Journal: bioRxiv
Article Title: A Preoptic Neurocircuit That Modulates Metabolic Flexibility
doi: 10.64898/2026.01.15.699760
Figure Lengend Snippet: (A) Immunostaining of axonal fibers in Vglut2-IRES-Cre mice transduced with Gq-DREADD (AAV-hSyn-DIO-hM3D(Gq)-mCherry). Axonal projections were identified in a variety of downstream regions, including the paraventricular thalamus (PVT), paraventricular hypothalamus (PVH), dorsomedial hypothalamus (DMH), periacqueductal grey (PAG), locus coeruleus (LC), and raphe pallidus (RPa). ( B) Optogenetic branch stimulation of hindbrain-projecting avPOA Vglut2 neurons. Blue bars correspond to regions targeted for optogenetic stimulation via cannula implantation and subsequent exposure to 455 nm blue light. (C) ( Left) Glucose tolerance testing of Vglut2-IRES-Cre animals transduced with a Cre-dependent ChR2 (AAV-DIO-ChR2(H134R)-EYFP) or EYFP (AAV-DIO-EYFP) with optic fibers placed unilaterally above either the POA, POA→DMH-projecting neurons, or POA→RPA-projecting neurons. (Right) AUC of GTT for each cohort described (ns = p > 0.05, ** = p < 0.005, student’s two-tailed t-test, mean ±SEM, n = 4 EYFP, n = 4 POA, n = 3 POA→DMH, n = 3 POA→RPa Vglut2-IRES-Cre animals). (D) UMAP plot and Leiden clustering of all avPOA Vglut2 neurons identified in Hrvatin et al., 2020. (E) UMAP plot of avPOA Vglut2 neurons colored by Adcyap1 and LepR expression. (F) Schematic showing four groups of animals: mCherry (mCherry-transduced Vglut2-IRES-Cre, n = 11), Vglut2 (Gq-DREADD transduced Vglut2-IRES-Cre ) (n=10), PACAP (Gq-DREADD-transduced PACAP-2A-Cre ) (n=11), and LepR (Gq-DREADD-transduced LepR-IRES-Cre (n=3) animals injected with 1 mg/kg CNO. (G) Glucose tolerance test of mCherry (avPOA Vglut2-mCherry ), Vglut2 (avPOA Vglut2-Gq ), PACAP (avPOA PACAP-Gq ), and LepR (avPOA LepR-Gq ), animals from (F). (H) AUC analysis for (G) (** = p < 0.05, student’s two-tailed t-test, mean ±SEM, n=7 avPOA Vglut2-mCherry , n = 10 avPOA Vglut2-Gq , n = 11 avPOA PACAP-Gq , n = 3 avPOA LepR-Gq animals). ( I) Volcano Plot of 120-minute polar metabolomics of PACAP-2A-Cre animals transduced with a Cre-dependent Gq-DREADD and injected with either PBS or CNO (1 mg/kg). Animals were treated as in . Selected overrepresented and underrepresented metabolites labeled in red and blue, respectively. p-values calculated using two-tailed t-test; red lines corresponding to p-value and Log 2 -Fold changes set at p < 0.05 and |Log 2 -Fold change| > 0.6.
Article Snippet: For natural, chemogenetic, and optogenetic torpor experiments, we used 8-12-week-old Vglut2-IRES-Cre (The Jackson Laboratory, Stock 028863), Adcyap1-2A-Cre (The Jackson Laboratory, Stock 030155), and Lepr-IRES-Cre (The Jackson Laboratory, Strain 008320) mice.
Techniques: Immunostaining, Transduction, Two Tailed Test, Expressing, Injection, Labeling
