Glutamate receptor requirement for neuronal death from anoxia-reoxygenation: an in Vitro model for assessment of the neuroprotective effects of estrogens.
1. Previous studies demonstrated that estrogens, specifically 17 beta-estradiol, the potent, naturally occurring estrogen, are neuroprotective in a variety of models including glutamate toxicity. The aim of the present study is twofold: (1) to assess the requirement for glutamate receptors in neuronal cell death associated with anoxia-reoxygenation in three cell types, SK-N-SH and HT-22 neuronal cell lines and primary rat cortical neuronal cultures, and (2) to evaluate the neuroprotective activity of both 17 beta-estradiol and its weaker isomer, 17 alpha-estradiol, in both anoxia-reoxygenation and glutamate toxicity. 2. SK-N-SH and HT-22 cell lines, both of which lack NMDA receptors as assessed by MK-801 binding assays, were resistant to both anoxia-reoxygenation and glutamate-induced cell death. In contrast, primary rat cortical neurons, which exhibit both NMDA and AMPA receptors, were sensitive to brief periods of exposure to anoxia-reoxygenation or glutamate. As such, there appears to be an obligatory requirement for NMDA and/or AMPA receptors in neuronal cell death resulting from brief periods of anoxia followed by reoxygenation. 3. Using primary rat cortical neuronal cultures, we evaluated the neuroprotective activity of 17 beta-estradiol (1.3 or 133 nM) and 17 alpha-estradiol (133 nM) in both anoxia-reoxygenation and excitotoxicity models of cell death. We found that the 133 nM but not the 1.3 nM dose of the potent estrogen, 17 beta-estradiol, protected 58.0, 57.5, and 85.3% of the primary rat cortical neurons from anoxia-reoxygenation, glutamate, or AMPA toxicity, respectively, and the 133 nM dose of the weak estrogen, 17 alpha-estradiol, protected 74.6, 81.7, and 85.8% of cells from anoxia-reoxygenation, glutamate, or AMPA toxicity, respectively. These data demonstrate that pretreatment with estrogens can attenuate glutamate excitotoxicity and that this protection is independent of the ability of the steroid to bind the estrogen receptor.
Zaulyanov
L L
LL
Department of Pharmacodynamics, College of Pharmacy, University of Florida, Gainesville 32610, USA.
Green
P S
PS
Simpkins
J W
JW
D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D015687:Cell Hypoxia; D002540:Cerebral Cortex / Q000166:cytology; D003505:Cyclodextrins / Q000494:pharmacology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000494:pharmacology*; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005260:Female; D018698:Glutamic Acid / Q000633:toxicity*; D005998:Glycine / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D017136:Ion Transport; D051379:Mice; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000502:physiology; D009447:Neuroblastoma / Q000473:pathology; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009498:Neurotoxins / Q000633:toxicity*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000187:drug effects / Q000502:physiology; D011960:Receptors, Estrogen / Q000378:metabolism; D017470:Receptors, Glutamate / Q000187:drug effects* / Q000502:physiology; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000502:physiology; D014407:Tumor Cells, Cultured / Q000187:drug effects; D047391:alpha-Cyclodextrins; D047392:beta-Cyclodextrins
D003505:Cyclodextrins; D018691:Excitatory Amino Acid Antagonists; D009419:Nerve Tissue Proteins; D018696:Neuroprotective Agents; D009498:Neurotoxins; D018091:Receptors, AMPA; D011960:Receptors, Estrogen; D017470:Receptors, Glutamate; D016194:Receptors, N-Methyl-D-Aspartate; D047391:alpha-Cyclodextrins; D047392:beta-Cyclodextrins; C479658:hydroxypropyl-alpha-cyclodextrin; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D018698:Glutamic Acid; D004958:Estradiol; D016291:Dizocilpine Maleate; D005998:Glycine