antibodies against na v 1 7  (Alomone Labs)

Journal: The Journal of Comparative Neurology

Article Title: Na V 1.7 mRNA and Protein Expression in Resident Neurons of the Human Spinal Dorsal Horn

doi: 10.1002/cne.70168

Figure Lengend Snippet: Distribution of SCN9A (Na V 1.7) and TACR1 (NK1R) mRNAs in the human spinal dorsal horn using RNAscope. (A) Mosaic image of a human spinal cord (lumbar 5/sacral 1) labeled with RNAscope in situ hybridization for SCN9A (red) and TACR1 (green) mRNAs and co‐stained with DAPI (cyan). The 488 channel was left unstained (green) to reveal background autofluorescence and lipofuscin, which is present in all human neurons. Magenta line outlines gray matter. Higher magnification images for each channel are shown for (B) lamina I, (C) lamina II, (D) lamina III, (E) lamina IV, and (F) lamina V. (G) Percentage of SCN9A + nuclei that coexpressed TACR1 (red bar) and percentage of TACR1 + nuclei that coexpressed SCN9A (blue bar) for each lamina (LI–LV). (H) Percentage of TACR1 + nuclei that were copositive for SCN9A (blue bar) and percentage of SCN9A + nuclei that were copositive for TACR1 (red bar) for the entire dorsal horn (combined data of H). (I) S CN9A mRNA was also detected in (I) nuclei lining the central canal, likely ependymal cells, (J) preganglionic parasympathetic neurons in the sacral parasympathetic nucleus (SPSy), (K) motor neurons in the ventral horn, particularly Pes9, which contains motor neurons associated with the feet, and (L) preganglionic sympathetic neurons in the intermediolateral column (IML) from a lumbar 1 spinal cord section. LI–LV, lamina I–V; SPSy, sacral parasympathetic nucleus; Pes9, motor neurons of the foot; CC, central canal. Sample size: n = 4. Scale bars: A = 500 µm; B–F = 20 µm; I–L = 50 µm.

Article Snippet: Several Na V 1.7 inhibitors have been developed, and a series of clinical trials have been conducted with mixed reports on pain outcomes and cardiovascular safety (Alles and Smith ; Biogen ; Dormer et al. ; Eagles et al. ; Kingwell ; McDonnell et al. ; Price et al. ).

Techniques: RNAscope, Labeling, In Situ Hybridization, Staining

Journal: The Journal of Comparative Neurology

Article Title: Na V 1.7 mRNA and Protein Expression in Resident Neurons of the Human Spinal Dorsal Horn

doi: 10.1002/cne.70168

Figure Lengend Snippet: Distribution of SCN9A (Na V 1.7) and GPR83 mRNAs in the human spinal dorsal horn using RNAscope. (A) Mosaic image of a human spinal cord section (lumbar 3) labeled with RNAscope in situ hybridization for SCN9A (red) and GPR83 (green) mRNAs and co‐stained with DAPI (cyan). The 488 channel was left unstained (green) to reveal background autofluorescence and lipofuscin, which is present in all human neurons. Magenta line outlines gray matter. Higher magnification images for each channel are shown for (B) lamina I, (C) lamina II, (D) lamina III, (E) lamina IV, and (F) lamina V. (G) Percentage of SCN9A + nuclei in that coexpressed GPR83 (red bar) and percentage of GPR83 + nuclei that coexpressed SCN9A (blue bar) for each lamina (LI–LV). (H) Percentage of GPR83 + nuclei that were copositive for SCN9A (blue bar) and percentage of SCN9A + nuclei that were copositive for GPR83 (red bar) for the entire dorsal horn (combined data of H). LI–LV, lamina I–V; D, dorsal nucleus; CC, central canal. Sample size: n = 4. Scale bars: A = 500 µm; B–F = 20 µm.

Article Snippet: Several Na V 1.7 inhibitors have been developed, and a series of clinical trials have been conducted with mixed reports on pain outcomes and cardiovascular safety (Alles and Smith ; Biogen ; Dormer et al. ; Eagles et al. ; Kingwell ; McDonnell et al. ; Price et al. ).

Techniques: RNAscope, Labeling, In Situ Hybridization, Staining

Journal: The Journal of Comparative Neurology

Article Title: Na V 1.7 mRNA and Protein Expression in Resident Neurons of the Human Spinal Dorsal Horn

doi: 10.1002/cne.70168

Figure Lengend Snippet: Na V 1.7 protein expression in the human lumbar spinal cord. Mosaic image of Na V 1.7 (red) protein staining in the human lumbar spinal cord (lumbar 5), co‐stained with DAPI (blue). White outline demarcates the gray matter. (B) Na V 1.7 protein gave an axonal and neuropil (synaptic) pattern in the spinal dorsal horn, mostly localized to lamina I–II (LI–LII). (C) Nav1.7 protein was also detected in the cytoplasm of large motor neurons in the ventral horn and (D) in axons in the anterior commissure (white arrows), as well as the cytoplasm of ependymal cells (cells outlining the central canal). (E) Representative 10x magnification confocal image of Na V 1.7 protein (red) co‐stained with the nociceptive presynaptic marker CGRP (blue), the presynaptic active zone marker Bassoon (green), and DAPI (cyan) in LI–LII of the spinal dorsal horn. (F) Overlay image (488, 555, 647, DAPI) of LI–LII in the negative control that was exposed to all of the same reagents except primary antibody and imaged and adjusted to the same settings as shown in E. (G) A higher magnification (100x) confocal view of the area outlined in yellow in the overlay image in C. (F) A digitally magnified view of the area outlined in white in E showing Na V 1.7 signal colocalized with CGRP and/or Bassoon (white arrows) or in close proximity to these proteins. LI–LV, lamina I–V; CC, central canal. Sample size: n = 4. Scale bars: A = 1 mm; B–C = 500 µm; D–F = 200 µm; G = 20 µm; H = 10 µm.

Article Snippet: Several Na V 1.7 inhibitors have been developed, and a series of clinical trials have been conducted with mixed reports on pain outcomes and cardiovascular safety (Alles and Smith ; Biogen ; Dormer et al. ; Eagles et al. ; Kingwell ; McDonnell et al. ; Price et al. ).

Techniques: Expressing, Staining, Marker, Negative Control

Journal: The Journal of Comparative Neurology

Article Title: Na V 1.7 mRNA and Protein Expression in Resident Neurons of the Human Spinal Dorsal Horn

doi: 10.1002/cne.70168

Figure Lengend Snippet: Investigation of Na V 1.7 protein in the soma of resident neurons in the dorsal horn. (A) Mosaic image of a human spinal cord section (lumbar 5–sacral 1) immunolabeled with Na V 1.7 (red), NeuN (green, soma and nucleus of neurons), and DAPI (blue, nuclei). Magenta line outlines the gray matter. Representative 40x images of Na V 1.7 (red), NeuN (green), and DAPI (blue) in (B) lamina I, (C) lamina II, (D) lamina III, (E) lamina IV, and (F) lamina V and their corresponding negative control that was exposed to all of the same reagents except primary antibody and imaged and adjusted with the same settings of each subregion. Na V 1.7 was not detected in the soma of any of the dorsal horn (lamina I–V) neurons. However, it was detected in the cell bodies of motor neurons and in preganglionic parasympathetic neurons. LI–LV, lamina I–V; SPSy, sacral parasympathetic nucleus; Pes9, motor neurons of the foot; CC, central canal. Sample size: n = 4. Scale bars: A = 500 µm; B–F = 100 µm.

Article Snippet: Several Na V 1.7 inhibitors have been developed, and a series of clinical trials have been conducted with mixed reports on pain outcomes and cardiovascular safety (Alles and Smith ; Biogen ; Dormer et al. ; Eagles et al. ; Kingwell ; McDonnell et al. ; Price et al. ).

Techniques: Immunolabeling, Negative Control

Journal: The Journal of Comparative Neurology

Article Title: Na V 1.7 mRNA and Protein Expression in Resident Neurons of the Human Spinal Dorsal Horn

doi: 10.1002/cne.70168

Figure Lengend Snippet: Evidence for postsynaptic Na V 1.7 expression in the human spinal cord. (A) Representative 10x image of Na V 1.7 (red), MAP2 (green), and DAPI (blue) staining in the human lumbar dorsal horn. The white outlines lamina I and lamina II. Representative 40x images of Na V 1.7 (red), MAP2 (green), and DAPI (blue) in (B) lamina I and (C) lamina II. White arrows point to the MAP2 signal that is localized around resident neurons (appears to be the plasma membrane) but is absent of Na V 1.7 signal. The image inset in panel C shows a 100x image of a large, LII neuron with a large apical dendrite that is devoid of Na V 1.7 signal. (D) Representative 20x image of Na V 1.7‐positive axonal fibers in the deeper lamina around LIV–LV. The white arrow points to Na V 1.7 and MAP2 copositive signal (yellow in overlay) that does not have a nucleus and is not a cell body. (E) Representative 20x image of Na V 1.7 staining in the anterior commissure (ac), where intensely labeled Na V 1.7‐positive axons are highlighted (white arrow). (F) A 100x image of Na V 1.7 (red), Ankyrin‐G (green), and DAPI (blue) staining in a motor neuron in the ventral horn. (G) A cropped, zoomed‐in image of Na V 1.7 (red), Ankyrin‐G (green), and DAPI (blue) signal in a lamina II dorsal horn neuron. LI–LV, lamina I–V; ac, anterior commissure. Sample size: n = 3–4. Scale bars: A = 200 µm; B and C = 50 µm; D and E = 100 µm; F = 20 µm; G = 5 µm.

Article Snippet: Several Na V 1.7 inhibitors have been developed, and a series of clinical trials have been conducted with mixed reports on pain outcomes and cardiovascular safety (Alles and Smith ; Biogen ; Dormer et al. ; Eagles et al. ; Kingwell ; McDonnell et al. ; Price et al. ).

Techniques: Expressing, Staining, Clinical Proteomics, Membrane, Labeling

Journal: The Journal of Comparative Neurology

Article Title: Distribution of Voltage‐Gated Sodium Channels and Scaffolding Proteins on Vestibular Calyx Ending Delineates the Axon Initial Segment

doi: 10.1002/cne.70127

Figure Lengend Snippet: Nodal protein‐like immunoreactivity reveals heminodes and first nodes on the distal process of vestibular afferents. (A) Schematic of calyceal microdomains (modified from Lysakowski et al. ). (B) β‐IV spectrin ( B‐IV Spec , red ) labels heminodes (Domain 4) and nodes in vestibular afferents and the upper portion of the calyx (Domain 2, asterisks ) in dimorphic afferents (indicated by a lack of calretinin ( Calret ) immunolabel, green ). (C,D) Neurofascin‐186 ( NF‐186 , red) labels heminodes ( arrows ) and nodes in crista ( C ) and otolith ( D ) organs. (E) Na V 1.6 ( green ), the most common nodal Na channel isoform, is present at the heminode, while Caspr ( red ) labels the hemi‐paranode ( brackets ). In the right half of this panel, myelin basic protein ( MBP , blue ) extends over the hemi‐paranode to the heminode. (F) Two different examples of ezrin labeling ( red ). Ezrin labels the heminode, MBP ( blue ) labels the internode, and there is a small gap between them as the myelin attenuates, the hemi‐paranode ( brackets , not immunolabeled here, but seen in panel ( E ), labeled with Caspr, red ). (G, H) qPCR demonstrates the presence of Na V 1.6 ( G ) and Na V 1.5 ( H ) in vestibular ganglion ( VG ) cells ( G, H ), but to a lesser extent in the vestibular ( VO ) and cochlear ( CO ) sensory organs. Brain ( Br ) and heart ( H ) served as controls for Na V 1.6 and Na V 1.5, respectively. Values are means ± SEM. (I, I’) Ezrin ( red ) and AnkG ( green ) are seen to be co‐extensive in a rat vestibular nerve node, while ezrin and AnkB (J, J’) are complementary, with ezrin ( red ) labeling the node and AnkB ( green ) the paranode. (K) Vestibular ganglion cell labeled with calretinin ( green ) and Ank G ( red ) labeling the first proximal node of Ranvier ( arrow ). Scale bars: in B‐D, K = 10 µm; in E (also applies to F, I, J) = 2 µm.

Article Snippet: Na V 1.6 (rb)/(ms) , Rasband/NeuroMab , gift, 75‐026 RRID:AB_2184197 , X (if) , X (ON, Br) , , X , , Rasband et al. ( ); datasheet .

Techniques: Modification, Immunolabeling, Labeling

Journal: The Journal of Comparative Neurology

Article Title: Distribution of Voltage‐Gated Sodium Channels and Scaffolding Proteins on Vestibular Calyx Ending Delineates the Axon Initial Segment

doi: 10.1002/cne.70127

Figure Lengend Snippet: Na V 1.2 labels Domain 3 in majority of afferents, first nodes, and stromal paranodes. (A, A’) Na V 1.2 ( green ) labels the calyx outer surfaces (Domain 3, arrows ) of D afferents in the extrastriolar zone of the utricular macula from an adult female rat. The calyx inner surfaces are marked with Caspr1 ( Caspr, red in A). The beginning of two parent axons (arrowheads) can be seen in the neuroepithelium, but could not be traced into the stroma, where the heminodes of most D units are located, presumably because they go out of the plane of this single section. Inset in A’, upper right , schematic of entire calyx, showing Domain 3 in red. (B) Na V 1.2 antibody ( green ) labels mostly stromal nodes ( arrowheads ), but not heminodes ( arrows ), both of which are marked with an ezrin antibody ( red ). (C) Deeper in the stroma of the crista in a maximum intensity projection, many nodes can be observed. The Na V 1.2 ( green ) extends beyond the boundaries of the node, marked by ezrin ( red ). C insets . Na V 1.6 antibody (in this case red ) immunostains two nodes of Ranvier in the stroma of a rat saccular macula. Similar to (C), Na V 1.2 ( green ) also stains the nodes (seen after removal of the Na V 1.6 immunolabel ( red ) in the bottom half of each inset) and extends well beyond each node into the paranode. Scale bars: A–C = 10 µm; C inset = 5 µm.

Article Snippet: Na V 1.6 (rb)/(ms) , Rasband/NeuroMab , gift, 75‐026 RRID:AB_2184197 , X (if) , X (ON, Br) , , X , , Rasband et al. ( ); datasheet .

Techniques: Immunolabeling

Journal: The Journal of Comparative Neurology

Article Title: Distribution of Voltage‐Gated Sodium Channels and Scaffolding Proteins on Vestibular Calyx Ending Delineates the Axon Initial Segment

doi: 10.1002/cne.70127

Figure Lengend Snippet: Na V 1.6 marks both nodes and heminodes, except for heminodes in calyx‐only afferents. Caspr1 ( Caspr, red ) labels the inner surface of calyces, the proximal (central) sides of heminodes (Domain 4, red in schematic, top left ) and in the paranodes on both sides of nodes in this maximum intensity projection image of a utricular macula. Note that all the nodes and most of the heminodes are labeled with Na V 1.6 ( green ), except for two heminodes ( arrows ), sitting just above the Caspr‐labeled hemi‐paranodes, presumably belonging to C fibers located above the basement membrane ( dashed line ). Scale bar = 5 µm.

Article Snippet: Na V 1.6 (rb)/(ms) , Rasband/NeuroMab , gift, 75‐026 RRID:AB_2184197 , X (if) , X (ON, Br) , , X , , Rasband et al. ( ); datasheet .

Techniques: Labeling, Membrane

Journal: The Journal of Comparative Neurology

Article Title: Distribution of Voltage‐Gated Sodium Channels and Scaffolding Proteins on Vestibular Calyx Ending Delineates the Axon Initial Segment

doi: 10.1002/cne.70127

Figure Lengend Snippet: Na V 1.8 antibody labels vestibular afferents and has three different patterns. (A) The heminodes of calyx‐only afferents ( green , color conventions are the same in all panels) are densely labeled with Na V 1.8‐like immunoreactivity ( red ), as seen in both longitudinal section (A) and (B) cross‐section of calyces labeled with calretinin. (C) Occasionally, the inner membrane of a calyx surrounding a type I hair cell, most likely belonging to a dimorphic afferent since these were found in the periphery, is labeled with Na V 1.8. However, because confocal lacks the resolution of electron microscopy, this could also be a type I hair cell membrane. In (D) Na V 1.8 is seen to label the calyx endings of calyces ( arrows ) with relatively light calretinin label (D’). Scale bars: A, C, D = 5 µm, B = 10 µm.

Article Snippet: Na V 1.8 , NeuroMab , 75‐166 RRID:AB_2183861 , , , , X , , Datasheet.

Techniques: Labeling, Membrane, Electron Microscopy

Journal: The Journal of Comparative Neurology

Article Title: Distribution of Voltage‐Gated Sodium Channels and Scaffolding Proteins on Vestibular Calyx Ending Delineates the Axon Initial Segment

doi: 10.1002/cne.70127

Figure Lengend Snippet: Na V 1.1 labels heminodes of both calyx‐only and dimorphic afferents. This maximum intensity projection of an image stack was taken from the central zone of an adult female rat crista. The tissue was fixed by intra‐labyrinthine perfusion with a methanol‐based fixative. (A) Calretinin (green) stains C axons; calyx terminals were not preserved because of the weak fixation. Na V 1.1 antibody (red) stains heminodes in five calretinin‐positive C fibers (numbered 1 ‐ 5) and three calretinin‐negative D fibers (arrows) . The basement membrane (BM) is indicated ( dashed line ). (A’) The channel containing calretinin labeling has been removed to reveal the Na V 1.1 labeling. Heminodes of D units appear above the BM because the section from which the stack was obtained was cut at a slightly oblique angle. (Insets, right) . Higher‐magnification of the five C ‐fiber heminodes, each stained for Na V 1.1 (red) and calretinin (green) ; the intensity of the calretinin label has been reduced to visualize the Na V 1.1 labeling more easily. (B) Higher magnification single image of both the calretinin‐labeled C fiber (left) numbered “5” and the upper right non‐calretinin‐labeled D fiber (right) in (A, A’) Scale bars: in A = 5 mm (also applies to A’); in A insets = 1 µm; and in B = 2 µm.

Article Snippet: Na V 1.1 (ms) , NeuroMab , 75‐023 RRID:AB_2238842 , , X (Br) , , X , , Datasheet.

Techniques: Membrane, Labeling, Staining