Journal: bioRxiv

Article Title: Ca V 1.2-dependent excitation-transcription coupling modulates nociception

doi: 10.1101/2025.06.25.661480

Figure Lengend Snippet: (A) Conditional Cav1.2 knockout mice (cKO, SNS-Cacna1c -/- ) were generated by crossing Cacna1c fl/fl (Ctrl) mice with mice expressing Cre under control of the sensory neuron-specific (SNS) Scn10a gene promoter. Single cell RNA-Seq data indicate that Scn10a encoding Nav1.8 is coexpressed with Cacna1c in peptidergic (PEP) and non-peptidergic nociceptors (NP), but not neurofilament-expressing mechano- and proprioceptors (NF). (B) Bulk RNA-Seq results for Cacna1c mRNA of DRGs from Cacna1c fl/fl versus SNS-Cacna1c -/- mice. TPM; transcripts per million. (C) HC imaging single cell data of DRG neurons indicating downregulation of Cav1.2 protein in neurons of SNS-Cacna1c -/- mice. (D) Mean values of replicate wells form n = 4 (2 male, 2 female) mice of the data shown in (C). Values of males are colored in blue, females in red. (E) Number of analyzable DRG neurons per isolation from n = 9 female (red) and male (blue) mice per genotype. (F-G) UCHL1 and RIIβ expression pattern in N = 9 mice (female in red and male in blue) of n = 300.000 DRG neurons of per genotype.

Article Snippet: The following antibodies were used in this study: chicken polyclonal antibodies anti-UCHL1 (1:2000, Novus, #NB110-58872), rabbit monoclonal antibody anti-phospho-CREB pS133 (1:500, clone F.959.4, Thermo Fisher, #MA5-11192), rabbit monoclonal antibody anti-phospho-RIIα (pS96) (1:1000, clone 151, Abcam, #ab32390), mouse monoclonal antibody anti-RIIβ (1:2000, BD Transduction Laboratories, #610625), mouse monoclonal anti-CaMKIIα (1:1000, Thermo Scientific, clone 6G9, #MA1-048), rabbit anti-CGRP antibody (1:1000, Bachem, #T-4032), mouse monoclonal anti-NF200 (1:1000, Sigma-Aldrich, clone N52, #N0142), mouse monoclonal IgG2a anti CGRP (1:500, Biorbyt/ Biozol, clone 4901, #orb319478), mouse monoclonal anti Nav1.8 (1:500, NeuroMAB, clone N134/12 75-166), mo-Cav1.2_CloneN263/31: mouse monoclonal anti Cav1.2 (1:200, NeuroMAB, clone N263/31, #73-257), rabbit polyclonal anti-TRPV1 (1:1000, Alomone, #ACC-030), and highly cross-adsorbed Alexa 647, 594, and 488 conjugated secondary antibodies (Invitrogen, Carlsbad, CA).

Techniques: Knock-Out, Generated, Expressing, Control, RNA Sequencing, Imaging, Isolation