Extended Data Fig. 1: VMPO neurons responding with a delay to a heat stimulus overlap with LepR-positive neurons and are receptive to becoming activated by heat acclimation.
From: Thermally induced neuronal plasticity in the hypothalamus mediates heat tolerance
a, Left: Brain sections revealing the VMPO of FosTRAP2;HTB mice that received z-4-hydroxytamoxifen (4-OHT; 50mg/kg i.p.) at room temperature (TRAP@RT) and after 2 hours of a 4-hour (TRAP@36 °C (4h)) or 8-hour (TRAP@36 °C (8h)) exposure to 36 °C,. Scale bar: 100 μm. Right top: Increasing the time of exposure to 36 °C increases the number of TRAPped GFP-positive cells in the VMPO. Kruskal-Wallis test, P <0.0001; Dunn’s multiple comparisons test, *P = 0.0110 (TRAP@RT: TRAP@36˚C (4h)), ***P < 0.0001 (TRAP@RT : TRAP@36˚C (8h)), **P = 0.0047 (TRAP@36˚C (4h) : TRAP@36˚C (8h)). n = 7 sections / 3 animals for TRAP@RT, n = 22/5 for TRAP@36˚C (4h) and n = 18/4 for TRAP@36˚C (8h). Right bottom: TRAPped neurons highly overlapped with Fos protein in animals exposed to warmth.; n = 22/5 for TRAP@36˚C (4h) and n = 18/4 for TRAP@36˚C (8h). b, Warm-stimulated (4 hours) and RT control brain sections of LepR-Cre;HTB mice show the rostral POA and MnPO, stained with GFP (LepR expression) and cFos. The overlap of LepR and cFos due to warm temperature exposure is quantified in the right panel graph (% of LepR-positive neurons, from 11 and 10 sections of N = 2 animals for RT and 36 °C, respectively; two-tailed T-test, ***P < 0.0001; scale bars: 100 μm). c, Expression of Pacap (Adcyap) and Bdnf transcripts assessed by bulk mRNA sequencing of FACS sorted LepR+ and LepR- cells obtained from POA tissue isolated from LepR-Cre;HTB mice. DWilcoxon test, ***P < 0.0001 (LepR+ : LepR-). n=18/3 (samples/mice); each data point represents expression of the respective gene in each sample plotted as a log2 normalized value. d, Core body temperature, BAT and tail temperature of ChR2-expressing and control mice before, during (blue shading) and after blue light stimulation (20 Hz, 10 msec pulses, 1min ON/3min OFF). n=4 mice per group. Data represent mean ± s.e.m. e, Right: Quantification of the number of neurons expressing cFos in VMPO of LepR-Cre;HTB kept at room temperature (RT), and stimulated at 36 °C and displayed as % of all cFos-positive neurons. Kruskal-Wallis test, P < 0.0001; Dunn’s multiple comparisons test, **P = 0.0047 (RT : 2h @ 36 °C), ***P < 0.0001 (RT : 4h @ 36 °C), **P = 0.0034 (2h @ 36 °C : 4h @ 36 °C). n = 7 sections / 2 animals for RT, 8 sections / 3 animals for 4h @ 36 °C, and 6 sections / 2 animals for 8h @ 36 °C. Left: Quantification of the absolute number cFos-positive cells. Kruskal-Wallis test, P = 0.0442; Dunn’s multiple comparisons test, P = 0.7224 (RT : 2h @ 36^C), P = 0.1709 (RT : 4h @ 36 °C), *P = 0.0381 (2h @ 36 °C : 4h @ 36 °C). f, Extent of VMPOLepR population. Representative images of 250 µm acute slices from LepR-Cre;HTB mice used for ex vivo electrophysiological experiments. Scale bar: 100 μm. Boxplots show median and interquartile range.
