title: Estrogen receptor beta-selective agonists stimulate calcium oscillations in human and mouse embryonic stem cell-derived neurons.
abstract: Estrogens are used extensively to treat hot flashes in menopausal women. Some of the beneficial effects of estrogens in hormone therapy on the brain might be due to nongenomic effects in neurons such as the rapid stimulation of calcium oscillations. Most studies have examined the nongenomic effects of estrogen receptors (ER) in primary neurons or brain slices from the rodent brain. However, these cells can not be maintained continuously in culture because neurons are post-mitotic. Neurons derived from embryonic stem cells could be a potential continuous, cell-based model to study nongenomic actions of estrogens in neurons if they are responsive to estrogens after differentiation. In this study ER-subtype specific estrogens were used to examine the role of ERalpha and ERbeta on calcium oscillations in neurons derived from human (hES) and mouse embryonic stem cells. Unlike the undifferentiated hES cells the differentiated cells expressed neuronal markers, ERbeta, but not ERalpha. The non-selective ER agonist 17beta-estradiol (E(2)) rapidly increased [Ca2+]i oscillations and synchronizations within a few minutes. No change in calcium oscillations was observed with the selective ERalpha agonist 4,4',4''-(4-Propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT). In contrast, the selective ERbeta agonists, 2,3-bis(4-Hydroxyphenyl)-propionitrile (DPN), MF101, and 2-(3-fluoro-4-hydroxyphenyl)-7-vinyl-1,3 benzoxazol-5-ol (ERB-041; WAY-202041) stimulated calcium oscillations similar to E(2). The ERbeta agonists also increased calcium oscillations and phosphorylated PKC, AKT and ERK1/2 in neurons derived from mouse ES cells, which was inhibited by nifedipine demonstrating that ERbeta activates L-type voltage gated calcium channels to regulate neuronal activity. Our results demonstrate that ERbeta signaling regulates nongenomic pathways in neurons derived from ES cells, and suggest that these cells might be useful to study the nongenomic mechanisms of estrogenic compounds.
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences and Center for Reproductive Sciences, University of California San Francisco, San Francisco, California, United States of America.
authors.forename: Lili
authors.initials: L
authors.lastname: Zhang
authors.forename: Brigitte E
authors.initials: BE
authors.lastname: Blackman
authors.forename: Marcus D
authors.initials: MD
authors.lastname: Schonemann
authors.forename: Tatjana
authors.initials: T
authors.lastname: Zogovic-Kapsalis
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Pan
authors.forename: Mary
authors.initials: M
authors.lastname: Tagliaferri
authors.forename: Heather A
authors.initials: HA
authors.lastname: Harris
authors.forename: Isaac
authors.initials: I
authors.lastname: Cohen
authors.forename: Renee A Reijo
authors.initials: RA
authors.lastname: Pera
authors.forename: Synthia H
authors.initials: SH
authors.lastname: Mellon
authors.forename: Richard I
authors.initials: RI
authors.lastname: Weiner
authors.forename: Dale C
authors.initials: DC
authors.lastname: Leitman
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D047629:Estrogen Receptor beta / Q000819:agonists*; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D051379:Mice; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism*; D009543:Nifedipine / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D010936:Plant Extracts / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D011422:Propionates
chemical_list: D002118:Calcium; D047629:Estrogen Receptor beta; D009543:Nifedipine; D009570:Nitriles; D010080:Oxazoles; D010936:Plant Extracts; C486182:2,3-bis(4-hydroxyphenyl)-propionitrile; D011422:Propionates; C478102:ERB 041; C517398:MF101 extract