Fig. 6: Downstream neurons of VLM-CA in DVC and MPA regulate body temperature.

a, b Anterograde tracing of VLM-CA neurons. a Schematic of anterograde tracing (upper) and labeled VLM-CA neurons (bottom). b Representative images of VLM-derived projections in DVC, DMH, MPA, and PVN (n = 4 regions per 7 independent biological replicate). Scale bars: 200 µm. c Representative images of Fos signals in DVC, DMH, MPA, and PVN of Dbh^VLM-hM3Dq (bottom panels, n = 3 animals) and Dbh^VLM-mCherry control (upper panels, n = 2 animals) mice. Fos immunostaining was performed 2 h after CNO injections. Scale bars: 100 µm. Schematic of the Fos-Triggered Downstream Neuronal Activation (FTDNA) strategy (d) and representative results (e) of selective expression of SSFO in VLM-activated neurons (Fos⁺) in the MPA (n = 521 cells in 3 MPA area sections from 3 animals). FTDNA-based optogenetic activation (indicated by blue bars, 20 Hz, 30 s) of the VLM^M3 → PVN^SSFO (f, P = 0.0028), VLM^M3 → DMH^SSFO (g, P = 0.0012), VLM^M3 → MPA^SSFO (h, P = 0.0042), and VLM^M3 → DVC^SSFO (i, P = 0.0019) neural circuits, respectively. The left panels show SSFO expression and fiber implantation, and the right panels show T_core. Two-way ANOVA analysis in (f, g, h, i). Data are presented as mean ± SEM. *P < 0.05; **P ≤ 0.01. Source data are provided as a Source Data file.