Extended Data Fig. 3: GnRHOB neurons express olfactory/vomeronasal receptors and mechanosensitive ion channels.
a, Schematic strategy for sorting GnRHOB and GnRHPOA cells from Gnrh::Gfp naïve male mice. b, mRNA expression level of Olfr113, Olfr323, Vmn1r209, Vmn2r1 in GnRH-positive (GFP + ) and GnRH-negative (GFP-) cells isolated from the OB. Values are mean ± s.e.m (n = 4; 12-week-old males; *P = 0.0286; ns P > 0.05; two-sided paired Mann-Whitney U test). c, Schematic of the murine olfactory epithelium and vomeronasal organ. d, Representative coronal section of a VNO from an adult male mouse (P90) showing expression of Vmn2r1 mRNA (white staining) by smFISH. The red dotted-line indicates the boundary between the basal and apical layer of the VNO. e, Representative coronal section of an OE from an adult male mouse (P90) showing expression of Olfr323 mRNA (red) by smFISH. f, Schematic representation of the OB. GnRH cells located in the OB granular layer (GrL) are depicted in green. g-i, Representative coronal section of the OB from adult male mice (P90) showing expression of Gnrh1 (green), Vmn2r1 (white) and Olfr323 (red) in the MOB granular layer (MOB GrL). Arrows point to a Gnrh1-positive neuron co-expressing Vmn2r1 and Olfr323, while the arrowhead in j point to a GnRH neuron which does not express Vmn2r1 nor Olfr323. n = 3 mice used for the smFISH. Scale bars: d, 50 µm; e, 100 µm; g-j, 10 µm; inset in i, 5 µm. k, Schematic representing single-nucleus RNA sequencing protocol. l, UMAP visualization of the OB representing the distribution of Piezo1 and Piezo2 across the different cell clusters. m, Heatmaps indicating the relative expression of Piezo1 and Piezo2 within each neuronal cluster for Gnrh1-positive and Gnrh1-negative cells of the OB. n, Percentage expression profile of Piezo receptors in Gnrh1+ compared to Gnrh1- of the OB. Abbreviations: Oligo, Oligodendrocytes; PG, Periglomerular cells; GC, Granule cells; M/T, Mitral/Tufted cells; OEC, Olfactory ensheathing cells; IN, Inhibitory neurons; sSA, Superficial short axon cells; ET, endothelial cells; MES, mesenchymal cells; dSA, Deep short axon cells.
