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Fig. 6
Chemogenetic ablation of NPVF neurons promotes wakefulness.
(A) mTagYFP and npvf coexpression in a 5-dpf Tg(npvf:mTagYFP-T2A-eNTR) zebrafish shown using YFP IHC and npvf FISH. (B–G) Tg(npvf:mTagYFP-T2A-eNTR) and WT siblings were treated with 5 mM MTZ for 60 h. Behavior was monitored from 6 to 8 dpf. (B) npvf ISH in 7-dpf Tg(npvf:mTagYFP-T2A-eNTR) zebrafish treated with DMSO or MTZ. Scale: 10 μm. NPVF neuronal loss is quantified as mean ± SEM in (C). (D–G) Transgenic animals treated with MTZ were more active (D,E) and slept less (F,G) than their identically treated WT siblings. Mean ± SEM for one representative experiment (D,F), or three pooled experiments (E,G) are shown. White and black bars under behavioral traces indicate day (14 h) and night (10 h), respectively. n = number of animals. ***p<0.005 by Two-way ANOVA with Holm-Sidak test (C) or Student’s t-test (E,G). See also Table 1 and Figure 6—figure supplement 1.