Journal: bioRxiv
Article Title: Mouse behavioral genomics identifies Creld1 as a gatekeeper of somatosensation
doi: 10.64898/2026.03.30.715210
Figure Lengend Snippet: a , Schematic diagram of the CRISPR/Cas9-mediated knockout of target genes expressed in DRG neurons via intraventricular injection of the AAV9 virus carrying the construct containing the sgRNA against the target gene and EGFP for indication of infection efficiency. 6-8 weeks post injection, the mice were subjected to experimental tests. b , Representative single-cell Ca 2+ imaging of DRG neurons derived from control sgRNA- or sgTrpv1-targeted mice in response to 10 μM capsaicin and 50 mM KCl. Individual neurons are color-coded. c , Scatterplot of the percentage of cultured GFP + DRG neurons showing capsaicin-induced Ca 2+ response. d , Representative traces of mechanically evoked whole-cell currents of rapidly inactivating (RA), intermediate inactivating (IA), and slowly inactivating (SA) kinetics recorded from cultured control DRG neurons. e , Proportion of control sgRNA- and sgPiezo2-targted GFP + DRG neurons showing the indicated responding properties. f , The peak current density of tetradotoxin (TTX)-sensitive Na v current of DRG neurons derived from the control sgRNA-and sgNa v 1.7-targeted mice. g , Representative trace of evoked action potential (APs) of the control sgRNA- and sgNa v 1.7-targeted DRG neurons. h , Firing frequency of the control sgRNA- and sgNa v 1.7-targeted DRG neurons in response to the injected currents. i , Rheobase of sgNa v 1.7-targeted DRG neurons compared with the control sgRNA-targeted neurons. j , The percentage of paw withdrawals in response to the indicated series of Von Frey filament stimulation. k , Scatterplot of the percentage of paw withdrawal in response to the pinprick test. l , Scatterplot of the threshold in response to the Randall-Selitto test. m , Scatterplot of the paw withdrawal latency in response to the hot plate test. n , Scatterplot of the paw withdrawal latency in response to the cold plate test. o-q , Scatterplot of the scratch numbers within 30 minutes in mice injected with histamine ( o ), chloroquine ( p ) or compound 48/80 ( q ). Data are presented as means ± SEM; sample sizes are indicated. Statistical significance is determined by Unpaired Student’s t test for c , f , i . Two-way ANOVA with Bonferroni’s multiple comparisons test for j . One-way ANOVA with Bonferroni’s multiple comparisons test for k - q , * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Article Snippet: In brief, cells were washed with the Ca 2+ imaging buffer (1 x HBSS buffer with 1.3 mM Ca 2+ , 10 mM HEPES, pH 7.2) and then loaded with the Ca 2+ imaging buffer containing 2.5 mM Ca 2+ indicator dye Fura-2, AM (ThermoFisher scientific, F1225) and 0.05% Pluronic F-127 (Beyotime, ST501).
Techniques: CRISPR, Knock-Out, Injection, Virus, Construct, Infection, Single Cell, Imaging, Derivative Assay, Control, Cell Culture, Randall–Selitto Test, Hot Plate Test
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