title: Estrogen receptor alpha and beta differentially regulate intracellular Ca(2+) dynamics leading to ERK phosphorylation and estrogen neuroprotection in hippocampal neurons.
abstract: Our previous analyses indicated that both estrogen receptor (ER) subtypes, ERalpha and ERbeta, contribute to estrogen neuroprotection [Zhao, L., Wu, T.-W., Brinton, R.D., 2004. Estrogen receptor subtypes alpha and beta contribute to neuroprotection and increased Bcl-2 expression in primary hippocampal neurons. Brain Res. 1010, 22-34]. In the present study, we sought to determine the underlying mechanisms by which ERalpha and ERbeta promote neuronal function, with a focus on neuroprotection, and whether these mechanisms are consistent with a classical nuclear or membrane ER-mediated response. Results of these analyses demonstrated that both the ERalpha-selective agonist, PPT (100 pM), and the ERbeta-selective agonist, DPN (100 pM), were effective in dynamically but differentially regulating intracellular calcium (Ca(2+)) signaling in hippocampal neurons. Consistent with the direct measurement of neuroprotective outcomes [Zhao, L., Wu, T.-W., Brinton, R.D., 2004. Estrogen receptor subtypes alpha and beta contribute to neuroprotection and increased Bcl-2 expression in primary hippocampal neurons. Brain Res. 1010, 22-34], PPT and DPN exerted comparable efficacy in attenuating excitotoxic glutamate (200 microM)-induced intracellular Ca(2+) rise. In contrast, DPN was more efficacious than PPT in potentiating a physiological concentration of glutamate (25 microM)-induced intracellular Ca(2+) rise in these neurons. Further analyses revealed that both PPT and DPN increased ERK phosphorylation, however, the temporal profile and magnitude of response were unique to each molecule. The presence of the L-type Ca(2+) channel inhibitor, nifedipine (10 microM), partially inhibited 17beta-estradiol- and PPT-induced increase in phosphorylated ERK expression, whereas it induced a complete inhibition of DPN-induced increase in ERK phosphorylation. Additional neuroprotective experiments demonstrated that the MAPK inhibitor, PD 98059 (5 microM), partially blocked 17beta-estradiol-induced promotion of neuronal survival against excitotoxic glutamate (200 microM)-induced neurotoxicity, whereas it completely blocked both PPT- and DPN-induced neuroprotection. The presence of the nuclear ER antagonist, ICI 182,780 (1 microM), not only failed to block all 3 molecule-induced neuroprotection, but coadministration of ICI 182,780 and each single molecule exerted a comparable or even greater neuroprotection. Taken together, as an expansion of our previous analyses, these data indicate that both ERalpha and ERbeta contribute to neuronal mechanisms leading to estrogen promotion of neuronal function but with unique signaling profiles. Activation of ERbeta and induction of intracellular Ca(2+) influx via the L-type channels appears to be more closely associated with estrogen promotion of memory mechanisms. However, ERalpha and ERbeta play an equivalently important role in mediating estrogen neuroprotection, and, which is dependent upon the activation of the MAPK signaling. Further, the present analyses suggest that separate from a classical nuclear ER-mediated response, estrogen promotes neuronal survival likely through a non-nuclear cytoplasm or membrane-associated ER-mediated rapid signaling cascade.
authors.affiliation: Department of Pharmacology and Pharmaceutical Sciences, Norris Foundation Laboratory for Neuroscience Research, University of Southern California, Los Angeles, CA 90089, USA.
authors.forename: Liqin
authors.initials: L
authors.lastname: Zhao
authors.forename: Roberta Diaz
authors.initials: RD
authors.lastname: Brinton
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000819:agonists / Q000502:physiology*; D047629:Estrogen Receptor beta / Q000819:agonists / Q000502:physiology*; D004967:Estrogens / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects*; D009543:Nifedipine / Q000494:pharmacology; D010636:Phenols / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors