Functional identity of hypothalamic melanocortin neurons depends on Tbx3

Abstract

Heterogeneous populations of hypothalamic neurons orchestrate energy balance via the release of specific signatures of neuropeptides. However, how specific intracellular machinery controls peptidergic identities and function of individual hypothalamic neurons remains largely unknown. The transcription factor T-box 3 (Tbx3) is expressed in hypothalamic neurons sensing and governing energy status, whereas human TBX3 haploinsufficiency has been linked with obesity. Here, we demonstrate that loss of Tbx3 function in hypothalamic neurons causes weight gain and other metabolic disturbances by disrupting both the peptidergic identity and plasticity of Pomc/Cart and Agrp/Npy neurons. These alterations are observed after loss of Tbx3 in both immature hypothalamic neurons and terminally differentiated mouse neurons. We further establish the importance of Tbx3 for body weight regulation in Drosophila melanogaster and show that TBX3 is implicated in the differentiation of human embryonic stem cells into hypothalamic Pomc neurons. Our data indicate that Tbx3 directs the terminal specification of neurons as functional components of the melanocortin system and is required for maintaining their peptidergic identity. In summary, we report the discovery of a key mechanistic process underlying the functional heterogeneity of hypothalamic neurons governing body weight and systemic metabolism.

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Fig. 1: Loss of Tbx3 in hypothalamic neurons promotes obesity.
Fig. 2: Loss of Tbx3 in Pomc but not Agrp neurons triggers obesity.
Fig. 3: Loss of Tbx3 impairs the postnatal melanocortin system.
Fig. 4: Tbx3 is critical for the differentiation of Pomc neurons.
Fig. 5: Tbx3 functions in Drosophila and human neurons.

Data availability

The authors declare that all data supporting the findings of this study are available within the paper and its supplementary information files. The RNA-seq database generated in our paper has been made publicly available through Gene Expression Omnibus (GEO accession number GSE119883).

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Acknowledgements

We thank A. Kispert and M.-O. Trowe (Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany) for kindly providing Tbx3-deficient embryos, J. Friedman (The Rockefeller University, Howard Hughes Medical Institute, New York, NY, USA) for scientific guidance and for graciously providing access to data shown in ref. 5, M. Guzmán (Complutense University, Madrid, Spain) for assistance with the generation of AAV-GFP and AAV-Cre viral particles, the Bloomington Drosophila Stock Center (BDSC) (NIH P40OD018537) for fly stocks, and C. Layritz, H. Hoffmann, N. Wiegert and C. L. Holleman for technical assistance and assistance with animal studies. A.F. is supported by a postoctoral fellowship from the Canadian Institutes of Health Research (Funding reference no. 152588). V.V.T. is supported by NIH-NIDDK grant 5K23DK110539 and in part by the Baylor-Hopkins Center for Mendelian Genomics through NHGRI grant 5U54HG006542. C.A.D. is supported by funding from the NIH (R01 DK52431, R01 DK110113 and P30 DK26687) and Columbia Stem Cell Initiative Seed Fund Program. We thank the Fondation Recherche Medicale (ARF20140129235, L.B.). This work was strongly supported by the Helmholtz Alliance ICEMED & the Helmholtz Initiative on Personalized Medicine iMed by Helmholtz Association. This work was supported in part by the Helmholtz cross-program topic ‘Metabolic Dysfunction’, the European Research Council ERC (AdG HypoFlam no. 695054) and in part by funding to M.H.T., Y.L., B.L. and V.K. from the Alexander von Humboldt Foundation.

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C.Q. and A.F. designed and performed the experiments and interpreted the data. Y.X., G.C., B.L. Y.-T.T., A.R., M.W., M.C.D., V.K., R.R., V.V.T., E.G., T.M.S., A.-L.P., T.G., O.L., A.C.-S., D.K., L.B., S.C.W., G.O.P., R.N., L.Z., I.C.G.K., A.M., C.G.-C., M.M., M.T. and C.A.D. performed experiments and/or edited the manuscript. M.H.T. conceptualized the project, interpreted the data, and cowrote the manuscript together with C.Q. and A.F.

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Correspondence to Matthias H. Tschöp.

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Supplementary Information

Supplementary Figures 1–6

Reporting Summary

Supplementary Table 1

qRT–PCR primer sequences

Supplementary Table 2

Reference proteins and proteins enriched with Tbx3 immunoprecipitation

Supplementary Table 3

Lineage tracing and neuropeptide expression.

Supplementary Table 4

Full vector sequence of pCas9_CD4

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Quarta, C., Fisette, A., Xu, Y. et al. Functional identity of hypothalamic melanocortin neurons depends on Tbx3. Nat Metab 1, 222–235 (2019). https://doi.org/10.1038/s42255-018-0028-1

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