The Progestin Receptor Interactome in the Female Mouse Hypothalamus: Interactions with Synaptic Proteins Are Isoform Specific and Ligand Dependent
Autor: | Cheryl F. Lichti, Marc J. Tetel, Didem Vardar-Ulu, Sabin A. Nettles, Larry Denner, Katherine J. Sellers, Shixia Huang, Moriah Harling, Dana D. Im, Dean P. Edwards, Cassandra Pattanayak, Deepak Srivastava, Kalpana D. Acharya |
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Rok vydání: | 2017 |
Předmět: |
Transcription
Genetic Plasma protein binding Ligands Proteomics Interactome Mice 0302 clinical medicine synapse estrogen Protein Isoforms Receptor Cells Cultured Glutathione Transferase Cerebral Cortex Neurons 0303 health sciences Estradiol General Neuroscience General Medicine Synapsin 5.1 New Research Integrative Systems Cell biology Cortex Female Signal transduction Receptors Progesterone Protein Binding Signal Transduction Gene isoform medicine.medical_specialty Ovariectomy Hypothalamus Nerve Tissue Proteins progesterone Biology 03 medical and health sciences proteomics Internal medicine medicine Animals 030304 developmental biology synapsin Embryo Mammalian Mice Inbred C57BL Endocrinology Gene Expression Regulation Synapses Synaptic plasticity 030217 neurology & neurosurgery |
Zdroj: | eNeuro |
ISSN: | 2373-2822 |
Popis: | 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. |
Databáze: | OpenAIRE |
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