Abstract
Glucagon-like peptide 1 (GLP-1) and its agonists exert anorexigenic effect at least partly via acting on GLP-1 receptors (GLP-1R) in the arcuate nucleus (ARC). While the anorexigenic, proopiomelanocortin (POMC) neurons of the ARC were shown previously to express GLP-1R, the putative GLP-1R-content of the orexigenic, neuropeptide Y (NPY) neurons remained so far undetected. As GLP-1R is abundant in the ventromedial ARC, where NPY neurons are located; here, we address the possibility that GLP-1 can act directly on the orexigenic NPY system via GLP-1R. Double-labeling immunocytochemistry and in situ hybridization were performed on tissues of adult male mice to detect GLP-1R in NPY neurons. In double-immunolabeled preparations, GLP-1R-immunoreactivity was observed in NPY neurons and in axons ensheathing the majority of NPY neurons. Ultrastructural studies confirmed that GLP-1R-immunoreactivity is associated with the outer membrane of NPY perikarya as well as with axons forming symmetric type, inhibitory synapses on NPY-containing neurons. Double-labeling in situ hybridization experiments demonstrated the expression of GLP-1R mRNA in approximately 20% of NPY mRNA-containing neurons of the ARC. In summary, our data demonstrate the presence of GLP-1R protein and mRNA in NPY neurons of ARC and also reveal the innervation of NPY neurons by GLP-1R-containing inhibitory neurons. These observations suggest that GLP-1 signaling can influence NPY neurons both directly and indirectly. Furthermore, GLP-1 signaling on energy homeostasis appears to involve both direct and indirect effects of GLP-1 on the orexigenic NPY neurons, in addition to the previously known effects via the anorexigenic POMC neuronal system.
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Acknowledgements
The authors would like to express their appreciation to Andrea Juhász and Veronika Penksza for their great technical assistance.
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This work was supported by Grants from the Hungarian Science Foundation (OTKA K124767 and K128317), Hungarian National Brain Research Program (2017–1.2.1-NKP-2017–00002), EU H2020 THYRAGE no. 666869.
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YR performed immunohistochemical, ultrastructural and in situ hybridization studies, contributed to data analysis. AS-S performed ultrastructural studies, analyzed data and was involved in the preparation of manuscript. DK was involved in the in situ hybridization studies. AK was involved in the ultrastructural studies. LM, RS, EH and BG generated the probes for in situ hybridization. CF planned and oversaw the experiments, interpreted data and was involved in the preparation of the manuscript.
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C.F. received funding from Novo Nordisk A/S. Y.R., A.Sz-Sz., D.K., A.K., L.M., R.S., E.H., and B.G. declare no competing interest.
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All experimental protocols were reviewed and approved by the Animal Welfare Committee at the Institute of Experimental Medicine. The study was performed in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments.
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Ruska, Y., Szilvásy-Szabó, A., Kővári, D. et al. Expression of glucagon-like peptide 1 receptor in neuropeptide Y neurons of the arcuate nucleus in mice. Brain Struct Funct 227, 77–87 (2022). https://doi.org/10.1007/s00429-021-02380-y
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DOI: https://doi.org/10.1007/s00429-021-02380-y