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

Creators: Jovanovic, Predrag; Pool, Allan-Hermann; Morones, Nancy; Wang, Yidan; Novinbakht, Edward; Keshishian, Nareg; Jang, Kaitlyn; Oka, Yuki¹; Riera, Celine E.

1. California Institute of Technology

Abstract

Olfactory cues are vital for prey animals like rodents to perceive and evade predators. Stress-induced hyperthermia, via brown adipose tissue (BAT) thermogenesis, boosts physical performance and facilitates escape. However, many aspects of this response, including thermogenic control and sex-specific effects, remain enigmatic. Our study unveils that the predator odor trimethylthiazoline (TMT) elicits BAT thermogenesis, suppresses feeding, and drives glucocorticoid release in female mice. Chemogenetic stimulation of olfactory bulb (OB) mitral cells recapitulates the thermogenic output of this response and associated stress hormone corticosterone release in female mice. Neuronal projections from OB to medial amygdala (MeA) and dorsomedial hypothalamus (DMH) exhibit female-specific cFos activity toward odors. Cell sorting and single-cell RNA-sequencing of DMH identify cholecystokinin (CCK)-expressing neurons as recipients of predator odor cues. Chemogenetic manipulation and neuronal silencing of DMH^(CCK) neurons further implicate these neurons in the propagation of predator odor-associated thermogenesis and food intake suppression, highlighting their role in female stress-induced hyperthermia.

Copyright and License

© The Author(s) 2023. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Acknowledgement

We thank the Applied Genomics core at CSMC for their assistance in processing RNA-seq, the Biobehavioral core for assisting with open field testing and the Flow Cytometry core for cell sorting. We thank Joshua Breunig for his assistance with light-sheet imaging, Michael Ramos and Ritchie Ho for their advice toward scRNA analysis. This work was supported by the American Diabetes Association Pathway to Stop Diabetes Grant 1-15-INI-12 (C.E.R.), the Klingenstein-Simons foundation (C.E.R.), the Larry L Hillblom Foundation fellowship 2019-D-014-FEL (P.J.) and the Cedars-Sinai Center for Research in Women's health and Sex Differences pilot award (C.E.R). Schematic cartoons were created with BioRender.com.

Contributions

C.E.R. and P.J. conceived the studies. P.J. performed experiments with assistance from Y.W., E.N., N.K. and K.J. A.H.P. and Y.O. executed the scRNAseq experiments, and bioinformatics analysis was conducted by P.J. N.M. and A.H.P. C.E.R. and P.J. wrote the manuscript with assistance from Y.O. and A.H.P. C.E.R. supervised the entire study.

Data Availability

All data generated and analyzed during this study are included in this paper and its supplementary information files. All sc-RNA seq data of mouse dorsomedial hypothalamus that are used in this study have been deposited in the the National Center for Biotechnology Information Gene Expression Omnibus (GEO) and are accessible through the GEO Series accession number: GSE232230. Mouse reference transcriptome v.2 used in scRNA-seq analysis is available for download at www.thepoollab.org/resources and custom mouse reference genome GRCm38.p5 used for bulk RNA-seq can be found under RefSeq assembly accession: GCF_000001635.25. Source data are provided with this paper.

Code Availability

The R code used to perform the scRNA-seq analysis is available from the corresponding author on request.

Conflict of Interest

The authors declare no competing interests.

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