Fig. 6: Indirect synaptic connections allow neuronal communication between MOB and DMH nuclei. | Nature Communications

Fig. 6: Indirect synaptic connections allow neuronal communication between MOB and DMH nuclei.

From: A sex-specific thermogenic neurocircuit induced by predator smell recruiting cholecystokinin neurons in the dorsomedial hypothalamus

Fig. 6

a Approach to identify direct synaptic connections with MOB using the presynaptic tracer AAV-hSyn-DIO-mCherry delivered to the MOB of Tbx21-Cre+ mice. mCherry positive cells were solely present in olfactory processing centers, Two independent experiments, N = 6. b Approach to identify indirect synaptic connections with MOB using the polysynaptic tracer AAV-hSyn-Cre injected into MOB of Ai14 mice (tdTomato reporter) and identification of projection neurons in DMH and LH, Two independent experiments, N = 5. c Summary of tdTomato signal localization in brain sections using QuckNII alignment (Allen Brain Atlas reference), Two independent experiments, N = 5. d Cartoon representation of cell sorting analysis to collect hypothalamic TdTomato-positive neurons and RNA-sequencing. e tdTomato+ hypothalamic neurons transcriptome levels of GABAergic, glutamatergic and other enriched genes, Two independent experiments, N = 14. f Experimental design to isolate single cells from microdissected DMH and scRNA-seq. g Uniform Manifold Approximation and Projection (UMAP) clustering of neuronal populations in the DMH identified by single cell RNA sequencing (10, 028 neurons, 6 pooled mice, 41 clusters). h UMAP highlighting GABAergic (Slc32a1) and glutamatergic (Slc17a6) neuronal clusters. i,j Subclustering into inhibitory and excitatory clusters revealed cluster GABA 14 as one coexpressing Ttr and Cck, color scheme of clusters corresponds to color scheme of UMAPs in Supplementary Fig. 7k Immunohistochemical and in situ hybridization co-staining of Cck (green) and Ttr mRNA (magenta) in DMH, white represent Cck/Ttr co-expression, scale 100 µm. l Violin plot of Cck expression in DMH neurons, Model-based Analysis of Single-cell Transcriptomics (MAST) analysis. Table S6 contains the detailed results of the statistical analysis. ****p < 0.0001. Source data are provided as a Source Data file. main olfactory bulb (MOB), dorsomedial hypothalamus (DMH), lateral hypothalamus (LH).

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