ip3r1  (Alomone Labs)


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

    Structured Review

    Alomone Labs ip3r1
    GFP expression in the brains of adult Tg( eno2:GFP ) zebrafish. ( A ) Parasagittal sections of Tg( eno2:GFP ) Pt404 (upper panel) and wild-type (lower panel) brain were labelled with an antibody to GFP. Bound antibody was detected using a histochemical reaction with a red product and nuclei were labelled with a blue counterstain. GFP expression was apparent throughout the brain of transgenic adults. ( B ) High-power view of medullary reticular formation of Tg( eno2:GFP ) Pt404 zebrafish brain, showing GFP expression in cells with neuronal morphology. The arrows show GFP-expressing neurons with axons in the plane of the section. ( C – F ) Double label confocal images of Tg( eno2:GFP ) Pt404 adult brains. Each panel consists of a set of three images: (i) Upper image; red: a cell type-specific marker of interest was localized using specific antibodies. C: Medulla; ChAT, choline acetyltransferase; cholinergic neurons; D: Olfactory bulb; TH, tyrosine hydroxylase; dopaminergic neurons; E: cerebellum; <t>IP3R1,</t> IP3 receptor 1; cerebellar Purkinje cells; D: thalamus; GABA, γ-amino butyric acid; inhibitory neurons; (ii) Middle image, green: GFP expression was localized in the same sections using a GFP antibody; (iii) Lower image: the merged images show co-localization of GFP and the cell type-specific marker (yellow), and were counterstained with DAPI to show nuclei (blue). The scale bar (10 µm) for each set of images is shown in the lower panel.
    Ip3r1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ip3r1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ip3r1 - by Bioz Stars, 2022-05
    93/100 stars

    Images

    1) Product Images from "Generation of a transgenic zebrafish model of Tauopathy using a novel promoter element derived from the zebrafish eno2 gene"

    Article Title: Generation of a transgenic zebrafish model of Tauopathy using a novel promoter element derived from the zebrafish eno2 gene

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkm608

    GFP expression in the brains of adult Tg( eno2:GFP ) zebrafish. ( A ) Parasagittal sections of Tg( eno2:GFP ) Pt404 (upper panel) and wild-type (lower panel) brain were labelled with an antibody to GFP. Bound antibody was detected using a histochemical reaction with a red product and nuclei were labelled with a blue counterstain. GFP expression was apparent throughout the brain of transgenic adults. ( B ) High-power view of medullary reticular formation of Tg( eno2:GFP ) Pt404 zebrafish brain, showing GFP expression in cells with neuronal morphology. The arrows show GFP-expressing neurons with axons in the plane of the section. ( C – F ) Double label confocal images of Tg( eno2:GFP ) Pt404 adult brains. Each panel consists of a set of three images: (i) Upper image; red: a cell type-specific marker of interest was localized using specific antibodies. C: Medulla; ChAT, choline acetyltransferase; cholinergic neurons; D: Olfactory bulb; TH, tyrosine hydroxylase; dopaminergic neurons; E: cerebellum; IP3R1, IP3 receptor 1; cerebellar Purkinje cells; D: thalamus; GABA, γ-amino butyric acid; inhibitory neurons; (ii) Middle image, green: GFP expression was localized in the same sections using a GFP antibody; (iii) Lower image: the merged images show co-localization of GFP and the cell type-specific marker (yellow), and were counterstained with DAPI to show nuclei (blue). The scale bar (10 µm) for each set of images is shown in the lower panel.
    Figure Legend Snippet: GFP expression in the brains of adult Tg( eno2:GFP ) zebrafish. ( A ) Parasagittal sections of Tg( eno2:GFP ) Pt404 (upper panel) and wild-type (lower panel) brain were labelled with an antibody to GFP. Bound antibody was detected using a histochemical reaction with a red product and nuclei were labelled with a blue counterstain. GFP expression was apparent throughout the brain of transgenic adults. ( B ) High-power view of medullary reticular formation of Tg( eno2:GFP ) Pt404 zebrafish brain, showing GFP expression in cells with neuronal morphology. The arrows show GFP-expressing neurons with axons in the plane of the section. ( C – F ) Double label confocal images of Tg( eno2:GFP ) Pt404 adult brains. Each panel consists of a set of three images: (i) Upper image; red: a cell type-specific marker of interest was localized using specific antibodies. C: Medulla; ChAT, choline acetyltransferase; cholinergic neurons; D: Olfactory bulb; TH, tyrosine hydroxylase; dopaminergic neurons; E: cerebellum; IP3R1, IP3 receptor 1; cerebellar Purkinje cells; D: thalamus; GABA, γ-amino butyric acid; inhibitory neurons; (ii) Middle image, green: GFP expression was localized in the same sections using a GFP antibody; (iii) Lower image: the merged images show co-localization of GFP and the cell type-specific marker (yellow), and were counterstained with DAPI to show nuclei (blue). The scale bar (10 µm) for each set of images is shown in the lower panel.

    Techniques Used: Expressing, Transgenic Assay, Marker

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Alomone Labs goat anti kv3 4 antibody
    Quantification of <t>Kv3.4</t> plasma membrane immunostaining from 2 weeks after laminectomy and SCI. A , Representative immunofluorescence sections stained with anti-Kv3.4 and pan-cadherin antibodies from laminectomy ( N = 3) and SCI ( N = 3) animals. B , Histogram
    Goat Anti Kv3 4 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti kv3 4 antibody/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti kv3 4 antibody - by Bioz Stars, 2022-05
    94/100 stars
      Buy from Supplier

    91
    Alomone Labs rabbit anti trkb
    Effect of 8-OH-DPAT on the protein expression of molecules from the <t>BDNF</t> signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) <t>TrkB.</t> (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p
    Rabbit Anti Trkb, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti trkb/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti trkb - by Bioz Stars, 2022-05
    91/100 stars
      Buy from Supplier

    Image Search Results


    Quantification of Kv3.4 plasma membrane immunostaining from 2 weeks after laminectomy and SCI. A , Representative immunofluorescence sections stained with anti-Kv3.4 and pan-cadherin antibodies from laminectomy ( N = 3) and SCI ( N = 3) animals. B , Histogram

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Quantification of Kv3.4 plasma membrane immunostaining from 2 weeks after laminectomy and SCI. A , Representative immunofluorescence sections stained with anti-Kv3.4 and pan-cadherin antibodies from laminectomy ( N = 3) and SCI ( N = 3) animals. B , Histogram

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Immunostaining, Immunofluorescence, Staining

    Kv3.4 current expression and inactivation from naive, laminectomy, and SCI. A , Scatter plot of I p for all data on a logarithmic scale. Dashed gray line is 1 SD from the center of the naive distribution (16.1 pA). B , I p / I 500 ratio for all groups examined.

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Kv3.4 current expression and inactivation from naive, laminectomy, and SCI. A , Scatter plot of I p for all data on a logarithmic scale. Dashed gray line is 1 SD from the center of the naive distribution (16.1 pA). B , I p / I 500 ratio for all groups examined.

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Expressing

    Kv3.4 current phenotypes after laminectomy and SCI. A , B , Representative Kv3.4 currents from laminectomy (Lam) and SCI neurons at 1 week and 2 weeks after surgery. Pulse protocol and scale bars are as in A . C , Pie charts showing the percentages

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Kv3.4 current phenotypes after laminectomy and SCI. A , B , Representative Kv3.4 currents from laminectomy (Lam) and SCI neurons at 1 week and 2 weeks after surgery. Pulse protocol and scale bars are as in A . C , Pie charts showing the percentages

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Laser Capture Microdissection

    Kv3.4 channel expression and current phenotypes in DRG neurons. A , DRG section from a naive animal immunostained as indicated. B , Distribution of peak currents from naive neurons and best fit Gaussian distribution (solid line; inset: x c = 33.8 pA, w =

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Kv3.4 channel expression and current phenotypes in DRG neurons. A , DRG section from a naive animal immunostained as indicated. B , Distribution of peak currents from naive neurons and best fit Gaussian distribution (solid line; inset: x c = 33.8 pA, w =

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Expressing

    Dynamic clamping of a spiking DRG neuron in the SCI group. A , Cell-attached Kv3.4 currents evoked by 150 pulses to +100 mV (black traces) and the mean current (red trace). Pulse protocol details in legend. Dashed line represents 0 pA. B , Variance

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Dynamic clamping of a spiking DRG neuron in the SCI group. A , Cell-attached Kv3.4 currents evoked by 150 pulses to +100 mV (black traces) and the mean current (red trace). Pulse protocol details in legend. Dashed line represents 0 pA. B , Variance

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques:

    Early changes: laminectomy and SCI alter the inactivation properties and expression of Kv3.4 channels in DRG neurons

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Early changes: laminectomy and SCI alter the inactivation properties and expression of Kv3.4 channels in DRG neurons

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Expressing

    Expression of Kv3.4 mRNA and protein from naive and 2 weeks after laminectomy and SCI. A–C , Box plots of single-cell mRNA copies of Kv3.4 mRNA (black = naive; blue = laminectomy; red = SCI). Box is 25–75 percentiles with median marked

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Expression of Kv3.4 mRNA and protein from naive and 2 weeks after laminectomy and SCI. A–C , Box plots of single-cell mRNA copies of Kv3.4 mRNA (black = naive; blue = laminectomy; red = SCI). Box is 25–75 percentiles with median marked

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Expressing

    Kv3.4 DRG immunostaining from naive and 2 weeks laminectomy and SCI. A – C , Representative immunofluorescence sections stained with anti-Kv3.4 antibody from naive, laminectomy and SCI animals. White arrows show neurons with significant staining

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Kv3.4 DRG immunostaining from naive and 2 weeks laminectomy and SCI. A – C , Representative immunofluorescence sections stained with anti-Kv3.4 antibody from naive, laminectomy and SCI animals. White arrows show neurons with significant staining

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques: Immunostaining, Immunofluorescence, Staining

    Kv3.4 channel biophysical properties from naive, laminectomy and SCI. A , B , Normalized G p - V relation and steady-state inactivation relation for naive (black) and 1 week after laminectomy (blue) and SCI (red). displays mean ± SEM of best-fit

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Kv3.4 channel biophysical properties from naive, laminectomy and SCI. A , B , Normalized G p - V relation and steady-state inactivation relation for naive (black) and 1 week after laminectomy (blue) and SCI (red). displays mean ± SEM of best-fit

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques:

    Kv3.4 currents in small-diameter DRG neurons undergo dynamic changes after laminectomy and SCI

    Journal: The Journal of Neuroscience

    Article Title: Dysregulation of Kv3.4 Channels in Dorsal Root Ganglia Following Spinal Cord Injury

    doi: 10.1523/JNEUROSCI.1594-14.2015

    Figure Lengend Snippet: Kv3.4 currents in small-diameter DRG neurons undergo dynamic changes after laminectomy and SCI

    Article Snippet: Sections were then incubated with primary goat anti-Kv3.4 antibody (1:1000, Alomone Labs Catalog #APC-019 RRID:AB_2040172) overnight at 4°C.

    Techniques:

    Representative immunostaining images of K v 3.4 in neurons expressing either tdTomato or eYFP (A B). For comparison purposes, tdTomato and eYFP cell bodies are visualised in grey pseudocolour and Kv3.4 staining in magenta. Compared to the tdTomato-filled neuron, the eYFP-filled neuron shows lower K v 3.4 immunoreactivity. Analysis of K v 3.4 immunostaining intensity revealed a 21% reduction in staining intensity in eYFP-filled cells relative to tdTomato-filled cells (C), confirming the action of U6.sgRNA(mK v 3.4).CMV.saCas9 at knocking down K v 3.4 levels. * p

    Journal: bioRxiv

    Article Title: Reducing voltage-dependent potassium channel Kv3.4 levels ameliorates synapse loss in a mouse model of Alzheimer’s disease

    doi: 10.1101/2021.11.24.469829

    Figure Lengend Snippet: Representative immunostaining images of K v 3.4 in neurons expressing either tdTomato or eYFP (A B). For comparison purposes, tdTomato and eYFP cell bodies are visualised in grey pseudocolour and Kv3.4 staining in magenta. Compared to the tdTomato-filled neuron, the eYFP-filled neuron shows lower K v 3.4 immunoreactivity. Analysis of K v 3.4 immunostaining intensity revealed a 21% reduction in staining intensity in eYFP-filled cells relative to tdTomato-filled cells (C), confirming the action of U6.sgRNA(mK v 3.4).CMV.saCas9 at knocking down K v 3.4 levels. * p

    Article Snippet: Membranes were incubated with primary antibody (anti-KCNC4; Alomone #APC-019; 1:200) overnight at 4 °C in blocking buffer.

    Techniques: Immunostaining, Expressing, Staining

    A schematic summary of Kv3.4 function in the growth cone. A , In normal growing axons, the growth cone membrane is depolarized by spontaneous electrical activity (1) or after the binding of an attractive guidance cue (such as netrin-1) to its receptor (such as DCC) (2). Membrane depolarization allows Ca 2+ influx through T-type and L-type Cav channels sequentially. Then, Kv3.4 channels are activated and Kv3.4-mediated A-type K + outward currents reduce membrane excitability. B , After Kv3.4 knockdown by Kv3.4shRNA or Kv3.4 blockade by BDSII, excessive extracellular Ca 2+ ions enter the growth cone, which leads to axon growth inhibition. BDSII-induced Ca 2+ influx does not require the release of intracellular Ca 2+ from the ER (endoplasmic reticulum). C , The membrane potential of growth cones can be depolarized by spontaneous electrical activity or by the binding of attractive guidance cues. Slight membrane depolarization induces sustained Ca 2+ elevation, and the opening of Kv3.4 channels quickly reduces membrane excitability, which can inhibit the generation of action potentials. Substantial membrane depolarization evokes Ca 2+ -dependent action potentials to generate Ca 2+ transients, and the activation of Kv3.4 channels repolarizes the membrane to reduce the amplitudes of action potentials, resulting in Ca 2+ transients with smaller amplitudes. Thus, by controlling growth cone membrane excitability, Kv3.4 acts to maintain [Ca 2+ ] i at an optimal concentration for normal axon growth. AP, Action potential; Cav, voltage-gated calcium channel; DCC, deleted in colorectal cancer; IP 3 R, inositol 1,4,5-triphosphate receptor.

    Journal: The Journal of Neuroscience

    Article Title: K+ Channel Kv3.4 Is Essential for Axon Growth by Limiting the Influx of Ca2+ into Growth Cones

    doi: 10.1523/JNEUROSCI.1076-16.2017

    Figure Lengend Snippet: A schematic summary of Kv3.4 function in the growth cone. A , In normal growing axons, the growth cone membrane is depolarized by spontaneous electrical activity (1) or after the binding of an attractive guidance cue (such as netrin-1) to its receptor (such as DCC) (2). Membrane depolarization allows Ca 2+ influx through T-type and L-type Cav channels sequentially. Then, Kv3.4 channels are activated and Kv3.4-mediated A-type K + outward currents reduce membrane excitability. B , After Kv3.4 knockdown by Kv3.4shRNA or Kv3.4 blockade by BDSII, excessive extracellular Ca 2+ ions enter the growth cone, which leads to axon growth inhibition. BDSII-induced Ca 2+ influx does not require the release of intracellular Ca 2+ from the ER (endoplasmic reticulum). C , The membrane potential of growth cones can be depolarized by spontaneous electrical activity or by the binding of attractive guidance cues. Slight membrane depolarization induces sustained Ca 2+ elevation, and the opening of Kv3.4 channels quickly reduces membrane excitability, which can inhibit the generation of action potentials. Substantial membrane depolarization evokes Ca 2+ -dependent action potentials to generate Ca 2+ transients, and the activation of Kv3.4 channels repolarizes the membrane to reduce the amplitudes of action potentials, resulting in Ca 2+ transients with smaller amplitudes. Thus, by controlling growth cone membrane excitability, Kv3.4 acts to maintain [Ca 2+ ] i at an optimal concentration for normal axon growth. AP, Action potential; Cav, voltage-gated calcium channel; DCC, deleted in colorectal cancer; IP 3 R, inositol 1,4,5-triphosphate receptor.

    Article Snippet: The specificity of anti-Kv3.2 (Alomone Labs catalog #APC-011, RRID:AB_2040168), anti-Kv3.3 (Alomone Labs catalog #APC-102, RRID:AB_2040170), and anti-Kv3.4 (Alomone Labs catalog #APC-019, RRID:AB_ 2040172) has been described previously ( ).

    Techniques: Activity Assay, Binding Assay, Droplet Countercurrent Chromatography, Inhibition, Activation Assay, Concentration Assay

    Knockdown of Kv3.4 inhibits axon elongation, pathfinding, and fasciculation in vivo . A–E , The right side spinal cord of chick embryo at HH15-HH17 was electroporated with constructs encoding EYFP (control, B ), EYFP/LacZshRNA ( C ), EYFP/Kv3.4shRNA ( D ), or EYFP/ Kv3.4shRNA/Kv3.4shRNA-resistant Kv3.4 (resKv3.4) ( E ). Embryos were fixed at HH22-HH23, and their spinal cords in open-book configurations show the trajectories of EYFP + commissural axons. A, Anterior; D, dorsal; P, posterior; V, ventral. B–E , Arrows indicate the bundle of commissural axons (ventral funiculus, VF). D , Arrowheads indicate stalling axons at the floor plate (FP). Asterisks indicate misguided axons. F , Summary of projection errors of spinal commissural axons. G , The percentage of EYFP + axons with projection errors. H , The width of the ventral funiculus. I , Western blotting was performed using lysate of HEK-293 cells transfected with constructs encoding Kv3.4/LacZshRNA/EYFP, Kv3.4/Kv3.4shRNA/EYFP, or Kv3.4shRNA/resKv3.4/EYFP. The major protein band of Kv3.4 at position of 100 kDa was shown, and GAPDH was as used as a loading control. J–M , In E15.5 rat brain, the ventricular zone (green) adjacent to the lateral ventricle (LV, blue) was electroporated with constructs encoding EYFP/LacZshRNA ( K ), EYFP/Kv3.4shRNA ( L ), or EYFP/Kv3.4shRNA/resKv3.4 ( M ). The positive electrode paddle was located on the left side of brain. Coronal sections of E20.5 rat brain were analyzed after embryos were grown in utero . EYFP + callosal axons, which project from the cingulate cortex (CgC) and frontal cortex (FC), only reach the contralateral cingulate cortex in the Kv3.4shRNA-expressing brain. PC, Parietal cortex. N , Measurement of axon projection to the contralateral side. Relative intensity in each region ( J , −2, −1, 0, 1, 2) is obtained by normalizing its fluorescence intensity with that in region 2. G , H , N , Numbers in parentheses indicate the total number of embryos analyzed. Data are mean ± SEM. * p

    Journal: The Journal of Neuroscience

    Article Title: K+ Channel Kv3.4 Is Essential for Axon Growth by Limiting the Influx of Ca2+ into Growth Cones

    doi: 10.1523/JNEUROSCI.1076-16.2017

    Figure Lengend Snippet: Knockdown of Kv3.4 inhibits axon elongation, pathfinding, and fasciculation in vivo . A–E , The right side spinal cord of chick embryo at HH15-HH17 was electroporated with constructs encoding EYFP (control, B ), EYFP/LacZshRNA ( C ), EYFP/Kv3.4shRNA ( D ), or EYFP/ Kv3.4shRNA/Kv3.4shRNA-resistant Kv3.4 (resKv3.4) ( E ). Embryos were fixed at HH22-HH23, and their spinal cords in open-book configurations show the trajectories of EYFP + commissural axons. A, Anterior; D, dorsal; P, posterior; V, ventral. B–E , Arrows indicate the bundle of commissural axons (ventral funiculus, VF). D , Arrowheads indicate stalling axons at the floor plate (FP). Asterisks indicate misguided axons. F , Summary of projection errors of spinal commissural axons. G , The percentage of EYFP + axons with projection errors. H , The width of the ventral funiculus. I , Western blotting was performed using lysate of HEK-293 cells transfected with constructs encoding Kv3.4/LacZshRNA/EYFP, Kv3.4/Kv3.4shRNA/EYFP, or Kv3.4shRNA/resKv3.4/EYFP. The major protein band of Kv3.4 at position of 100 kDa was shown, and GAPDH was as used as a loading control. J–M , In E15.5 rat brain, the ventricular zone (green) adjacent to the lateral ventricle (LV, blue) was electroporated with constructs encoding EYFP/LacZshRNA ( K ), EYFP/Kv3.4shRNA ( L ), or EYFP/Kv3.4shRNA/resKv3.4 ( M ). The positive electrode paddle was located on the left side of brain. Coronal sections of E20.5 rat brain were analyzed after embryos were grown in utero . EYFP + callosal axons, which project from the cingulate cortex (CgC) and frontal cortex (FC), only reach the contralateral cingulate cortex in the Kv3.4shRNA-expressing brain. PC, Parietal cortex. N , Measurement of axon projection to the contralateral side. Relative intensity in each region ( J , −2, −1, 0, 1, 2) is obtained by normalizing its fluorescence intensity with that in region 2. G , H , N , Numbers in parentheses indicate the total number of embryos analyzed. Data are mean ± SEM. * p

    Article Snippet: The specificity of anti-Kv3.2 (Alomone Labs catalog #APC-011, RRID:AB_2040168), anti-Kv3.3 (Alomone Labs catalog #APC-102, RRID:AB_2040170), and anti-Kv3.4 (Alomone Labs catalog #APC-019, RRID:AB_ 2040172) has been described previously ( ).

    Techniques: In Vivo, Construct, Western Blot, Transfection, In Utero, Expressing, Fluorescence

    Knockdown of Kv3.4 inhibits neurite protrusion and axon elongation. A–D , The spinal cord of chick embryos at HH15-HH17 was electroporated with constructs encoding EYFP alone (control, A ), LacZshRNA/EYFP (LacZshRNA, B ), Kv3.4shRNA/EYFP (Kv3.4shRNA, C ), or Kv3.4shRNA/resKv3.4[Kv3.4shRNA-resistant Kv3.4]/EYFP (Kv3.4shRNA + resKv3.4, D ). The dorsal spinal cord was dissociated at HH21-HH23 and cultured for 20 h before immunolabeling Kv3.4 (red). A–D , A′–D′ , Top, Neurons without neurites. Bottom, Axon-bearing neurons. Nontransfected neurons in each culture were used for comparison, and their nuclei were labeled by DAPI (blue). Scale bars: Top, 17 μm; Bottom, 20 μm. A , B , In the control or LacZshRNA + neurons, Kv3.4-IR was strong in the somatic surfaces of neurons without neurites, but it became more evident in the growth cones of axon-bearing neurons. LacZshRNA did not suppress Kv3.4 expression. C , Kv3.4shRNA strongly reduced Kv3.4-IR in neurons without neurites (arrowhead) but had a weaker effect in axon-bearing neurons. D , Cotransfection of resKv3.4 rescued the knockdown effect caused by Kv3.4shRNA. E , After measuring the fluorescence intensity of neurons with or without neurites (30 neuron counts for each; total 60 counts), the relative Kv3.4 protein level was obtained by dividing the Kv3.4 fluorescent intensity of EYFP + neurons by that of EYFP − neurons. F , The percentage of protrusion-bearing neurons. G , The percentage of axon-bearing neurons. H , The average of axon length. I , The cumulative distribution of axon length (Kolmogorov–Smirnov test) shows that the axons of Kv3.4shRNA-transfected neurons are shorter. Numbers in parentheses indicate total EYFP + neuron counts pooled from three independent experiments. Data are mean ± SEM. * p

    Journal: The Journal of Neuroscience

    Article Title: K+ Channel Kv3.4 Is Essential for Axon Growth by Limiting the Influx of Ca2+ into Growth Cones

    doi: 10.1523/JNEUROSCI.1076-16.2017

    Figure Lengend Snippet: Knockdown of Kv3.4 inhibits neurite protrusion and axon elongation. A–D , The spinal cord of chick embryos at HH15-HH17 was electroporated with constructs encoding EYFP alone (control, A ), LacZshRNA/EYFP (LacZshRNA, B ), Kv3.4shRNA/EYFP (Kv3.4shRNA, C ), or Kv3.4shRNA/resKv3.4[Kv3.4shRNA-resistant Kv3.4]/EYFP (Kv3.4shRNA + resKv3.4, D ). The dorsal spinal cord was dissociated at HH21-HH23 and cultured for 20 h before immunolabeling Kv3.4 (red). A–D , A′–D′ , Top, Neurons without neurites. Bottom, Axon-bearing neurons. Nontransfected neurons in each culture were used for comparison, and their nuclei were labeled by DAPI (blue). Scale bars: Top, 17 μm; Bottom, 20 μm. A , B , In the control or LacZshRNA + neurons, Kv3.4-IR was strong in the somatic surfaces of neurons without neurites, but it became more evident in the growth cones of axon-bearing neurons. LacZshRNA did not suppress Kv3.4 expression. C , Kv3.4shRNA strongly reduced Kv3.4-IR in neurons without neurites (arrowhead) but had a weaker effect in axon-bearing neurons. D , Cotransfection of resKv3.4 rescued the knockdown effect caused by Kv3.4shRNA. E , After measuring the fluorescence intensity of neurons with or without neurites (30 neuron counts for each; total 60 counts), the relative Kv3.4 protein level was obtained by dividing the Kv3.4 fluorescent intensity of EYFP + neurons by that of EYFP − neurons. F , The percentage of protrusion-bearing neurons. G , The percentage of axon-bearing neurons. H , The average of axon length. I , The cumulative distribution of axon length (Kolmogorov–Smirnov test) shows that the axons of Kv3.4shRNA-transfected neurons are shorter. Numbers in parentheses indicate total EYFP + neuron counts pooled from three independent experiments. Data are mean ± SEM. * p

    Article Snippet: The specificity of anti-Kv3.2 (Alomone Labs catalog #APC-011, RRID:AB_2040168), anti-Kv3.3 (Alomone Labs catalog #APC-102, RRID:AB_2040170), and anti-Kv3.4 (Alomone Labs catalog #APC-019, RRID:AB_ 2040172) has been described previously ( ).

    Techniques: Construct, Cell Culture, Immunolabeling, Labeling, Expressing, Cotransfection, Fluorescence, Transfection

    Kv3.4 in the axonal growth cones of motoneurons, DRG neurons, RGCs, and callosal projection neurons. A , B , Transverse sections of the spinal cord (SC) of HH23 chick embryos were immunostained, showing Kv3.4-IR in the axonal bundle (arrowhead) of motoneurons (MN) ( A ) and the bifurcation zone (BZ) of DRG neuron afferents ( B ). C , D , Motoneurons and DRG neurons of HH21-HH23 chick embryos were dissociated, cultured for 20 h, and double immunostained for Kv3.4 and Islet1/2. E , F , RGCs and callosal projection neurons (CPNs) were dissociated from the retina and cingulate/frontal cortices of E18.5 rat embryo, respectively. After 16 h of culture, cells were double immunostained for Kv3.4 and Islet1/2 ( E′ ) or TAG-1 ( F′ ). Kv3.4-IR is evident in axonal growth cones ( C″–F″ , arrows). Scale bars: A , 38 μm; B , 25 μm; C , 16 μm; D , 13 μm; E , F , 16 μm.

    Journal: The Journal of Neuroscience

    Article Title: K+ Channel Kv3.4 Is Essential for Axon Growth by Limiting the Influx of Ca2+ into Growth Cones

    doi: 10.1523/JNEUROSCI.1076-16.2017

    Figure Lengend Snippet: Kv3.4 in the axonal growth cones of motoneurons, DRG neurons, RGCs, and callosal projection neurons. A , B , Transverse sections of the spinal cord (SC) of HH23 chick embryos were immunostained, showing Kv3.4-IR in the axonal bundle (arrowhead) of motoneurons (MN) ( A ) and the bifurcation zone (BZ) of DRG neuron afferents ( B ). C , D , Motoneurons and DRG neurons of HH21-HH23 chick embryos were dissociated, cultured for 20 h, and double immunostained for Kv3.4 and Islet1/2. E , F , RGCs and callosal projection neurons (CPNs) were dissociated from the retina and cingulate/frontal cortices of E18.5 rat embryo, respectively. After 16 h of culture, cells were double immunostained for Kv3.4 and Islet1/2 ( E′ ) or TAG-1 ( F′ ). Kv3.4-IR is evident in axonal growth cones ( C″–F″ , arrows). Scale bars: A , 38 μm; B , 25 μm; C , 16 μm; D , 13 μm; E , F , 16 μm.

    Article Snippet: The specificity of anti-Kv3.2 (Alomone Labs catalog #APC-011, RRID:AB_2040168), anti-Kv3.3 (Alomone Labs catalog #APC-102, RRID:AB_2040170), and anti-Kv3.4 (Alomone Labs catalog #APC-019, RRID:AB_ 2040172) has been described previously ( ).

    Techniques: Cell Culture

    Kv3.4 in the axonal growth cones of dorsal spinal commissural neurons. A–F , Transverse sections of the spinal cord of chick embryos were immunostained for Kv3.4. A , Absence of Kv3.4-IR in the dorsal spinal cord at HH17. Kv3.4-IR in precrossing commissural axons ( B–F , arrowheads) is evident during HH19-HH25 but disappears at HH27. D , Arrows indicate postcrossing commissural axons projecting from the other side of spinal cord. FP, Floor plate. G–L , Transverse sections of the spinal cord at HH23 were immunostained as indicated. G , Absence of Kv1.5-IR. H , Kv4.2-IR in the somata and dendrites of motoneurons (MN). I , Kv4.3-IR in the bifurcation zone (BZ). In addition to the BZ, Kv3.1b-IR is strong in postcrossing commissural axons ( J , arrow) but weak in precrossing commissural axons ( J , arrowhead). K , Absence of Kv3.2-IR. L , Kv3.3 in motoneurons. M–M″ , Double staining in transverse sections of the spinal cord at HH21 shows colocalization of Kv3.4 and axonin-1 in the growth cones (arrowheads) of commissural axons. N–N″ , Colocalization of Kv3.4 and axonin-1 in cultured dorsal spinal neurons isolated from HH21-HH23 chick embryos. O–P″ , Red fluorescence-tagged phalloidin colabeling reveals enrichment of Kv3.4 in the growth cone ( O–O″ ) and Kv3.1b in the soma/axon shaft ( P–P″ ) of cultured dorsal spinal neurons. Q–Q″ , Kv3.4 and DiI colabeling. White represents Kv3.4-abundant regions. Blue represents Kv3.4-sparse regions ( Q″ ). R , Ratio of Kv3.4/DiI in the soma, axon shaft, or growth cone of each neuron was obtained by dividing the fluorescence intensity of Kv3.4 by that of DiI. Data are mean ± SEM ( n = 8 neurons, pooled from three independent experiments done on different days). *** p

    Journal: The Journal of Neuroscience

    Article Title: K+ Channel Kv3.4 Is Essential for Axon Growth by Limiting the Influx of Ca2+ into Growth Cones

    doi: 10.1523/JNEUROSCI.1076-16.2017

    Figure Lengend Snippet: Kv3.4 in the axonal growth cones of dorsal spinal commissural neurons. A–F , Transverse sections of the spinal cord of chick embryos were immunostained for Kv3.4. A , Absence of Kv3.4-IR in the dorsal spinal cord at HH17. Kv3.4-IR in precrossing commissural axons ( B–F , arrowheads) is evident during HH19-HH25 but disappears at HH27. D , Arrows indicate postcrossing commissural axons projecting from the other side of spinal cord. FP, Floor plate. G–L , Transverse sections of the spinal cord at HH23 were immunostained as indicated. G , Absence of Kv1.5-IR. H , Kv4.2-IR in the somata and dendrites of motoneurons (MN). I , Kv4.3-IR in the bifurcation zone (BZ). In addition to the BZ, Kv3.1b-IR is strong in postcrossing commissural axons ( J , arrow) but weak in precrossing commissural axons ( J , arrowhead). K , Absence of Kv3.2-IR. L , Kv3.3 in motoneurons. M–M″ , Double staining in transverse sections of the spinal cord at HH21 shows colocalization of Kv3.4 and axonin-1 in the growth cones (arrowheads) of commissural axons. N–N″ , Colocalization of Kv3.4 and axonin-1 in cultured dorsal spinal neurons isolated from HH21-HH23 chick embryos. O–P″ , Red fluorescence-tagged phalloidin colabeling reveals enrichment of Kv3.4 in the growth cone ( O–O″ ) and Kv3.1b in the soma/axon shaft ( P–P″ ) of cultured dorsal spinal neurons. Q–Q″ , Kv3.4 and DiI colabeling. White represents Kv3.4-abundant regions. Blue represents Kv3.4-sparse regions ( Q″ ). R , Ratio of Kv3.4/DiI in the soma, axon shaft, or growth cone of each neuron was obtained by dividing the fluorescence intensity of Kv3.4 by that of DiI. Data are mean ± SEM ( n = 8 neurons, pooled from three independent experiments done on different days). *** p

    Article Snippet: The specificity of anti-Kv3.2 (Alomone Labs catalog #APC-011, RRID:AB_2040168), anti-Kv3.3 (Alomone Labs catalog #APC-102, RRID:AB_2040170), and anti-Kv3.4 (Alomone Labs catalog #APC-019, RRID:AB_ 2040172) has been described previously ( ).

    Techniques: Double Staining, Cell Culture, Isolation, Fluorescence

    Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival

    doi: 10.3389/fncel.2019.00384

    Figure Lengend Snippet: Effect of 8-OH-DPAT on the protein expression of molecules from the BDNF signaling pathway in the HC, determined by Western Blot. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice receiving the 5-HT 1 A agonist 8-OH-DPAT (striped bars) or NaCl (white bars). (E) Representative membrane showing signal for p75 (75 kDa) and tubuline (52 kDa). 1: Control-NaCl. 2: Control-DPAT. 3: PCPA-NaCl. 4: PCPA-DPAT. Data are expressed as mean ± S.E.M., n = 6 per experimental group. ∗ p

    Article Snippet: Western Blotting Membranes were incubated for 1 h with blocking solution (5% milk in TBST) and then probed overnight at 4°C with mouse anti-BDNF (1:2000; Icosagen; 327-100 clone 3C11), rabbit anti-p75 (1:700; Alomone Labs; ANT-007), rabbit anti-TrkB (1:700; Alomone Labs; ANT-019), and rabbit anti-proBDNF (1:250; Abcam; ab72440) in TBST. β-III Tubulin was used as a loading control (1:2500; R & D Systems).

    Techniques: Expressing, Western Blot, Mouse Assay

    Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Differential Hippocampal Expression of BDNF Isoforms and Their Receptors Under Diverse Configurations of the Serotonergic System in a Mice Model of Increased Neuronal Survival

    doi: 10.3389/fncel.2019.00384

    Figure Lengend Snippet: Protein expression of molecules from the BDNF signaling pathway determined by Western Blot and representative membranes. (A) mBDNF. (B) TrkB. (C) proBDNF. (D) p75 in hyposerotonergic (PCPA-treated) or control (vehicle-treated) mice for 4 weeks. (E–H) Representative membrane showing signal for TrkB, p75, pro-BDNF, and mBDNF in Control and PCPA-treatd mice. Data are expressed as mean ± S.E.M., n = 11 (control) and 12 (PCPA). ∗ p

    Article Snippet: Western Blotting Membranes were incubated for 1 h with blocking solution (5% milk in TBST) and then probed overnight at 4°C with mouse anti-BDNF (1:2000; Icosagen; 327-100 clone 3C11), rabbit anti-p75 (1:700; Alomone Labs; ANT-007), rabbit anti-TrkB (1:700; Alomone Labs; ANT-019), and rabbit anti-proBDNF (1:250; Abcam; ab72440) in TBST. β-III Tubulin was used as a loading control (1:2500; R & D Systems).

    Techniques: Expressing, Western Blot, Mouse Assay