Fig. 2: Acute restraint stress activates MeA→VMH neurons but not MeA→BNST neurons.
From: Amygdala–liver signalling orchestrates glycaemic responses to stress
a,b, Quantification (a) and imaging (b) of mCherry-labelled synaptophysin with anterograde tracing from MeA neurons. Scale bars, 100 μm. c, Schematic of dual retrograde tracing with injection of AAVretro-RFP and AAVretro-GFP into BNST and VMH, respectively. d, RFP (BNST-projecting neurons) and GFP (VMH-projecting neurons) expression in MeA, with colocalization shown in white. Scale bar, 200 μm. e, Quantification of BNST-projecting and VMH-projecting MeA neurons. f, Number of FOS+ cells in VMH-projecting and BNST-projecting MeA neurons with and without (control) 30 min of restraint stress. g, Colocalization of FOS expression with GFP-labelled VMH-projecting MeA neurons (white arrows). Scale bar, 100 μm. h, Schematic of fibre photometry experiment (right) and axonal GCaMP8s expression and VMH fibre placement (left). 3V, third ventricle. Scale bar, 200 μm. i, GCaMP8s z-score in MeAVMH axons before, during and after 30 min of restraint. Shade bars indicate capture (pink), restraint (red) and release (grey). j, GCaMP8s z-score in MeA→VMH axons aligned to start of the capture period. k, Mean GCaMP8s z-score in MeA→VMH axons for 5 min baseline, 30 s capture, 30 min restraint and 5 min release periods. l, Schematic of fibre photometry (right) and axonal GCaMP8s expression and BNST fibre placement (left). ac, anterior commissure. Scale bar, 200 μm. m, GCaMP8s z-score in MeA→BNST axons before, during and after 30 min of restraint. Bars at the top indicate capture (pink), restraint (red) and release (grey). n, GCaMP8s z-score in MeA→BNST axons aligned to start of the capture period. o, Mean GCaMP8s z-score in MeA→BNST axons for 5 min baseline, 30 s capture, 30 min restraint and 5 min release periods. Data are mean ± s.e.m. Individual data points represent individual mice. Sample size and statistical analysis in Supplementary Data Table 1.
