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
View: Text | PDF
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

×

Figure 5

Translational profiling of POMCDrd2+ neurons indicates enriched Sst responsiveness compared with Drd2– POMC neurons.

Options: View larger image (or click on image) Download as PowerPoint
Translational profiling of POMCDrd2+ neurons indicates enriched Sst resp...
(A and B) Immunofluorescence staining in ARC of POMCDre Drd2Cre R26-lx-rx-EGFP-L10a mice. (A) Scale bar: 50 μm. (B) Percentage of GFP+ POMC neurons. Data are represented as mean ± SEM; n = 4. P values were calculated using unpaired 2-tailed Student’s t tests. **P < 0.01. (C) Differentially enriched genes in immunoprecipitation (IP) versus input (IN) of bacTRAP hypothalamus samples of POMCDre Drd2Cre R26-lx-rx-EGFP-L10a mice. n = 3 replicates from N = 56 mice. Differential gene expression analysis was performed using the DESeq2 1.28.0 (62) R package. (D) Overlap of differentially enriched genes in bacterial artificial chromosome — translating ribosome affinity purification (bacTRAP) samples of POMCDre Drd2Cre R26-lx-rx-EGFP-L10a and POMCDre R26-rx-EGFP-L10a mice. (E) Gene enrichment in POMCDrd2+ versus whole POMC population. Genes belonging to GO term neuropeptide signaling pathway (GO:0007218) are highlighted. For POMCDrd2+: n = 3 replicates from N = 56 mice. For POMC: n = 3 replicates from N = 9 mice. (F–H) RNA in situ hybridization in ARC of C57BL/6N mice. (F) POMCDrd2– neurons are outlined in white, POMCDrd2+ neurons in magenta. Scale bars: 50 μm (left), 20 μm (right). (G and H) Signal intensity quantification of Drd2 (G) and Sstr1 (H). Data are represented as median ± SEM. n = 1,301–2,961 POMCDrd2– and n = 674–1,468 POMCDrd2+ cells from N = 5 mice. P values were calculated using 2-tailed Mann-Whitney rank tests. ****P < 0.0001. (I and J) Perforated patch clamp recordings of Sst-treated POMC neurons (I) or POMCDrd2+ neurons (J). A neuron was considered responsive if the change in firing frequency induced by drug application was 3 times larger than the SD. (K and L) Reclustering of Sst neurons from Campbell et al. (2) (K) and corresponding Th RNA expression levels (L). (M and N) RNA in situ hybridization in C57BL/6N mice. (M) Percentages of Sst and Th coexpression in hypothalamic and subthalamic dopaminergic brain regions. Data are represented as mean ± SEM; n = 5. No statistical tests were applied. mt, mammillothalamic tract; A11, A11 dopamine cells. (N) Representative image of A11 dopaminergic cell group. Scale bar: 50 μm.