Fig. 4
PA-FdCre–mediated Cav3.1 knockdown in the MS increases object exploration. a Injection of a mixture of AAVs expressing PA-Flp, LF-FdCd and flox-shCav3.1 (or flox-shControl) into the MS of a 8-wk-old mice brain, with (w/) or without (w/o) subsequent noninvasive white LED (2 mW mm−2) illumination 2 wk after infection. b Schematic depicting object-exploration behavior in PA-FdCre–mediated, MS-specific, Cav3.1-knockdown mice. c Immunohistochemical detection of Cav3.1 in the MS following co-infection with PA-Flp-, LF-FdCd- and flox-shCav3.1-expressing AAVs. PA-Flp (mCh) and flox-shCav3.1 (GFP) fluorescence, Cav3.1 immunofluorescence (violet), and merged images are presented. Arrowheads indicate Cav3.1-positive cells. Arrows indicate PA-Flp/flox-shCav3.1–positive, Cav3.1-negative cells. Scale bar: 50 μm. d Cumulative traces of representative navigation pathways of shCav3.1 with LED and shCav3.1 without LED mice during object-exploration behavior. e Increased object-exploration time for shCav3.1 w/ LED mice (n = 9/group) compared with shCav3.1 w/o LED mice (n = 8; P = 0.003, two-way repeated measures ANOVA; P = 0.004, Bonferroni post-hoc test), shControl w/ LED mice (n = 5; P = 0.003, two-way repeated measures ANOVA; P = 0.028, Bonferroni post-hoc test), and shControl w/o LED (n = 5, P = 0.003, two-way repeated measures ANOVA; P = 0.009, Bonferroni post-hoc test). f Increased total object-exploration time for shCav3.1 w/ LED mice (n = 9) compared with that for shCav3.1 w/o LED mice (n = 8; P = 0.003, one-way ANOVA; P = 0.004, Bonferroni post-hoc test), shControl w/ LED mice (n = 5; P = 0.003, one-way ANOVA; P = 0.029, Bonferroni post-hoc test), and shControl w/o LED mice (n = 5; P = 0.003, one-way ANOVA; P = 0.008, Bonferroni post-hoc test). g Locomotor activity of shCav3.1 w/ LED (n = 9), shCav3.1 w/o LED (n = 8), shControl w/ LED mice (n = 5), and shControl w/o LED (n = 5) mice in the experimental cage during the 60-min habituation period of the object-exploration task (P = 0.612, one-way ANOVA). e–g Data represent means ± s.e.m. (*P < 0.05, **P < 0.01)
