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Estrogen inhibits NPY secretion through membrane-associated estrogen receptor (ER)-α in clonal, immortalized hypothalamic neurons

International Journal of Obesity volume 35, pages 198–207 (2011)Cite this article

Subjects

Abstract

Objective:

Estrogen (E2) has an inhibitory effect on food intake by acting centrally in the hypothalamus, although it is not clear which hypothalamic neurons are involved in this process. Earlier studies from our lab and others have implicated neuropeptide Y (NPY) as an important central anorexigenic target of E2. This study was designed to investigate whether E2 can directly regulate NPY secretion and examine the cellular mechanisms and receptors responsible for this anorexigenic action of E2.

Design:

Clonal, murine, hypothalamic neuronal cell models, mHypoE-42 and mHypoA-2/12, were investigated for NPY secretory responses to 17β-estradiol (E2) in the presence or absence of pharmacological inhibitors directed against the phosphatidylinositol-3-kinase (PI3K), mitogen-activated protein kinase (MAPK) and AMP-activated kinase (AMPK) pathways or to estrogen receptor (ER) specific agonists/antagonists.

Measurements:

The presence of hypothalamic markers and characterization of neuronal cell lines was completed with polymerase chain reaction. NPY levels were measured using an enzyme immunoassay (EIA). The expression of ER-α and caveolin-1 was analyzed using immunocytochemistry.

Results:

E2 significantly decreased NPY secretion in both the mHypoE-42 and mHypoA-2/12 neurons. The E2-mediated repression of NPY secretion in the mHypoE-42 and mHypoA-2/12 neurons required ER-α, but not ER-β, as shown by studies using an ER-specific agonist/antagonists. Additionally, using immunocytochemistry we detected colocalization of ER-α and the membrane-associated signaling protein caveolin-1. Importantly, using E2-conjugated bovine serum albumin (E2-BSA) and ER antagonists, we were able to show that the E2-mediated decrease in NPY secretion occurred through membrane-bound ER-α. Finally, using a combination of pharmacological inhibitors, we found that inhibition of the PI3K or AMPK pathway blocked the E2-mediated decrease in NPY secretion.

Conclusion:

These findings indicate that the central anorexigenic action of E2 occurs at least partially through hypothalamic NPY-synthesizing neurons. This regulation of NPY secretion occurs through rapid signaling mechanisms through membrane bound ER-α.

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Acknowledgements

Support: This work was supported by the Canadian Institutes for Health Research (CIHR) Operating Grants. D.D.B. holds a Canada Research Chair in Neuroendocrinology.

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Authors and Affiliations

  1. Department of Physiology, Toronto General Hospital Research Institute, University Health Network, Toronto, ON, Canada

    S S Dhillon & D D Belsham

  2. Department of Obstetrics and Gynaecology and Medicine, University of Toronto and Division of Cellular and Molecular Biology, Toronto General Hospital Research Institute, University Health Network, Toronto, ON, Canada

    D D Belsham

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Correspondence to D D Belsham.

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Dhillon, S., Belsham, D. Estrogen inhibits NPY secretion through membrane-associated estrogen receptor (ER)-α in clonal, immortalized hypothalamic neurons. Int J Obes 35, 198–207 (2011). https://doi.org/10.1038/ijo.2010.124

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