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. 2021 Aug 23;12:5080. doi: 10.1038/s41467-021-25368-y

Fig. 2. The PVT provides a dense glutamatergic projection to the BNST.

Fig. 2

a Viral unilateral retrograde tracing strategy in the BNST of VGLUT2-Cre mice (N’s = 4 M, 4 F). b % cells in the basolateral amygdala (BLA) and paraventricular nucleus of the thalamus (PVT) sections that project to the BNST (the densest sources of glutamatergic input identified), calculated as (total # of retrogradely-labeled cells/total DAPI+) × 100 and presented as mean values ± SEM. Mixed-effects model: main effect of subregion (F4,23 = 32.01, P < 0.0001, not indicated) but no effect of sex (P = 0.967) or interaction (P = 0.524); post-hoc two-tailed t-tests with Holm-Sidak corrections between subregions: BLA has fewer BNST-projecting neurons than the PVT across all A/P coordinates (all $$$$Ps < 0.0001), and that the amPVT has more than the mPVT (t23 = 2.94, *P = 0.037) and pPVT (t23 = 3.15, *P = 0.027), with no other differences: aPVT vs. amPVT (P = 0.085); aPVT vs. mPVT (P = 0.873); aPVT vs. pPVT (P = 0.873); mPVT vs. pPVT (P = 0.873). aPVT, anterior PVT; amPVT anterior-mid PVT; mPVT, mid PVT: pPVT, posterior PVT. c Proportion of VGLUT2-Cre+ (EGFP-labeled) and VGLUT2-Cre− (tdTomato-labeled) BNST-projecting PVT (PVTBNST) neurons in males and females, showing that almost all PVTBNST neurons are VGLUT2+ in both males and females with no difference between sexes (two-tailed unpaired t-test: t6 = 1.24, P = 0.261). d Representative images of coronal brain slices from a virus-injected mouse illustrating the expression of DAPI (blue) and all BNST-projecting cells (both VGLUT2-Cre + and - in green) in the BLA and across the A/P extent of the PVT. e Density heat maps illustrating the average number of BNST-projecting cell bodies within a 50 µM radius of each identified projection neuron for samples from each sex, matched and scaled similarly to representative images in d (top: males, bottom: females).