Functionally distinct POMC-expressing neuron subpopulations in hypothalamus revealed by intersectional targeting

Nasim Biglari,Jonas Schumacher,Jens C Brüning,Tim Klöckener,Henning Fenselau,Svenja Corneliussen,Claudia M Wunderlich,Weiyi Chen,Lars Paeger,Stefan Vollmar,Fiona M Gribble,Tamara Sotelo-Hitschfeld,Jan Radermacher,Peter Kloppenburg,Frank Reimann,Isabella Gaziano,Frank Wunderlich ,Paul Klemm,Frank Edenhofer,Amin Abbasloo,Michael Sué

doi:10.1038/s41593-021-00854-0

Abstract

Pro-opiomelanocortin (POMC)-expressing neurons in the arcuate nucleus of the hypothalamus represent key regulators of metabolic homeostasis. Electrophysiological and single-cell sequencing experiments have revealed a remarkable degree of heterogeneity of these neurons. However, the exact molecular basis and functional consequences of this heterogeneity have not yet been addressed. Here, we have developed new mouse models in which intersectional Cre/Dre-dependent recombination allowed for successful labeling, translational profiling and functional characterization of distinct POMC neurons expressing the leptin receptor (Lepr) and glucagon like peptide 1 receptor (Glp1r). Our experiments reveal that POMCLepr+ and POMCGlp1r+ neurons represent largely nonoverlapping subpopulations with distinct basic electrophysiological properties. They exhibit a specific anatomical distribution within the arcuate nucleus and differentially express receptors for energy-state communicating hormones and neurotransmitters. Finally, we identify a differential ability of these subpopulations to suppress feeding. Collectively, we reveal a notably distinct functional microarchitecture of critical metabolism-regulatory neurons.

Highlights

Pro-opiomelanocortin (POMC)-expressing neurons in the arcuate nucleus of the hypothalamus represent key regulators of metabolic homeostasis
30% of POMC neurons increase firing in response to leptin in brain slice electrophysiology, and this response does not overlap with POMC cells in which insulin modulates firing[9]
Having confirmed the specific expression of the EGFPL10a fusion protein using the ROSA26lSlrSrEGFPL10a mouse line (Extended Data Fig. 5c) and validated the successful, specific pulldown of RNA of subpopulations of POMC neurons, our aim was to compare the ribosome-associated transcriptome of POMCLepr+ neurons with POMCGlp1r+ neurons

Summary

Introduction

Pro-opiomelanocortin (POMC)-expressing neurons in the arcuate nucleus of the hypothalamus represent key regulators of metabolic homeostasis. Our experiments reveal that POMCLepr+ and POMCGlp1r+ neurons represent largely nonoverlapping subpopulations with distinct basic electrophysiological properties They exhibit a specific anatomical distribution within the arcuate nucleus and differentially express receptors for energy-state communicating hormones and neurotransmitters. The melanocortin circuitry comprising agouti-related peptide (AgRP)-expressing neurons and POMC-expressing neurons in the arcuate nucleus of the hypothalamus (ARC) represents a prototypic homeostatic regulatory neurocircuit of metabolic homeostasis[1] These targets of insulin, leptin and glucagon like peptide (Glp1) integrate multiple inputs to compute the energy state of the organism and adapt feeding behavior[2]. Cre-loxP-mediated recombination provides a critical repertoire for cell-type-specific inactivation or activation of genes or transgenes[13] These transgenes may represent genetically encoded fluorophores, expression of chemogenetically modifiable designer G-protein-coupled receptors, hM3Dq or light-regulated ion channels for remotely controlled activation or inhibition of Cre-expressing neurons in vivo[14,15]. An additional, complementary recombinase system that builds on We have dedicated our efforts toward further developing a toolexpression of Dre-recombinase, an alternative and complementary box of transgenic mice that offers the opportunity to genetically approach has been defined[17]

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