asc 009 rrid ab 2040202  (Alomone Labs)

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) , Alomone , ASC‐009 RRID:AB_2040202 , , X (SN, Br, DRG) , , X , X (w, if) , Black et al. ( ) .

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) , Alomone , ASC‐009 RRID:AB_2040202 , , X (SN, Br, DRG) , , X , X (w, if) , Black et al. ( ) .

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) , Alomone , ASC‐009 RRID:AB_2040202 , , X (SN, Br, DRG) , , X , X (w, if) , Black et al. ( ) .

Techniques: Labeling, Membrane