Fig. 3

Cholinergic transmission regulates the dNGIs survival. a Experimental schedule shows that mice at 120 ± 2 days old of age were administered with a single dose of BrdU. 7 days after the BrdU administration (DAI), TBS was applied for 16 consecutive days. BrdU staining was performed at 1, 3, 7, 11, 21, or 28DAI. Representative theta waves recorded from dDG region of vChATs^ChR2+ mice in response to TBS. b, c Representative images b, and summarized numbers c, of BrdU⁺ cells in the vChATs^ChR2+ and vChATs^ChR2− mice treated without (non-TBS) or with TBS (mean ± SEM, n = 9 mice/group, *p < 0.01, two-way ANOVA with Bonferroni post-hoc test). d, e Representative images d and a plot e shows the actual (blue circles) and the averaged numbers (red triangles) of GFP⁺BrdU⁺ cells in the vChATs^ChR2+ and vChATs^ChR2− mice treated without (non-TBS) or with TBS (mean ±SEM, n = 11 mice/group, *p < 0.01, two-way ANOVA with Bonferroni post-hoc test). f Experimental schedule (top) for the BrdU administration and ALL application in both vChATs^NpHR+ and vChATs^NpHR− mice in vivo. BrdU staining was performed at 1, 3, 7, 11, 21, or 28DAI. Representative recordings (middle) indicate that the application of ALL (yellow light) blocked spontaneous excitatory currents in vChATs^NpHR+ cells at a holding potential of −60 mV. Bar graph (bottom) indicates the numbers of BrdU⁺ cells in the vChATs^NpHR+ and vChATs^NpHR− mice treated without (non-ALL) or with ALL (ALL). BrdU labeling was performed at 1, 3, 7, 11, 21, or 28 DAI (mean ± SEM, n = 9 mice/group, *p < 0.01, two-way ANOVA with Bonferroni post-hoc test). g Experimental arrangements show the schedules of rAAV1/2 virus injection and BrdU administration. 7 days after the injection, TBS was applied for 16 consecutive days. Western blots and RT-PCR analysis were performed 5 days after the end of TBS. Representative images of mRNAs and proteins show 3 replicates (lanes 1, 2 and 3) in each condition; no virus (I), sM1siRNA (II) and M1siRNA virus injections (III). Similar results were seen in each of the four experiments