17β-estradiol rapidly activates calcium release from intracellular stores via the GPR30 pathway and MAPK phosphorylation in osteocyte-like MLO-Y4 cells

Handle URI:
http://hdl.handle.net/10754/562124
Title:
17β-estradiol rapidly activates calcium release from intracellular stores via the GPR30 pathway and MAPK phosphorylation in osteocyte-like MLO-Y4 cells
Authors:
Ren, Jian; Wu, Junhua
Abstract:
Estrogen regulates critical cellular functions, and its deficiency initiates bone turnover and the development of bone mass loss in menopausal females. Recent studies have demonstrated that 17β-estradiol (E 2) induces rapid non-genomic responses that activate downstream signaling molecules, thus providing a new perspective to understand the relationship between estrogen and bone metabolism. In this study, we investigated rapid estrogen responses, including calcium release and MAPK phosphorylation, in osteocyte-like MLO-Y4 cells. E 2 elevated [Ca 2+] i and increased Ca 2+ oscillation frequency in a dose-dependent manner. Immunolabeling confirmed the expression of three estrogen receptors (ERα, ERβ, and G protein-coupled receptor 30 [GPR30]) in MLO-Y4 cells and localized GPR30 predominantly to the plasma membrane. E 2 mobilized calcium from intracellular stores, and the use of selective agonist(s) for each ER showed that this was mediated mainly through the GPR30 pathway. MAPK phosphorylation increased in a biphasic manner, with peaks occurring after 7 and 60 min. GPR30 and classical ERs showed different temporal effects on MAPK phosphorylation and contributed to MAPK phosphorylation sequentially. ICI182,780 inhibited E 2 activation of MAPK at 7 min, while the GPR30 agonist G-1 and antagonist G-15 failed to affect MAPK phosphorylation levels. G-1-mediated MAPK phosphorylation at 60 min was prevented by prior depletion of calcium stores. Our data suggest that E 2 induces the non-genomic responses Ca 2+ release and MAPK phosphorylation to regulate osteocyte function and indicate that multiple receptors mediate rapid E 2 responses. © 2012 Springer Science+Business Media, LLC.
KAUST Department:
Advanced Nanofabrication, Imaging and Characterization Core Lab; Core Labs
Publisher:
Springer Verlag
Journal:
Calcified Tissue International
Issue Date:
6-Mar-2012
DOI:
10.1007/s00223-012-9581-x
PubMed ID:
22392527
Type:
Article
ISSN:
0171967X
Sponsors:
We are indebted to Dr. Lynda Bonewald for her kind gift of MLO Y4 cells and thank Ms. Jennifer Rosser for help in arranging the transportation and logistics. This work was supported by the Nature Science Foundation of Shanghai, China (Grant 11ZR1440700).
Appears in Collections:
Articles; Advanced Nanofabrication, Imaging and Characterization Core Lab

Full metadata record

DC FieldValue Language
dc.contributor.authorRen, Jianen
dc.contributor.authorWu, Junhuaen
dc.date.accessioned2015-08-03T09:45:22Zen
dc.date.available2015-08-03T09:45:22Zen
dc.date.issued2012-03-06en
dc.identifier.issn0171967Xen
dc.identifier.pmid22392527en
dc.identifier.doi10.1007/s00223-012-9581-xen
dc.identifier.urihttp://hdl.handle.net/10754/562124en
dc.description.abstractEstrogen regulates critical cellular functions, and its deficiency initiates bone turnover and the development of bone mass loss in menopausal females. Recent studies have demonstrated that 17β-estradiol (E 2) induces rapid non-genomic responses that activate downstream signaling molecules, thus providing a new perspective to understand the relationship between estrogen and bone metabolism. In this study, we investigated rapid estrogen responses, including calcium release and MAPK phosphorylation, in osteocyte-like MLO-Y4 cells. E 2 elevated [Ca 2+] i and increased Ca 2+ oscillation frequency in a dose-dependent manner. Immunolabeling confirmed the expression of three estrogen receptors (ERα, ERβ, and G protein-coupled receptor 30 [GPR30]) in MLO-Y4 cells and localized GPR30 predominantly to the plasma membrane. E 2 mobilized calcium from intracellular stores, and the use of selective agonist(s) for each ER showed that this was mediated mainly through the GPR30 pathway. MAPK phosphorylation increased in a biphasic manner, with peaks occurring after 7 and 60 min. GPR30 and classical ERs showed different temporal effects on MAPK phosphorylation and contributed to MAPK phosphorylation sequentially. ICI182,780 inhibited E 2 activation of MAPK at 7 min, while the GPR30 agonist G-1 and antagonist G-15 failed to affect MAPK phosphorylation levels. G-1-mediated MAPK phosphorylation at 60 min was prevented by prior depletion of calcium stores. Our data suggest that E 2 induces the non-genomic responses Ca 2+ release and MAPK phosphorylation to regulate osteocyte function and indicate that multiple receptors mediate rapid E 2 responses. © 2012 Springer Science+Business Media, LLC.en
dc.description.sponsorshipWe are indebted to Dr. Lynda Bonewald for her kind gift of MLO Y4 cells and thank Ms. Jennifer Rosser for help in arranging the transportation and logistics. This work was supported by the Nature Science Foundation of Shanghai, China (Grant 11ZR1440700).en
dc.publisherSpringer Verlagen
dc.subjectCalcium ionen
dc.subjectEstrogenen
dc.subjectEstrogen receptoren
dc.subjectGPR30en
dc.subjectMAPKen
dc.subjectOsteocyteen
dc.title17β-estradiol rapidly activates calcium release from intracellular stores via the GPR30 pathway and MAPK phosphorylation in osteocyte-like MLO-Y4 cellsen
dc.typeArticleen
dc.contributor.departmentAdvanced Nanofabrication, Imaging and Characterization Core Laben
dc.contributor.departmentCore Labsen
dc.identifier.journalCalcified Tissue Internationalen
dc.contributor.institutionSchool of Stomatology, Tongji University, No. 399, Yanchang Middle Road, Shanghai 200072, Chinaen
kaust.authorRen, Jianen
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