv4.2 with intensities similar to controls where some were strongly labeled ( arrow ) and others weakly labeled ( arrowhead ). f, Double exposure of Kv4.2-positive neurons, yellow ; NF200-positive neurons, red . g, Neurons from the basal cochlea stained similarly to those from the apex. h, The double exposure shows that not all of the population of neurons identified with NF200 stained with anti-Kv4.2. i–l , Cultures of spiral ganglion neurons supplemented with 5 ng/ml NT-3. The distribution of Kv4.2 protein in neurons isolated from both the base and the apex was significantly increased from controls, and the apex–base difference was preserved. i, Neurons isolated from the apex of the cochlea labeled strongly with the Kv4.2 antibody. j, The double exposure shows that most of the neurons identified with anti-NF200 labeled with anti-Kv4.2. k, The addition of NT-3 significantly increased the amount of Kv4.2 staining in basal neurons compared with controls; however, a large population of neurons remained only lightly labeled. l, The double exposure of Kv4.2-positive neurons ( yellow ) and unlabeled NF200-positive neurons ( red ). For a–l , spiral ganglia were isolated from postnatal day 4 CBA/CaJ mice and maintained for 7 d in vitro . Tissues were incubated in a 1:400 dilution of anti-Kv4.2 overnight at 4°C. m, Histogram of the weighted percentage of Kv4.2 antibody staining of apical and basal spiral ganglion neurons in each condition ( Control, BDNF , and NT-3 ) for four experiments. ** p
    Figure Legend Snippet: Exposure to NT-3 increased Kv4.2 antibody staining, whereas BDNF had relatively little effect. a–l , Representative images of spiral ganglion neurons from the apex and base in control, BDNF, and NT-3 conditions. The top panels ( a, c, e, g, i, k ) show neurons labeled with FITC-conjugated Kv4.2 antibody, and the bottom panels ( b, d, f, h, j, l ) show the overlay of the green FITC-conjugated Kv4.2 antibody and the red TRITC-conjugated NF200 antibody. a–d , Spiral ganglion neurons cultured without exogenously added neurotrophins. Apical neurons showed significantly more anti-Kv4.2 label than basal neurons. a, Anti-Kv4.2 stained a population of neurons removed from the apex of the cochlea. The intensity of Kv4.2 label, however, was not uniform. Neurons from the same cochlear location were either intensely labeled ( arrow ) or weakly labeled ( arrowhead ). b, A double exposure of Kv4.2 ( green ) and NF200 ( red ) staining revealed that not all neurons observed in b labeled with Kv4.2 (compare with neurons in a ). c, Spiral ganglion neurons labeled lightly with anti-Kv4.2. d, The double exposure shows NF200-positive labeling, indicating that neurons were present in the culture, however, the intensity of label with the Kv4.2 antibody ( green ) was lower in basal ( c ) than apical neurons ( a ). e–h , Neurons isolated from the cochlea and grown in media supplemented with 5 ng/ml BDNF. Kv4.2 antibody staining was increased in neurons isolated from the basal cochlea but not the apex, therefore eliminating the apex–base difference observed in control cultures. e, The majority of neurons from the apex stained with anti-Kv4.2 with intensities similar to controls where some were strongly labeled ( arrow ) and others weakly labeled ( arrowhead ). f, Double exposure of Kv4.2-positive neurons, yellow ; NF200-positive neurons, red . g, Neurons from the basal cochlea stained similarly to those from the apex. h, The double exposure shows that not all of the population of neurons identified with NF200 stained with anti-Kv4.2. i–l , Cultures of spiral ganglion neurons supplemented with 5 ng/ml NT-3. The distribution of Kv4.2 protein in neurons isolated from both the base and the apex was significantly increased from controls, and the apex–base difference was preserved. i, Neurons isolated from the apex of the cochlea labeled strongly with the Kv4.2 antibody. j, The double exposure shows that most of the neurons identified with anti-NF200 labeled with anti-Kv4.2. k, The addition of NT-3 significantly increased the amount of Kv4.2 staining in basal neurons compared with controls; however, a large population of neurons remained only lightly labeled. l, The double exposure of Kv4.2-positive neurons ( yellow ) and unlabeled NF200-positive neurons ( red ). For a–l , spiral ganglia were isolated from postnatal day 4 CBA/CaJ mice and maintained for 7 d in vitro . Tissues were incubated in a 1:400 dilution of anti-Kv4.2 overnight at 4°C. m, Histogram of the weighted percentage of Kv4.2 antibody staining of apical and basal spiral ganglion neurons in each condition ( Control, BDNF , and NT-3 ) for four experiments. ** p

    Techniques Used: Staining, Labeling, Cell Culture, Isolation, Crocin Bleaching Assay, Mouse Assay, In Vitro, Incubation

    17) Product Images from "Unique somato-dendritic distribution pattern of Kv4.2 channels on hippocampal CA1 pyramidal cells"

    Article Title: Unique somato-dendritic distribution pattern of Kv4.2 channels on hippocampal CA1 pyramidal cells

    Journal: The European Journal of Neuroscience

    doi: 10.1111/j.1460-9568.2011.07907.x

    Densities of Kv4.2 immunogold particles in different somato-dendritic compartments of rat CA1 PCs. Bar graphs illustrate the background subtracted densities (mean ± SD, in gold/μm 2 ) of immunogold particles in the somato-dendritic compartments. The bars are colour coded to different subcellular compartments as indicated in the schematic drawing of a CA1 PC. Note the moderate increase in the density along the proximo-distal axis of PCs within the stratum radiatum and the subsequent decrease in the stratum lacunosum-moleculare.
    Figure Legend Snippet: Densities of Kv4.2 immunogold particles in different somato-dendritic compartments of rat CA1 PCs. Bar graphs illustrate the background subtracted densities (mean ± SD, in gold/μm 2 ) of immunogold particles in the somato-dendritic compartments. The bars are colour coded to different subcellular compartments as indicated in the schematic drawing of a CA1 PC. Note the moderate increase in the density along the proximo-distal axis of PCs within the stratum radiatum and the subsequent decrease in the stratum lacunosum-moleculare.

    Techniques Used:

    The Kv4.2 subunit is excluded from postsynaptic membrane specializations. (A) Electron micrograph illustrating an excitatory synapse, revealed by the accumulation of the postsynaptic density marker PSD-95 (15-nm gold). Arrowheads point to gold particles labelling the Kv4.2 subunit (10-nm gold) around the PSD. (B) High-magnification image of an inhibitory synapse identified by the enrichment of gold particles (5-nm gold) labelling the γ-aminobutyric acid (GABA) A receptor β3 subunit. Note the clustering of gold particles labelling the Kv4.2 subunit (10-nm gold) in the extrasynaptic membrane, but not within the inhibitory synapse. Scale bars: 100 nm (A); 50 nm (B).
    Figure Legend Snippet: The Kv4.2 subunit is excluded from postsynaptic membrane specializations. (A) Electron micrograph illustrating an excitatory synapse, revealed by the accumulation of the postsynaptic density marker PSD-95 (15-nm gold). Arrowheads point to gold particles labelling the Kv4.2 subunit (10-nm gold) around the PSD. (B) High-magnification image of an inhibitory synapse identified by the enrichment of gold particles (5-nm gold) labelling the γ-aminobutyric acid (GABA) A receptor β3 subunit. Note the clustering of gold particles labelling the Kv4.2 subunit (10-nm gold) in the extrasynaptic membrane, but not within the inhibitory synapse. Scale bars: 100 nm (A); 50 nm (B).

    Techniques Used: Marker

    Specificity test for Kv4.2 subunit labelling on the axo-somato-dendritic surface of CA1 PCs using SDS-FRL. (A) Electron micrograph illustrating the P-face of a spiny CA1 PC dendrite of wild-type mouse immunolabelled for the Kv4.2 subunit. Note that the density and distribution pattern of the immunogold labelling for the Kv4.2 subunit in mouse is similar to that seen in rat. (B) The lack of immunogold labelling for the Kv4.2 subunit in a spiny dendrite of a Kv4.2 −/− mouse demonstrates the specificity of the labelling using SDS-FRL. (C) Axon terminals in rat identified by immunolabelling for SNAP-25 (15-nm gold) contain a low number of immunogold particles (arrows) for the Kv4.2 subunit (10-nm gold). (D and E) Immunogold particles for the Kv4.2 subunit were present in both Kv4.2 +/+ (D) and Kv4.2 −/− mice (E) at approximately the same density. dendr, dendrite; sp, spine. Scale bars: 250 nm (A–E).
    Figure Legend Snippet: Specificity test for Kv4.2 subunit labelling on the axo-somato-dendritic surface of CA1 PCs using SDS-FRL. (A) Electron micrograph illustrating the P-face of a spiny CA1 PC dendrite of wild-type mouse immunolabelled for the Kv4.2 subunit. Note that the density and distribution pattern of the immunogold labelling for the Kv4.2 subunit in mouse is similar to that seen in rat. (B) The lack of immunogold labelling for the Kv4.2 subunit in a spiny dendrite of a Kv4.2 −/− mouse demonstrates the specificity of the labelling using SDS-FRL. (C) Axon terminals in rat identified by immunolabelling for SNAP-25 (15-nm gold) contain a low number of immunogold particles (arrows) for the Kv4.2 subunit (10-nm gold). (D and E) Immunogold particles for the Kv4.2 subunit were present in both Kv4.2 +/+ (D) and Kv4.2 −/− mice (E) at approximately the same density. dendr, dendrite; sp, spine. Scale bars: 250 nm (A–E).

    Techniques Used: Mouse Assay

    Quantitative analysis of immunogold labelling for the Kv4.2 subunit
    Figure Legend Snippet: Quantitative analysis of immunogold labelling for the Kv4.2 subunit

    Techniques Used:

    Distribution of immunogold particles for the Kv4.2 subunit in oblique dendrites and spines of CA1 PCs in the stratum radiatum. (A) Immunogold particles are homogenously distributed along the P-face of an oblique dendrite (obl.d.) in the proximal stratum radiatum (prox. sr). (B) Slightly higher immunogold particle density for the Kv4.2 subunit can be seen on an oblique dendrite from the middle stratum radiatum (mid. sr). Note that a spine (sp) with high density of immunogold particles emerges from the dendrite and faces an axon terminal (at) E-face. (C) A spiny oblique dendrite in the distal stratum radiatum (dist. sr) contains a large number of immunogold particles for the Kv4.2 subunit. Scale bars: 250 nm (A–C).
    Figure Legend Snippet: Distribution of immunogold particles for the Kv4.2 subunit in oblique dendrites and spines of CA1 PCs in the stratum radiatum. (A) Immunogold particles are homogenously distributed along the P-face of an oblique dendrite (obl.d.) in the proximal stratum radiatum (prox. sr). (B) Slightly higher immunogold particle density for the Kv4.2 subunit can be seen on an oblique dendrite from the middle stratum radiatum (mid. sr). Note that a spine (sp) with high density of immunogold particles emerges from the dendrite and faces an axon terminal (at) E-face. (C) A spiny oblique dendrite in the distal stratum radiatum (dist. sr) contains a large number of immunogold particles for the Kv4.2 subunit. Scale bars: 250 nm (A–C).

    Techniques Used:

    Distribution of the Kv4.2 subunit immunoreactivity in the hippocampus and the specificity of immunoreactions. (A) The immunfluorescent reaction shows strong, homogenous neuropil labelling for the Kv4.2 subunit in strata oriens (so) and radiatum (sr) of rat CA1 region, with a slight reduction of the immunolabelling in stratum lacunosum-moleculare (slm). (B) A very similar labelling pattern was observed in the mouse hippocampus. (C) The labelling was absent in the CA1 area of Kv4.2 −/− mice, demonstrating the specificity of the immunoreaction. sp, stratum pyramidale. All images are single confocal sections. Scale bars: 50 μm (A–C).
    Figure Legend Snippet: Distribution of the Kv4.2 subunit immunoreactivity in the hippocampus and the specificity of immunoreactions. (A) The immunfluorescent reaction shows strong, homogenous neuropil labelling for the Kv4.2 subunit in strata oriens (so) and radiatum (sr) of rat CA1 region, with a slight reduction of the immunolabelling in stratum lacunosum-moleculare (slm). (B) A very similar labelling pattern was observed in the mouse hippocampus. (C) The labelling was absent in the CA1 area of Kv4.2 −/− mice, demonstrating the specificity of the immunoreaction. sp, stratum pyramidale. All images are single confocal sections. Scale bars: 50 μm (A–C).

    Techniques Used: Mouse Assay

    High-resolution immunogold localization of the Kv4.2 subunit in somata and apical dendrites of CA1 PCs. (A–E) Low-magnification images of P-faces of a PC soma (A) and spiny apical dendrites from the proximal (B), middle (C) and distal (D) stratum radiatum and stratum lacunosum-moleculare (E) show a rather uniform immunogold labelling pattern. (A′–E′) High-magnification images of the boxed region in (A–E). (F) Immunogold particles for the Kv4.2 subunit are homogenously distributed around a branch point on a PC dendrite. (F′) High-magnification view of the boxed region in (F). dist. sr, distal stratum radiatum; mid. sr, middle stratum radiatum; prox. sr, proximal stratum radiatum; slm, stratum lacunosum-moleculare; sp, stratum pyramidale. Scale bars: 1 μm (A); 500 nm (B–F); 100 nm (A′–F′).
    Figure Legend Snippet: High-resolution immunogold localization of the Kv4.2 subunit in somata and apical dendrites of CA1 PCs. (A–E) Low-magnification images of P-faces of a PC soma (A) and spiny apical dendrites from the proximal (B), middle (C) and distal (D) stratum radiatum and stratum lacunosum-moleculare (E) show a rather uniform immunogold labelling pattern. (A′–E′) High-magnification images of the boxed region in (A–E). (F) Immunogold particles for the Kv4.2 subunit are homogenously distributed around a branch point on a PC dendrite. (F′) High-magnification view of the boxed region in (F). dist. sr, distal stratum radiatum; mid. sr, middle stratum radiatum; prox. sr, proximal stratum radiatum; slm, stratum lacunosum-moleculare; sp, stratum pyramidale. Scale bars: 1 μm (A); 500 nm (B–F); 100 nm (A′–F′).

    Techniques Used:

    18) Product Images from "Kv4.2 potassium channels segregate to extrasynaptic domains and influence intrasynaptic NMDA receptor NR2B subunit expression"

    Article Title: Kv4.2 potassium channels segregate to extrasynaptic domains and influence intrasynaptic NMDA receptor NR2B subunit expression

    Journal: Brain Structure & Function

    doi: 10.1007/s00429-012-0450-1

    Modulatory action of Kv4.2 on the synaptic expression of NR2B. a In a double-immunolabeling experiment for NMDA-type glutamate receptor subunit 1 (NR1) and subunit 2B (NR2B), both NR1 (10 nm gold) and NR2B (5 nm gold; arrowheads ) are observed on the P-face of the postsynaptic membrane specialization of an excitatory synapse in an ITC neuron from a Kv4.2 +/+ mouse. b In an ITC neuron from a Kv4.2 −/− mouse, NR1 (10 nm gold) and NR2B immunolabeling (5 nm gold; arrowheads ) are observed in the postsynaptic membrane of an excitatory synapse at a density similar to that observed in Kv4.2 +/+ neurons. Yet, the frequency of such synapses, co-immunolabeled for both the NR1 and NR2B subunits, was increased in ITC neurons of Kv4.2 −/− mice. Scale bar 150 nm ( a , b )
    Figure Legend Snippet: Modulatory action of Kv4.2 on the synaptic expression of NR2B. a In a double-immunolabeling experiment for NMDA-type glutamate receptor subunit 1 (NR1) and subunit 2B (NR2B), both NR1 (10 nm gold) and NR2B (5 nm gold; arrowheads ) are observed on the P-face of the postsynaptic membrane specialization of an excitatory synapse in an ITC neuron from a Kv4.2 +/+ mouse. b In an ITC neuron from a Kv4.2 −/− mouse, NR1 (10 nm gold) and NR2B immunolabeling (5 nm gold; arrowheads ) are observed in the postsynaptic membrane of an excitatory synapse at a density similar to that observed in Kv4.2 +/+ neurons. Yet, the frequency of such synapses, co-immunolabeled for both the NR1 and NR2B subunits, was increased in ITC neurons of Kv4.2 −/− mice. Scale bar 150 nm ( a , b )

    Techniques Used: Expressing, Immunolabeling, Mouse Assay

    SDS-FRL confirms the localization of Kv4.2 in the somato-dendritic plasma membrane of ITC neurons. a Immunoparticles labeling Kv4.2 subunits (10 nm gold) are scattered on the plasma membrane of a large dendritic trunk of a rat ITC neuron. The immunolabeling is restricted to the plasma membrane P-face. b Scattered immunopartices are also observed in the plasma membrane P-face of a large dendritic trunk of a mouse ITC neuron. c In addition to the labeling of the dendritic trunk, Kv4.2 immunoparticles are localized to the head and neck of spines. d Both the plasma membrane E- and P-face of an ITC dendrite from a Kv4.2 −/− mouse are immunonegative when reacted with the Kv4.2 (209–225) antibody (10 nm gold). e An ITC dendritic trunk from a wild-type mouse (Kv4.2 +/+ ) is immunopositive for both Kv4.2 (209–225) (5 nm gold) and MOR (15 nm gold) in a double-immunolabeling experiment. f At higher resolution ( boxed area in e ), both 5 and 15 nm gold particles revealing Kv4.2 subunits and MOR, respectively, are visible on the P-face of the dendritic plasma membrane. MOR μ-opioid receptor. Scale bar 300 nm ( a , c ); 200 nm ( b , d ); 400 nm ( e ); 150 nm ( f )
    Figure Legend Snippet: SDS-FRL confirms the localization of Kv4.2 in the somato-dendritic plasma membrane of ITC neurons. a Immunoparticles labeling Kv4.2 subunits (10 nm gold) are scattered on the plasma membrane of a large dendritic trunk of a rat ITC neuron. The immunolabeling is restricted to the plasma membrane P-face. b Scattered immunopartices are also observed in the plasma membrane P-face of a large dendritic trunk of a mouse ITC neuron. c In addition to the labeling of the dendritic trunk, Kv4.2 immunoparticles are localized to the head and neck of spines. d Both the plasma membrane E- and P-face of an ITC dendrite from a Kv4.2 −/− mouse are immunonegative when reacted with the Kv4.2 (209–225) antibody (10 nm gold). e An ITC dendritic trunk from a wild-type mouse (Kv4.2 +/+ ) is immunopositive for both Kv4.2 (209–225) (5 nm gold) and MOR (15 nm gold) in a double-immunolabeling experiment. f At higher resolution ( boxed area in e ), both 5 and 15 nm gold particles revealing Kv4.2 subunits and MOR, respectively, are visible on the P-face of the dendritic plasma membrane. MOR μ-opioid receptor. Scale bar 300 nm ( a , c ); 200 nm ( b , d ); 400 nm ( e ); 150 nm ( f )

    Techniques Used: Labeling, Immunolabeling

    Kv4.2 does not reside within the postsynaptic membrane specializations of excitatory synapses as revealed by a double-replica approach. a Immunogold particles revealing NR1 subunits (10 nm gold) are localized to the E-face of an ITC dendrite. b On the mirror replica, immunogold particles revealing Kv4.2 subunits (10 nm gold) are observed on the P-face of the same dendrite. c At higher resolution ( boxed area in a ), immunolabeling for NR1 subunits is evident on the E-face of a postsynaptic membrane specialization of a glutamatergic synapse (shown in orange ), which is characterized by the clustering of intramembrane particles. d Immunolabeling for Kv4.2 subunits is present on the P-face of the extrasynaptic plasma membrane at higher resolution ( boxed area in b ). The postsynaptic membrane specialization by itself is free of any Kv4.2 immunolabeling. Sp spine. Scale bar 300 nm ( a , b ); 150 nm ( c , d )
    Figure Legend Snippet: Kv4.2 does not reside within the postsynaptic membrane specializations of excitatory synapses as revealed by a double-replica approach. a Immunogold particles revealing NR1 subunits (10 nm gold) are localized to the E-face of an ITC dendrite. b On the mirror replica, immunogold particles revealing Kv4.2 subunits (10 nm gold) are observed on the P-face of the same dendrite. c At higher resolution ( boxed area in a ), immunolabeling for NR1 subunits is evident on the E-face of a postsynaptic membrane specialization of a glutamatergic synapse (shown in orange ), which is characterized by the clustering of intramembrane particles. d Immunolabeling for Kv4.2 subunits is present on the P-face of the extrasynaptic plasma membrane at higher resolution ( boxed area in b ). The postsynaptic membrane specialization by itself is free of any Kv4.2 immunolabeling. Sp spine. Scale bar 300 nm ( a , b ); 150 nm ( c , d )

    Techniques Used: Immunolabeling

    Quantitative characterization of the distribution of Kv4.2 immunogold particles in ITC neurons and other central principal cells. a A sample SDS-FRL image of a portion of an ITC neuron soma. The area of the relevant profile is colored in blue . Immunogold particles labeling Kv4.2 subunits are marked with a black dot , a 20-nm radius circle around each particle is shown in yellow and the centroid of overlapping circles is marked with a red open circle . A tight cluster of particles is defined as particles residing within the overlapping yellow circles . b Cumulative probability curves for the nearest neighbor distances (NNDs) between individual immunogold particles (in black ) and tight cluster centers along with single particles (in red ), and NNDs of the calculated random distribution (in blue ). c A sample image of a portion of an ITC dendrite and d respective NND analysis. e – h Sample images and NND analyses for CA1 pyramidal neuron soma and dendrite. i – j Sample image and NND analysis for cerebellar granule cell soma. k Histogram showing the immunogold particle density in subcellular domains of ITC neurons (soma: 8 profiles with a mean area of 2.60 μm 2 ; dendrite: 22 profiles with a mean area of 2.05 μm 2 ), CA1 pyramidal neurons (soma: 6 profiles with a mean area of 2.52 μm 2 ; dendrite: 19 profiles with a mean area of 1.52 μm 2 ) and cerebellar granule cells (soma: 25 profiles with a mean area of 2.54 μm 2 ). Cb cerebellar granule cell, NND nearest neighbor distance. Error bars , SEM. * p
    Figure Legend Snippet: Quantitative characterization of the distribution of Kv4.2 immunogold particles in ITC neurons and other central principal cells. a A sample SDS-FRL image of a portion of an ITC neuron soma. The area of the relevant profile is colored in blue . Immunogold particles labeling Kv4.2 subunits are marked with a black dot , a 20-nm radius circle around each particle is shown in yellow and the centroid of overlapping circles is marked with a red open circle . A tight cluster of particles is defined as particles residing within the overlapping yellow circles . b Cumulative probability curves for the nearest neighbor distances (NNDs) between individual immunogold particles (in black ) and tight cluster centers along with single particles (in red ), and NNDs of the calculated random distribution (in blue ). c A sample image of a portion of an ITC dendrite and d respective NND analysis. e – h Sample images and NND analyses for CA1 pyramidal neuron soma and dendrite. i – j Sample image and NND analysis for cerebellar granule cell soma. k Histogram showing the immunogold particle density in subcellular domains of ITC neurons (soma: 8 profiles with a mean area of 2.60 μm 2 ; dendrite: 22 profiles with a mean area of 2.05 μm 2 ), CA1 pyramidal neurons (soma: 6 profiles with a mean area of 2.52 μm 2 ; dendrite: 19 profiles with a mean area of 1.52 μm 2 ) and cerebellar granule cells (soma: 25 profiles with a mean area of 2.54 μm 2 ). Cb cerebellar granule cell, NND nearest neighbor distance. Error bars , SEM. * p

    Techniques Used: Labeling

    Segregation of Kv4.2 to extrasynaptic domains. a In a double-immunolabeling experiment for PSD-95, visualized with 15 nm gold particles, and Kv4.2, visualized with 10 nm gold particles ( arrowheads ), Kv4.2 subunits are observed at extrasynaptic sites of an ITC dendritic spine in a wild-type mouse (Kv4.2 +/+ ). Conversely, PSD-95 labeling can be seen within the postsynaptic membrane specialization of an excitatory synapse. b In a Kv4.2 −/− mouse, labeling for PSD-95 (10 nm gold) can clearly be detected within the postsynaptic membrane specialization of a dendritic spine. No immunoreactivity for Kv4.2 (15 nm gold) is observed in Kv4.2 −/− mouse tissue. Scale bar 200 nm ( a , b )
    Figure Legend Snippet: Segregation of Kv4.2 to extrasynaptic domains. a In a double-immunolabeling experiment for PSD-95, visualized with 15 nm gold particles, and Kv4.2, visualized with 10 nm gold particles ( arrowheads ), Kv4.2 subunits are observed at extrasynaptic sites of an ITC dendritic spine in a wild-type mouse (Kv4.2 +/+ ). Conversely, PSD-95 labeling can be seen within the postsynaptic membrane specialization of an excitatory synapse. b In a Kv4.2 −/− mouse, labeling for PSD-95 (10 nm gold) can clearly be detected within the postsynaptic membrane specialization of a dendritic spine. No immunoreactivity for Kv4.2 (15 nm gold) is observed in Kv4.2 −/− mouse tissue. Scale bar 200 nm ( a , b )

    Techniques Used: Immunolabeling, Labeling

    Distribution of Kv4.2 in the mouse amygdala revealed by immunoperoxidase labeling. a Prominent Kv4.2 immunoreactivity is detected in all ITC clusters and the ITC nucleus in C57Bl/6 wild-type mice (Kv4.2 +/+ ). Only low to moderate levels of immunoreactivity are observed in other amygdaloid areas than the ITC. b Higher magnification of the Imp cluster ( boxed area in a ). Diffuse Kv4.2 immunoreactivity is primarily observed in the neuropil. c A similar immunostaining pattern for Kv4.2 is observed in the ITC nucleus ( boxed area in a ). d Specificity of Kv4.2 immunolabeling is confirmed on respective brain areas from a Kv4.2 −/− mouse. BL basolateral amygala, Ce central nucleus of amygdala, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, Is LA supralateral ITC cluster, La lateral amygdala. Scale bar 200 μm ( a , d ); 40 μm ( b , c )
    Figure Legend Snippet: Distribution of Kv4.2 in the mouse amygdala revealed by immunoperoxidase labeling. a Prominent Kv4.2 immunoreactivity is detected in all ITC clusters and the ITC nucleus in C57Bl/6 wild-type mice (Kv4.2 +/+ ). Only low to moderate levels of immunoreactivity are observed in other amygdaloid areas than the ITC. b Higher magnification of the Imp cluster ( boxed area in a ). Diffuse Kv4.2 immunoreactivity is primarily observed in the neuropil. c A similar immunostaining pattern for Kv4.2 is observed in the ITC nucleus ( boxed area in a ). d Specificity of Kv4.2 immunolabeling is confirmed on respective brain areas from a Kv4.2 −/− mouse. BL basolateral amygala, Ce central nucleus of amygdala, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, Is LA supralateral ITC cluster, La lateral amygdala. Scale bar 200 μm ( a , d ); 40 μm ( b , c )

    Techniques Used: Labeling, Mouse Assay, Immunostaining, Immunolabeling

    Kv4.2 immunolabeling in the rat amygdala using antibodies directed against different regions of the Kv4.2 protein. a – c The Kv4.2 (454–469) antibody reveals prominent immunolabeling of all ITC clusters and the ITC nucleus from rostral ( a ) to medial ( b ) and caudal ( c ) levels of the amygdala using an immunoperoxidase staining technique. Other amygdaloid areas are only faintly labeled. d – f An equivalent immunolabeling pattern is observed with the Kv4.2 (209–225) antibody. BL basolateral amygdala, Ce central nucleus of amygdala, Ii LA intralateral paracapsular ITC cluster, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral nucleus of amygdala. Scale bar 350 μm ( a – f )
    Figure Legend Snippet: Kv4.2 immunolabeling in the rat amygdala using antibodies directed against different regions of the Kv4.2 protein. a – c The Kv4.2 (454–469) antibody reveals prominent immunolabeling of all ITC clusters and the ITC nucleus from rostral ( a ) to medial ( b ) and caudal ( c ) levels of the amygdala using an immunoperoxidase staining technique. Other amygdaloid areas are only faintly labeled. d – f An equivalent immunolabeling pattern is observed with the Kv4.2 (209–225) antibody. BL basolateral amygdala, Ce central nucleus of amygdala, Ii LA intralateral paracapsular ITC cluster, Ilp lateral paracapsular ITC cluster, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral nucleus of amygdala. Scale bar 350 μm ( a – f )

    Techniques Used: Immunolabeling, Immunoperoxidase Staining, Labeling

    Subcellular localization of Kv4.2 to somato-dendritic domains of ITC neurons revealed by pre-embedding immunoperoxidase and immunometal electron microscopy. a A small dendritic trunk of a neuron in the rat ITC nucleus displays dense and diffuse immunolabeling for Kv4.2 [Kv4.2 (454–469) antibody]. The immunolabeling appears as an electron-opaque reaction product. A synaptic terminal, contacting this dendrite, is free of any immunolabeling. b A dendritic spine of a neuron in the mouse ITC nucleus shows dense and diffuse labeling for Kv4.2 [Kv4.2 (209–225) antibody]. An axon terminal contacting this spine, as well as other cellular profiles, is free of any immunolabeling. c A dendritic spine of a mouse Imp neuron shows the same dense and diffuse labeling pattern for Kv4.2 [Kv4.2 (454–469) antibody]. d In a double-labeling experiment, a Kv4.2 immunopositive spine of a rat Imp neuron (immunoperoxidase reaction) is also immunoreactive for MOR (immunometal reaction). e A small dendrite of a mouse Imp neuron, immunolabeled for Kv4.2 (immunoperoxidase reaction), is also immunoreactive for MOR (immunometal reaction). An axon terminal, contacting this dendrite, is free of any immunolabeling. f In a sample from a Kv4.2 −/− mouse, a large dendrite of an Imp neuron is labeled for MOR (immunometal reaction), but devoid of Kv4.2 immunolabeling (immunoperoxidase reaction). g Immunometal particles (indicated by arrowheads ) for Kv4.2, when applying the Kv4.2 (454–469) antibody, are observed at the intracellular side of the plasma membrane of a rat Imp neuron. The postsynaptic specialization of an excitatory synapse is free of any immunolabeling. Some immunoparticles appear localized perisynaptically. h Immunometal particles (indicated by arrowheads) for Kv4.2 appear at the intracellular side of an ITC neuron dendrite also when applying the Kv4.2 (209–225) antibody. Both the axon forming a synapse with this dendrite and the postsynaptic specialization are free of any immunolabeling. i The plasma membrane of an Imp dendrite is decorated with immunoparticles at the intracellular side applying the Kv4.2 (209–225) antibody. The postsynaptic specialization of an inhibitory synapse on this dendrite is also free of any immunolabeling. At axon terminal, MOR μ-opioid receptor. Scale bar 200 nm ( a – c , e , h , i ); 300 nm ( d , f , g )
    Figure Legend Snippet: Subcellular localization of Kv4.2 to somato-dendritic domains of ITC neurons revealed by pre-embedding immunoperoxidase and immunometal electron microscopy. a A small dendritic trunk of a neuron in the rat ITC nucleus displays dense and diffuse immunolabeling for Kv4.2 [Kv4.2 (454–469) antibody]. The immunolabeling appears as an electron-opaque reaction product. A synaptic terminal, contacting this dendrite, is free of any immunolabeling. b A dendritic spine of a neuron in the mouse ITC nucleus shows dense and diffuse labeling for Kv4.2 [Kv4.2 (209–225) antibody]. An axon terminal contacting this spine, as well as other cellular profiles, is free of any immunolabeling. c A dendritic spine of a mouse Imp neuron shows the same dense and diffuse labeling pattern for Kv4.2 [Kv4.2 (454–469) antibody]. d In a double-labeling experiment, a Kv4.2 immunopositive spine of a rat Imp neuron (immunoperoxidase reaction) is also immunoreactive for MOR (immunometal reaction). e A small dendrite of a mouse Imp neuron, immunolabeled for Kv4.2 (immunoperoxidase reaction), is also immunoreactive for MOR (immunometal reaction). An axon terminal, contacting this dendrite, is free of any immunolabeling. f In a sample from a Kv4.2 −/− mouse, a large dendrite of an Imp neuron is labeled for MOR (immunometal reaction), but devoid of Kv4.2 immunolabeling (immunoperoxidase reaction). g Immunometal particles (indicated by arrowheads ) for Kv4.2, when applying the Kv4.2 (454–469) antibody, are observed at the intracellular side of the plasma membrane of a rat Imp neuron. The postsynaptic specialization of an excitatory synapse is free of any immunolabeling. Some immunoparticles appear localized perisynaptically. h Immunometal particles (indicated by arrowheads) for Kv4.2 appear at the intracellular side of an ITC neuron dendrite also when applying the Kv4.2 (209–225) antibody. Both the axon forming a synapse with this dendrite and the postsynaptic specialization are free of any immunolabeling. i The plasma membrane of an Imp dendrite is decorated with immunoparticles at the intracellular side applying the Kv4.2 (209–225) antibody. The postsynaptic specialization of an inhibitory synapse on this dendrite is also free of any immunolabeling. At axon terminal, MOR μ-opioid receptor. Scale bar 200 nm ( a – c , e , h , i ); 300 nm ( d , f , g )

    Techniques Used: Electron Microscopy, Immunolabeling, Labeling

    Colocalization of Kv4.2 and μ-opioid receptors (MOR) in the ITCs. a Double-labeling immunofluorescence for Kv4.2 (in green ) and MOR (in red ) reveals a high degree of coexistence in the amygdala, which is evident in the merged images. b At higher resolution, immunoreactivity for Kv4.2 (in green ) as well as for MOR (in red ) appears dense and diffuse in ITC clusters such as the Imp. The distribution profile of these proteins is highly similar ( merge ). BL basolateral amygdala, Ce central nucleus of amygdala, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral amygdala. Scale bar 350 μm ( a ), 50 μm ( b )
    Figure Legend Snippet: Colocalization of Kv4.2 and μ-opioid receptors (MOR) in the ITCs. a Double-labeling immunofluorescence for Kv4.2 (in green ) and MOR (in red ) reveals a high degree of coexistence in the amygdala, which is evident in the merged images. b At higher resolution, immunoreactivity for Kv4.2 (in green ) as well as for MOR (in red ) appears dense and diffuse in ITC clusters such as the Imp. The distribution profile of these proteins is highly similar ( merge ). BL basolateral amygdala, Ce central nucleus of amygdala, Imp medial paracapsular ITC cluster, IN ITC nucleus, La lateral amygdala. Scale bar 350 μm ( a ), 50 μm ( b )

    Techniques Used: Labeling, Immunofluorescence

    19) Product Images from "Unique somato-dendritic distribution pattern of Kv4.2 channels on hippocampal CA1 pyramidal cells"

    Article Title: Unique somato-dendritic distribution pattern of Kv4.2 channels on hippocampal CA1 pyramidal cells

    Journal: The European Journal of Neuroscience

    doi: 10.1111/j.1460-9568.2011.07907.x

    Densities of Kv4.2 immunogold particles in different somato-dendritic compartments of rat CA1 PCs. Bar graphs illustrate the background subtracted densities (mean ± SD, in gold/μm 2 ) of immunogold particles in the somato-dendritic compartments. The bars are colour coded to different subcellular compartments as indicated in the schematic drawing of a CA1 PC. Note the moderate increase in the density along the proximo-distal axis of PCs within the stratum radiatum and the subsequent decrease in the stratum lacunosum-moleculare.
    Figure Legend Snippet: Densities of Kv4.2 immunogold particles in different somato-dendritic compartments of rat CA1 PCs. Bar graphs illustrate the background subtracted densities (mean ± SD, in gold/μm 2 ) of immunogold particles in the somato-dendritic compartments. The bars are colour coded to different subcellular compartments as indicated in the schematic drawing of a CA1 PC. Note the moderate increase in the density along the proximo-distal axis of PCs within the stratum radiatum and the subsequent decrease in the stratum lacunosum-moleculare.

    Techniques Used:

    The Kv4.2 subunit is excluded from postsynaptic membrane specializations. (A) Electron micrograph illustrating an excitatory synapse, revealed by the accumulation of the postsynaptic density marker PSD-95 (15-nm gold). Arrowheads point to gold particles labelling the Kv4.2 subunit (10-nm gold) around the PSD. (B) High-magnification image of an inhibitory synapse identified by the enrichment of gold particles (5-nm gold) labelling the γ-aminobutyric acid (GABA) A receptor β3 subunit. Note the clustering of gold particles labelling the Kv4.2 subunit (10-nm gold) in the extrasynaptic membrane, but not within the inhibitory synapse. Scale bars: 100 nm (A); 50 nm (B).
    Figure Legend Snippet: The Kv4.2 subunit is excluded from postsynaptic membrane specializations. (A) Electron micrograph illustrating an excitatory synapse, revealed by the accumulation of the postsynaptic density marker PSD-95 (15-nm gold). Arrowheads point to gold particles labelling the Kv4.2 subunit (10-nm gold) around the PSD. (B) High-magnification image of an inhibitory synapse identified by the enrichment of gold particles (5-nm gold) labelling the γ-aminobutyric acid (GABA) A receptor β3 subunit. Note the clustering of gold particles labelling the Kv4.2 subunit (10-nm gold) in the extrasynaptic membrane, but not within the inhibitory synapse. Scale bars: 100 nm (A); 50 nm (B).

    Techniques Used: Marker

    Specificity test for Kv4.2 subunit labelling on the axo-somato-dendritic surface of CA1 PCs using SDS-FRL. (A) Electron micrograph illustrating the P-face of a spiny CA1 PC dendrite of wild-type mouse immunolabelled for the Kv4.2 subunit. Note that the density and distribution pattern of the immunogold labelling for the Kv4.2 subunit in mouse is similar to that seen in rat. (B) The lack of immunogold labelling for the Kv4.2 subunit in a spiny dendrite of a Kv4.2 −/− mouse demonstrates the specificity of the labelling using SDS-FRL. (C) Axon terminals in rat identified by immunolabelling for SNAP-25 (15-nm gold) contain a low number of immunogold particles (arrows) for the Kv4.2 subunit (10-nm gold). (D and E) Immunogold particles for the Kv4.2 subunit were present in both Kv4.2 +/+ (D) and Kv4.2 −/− mice (E) at approximately the same density. dendr, dendrite; sp, spine. Scale bars: 250 nm (A–E).
    Figure Legend Snippet: Specificity test for Kv4.2 subunit labelling on the axo-somato-dendritic surface of CA1 PCs using SDS-FRL. (A) Electron micrograph illustrating the P-face of a spiny CA1 PC dendrite of wild-type mouse immunolabelled for the Kv4.2 subunit. Note that the density and distribution pattern of the immunogold labelling for the Kv4.2 subunit in mouse is similar to that seen in rat. (B) The lack of immunogold labelling for the Kv4.2 subunit in a spiny dendrite of a Kv4.2 −/− mouse demonstrates the specificity of the labelling using SDS-FRL. (C) Axon terminals in rat identified by immunolabelling for SNAP-25 (15-nm gold) contain a low number of immunogold particles (arrows) for the Kv4.2 subunit (10-nm gold). (D and E) Immunogold particles for the Kv4.2 subunit were present in both Kv4.2 +/+ (D) and Kv4.2 −/− mice (E) at approximately the same density. dendr, dendrite; sp, spine. Scale bars: 250 nm (A–E).

    Techniques Used: Mouse Assay

    Quantitative analysis of immunogold labelling for the Kv4.2 subunit
    Figure Legend Snippet: Quantitative analysis of immunogold labelling for the Kv4.2 subunit

    Techniques Used:

    Distribution of immunogold particles for the Kv4.2 subunit in oblique dendrites and spines of CA1 PCs in the stratum radiatum. (A) Immunogold particles are homogenously distributed along the P-face of an oblique dendrite (obl.d.) in the proximal stratum radiatum (prox. sr). (B) Slightly higher immunogold particle density for the Kv4.2 subunit can be seen on an oblique dendrite from the middle stratum radiatum (mid. sr). Note that a spine (sp) with high density of immunogold particles emerges from the dendrite and faces an axon terminal (at) E-face. (C) A spiny oblique dendrite in the distal stratum radiatum (dist. sr) contains a large number of immunogold particles for the Kv4.2 subunit. Scale bars: 250 nm (A–C).
    Figure Legend Snippet: Distribution of immunogold particles for the Kv4.2 subunit in oblique dendrites and spines of CA1 PCs in the stratum radiatum. (A) Immunogold particles are homogenously distributed along the P-face of an oblique dendrite (obl.d.) in the proximal stratum radiatum (prox. sr). (B) Slightly higher immunogold particle density for the Kv4.2 subunit can be seen on an oblique dendrite from the middle stratum radiatum (mid. sr). Note that a spine (sp) with high density of immunogold particles emerges from the dendrite and faces an axon terminal (at) E-face. (C) A spiny oblique dendrite in the distal stratum radiatum (dist. sr) contains a large number of immunogold particles for the Kv4.2 subunit. Scale bars: 250 nm (A–C).

    Techniques Used:

    Distribution of the Kv4.2 subunit immunoreactivity in the hippocampus and the specificity of immunoreactions. (A) The immunfluorescent reaction shows strong, homogenous neuropil labelling for the Kv4.2 subunit in strata oriens (so) and radiatum (sr) of rat CA1 region, with a slight reduction of the immunolabelling in stratum lacunosum-moleculare (slm). (B) A very similar labelling pattern was observed in the mouse hippocampus. (C) The labelling was absent in the CA1 area of Kv4.2 −/− mice, demonstrating the specificity of the immunoreaction. sp, stratum pyramidale. All images are single confocal sections. Scale bars: 50 μm (A–C).
    Figure Legend Snippet: Distribution of the Kv4.2 subunit immunoreactivity in the hippocampus and the specificity of immunoreactions. (A) The immunfluorescent reaction shows strong, homogenous neuropil labelling for the Kv4.2 subunit in strata oriens (so) and radiatum (sr) of rat CA1 region, with a slight reduction of the immunolabelling in stratum lacunosum-moleculare (slm). (B) A very similar labelling pattern was observed in the mouse hippocampus. (C) The labelling was absent in the CA1 area of Kv4.2 −/− mice, demonstrating the specificity of the immunoreaction. sp, stratum pyramidale. All images are single confocal sections. Scale bars: 50 μm (A–C).

    Techniques Used: Mouse Assay

    High-resolution immunogold localization of the Kv4.2 subunit in somata and apical dendrites of CA1 PCs. (A–E) Low-magnification images of P-faces of a PC soma (A) and spiny apical dendrites from the proximal (B), middle (C) and distal (D) stratum radiatum and stratum lacunosum-moleculare (E) show a rather uniform immunogold labelling pattern. (A′–E′) High-magnification images of the boxed region in (A–E). (F) Immunogold particles for the Kv4.2 subunit are homogenously distributed around a branch point on a PC dendrite. (F′) High-magnification view of the boxed region in (F). dist. sr, distal stratum radiatum; mid. sr, middle stratum radiatum; prox. sr, proximal stratum radiatum; slm, stratum lacunosum-moleculare; sp, stratum pyramidale. Scale bars: 1 μm (A); 500 nm (B–F); 100 nm (A′–F′).
    Figure Legend Snippet: High-resolution immunogold localization of the Kv4.2 subunit in somata and apical dendrites of CA1 PCs. (A–E) Low-magnification images of P-faces of a PC soma (A) and spiny apical dendrites from the proximal (B), middle (C) and distal (D) stratum radiatum and stratum lacunosum-moleculare (E) show a rather uniform immunogold labelling pattern. (A′–E′) High-magnification images of the boxed region in (A–E). (F) Immunogold particles for the Kv4.2 subunit are homogenously distributed around a branch point on a PC dendrite. (F′) High-magnification view of the boxed region in (F). dist. sr, distal stratum radiatum; mid. sr, middle stratum radiatum; prox. sr, proximal stratum radiatum; slm, stratum lacunosum-moleculare; sp, stratum pyramidale. Scale bars: 1 μm (A); 500 nm (B–F); 100 nm (A′–F′).

    Techniques Used:

    20) Product Images from "Cellular and Subcellular Localisation of Kv4-Associated KChIP Proteins in the Rat Cerebellum"

    Article Title: Cellular and Subcellular Localisation of Kv4-Associated KChIP Proteins in the Rat Cerebellum

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21176403

    Regional and cellular distribution of voltage-gated potassium (Kv) channel subunits Kv4.2 and Kv4.3 in the cerebellum. ( A – D ) Immunoreactivity for Kv4.2 and Kv4.3 in the rat cerebellar cortex using a pre-embedding immunoperoxidase method at the light microscopic level. Parasagittal photomicrographs of the cerebellar cortex. The strongest immunoreactivity for Kv4.2 and Kv4.3 was found in the granule cell layer (gcl). Strong immunoreactivity for Kv4.3 was also observed in the molecular layer (ml), but weaker for Kv4.2. The white matter (wm) was always devoid of any immunolabelling. Immunoreactivity for Kv4.2 and Kv4.3 in the molecular layer was mostly neuropilar, but Kv4.3 labelling was also detected in cell bodies and dendrites of basket cells (black arrows). In the granule cell layer, Kv4.2 and Kv4.3 immunolabelling particularly concentrated in glomeruli (white arrows) and surrounding GCs. Scale bars: ( A , B ), 50 µm; ( C , D ), 25 µm.
    Figure Legend Snippet: Regional and cellular distribution of voltage-gated potassium (Kv) channel subunits Kv4.2 and Kv4.3 in the cerebellum. ( A – D ) Immunoreactivity for Kv4.2 and Kv4.3 in the rat cerebellar cortex using a pre-embedding immunoperoxidase method at the light microscopic level. Parasagittal photomicrographs of the cerebellar cortex. The strongest immunoreactivity for Kv4.2 and Kv4.3 was found in the granule cell layer (gcl). Strong immunoreactivity for Kv4.3 was also observed in the molecular layer (ml), but weaker for Kv4.2. The white matter (wm) was always devoid of any immunolabelling. Immunoreactivity for Kv4.2 and Kv4.3 in the molecular layer was mostly neuropilar, but Kv4.3 labelling was also detected in cell bodies and dendrites of basket cells (black arrows). In the granule cell layer, Kv4.2 and Kv4.3 immunolabelling particularly concentrated in glomeruli (white arrows) and surrounding GCs. Scale bars: ( A , B ), 50 µm; ( C , D ), 25 µm.

    Techniques Used:

    Subcellular distribution of voltage-gated potassium (Kv) channel subunits Kv4.2 and Kv4.3 channels. Immunoreactivity for Kv4.2 and Kv4.3 in the cerebellar cortex as demonstrated by pre-embedding immunogold labelling. ( A – D ) Immunoparticles for both Kv4.2 and Kv4.3 in the molecular layer were found in the plasma membrane (arrows) and intracellular sites (crossed arrows) of PC dendrites (Den) and spines (s) establishing synapses with parallel fibres (pf). Although, at low frequency, Kv4.3 immunoparticles were also found in the plasma membrane (arrowheads) of parallel fibres (pf). ( E – J ) Immunoparticles for both Kv4.2 and Kv4.3 in the granule cell layer were found in the plasma membrane (arrows) of GC somata and GC dendrites (Den) in cerebellar glomeruli. In addition, immunolabelling was found intracellularly (crossed arrows). Few immunoparticles were also observed presynaptically along the plasma membrane (arrowheads) and intracellular sites (double arrowheads) in mossy fibre (mf) axon terminals. Scale bars: ( A – J ), 200 nm.
    Figure Legend Snippet: Subcellular distribution of voltage-gated potassium (Kv) channel subunits Kv4.2 and Kv4.3 channels. Immunoreactivity for Kv4.2 and Kv4.3 in the cerebellar cortex as demonstrated by pre-embedding immunogold labelling. ( A – D ) Immunoparticles for both Kv4.2 and Kv4.3 in the molecular layer were found in the plasma membrane (arrows) and intracellular sites (crossed arrows) of PC dendrites (Den) and spines (s) establishing synapses with parallel fibres (pf). Although, at low frequency, Kv4.3 immunoparticles were also found in the plasma membrane (arrowheads) of parallel fibres (pf). ( E – J ) Immunoparticles for both Kv4.2 and Kv4.3 in the granule cell layer were found in the plasma membrane (arrows) of GC somata and GC dendrites (Den) in cerebellar glomeruli. In addition, immunolabelling was found intracellularly (crossed arrows). Few immunoparticles were also observed presynaptically along the plasma membrane (arrowheads) and intracellular sites (double arrowheads) in mossy fibre (mf) axon terminals. Scale bars: ( A – J ), 200 nm.

    Techniques Used:

    Compartmentalisation of voltage-gated potassium (Kv) channel subunits Kv4.2 and Kv4.3 in cerebellar cells. ( A , B ) Bar graphs showing the percentage of immunoparticles for Kv4.2 and Kv4.3 in neuronal compartment in the molecular layer. Immunoparticles for Kv4.2 were mostly localised at the postsynaptic compartment (99% of all particles), while Kv4.3 was distributed at postsynaptic (88.4%) and presynaptic (11.6%) compartments. ( C , D ) Bar graphs showing the percentage of immunoparticles for Kv4.2 and Kv4.3 in the neuronal compartment in the granule cell layer. A total of 667 immunoparticles for Kv4.2 and 771 for Kv4.3 were analysed. Postsynaptically, immunoparticles were detected in dendrites of GCs (91.6% for Kv4.2; 82.2% for Kv4.3), distributed along the plasma membrane (46.3% for Kv4.2; 62.3% for Kv4.3) and at cytoplasmic sites (53.7% for Kv4.2; 37.7% for Kv4.3). Presynaptically, immunoparticles were detected in mossy fibre terminals (8.4% for Kv4.2; 17.8% for Kv4.3), distributed at cytoplasmic sites (87.5% for Kv4.2; 53.3% for Kv4.3) and along the plasma membrane (12.5% for Kv4.2; 46.7% for Kv4.3). ( E , F ) Histogram showing the distribution of immunoparticles for Kv4.2 and Kv4.3 in relation to glutamate release sites in dendritic spines of PCs. About 46% of immunolabelled Kv4.2 and 52% of immunolabelled Kv4.3 were located in a 60–300 nm wide band. These data show that Kv4.2 immunoparticles were more equally distributed along PC spines, while immunoparticles Kv4.3 were skewed toward the PSD of PC spines.
    Figure Legend Snippet: Compartmentalisation of voltage-gated potassium (Kv) channel subunits Kv4.2 and Kv4.3 in cerebellar cells. ( A , B ) Bar graphs showing the percentage of immunoparticles for Kv4.2 and Kv4.3 in neuronal compartment in the molecular layer. Immunoparticles for Kv4.2 were mostly localised at the postsynaptic compartment (99% of all particles), while Kv4.3 was distributed at postsynaptic (88.4%) and presynaptic (11.6%) compartments. ( C , D ) Bar graphs showing the percentage of immunoparticles for Kv4.2 and Kv4.3 in the neuronal compartment in the granule cell layer. A total of 667 immunoparticles for Kv4.2 and 771 for Kv4.3 were analysed. Postsynaptically, immunoparticles were detected in dendrites of GCs (91.6% for Kv4.2; 82.2% for Kv4.3), distributed along the plasma membrane (46.3% for Kv4.2; 62.3% for Kv4.3) and at cytoplasmic sites (53.7% for Kv4.2; 37.7% for Kv4.3). Presynaptically, immunoparticles were detected in mossy fibre terminals (8.4% for Kv4.2; 17.8% for Kv4.3), distributed at cytoplasmic sites (87.5% for Kv4.2; 53.3% for Kv4.3) and along the plasma membrane (12.5% for Kv4.2; 46.7% for Kv4.3). ( E , F ) Histogram showing the distribution of immunoparticles for Kv4.2 and Kv4.3 in relation to glutamate release sites in dendritic spines of PCs. About 46% of immunolabelled Kv4.2 and 52% of immunolabelled Kv4.3 were located in a 60–300 nm wide band. These data show that Kv4.2 immunoparticles were more equally distributed along PC spines, while immunoparticles Kv4.3 were skewed toward the PSD of PC spines.

    Techniques Used:

    21) Product Images from "Contribution of Kv4 channels toward the A-type potassium current in murine colonic myocytes"

    Article Title: Contribution of Kv4 channels toward the A-type potassium current in murine colonic myocytes

    Journal: The Journal of Physiology

    doi: 10.1113/jphysiol.2002.025163

    Kv4.2- and Kv4.3-like immunoreactivity in the tunica muscularis of murine colon and jejunum Haematoxylin counterstain. A and B , Kv4.2-like ( A ) and Kv4.3-like ( B ) immunoreactivity (in brown) throughout the circular (cm) and longitudinal (lm) muscle layers of the tunica muscularis in murine colon. Arrowheads indicate Kv4-like immunoreactivity found within myenteric ganglia. C and D , Kv4.2-like ( C ) and Kv4.3-like ( D ) immunoreactivity (in brown) throughout the circular (cm) and longitudinal (lm) layers of the tunica muscularis in murine jejunum. Scale bars, 20 μm.
    Figure Legend Snippet: Kv4.2- and Kv4.3-like immunoreactivity in the tunica muscularis of murine colon and jejunum Haematoxylin counterstain. A and B , Kv4.2-like ( A ) and Kv4.3-like ( B ) immunoreactivity (in brown) throughout the circular (cm) and longitudinal (lm) muscle layers of the tunica muscularis in murine colon. Arrowheads indicate Kv4-like immunoreactivity found within myenteric ganglia. C and D , Kv4.2-like ( C ) and Kv4.3-like ( D ) immunoreactivity (in brown) throughout the circular (cm) and longitudinal (lm) layers of the tunica muscularis in murine jejunum. Scale bars, 20 μm.

    Techniques Used:

    22) Product Images from "Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels"

    Article Title: Encephalitis and antibodies to DPPX, a subunit of Kv4.2 potassium channels

    Journal: Annals of neurology

    doi: 10.1002/ana.23756

    Immunoprecipitation of DPPX In cultures of dissociated rat hippocampal neurons, patients’ antibodies showed intense reactivity with the neuronal cell surface (A), bar = 10 μm. Immunoprecipitation of the antigen with serum of the index case is shown in B, where the precipitated proteins were run in a gel and subsequently stained with EZblue. Note that patient’s antibodies precipitated a protein (band close to 102 kDa in lane P), which was excised from the gel and analyzed by mass spectrometry, demonstrating sequences of DPPX. Lane N is the precipitate obtained from control serum. Immunoblot of these proteins with a rabbit polyclonal antibody against DPPX (1:1000, developed by BR) confirmed that the band corresponded to DPPX (C).
    Figure Legend Snippet: Immunoprecipitation of DPPX In cultures of dissociated rat hippocampal neurons, patients’ antibodies showed intense reactivity with the neuronal cell surface (A), bar = 10 μm. Immunoprecipitation of the antigen with serum of the index case is shown in B, where the precipitated proteins were run in a gel and subsequently stained with EZblue. Note that patient’s antibodies precipitated a protein (band close to 102 kDa in lane P), which was excised from the gel and analyzed by mass spectrometry, demonstrating sequences of DPPX. Lane N is the precipitate obtained from control serum. Immunoblot of these proteins with a rabbit polyclonal antibody against DPPX (1:1000, developed by BR) confirmed that the band corresponded to DPPX (C).

    Techniques Used: Immunoprecipitation, Staining, Mass Spectrometry

    Expression of DPPX in myenteric plexus Transverse section of small bowel of rat showing the longitudinal muscular layer (LM), circular muscular layer (CM), submucosal layer (SM), and glans (G). The myenteric plexus (Plex) is revealed as clusters of large neurons between the two muscular layers. In the 3 panels (A–C) the nuclei of the neurons (red) was labeled with anti-Hu (a highly specific neuronal marker). Panel A, shows in green the DPPX immunostaining using a rabbit polyclonal antibody (1:1000, developed by BD); panel B shows the DPPX reactivity of serum from one of the patients with encephalitis, and panel C shows the lack of reactivity of serum from a healthy subject. Note that DPPX is predominantly expressed in the cytoplasm-membrane of the large clustered neurons of the myenteric plexus, and is also detected in a fine longitudinal pattern in CM and SM where the submucosal plexus is located. Bar = 20μm.
    Figure Legend Snippet: Expression of DPPX in myenteric plexus Transverse section of small bowel of rat showing the longitudinal muscular layer (LM), circular muscular layer (CM), submucosal layer (SM), and glans (G). The myenteric plexus (Plex) is revealed as clusters of large neurons between the two muscular layers. In the 3 panels (A–C) the nuclei of the neurons (red) was labeled with anti-Hu (a highly specific neuronal marker). Panel A, shows in green the DPPX immunostaining using a rabbit polyclonal antibody (1:1000, developed by BD); panel B shows the DPPX reactivity of serum from one of the patients with encephalitis, and panel C shows the lack of reactivity of serum from a healthy subject. Note that DPPX is predominantly expressed in the cytoplasm-membrane of the large clustered neurons of the myenteric plexus, and is also detected in a fine longitudinal pattern in CM and SM where the submucosal plexus is located. Bar = 20μm.

    Techniques Used: Expressing, Labeling, Marker, Immunostaining

    Related Articles

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    Article Title: Development of Action Potential Waveform in Hippocampal CA1 Pyramidal Neurons.
    Article Snippet: .. CA1 pyramidal neurons undergo intense morphological and electrophysiological changes from the second to third postnatal weeks in rats throughout a critical period associated with the emergence of exploratory behavior.. Using whole cell current-clamp recordings in vitro and neurochemical methods, we studied the development of the somatic action potential (AP) waveform and some of the underlying channels in this critical period. ..

    Incubation:

    Article Title: Development of Action Potential Waveform in Hippocampal CA1 Pyramidal Neurons.
    Article Snippet: .. CA1 pyramidal neurons undergo intense morphological and electrophysiological changes from the second to third postnatal weeks in rats throughout a critical period associated with the emergence of exploratory behavior.. Using whole cell current-clamp recordings in vitro and neurochemical methods, we studied the development of the somatic action potential (AP) waveform and some of the underlying channels in this critical period. ..

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    Alomone Labs kv4 2
    Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, <t>Kv4.2,</t> Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P
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    Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, Kv4.2, Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P

    Journal: Nature neuroscience

    Article Title: G9a Is Essential for Epigenetic Silencing of K+ Channel Genes in Acute-to-Chronic Pain Transition

    doi: 10.1038/nn.4165

    Figure Lengend Snippet: Inhibition of G9a activity normalizes K + channel gene expression in the DRG diminished by nerve injury ( a-d) Effects of intrathecal treatments with vehicle (n = 10), SAHA (50 μg, n = 9), UNC0638 (10 μg, n = 8), GSK503 (5 μg, n = 10), SAHA plus GSK503 (n = 8), SAHA plus UNC0638 (n = 9) or UNC0638 plus GSK503 (n = 8) on the mRNA levels of Kcna4 ( a ), Kcnd2 ( b ), Kcnq2 ( c ), and Kcnma1 ( d ) in the DRG obtained from SNL rats 28 days after surgery. Data from sham control rats were plotted as the control (n = 6 rats). ( e,f ) Effects of nerve injury and UNC0638 on the protein levels of Kv1.4, Kv4.2, Kv7.2 and BKα1 in the L5 and L6 DRG (n = 6 rats in each group). ( g,h ) Effect of G9a-specific siRNA on the G9a and H3K9me2 protein levels in the DRG obtained from SNL rats 24 h after the last injection (n = 5 in each group). ( i,j ) Effects of G9a-specific siRNA on the mRNA levels of G9a, Ezh2, Kcna4, Kcnd2, Kcnq2 and Kcnma1 in the DRG obtained from SNL ( i ) and sham control ( j ) rats 24 h after the last injection (n = 10 in each group). Data are presented as mean ± s.e.m. Statistical analysis was performed using two-way ANOVA followed by Bonferroni’s post hoc tests ( a-d ), one-way ANOVA ( f,h,i ), or Mann-Whitney test ( j ). * P

    Article Snippet: The membranes were treated with 5% bovine serum albumin in Tris buffer containing Tween 20 (TNT) for 2 h and then incubated with one of the following primary antibodies overnight at 4°C: Kv1.4 (catalog #05-409, Upstate Biotechnology), Kv4.2 (catalog #APC-023, Alomone Labs), Kv7.2 (catalog #APC-050, Alomone Labs), BKα1 (catalog #APC-021, Alomone Labs), acetyl-H3 (catalog #06-599, Millipore), H3K9ac (catalog #39917, Active Motif), H3K27me3 (catalog #9733, Cell Signaling Technology), histone H3 (catalog #9715, Cell Signaling Technology), H3K9me2 (catalog #ab1220, Abcam), G9a (catalog #09-071, Millipore), HDAC1 (catalog #IMG-337, Imgenex), and HDAC2, HDAC4, HDAC5 and EZH2 (catalog #2540, #2072, #2082 and #5246, respectively; Cell Signaling Technology).

    Techniques: Inhibition, Activity Assay, Expressing, Injection, MANN-WHITNEY

    VE and LOC neurons express Kv4.3 and Kv 4.2 subunits in the mouse. Adult mouse immunofluorescently labeled against ChAT (red) and Kv4.2 or Kv4.3 α-subunits (green) using shock-frozen tissue. The ChAT labeling overlaps with the Kv4.3 labeling in the mouse VE neurons (A–C). Likewise, both the ChAT-positive LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) show Kv4.3 expression in the mouse (D–F). In a similar fashion, both the VE neurons (G–I) and the LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) (J–L) are immunolabeled against Kv4.2, which indicate that the Kv4 channels are hetero-meric in these auditory brainstem nuclei. Scale bars: 100 µm.

    Journal: PLoS ONE

    Article Title: Physiological Characterization of Vestibular Efferent Brainstem Neurons Using a Transgenic Mouse Model

    doi: 10.1371/journal.pone.0098277

    Figure Lengend Snippet: VE and LOC neurons express Kv4.3 and Kv 4.2 subunits in the mouse. Adult mouse immunofluorescently labeled against ChAT (red) and Kv4.2 or Kv4.3 α-subunits (green) using shock-frozen tissue. The ChAT labeling overlaps with the Kv4.3 labeling in the mouse VE neurons (A–C). Likewise, both the ChAT-positive LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) show Kv4.3 expression in the mouse (D–F). In a similar fashion, both the VE neurons (G–I) and the LOC efferent cells (arrows) and the principal cells of the lateral superior olive (arrowheads) (J–L) are immunolabeled against Kv4.2, which indicate that the Kv4 channels are hetero-meric in these auditory brainstem nuclei. Scale bars: 100 µm.

    Article Snippet: Primary antibodies used were AlexaFluor488-conjugated rabbit anti-GFP 1∶500 (Invitrogen, Corp., Carlsbad, CA), goat anti-ChAT 1∶100 (Millipore, Corp., Temecula, CA), rabbit anti-Kv4.2 1∶100 and rabbit anti-Kv4.3 1∶100 (Alomone Labs, Ltd., Jerusalem, Israel).

    Techniques: Labeling, Expressing, Immunolabeling

    Blockade of phosphorylation at the PKC sites increases the surface expression of Kv4.2 channels in COS-7 cells

    Journal: The Biochemical journal

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    doi: 10.1042/BJ20081213

    Figure Lengend Snippet: Blockade of phosphorylation at the PKC sites increases the surface expression of Kv4.2 channels in COS-7 cells

    Article Snippet: Briefly, after fixing cells with 4% paraformaldehyde for 30 min, and incubation in 0.3% Triton X-100 in PBS for 20 min at room temperature, the cells were blocked by 10% fetal bovine serum in PBS for 60 min at room temperature, and then incubated for 60 min at room temperature or overnight at 4°C with primary antibodies: polyclonal Kv4.2 antibody from Alomone Labs (Jerusalem, Israel) generated against residues 454-469 ( SNQLQSSEDEPAFVSK) on the C-terminal or a monoclonal Kv4.2 antibody against an ectodomain [ ].

    Techniques: Expressing

    PKC phosphorylates the Kv4.2 C-terminal but not the N-terminal cytoplasmic domains in vitro

    Journal: The Biochemical journal

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    doi: 10.1042/BJ20081213

    Figure Lengend Snippet: PKC phosphorylates the Kv4.2 C-terminal but not the N-terminal cytoplasmic domains in vitro

    Article Snippet: Briefly, after fixing cells with 4% paraformaldehyde for 30 min, and incubation in 0.3% Triton X-100 in PBS for 20 min at room temperature, the cells were blocked by 10% fetal bovine serum in PBS for 60 min at room temperature, and then incubated for 60 min at room temperature or overnight at 4°C with primary antibodies: polyclonal Kv4.2 antibody from Alomone Labs (Jerusalem, Israel) generated against residues 454-469 ( SNQLQSSEDEPAFVSK) on the C-terminal or a monoclonal Kv4.2 antibody against an ectodomain [ ].

    Techniques: In Vitro

    Generation and characterization of the Kv4.2 phospho-specific antibodies for the Ser447 and Ser537 site

    Journal: The Biochemical journal

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    doi: 10.1042/BJ20081213

    Figure Lengend Snippet: Generation and characterization of the Kv4.2 phospho-specific antibodies for the Ser447 and Ser537 site

    Article Snippet: Briefly, after fixing cells with 4% paraformaldehyde for 30 min, and incubation in 0.3% Triton X-100 in PBS for 20 min at room temperature, the cells were blocked by 10% fetal bovine serum in PBS for 60 min at room temperature, and then incubated for 60 min at room temperature or overnight at 4°C with primary antibodies: polyclonal Kv4.2 antibody from Alomone Labs (Jerusalem, Israel) generated against residues 454-469 ( SNQLQSSEDEPAFVSK) on the C-terminal or a monoclonal Kv4.2 antibody against an ectodomain [ ].

    Techniques:

    PKC phosphorylation of the Kv4.2 C-terminal augments ERK phosphorylation of the Kv4.2 C-terminal in vitro

    Journal: The Biochemical journal

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    doi: 10.1042/BJ20081213

    Figure Lengend Snippet: PKC phosphorylation of the Kv4.2 C-terminal augments ERK phosphorylation of the Kv4.2 C-terminal in vitro

    Article Snippet: Briefly, after fixing cells with 4% paraformaldehyde for 30 min, and incubation in 0.3% Triton X-100 in PBS for 20 min at room temperature, the cells were blocked by 10% fetal bovine serum in PBS for 60 min at room temperature, and then incubated for 60 min at room temperature or overnight at 4°C with primary antibodies: polyclonal Kv4.2 antibody from Alomone Labs (Jerusalem, Israel) generated against residues 454-469 ( SNQLQSSEDEPAFVSK) on the C-terminal or a monoclonal Kv4.2 antibody against an ectodomain [ ].

    Techniques: In Vitro

    Functional characterization of PKC sites within Kv4.2

    Journal: The Biochemical journal

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    doi: 10.1042/BJ20081213

    Figure Lengend Snippet: Functional characterization of PKC sites within Kv4.2

    Article Snippet: Briefly, after fixing cells with 4% paraformaldehyde for 30 min, and incubation in 0.3% Triton X-100 in PBS for 20 min at room temperature, the cells were blocked by 10% fetal bovine serum in PBS for 60 min at room temperature, and then incubated for 60 min at room temperature or overnight at 4°C with primary antibodies: polyclonal Kv4.2 antibody from Alomone Labs (Jerusalem, Israel) generated against residues 454-469 ( SNQLQSSEDEPAFVSK) on the C-terminal or a monoclonal Kv4.2 antibody against an ectodomain [ ].

    Techniques: Functional Assay

    Candidate PKC consensus sites within the Kv4.2 channel subunit

    Journal: The Biochemical journal

    Article Title: Kv4.2 is a locus for PKC and ERK/MAPK cross-talk

    doi: 10.1042/BJ20081213

    Figure Lengend Snippet: Candidate PKC consensus sites within the Kv4.2 channel subunit

    Article Snippet: Briefly, after fixing cells with 4% paraformaldehyde for 30 min, and incubation in 0.3% Triton X-100 in PBS for 20 min at room temperature, the cells were blocked by 10% fetal bovine serum in PBS for 60 min at room temperature, and then incubated for 60 min at room temperature or overnight at 4°C with primary antibodies: polyclonal Kv4.2 antibody from Alomone Labs (Jerusalem, Israel) generated against residues 454-469 ( SNQLQSSEDEPAFVSK) on the C-terminal or a monoclonal Kv4.2 antibody against an ectodomain [ ].

    Techniques:

    I to recovery from inactivation. A) The recovery kinetics was tested by a double-pulse protocol with interpulse time varying from 50 ms to 15 sec (n=12 RV and 7 LV cells from n=3 hearts). B) The amplitudes of the slow and fast inactivating components of I to (I to,si and I I to,fi ) as a function of inter-pulse interval were determined by fitting the time course of I to decay during the second pulse to a double exponential function. The x-axis of inter-pulse intervals is in a logarithmic scale. C) The amplitudes of I to,fi and I to,si from RV and LV. Fast and slow-inactivating components (I to,fi and I to,si ) of each I to,f and I to,s were calculated as described in Methods and represented as a stacked column plot. D) Western blots of Kv4.2, Kv1.4, and KChIP2 from LQT1 hearts. E). The accessory unit of I to , KChIP2, known to affect inactivation and recovery kinetics, was twofold higher in RV (ANOVA, p .

    Journal: Circulation. Arrhythmia and electrophysiology

    Article Title: Transient Outward K+ Current (Ito) Underlies the Right Ventricular Initiation of Polymorphic Ventricular Tachycardia in a Transgenic Rabbit Model of Long QT Type 1

    doi: 10.1161/CIRCEP.117.005414

    Figure Lengend Snippet: I to recovery from inactivation. A) The recovery kinetics was tested by a double-pulse protocol with interpulse time varying from 50 ms to 15 sec (n=12 RV and 7 LV cells from n=3 hearts). B) The amplitudes of the slow and fast inactivating components of I to (I to,si and I I to,fi ) as a function of inter-pulse interval were determined by fitting the time course of I to decay during the second pulse to a double exponential function. The x-axis of inter-pulse intervals is in a logarithmic scale. C) The amplitudes of I to,fi and I to,si from RV and LV. Fast and slow-inactivating components (I to,fi and I to,si ) of each I to,f and I to,s were calculated as described in Methods and represented as a stacked column plot. D) Western blots of Kv4.2, Kv1.4, and KChIP2 from LQT1 hearts. E). The accessory unit of I to , KChIP2, known to affect inactivation and recovery kinetics, was twofold higher in RV (ANOVA, p .

    Article Snippet: The antibodies for Kv1.4, Kv4.2, and KChIP2 were purchased from Alomone Labs (Jerusalem, Israel).

    Techniques: Mass Spectrometry, Size-exclusion Chromatography, Western Blot