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. 2023 Apr 12;6(6):e202301992. doi: 10.26508/lsa.202301992

Figure 3. Chemogenetic inhibition of Prdm13+ DMH neurons promotes excessive sleepiness during SD.

Figure 3.

(A) Experimental procedure for chemogenetic inhibition of Prdm13+ DMH neurons. AAV-hSyn-DIO-hM4Di-mCherry virus was injected into the DMH of Prdm13CreERT2/+ mice, and tamoxifen was intraperitoneally injected for three consecutive days (200 mg/kg body weight, i.p.). During SD between 6 am and 11 am, clozapine-N-oxide (CNO) was intraperitoneally injected at 8 am (1 mg/kg body weight). (B) A representative image of the injection site (mCherry, red) in the DMH, near the 3 V. Scale bar indicates 100 μm. (C, D) Numbers of sleep attempts during SD from 6 am to 8 am (6–8), 8 am to 9 am (8–9), 9 am to 10 am (9–10), and 10 am to 11 am (10–11) in mice with CNO or vehicle injection in Prdm13CreERT2/+(C left) and Prdm13+/+(D left) mice (n = 6–12). Values are shown as means ± S.E., ***P < 0.001 and non-significant (ns) by repeated measures ANOVA with Bonferroni’s post hoc test. The number of sleep attempts within 2 h after CNO or vehicle injection in individual Prdm13CreERT2/+ (C right) and Prdm13+/+ (D right) mice. ***P < 0.001 by paired t test was obtained for the mean number of sleep attempts within 2 h between CNO and vehicle injection in Prdm13CreERT2/+ mice (C right) (n = 7) but not in Prdm13+/+ mice (D right) (n = 6). (E) SWA after SD of Prdm13CreERT2/+ mice (n = 5). Normalized power is relative to the average of the 24-h baseline day. Values are shown as means ± S.E.