Dopamine-induced interactions of female mouse hypothalamic proteins with progestin receptor-A in the absence of hormone

Kalpana D Acharya; Sabin A Nettles; Cheryl F Lichti; Katherine Warre-Cornish; Lucia Dutan Polit; Deepak P Srivastava; Larry Denner; Marc J Tetel

doi:10.1111/jne.12904

Dopamine-induced interactions of female mouse hypothalamic proteins with progestin receptor-A in the absence of hormone

J Neuroendocrinol. 2020 Oct;32(10):e12904. doi: 10.1111/jne.12904. Epub 2020 Sep 30.

Authors

Kalpana D Acharya¹, Sabin A Nettles¹, Cheryl F Lichti², Katherine Warre-Cornish^{3

4}, Lucia Dutan Polit^{3

4}, Deepak P Srivastava^{3

4}, Larry Denner⁵, Marc J Tetel¹

Affiliations

¹ Neuroscience Department, Wellesley College, Wellesley, MA, USA.
² Department of Pathology and Immunology, Washington University School of Medicine, St Louis, MO, USA.
³ Department of Basic and Clinical Neuroscience, The Maurice Wohl Clinical Neuroscience Institute, Institute of Psychiatry Psychology and Neuroscience, King's College London, London, UK.
⁴ MRC Centre for Neurodevelopmental Disorders, King's College London, London, UK.
⁵ Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX, USA.

Abstract

Neural progestin receptors (PR) function in reproduction, neural development, neuroprotection, learning, memory and the anxiety response. In the absence of progestins, PR can be activated by dopamine (DA) in the rodent hypothalamus to elicit female sexual behaviour. The present study investigated mechanisms of DA activation of PR by testing the hypothesis that proteins from DA-treated hypothalami interact with PR in the absence of progestins. Ovariectomised, oestradiol-primed mice were infused with a D1-receptor agonist, SKF38393 (SKF), into the third ventricle 30 minutes prior to death. Proteins from SKF-treated hypothalami were pulled-down with glutathione S-transferase-tagged mouse PR-A or PR-B and the interactomes were analysed by mass spectrometry. The largest functional group to interact with PR-A in a DA-dependent manner was synaptic proteins. To test the hypothesis that DA activation of PR regulates synaptic proteins, we developed oestradiol-induced PR-expressing hypothalamic-like neurones derived from human-induced pluripotent stem cells (hiPSCs). Similar to progesterone (P4), SKF treatment of hiPSCs increased synapsin1/2 expression. This SKF-dependent effect was blocked by the PR antagonist RU486, suggesting that PR are necessary for this DA-induced increase. The second largest DA-dependent PR-A protein interactome comprised metabolic regulators involved in glucose metabolism, lipid synthesis and mitochondrial energy production. Interestingly, hypothalamic proteins interacted with PR-A, but not PR-B, in an SKF-dependent manner, suggesting that DA promotes the interaction of multiple hypothalamic proteins with PR-A. These in vivo and in vitro results indicate novel mechanisms by which DA can differentially activate PR isoforms in the absence of P4 and provide a better understanding of ligand-independent PR activation in reproductive, metabolic and mental health disorders in women.

Keywords: energy homeostasis; induced pluripotent stem cells; metabolism; progesterone; synapse.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Dopamine / pharmacology*
Female
Hypothalamus / drug effects
Hypothalamus / metabolism
Mice
Mice, Inbred C57BL
Nerve Tissue Proteins / metabolism*
Neurons / drug effects
Neurons / metabolism
Progesterone / pharmacology
Protein Binding / drug effects
Protein Isoforms / drug effects
Protein Isoforms / metabolism
Receptors, Progesterone / drug effects
Receptors, Progesterone / metabolism*
Signal Transduction / drug effects

Substances

Nerve Tissue Proteins
Protein Isoforms
Receptors, Progesterone
Progesterone
Dopamine

Abstract

Publication types

MeSH terms

Substances

Grants and funding