Fig. 6: Projection-specific transcriptomic analysis identifies DMH-projecting LPBSST neurons as cold-activated neurons.
From: A parabrachial-hypothalamic parallel circuit governs cold defense in mice
a Projection-specific transcriptomic analysis (retro-TRAP), where GFP-tagged translational ribosomes from DMH-projecting LPBVglut2 neurons were immunoprecipitated and associated mRNAs were sequenced. SST, somatostatin. b Volcano plots (q value versus log2 fold change) for LPB mRNAs after retro-TRAP sequencing. c Retro-TRAP fold enrichment (IP/Input) for PB-expressed genes downloaded from the Allen Institute. d, e Overlap between SST-IRES-Cre labeled neurons (Tdt+) in the LPB and the following immuno-positive neurons: warm-induced cFos (10oC, 2 h; d), and cold-induced cFos (38oC, 2 h; e). (n = 3 mice each). f Overlap between DMH-projecting LPBSST neurons and cold-induced cFos (10oC, 2 h) (n = 3 mice each). g Axonal tracing of LPBSST neurons by ChR2, where AAV9-hEF1a-DIO-hChR2-EYFP was injected into the LPB of SST-IRES-Cre mice. PVH, paraventricular hypothalamic. h, i Expression of ChR2 in LPBSST neural somas (h) and terminals in the POA or DMH (i). j Induction of EPSCs in DMH neurons by light stimulation of LPBSST & ChR2 terminals (blue, 6 mW, 10 ms, 10 hz). k EPSCs were blocked by GluR antagonists D-AP5 and CNQX. Latency and response rate (3 of 28 randomly recorded neurons from 3 mice) are shown on the right. l EPSCs were blocked after TTX treatment but were restored by 4-AP treatment. All data are the mean ± sem. Scale bars in the enlarged panel of (d–f) are 10 μm. Source data are provided as a Source data file.
