The progestin receptor interactome in the female mouse hypothalamus

Interactions with synaptic proteins are isoform specific and ligand dependent

Kalpana D. Acharya, Sabin A. Nettles, Katherine J. Sellers, Dana D. Im, Moriah Harling, Cassandra Pattanayak, Didem Vardar-Ulu, Cheryl F. Lichti, Shixia Huang, Dean P. Edwards, Deepak P. Srivastava, Larry Denner, Marc J. Tetel

Research output: Contribution to journalArticle

Abstract

Progestins bind to the progestin receptor (PR) isoforms, PR-A and PR-B, in brain to influence development, female reproduction, anxiety, and stress. Hormone-activated PRs associate with multiple proteins to form functional complexes. In the present study, proteins from female mouse hypothalamus that associate with PR were isolated using affinity pull-down assays with glutathione S-transferase–tagged mouse PR-A and PR-B. Using complementary proteomics approaches, reverse phase protein array (RPPA) and mass spectrometry, we identified hypothalamic proteins that interact with PR in a ligand-dependent and isoform-specific manner and were confirmed by Western blot. Synaptic proteins, including synapsin-I and synapsin-II, interacted with agonist-bound PR isoforms, suggesting that both isoforms function in synaptic plasticity. In further support, synaptogyrin-III and synapsin-III associated with PR-A and PR-B, respectively. PR also interacted with kinases, including c-Src, mTOR, and MAPK1, confirming phosphorylation as an integral process in rapid effects of PR in the brain. Consistent with a role in transcriptional regulation, PR associated with transcription factors and coactivators in a ligand-specific and isoform-dependent manner. Interestingly, both PR isoforms associated with a key regulator of energy homeostasis, FoxO1, suggesting a novel role for PR in energy metabolism. Because many identified proteins in this PR interactome are synaptic proteins, we tested the hypothesis that progestins function in synaptic plasticity. Indeed, progesterone enhanced synaptic density, by increasing synapsin-I–positive synapses, in rat primary cortical neuronal cultures. This novel combination of RPPA and mass spectrometry allowed identification of PR action in synaptic remodeling and energy homeostasis and reveals unique roles for progestins in brain function and disease.

Original languageEnglish (US)
Article numbere0272-17.2017
JournaleNeuro
Volume4
Issue number5
DOIs
StatePublished - Sep 1 2017

Fingerprint

Progesterone Receptors
Hypothalamus
Protein Isoforms
Ligands
Synapsins
Progestins
Protein Array Analysis
Proteins
Neuronal Plasticity
Synaptogyrins
Mass Spectrometry
Homeostasis
Brain
Brain Diseases
Proteomics
Synapses
Energy Metabolism

Keywords

  • Cortex
  • Estrogen
  • Progesterone
  • Proteomics
  • Synapse
  • Synapsin

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Acharya, K. D., Nettles, S. A., Sellers, K. J., Im, D. D., Harling, M., Pattanayak, C., ... Tetel, M. J. (2017). The progestin receptor interactome in the female mouse hypothalamus: Interactions with synaptic proteins are isoform specific and ligand dependent. eNeuro, 4(5), [e0272-17.2017]. https://doi.org/10.1523/ENEURO.0272-17.2017

The progestin receptor interactome in the female mouse hypothalamus : Interactions with synaptic proteins are isoform specific and ligand dependent. / Acharya, Kalpana D.; Nettles, Sabin A.; Sellers, Katherine J.; Im, Dana D.; Harling, Moriah; Pattanayak, Cassandra; Vardar-Ulu, Didem; Lichti, Cheryl F.; Huang, Shixia; Edwards, Dean P.; Srivastava, Deepak P.; Denner, Larry; Tetel, Marc J.

In: eNeuro, Vol. 4, No. 5, e0272-17.2017, 01.09.2017.

Research output: Contribution to journalArticle

Acharya, KD, Nettles, SA, Sellers, KJ, Im, DD, Harling, M, Pattanayak, C, Vardar-Ulu, D, Lichti, CF, Huang, S, Edwards, DP, Srivastava, DP, Denner, L & Tetel, MJ 2017, 'The progestin receptor interactome in the female mouse hypothalamus: Interactions with synaptic proteins are isoform specific and ligand dependent', eNeuro, vol. 4, no. 5, e0272-17.2017. https://doi.org/10.1523/ENEURO.0272-17.2017
Acharya, Kalpana D. ; Nettles, Sabin A. ; Sellers, Katherine J. ; Im, Dana D. ; Harling, Moriah ; Pattanayak, Cassandra ; Vardar-Ulu, Didem ; Lichti, Cheryl F. ; Huang, Shixia ; Edwards, Dean P. ; Srivastava, Deepak P. ; Denner, Larry ; Tetel, Marc J. / The progestin receptor interactome in the female mouse hypothalamus : Interactions with synaptic proteins are isoform specific and ligand dependent. In: eNeuro. 2017 ; Vol. 4, No. 5.
@article{2b75af1b522143d2a22623054e258a45,
title = "The progestin receptor interactome in the female mouse hypothalamus: Interactions with synaptic proteins are isoform specific and ligand dependent",
abstract = "Progestins bind to the progestin receptor (PR) isoforms, PR-A and PR-B, in brain to influence development, female reproduction, anxiety, and stress. Hormone-activated PRs associate with multiple proteins to form functional complexes. In the present study, proteins from female mouse hypothalamus that associate with PR were isolated using affinity pull-down assays with glutathione S-transferase–tagged mouse PR-A and PR-B. Using complementary proteomics approaches, reverse phase protein array (RPPA) and mass spectrometry, we identified hypothalamic proteins that interact with PR in a ligand-dependent and isoform-specific manner and were confirmed by Western blot. Synaptic proteins, including synapsin-I and synapsin-II, interacted with agonist-bound PR isoforms, suggesting that both isoforms function in synaptic plasticity. In further support, synaptogyrin-III and synapsin-III associated with PR-A and PR-B, respectively. PR also interacted with kinases, including c-Src, mTOR, and MAPK1, confirming phosphorylation as an integral process in rapid effects of PR in the brain. Consistent with a role in transcriptional regulation, PR associated with transcription factors and coactivators in a ligand-specific and isoform-dependent manner. Interestingly, both PR isoforms associated with a key regulator of energy homeostasis, FoxO1, suggesting a novel role for PR in energy metabolism. Because many identified proteins in this PR interactome are synaptic proteins, we tested the hypothesis that progestins function in synaptic plasticity. Indeed, progesterone enhanced synaptic density, by increasing synapsin-I–positive synapses, in rat primary cortical neuronal cultures. This novel combination of RPPA and mass spectrometry allowed identification of PR action in synaptic remodeling and energy homeostasis and reveals unique roles for progestins in brain function and disease.",
keywords = "Cortex, Estrogen, Progesterone, Proteomics, Synapse, Synapsin",
author = "Acharya, {Kalpana D.} and Nettles, {Sabin A.} and Sellers, {Katherine J.} and Im, {Dana D.} and Moriah Harling and Cassandra Pattanayak and Didem Vardar-Ulu and Lichti, {Cheryl F.} and Shixia Huang and Edwards, {Dean P.} and Srivastava, {Deepak P.} and Larry Denner and Tetel, {Marc J.}",
year = "2017",
month = "9",
day = "1",
doi = "10.1523/ENEURO.0272-17.2017",
language = "English (US)",
volume = "4",
journal = "eNeuro",
issn = "2373-2822",
publisher = "Society for Neuroscience",
number = "5",

}

TY - JOUR

T1 - The progestin receptor interactome in the female mouse hypothalamus

T2 - Interactions with synaptic proteins are isoform specific and ligand dependent

AU - Acharya, Kalpana D.

AU - Nettles, Sabin A.

AU - Sellers, Katherine J.

AU - Im, Dana D.

AU - Harling, Moriah

AU - Pattanayak, Cassandra

AU - Vardar-Ulu, Didem

AU - Lichti, Cheryl F.

AU - Huang, Shixia

AU - Edwards, Dean P.

AU - Srivastava, Deepak P.

AU - Denner, Larry

AU - Tetel, Marc J.

PY - 2017/9/1

Y1 - 2017/9/1

N2 - Progestins bind to the progestin receptor (PR) isoforms, PR-A and PR-B, in brain to influence development, female reproduction, anxiety, and stress. Hormone-activated PRs associate with multiple proteins to form functional complexes. In the present study, proteins from female mouse hypothalamus that associate with PR were isolated using affinity pull-down assays with glutathione S-transferase–tagged mouse PR-A and PR-B. Using complementary proteomics approaches, reverse phase protein array (RPPA) and mass spectrometry, we identified hypothalamic proteins that interact with PR in a ligand-dependent and isoform-specific manner and were confirmed by Western blot. Synaptic proteins, including synapsin-I and synapsin-II, interacted with agonist-bound PR isoforms, suggesting that both isoforms function in synaptic plasticity. In further support, synaptogyrin-III and synapsin-III associated with PR-A and PR-B, respectively. PR also interacted with kinases, including c-Src, mTOR, and MAPK1, confirming phosphorylation as an integral process in rapid effects of PR in the brain. Consistent with a role in transcriptional regulation, PR associated with transcription factors and coactivators in a ligand-specific and isoform-dependent manner. Interestingly, both PR isoforms associated with a key regulator of energy homeostasis, FoxO1, suggesting a novel role for PR in energy metabolism. Because many identified proteins in this PR interactome are synaptic proteins, we tested the hypothesis that progestins function in synaptic plasticity. Indeed, progesterone enhanced synaptic density, by increasing synapsin-I–positive synapses, in rat primary cortical neuronal cultures. This novel combination of RPPA and mass spectrometry allowed identification of PR action in synaptic remodeling and energy homeostasis and reveals unique roles for progestins in brain function and disease.

AB - Progestins bind to the progestin receptor (PR) isoforms, PR-A and PR-B, in brain to influence development, female reproduction, anxiety, and stress. Hormone-activated PRs associate with multiple proteins to form functional complexes. In the present study, proteins from female mouse hypothalamus that associate with PR were isolated using affinity pull-down assays with glutathione S-transferase–tagged mouse PR-A and PR-B. Using complementary proteomics approaches, reverse phase protein array (RPPA) and mass spectrometry, we identified hypothalamic proteins that interact with PR in a ligand-dependent and isoform-specific manner and were confirmed by Western blot. Synaptic proteins, including synapsin-I and synapsin-II, interacted with agonist-bound PR isoforms, suggesting that both isoforms function in synaptic plasticity. In further support, synaptogyrin-III and synapsin-III associated with PR-A and PR-B, respectively. PR also interacted with kinases, including c-Src, mTOR, and MAPK1, confirming phosphorylation as an integral process in rapid effects of PR in the brain. Consistent with a role in transcriptional regulation, PR associated with transcription factors and coactivators in a ligand-specific and isoform-dependent manner. Interestingly, both PR isoforms associated with a key regulator of energy homeostasis, FoxO1, suggesting a novel role for PR in energy metabolism. Because many identified proteins in this PR interactome are synaptic proteins, we tested the hypothesis that progestins function in synaptic plasticity. Indeed, progesterone enhanced synaptic density, by increasing synapsin-I–positive synapses, in rat primary cortical neuronal cultures. This novel combination of RPPA and mass spectrometry allowed identification of PR action in synaptic remodeling and energy homeostasis and reveals unique roles for progestins in brain function and disease.

KW - Cortex

KW - Estrogen

KW - Progesterone

KW - Proteomics

KW - Synapse

KW - Synapsin

UR - http://www.scopus.com/inward/record.url?scp=85032154941&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85032154941&partnerID=8YFLogxK

U2 - 10.1523/ENEURO.0272-17.2017

DO - 10.1523/ENEURO.0272-17.2017

M3 - Article

VL - 4

JO - eNeuro

JF - eNeuro

SN - 2373-2822

IS - 5

M1 - e0272-17.2017

ER -