Dopamine-inhibited POMCDrd2+ neurons in the ARC acutely regulate feeding and body temperature
Isabella Gaziano, Svenja Corneliussen, Nasim Biglari, René Neuhaus, Linyan Shen, Tamara Sotelo-Hitschfeld, Paul Klemm, Lukas Steuernagel, Alain J. De Solis, Weiyi Chen, F. Thomas Wunderlich, Peter Kloppenburg, Jens C. Brüning
Isabella Gaziano, Svenja Corneliussen, Nasim Biglari, René Neuhaus, Linyan Shen, Tamara Sotelo-Hitschfeld, Paul Klemm, Lukas Steuernagel, Alain J. De Solis, Weiyi Chen, F. Thomas Wunderlich, Peter Kloppenburg, Jens C. Brüning
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Research Article Endocrinology Neuroscience

Dopamine-inhibited POMCDrd2+ neurons in the ARC acutely regulate feeding and body temperature

Abstract

Dopamine acts on neurons in the arcuate nucleus (ARC) of the hypothalamus, which controls homeostatic feeding responses. Here we demonstrate a differential enrichment of dopamine receptor 1 (Drd1) expression in food intake–promoting agouti related peptide (AgRP)/neuropeptide Y (NPY) neurons and a large proportion of Drd2-expressing anorexigenic proopiomelanocortin (POMC) neurons. Owing to the nature of these receptors, this translates into a predominant activation of AgRP/NPY neurons upon dopamine stimulation and a larger proportion of dopamine-inhibited POMC neurons. Employing intersectional targeting of Drd2-expressing POMC neurons, we reveal that dopamine-mediated POMC neuron inhibition is Drd2 dependent and that POMCDrd2+ neurons exhibit differential expression of neuropeptide signaling mediators compared with the global POMC neuron population, which manifests in enhanced somatostatin responsiveness of POMCDrd2+ neurons. Selective chemogenetic activation of POMCDrd2+ neurons uncovered their ability to acutely suppress feeding and to preserve body temperature in fasted mice. Collectively, the present study provides the molecular and functional characterization of POMCDrd2+ neurons and aids our understanding of dopamine-dependent control of homeostatic energy-regulatory neurocircuits.

Authors

Isabella Gaziano, Svenja Corneliussen, Nasim Biglari, René Neuhaus, Linyan Shen, Tamara Sotelo-Hitschfeld, Paul Klemm, Lukas Steuernagel, Alain J. De Solis, Weiyi Chen, F. Thomas Wunderlich, Peter Kloppenburg, Jens C. Brüning

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Figure 4

Combinatorial use of Cre and Dre recombinases allows for intersectional targeting of POMCDrd2+ neurons.

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Combinatorial use of Cre and Dre recombinases allows for intersectional ...
(A) RNA in situ hybridization against Pomc, Th, and Drd2 in ARC of C57BL/6N mice. Scale bar: 50 μm. (B and C) Colocalization percentages of Drd2 with ARC TH (B) or POMC (C) neurons as quantified from RNA in situ hybridization in C57BL/6N mice (A). Overlap of TH neurons with POMC neurons is shown in C. Data are represented as mean ± SEM; n = 4. No statistical tests were applied. (D) Immunofluorescence staining against POMC and ZsGreen in ARC of POMCDre Drd2Cre R26-lx-rx-ZsGreen mice. Scale bar: 50 μm. (E) Percentage of ZsGreen+ neurons over total POMC neuronal population in male and female POMCDre Drd2Cre R26-lx-rx-ZsGreen mice as quantified from immunofluorescence staining (D). Data are represented as mean ± SEM; n = 4. P values were calculated using unpaired 2-tailed Student’s t tests. (F) Representative perforated patch-clamp recordings with rate histograms (top) and original traces (bottom) of a single POMCDrd2+ neuron with increasing dopamine concentrations (0.3 μM, 3 μM, 10 μM, 30 μM) and subsequent application of the Drd2 agonist quinpirole at 10 μM. Data are also shown as part of G at the indicated row. (G) Heatmap of 11 perforated patch-clamp recordings of POMCDrd2+ neurons in male and female POMCDre Drd2Cre R26-lx-rx-ZsGreen mice. Changes in action potential frequency from baseline and corresponding z scores with increasing dopamine concentrations (0.3 μM, 3 μM, 10 μM, 30 μM) are depicted on the left and right, respectively. (H and I) Statistical quantification of dopamine responses (H) from G or quinpirole responses (I) in POMCDrd2+ neurons. A neuron was considered responsive if the change in firing frequency induced by drug application was 3 times larger than the SD.