Fig. 5: Generation of mice with loss of diurnal rhythms of γ2 subunit expression in adult PVH.

a Diagram showing delivery of AAV-Cre-GFP to the PVH of γ2^flox/flox::γ2-TB^flox/flox to clamp γ2 expression at a constant level in adult mice. Cre-mediated deletion of the endogenous γ2 and expression of wild-type γ2 cDNA driven by the ROSA26 promoter lead to replacement of rhythmic expression of γ2 with a static expression level of γ2 driven by the ROSA26 promoter. b Validation of viral delivery to the PVH with GFP for vector delivery and red for immunostaining of γ2 expression. GFP expression (AAV-GFP or AAV-Cre-GFP) was verified in the PVH (top panels) and γ2 expression was verified by immunostaining (bottom panels): no expression of γ2 in Cre-mediated deletion (bottom middle, arrow) and re-expression by Cre-mediated γ2 expression driven by the ROSA26 promoter (bottom right, arrow), noticing consistency in the expression between Cre-GFP (top right) and γ2 (bottom right). c, d The amplitude (c, n = 15 each) and frequency (n = 12 for EPSCs and 14 for IPSCs) of sEPSCs and sIPSCs recorded during day and night periods from PVH neurons 3–4 weeks after AAV-Cre-GFP delivery. All data are represented as mean ± SEM and analyzed by unpaired Student’s t-tests. TB transcription blocker. Scale bars: 200 µm, 3V third ventricle. Source data are provided as a Source Data file.