Muscarinic receptor subtype determines vulnerability to oxidative stress in COS-7 cells.
Research has suggested that there may be increased brain-region selective vulnerability to oxidative stress in aging and that Vulnerability to oxidative stress may be important in determining regional differences in neuronal aging. We assessed whether one factor determining vulnerability to oxidative stress might involve qualitative/quantitative differences in receptor subtypes in various neuronal populations. COS-7 cells were transfected with one of five muscarinic receptor subtypes (M1-M5 AChR) to DA (1 mM for 4 h) and intracellular Ca2+ levels were examined via fluorescent imaging analysis prior to and following 750 microM oxotremorine (oxo). Results indicated that the ability of the cells to clear excess Ca2+ (i.e., Ca2+ Recovery) following oxo stimulation varied as a function of transfected mAChR subtype, with DA-treated M1, M2, or M4 cells showing greater decrements in Recovery than those transfected with M3 or M5 AChR. A similar pattern of results in M1- or M3-transfected DA-exposed cells was seen with respect to Viability. Viability of the untransfected cells was unaffected by DA. Pretreatment with Trolox (a Vitamin E analog) or PBN (a nitrone trapping agent) did not alter the DA effects on cell Recovery in the M1-transfected cells, but were effective in preventing the decrements in Viability. The calcium channel antagonists (L and N, respectively), Nifedipine and Conotoxin prevented both the DA-induced deficits in Recovery and Viability. Results are discussed in terms of receptor involvement in the regional differences in Vulnerability to oxidative stress with age, and that loss of neuronal function may not inevitably lead to cell death.
Joseph
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JA
USDA-Human Nutrition Research Center on Aging at Tufts University, Boston, MA 02111, USA. jjoseph@hnrc.tufts.edu
Fisher
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DR
Strain
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D000375:Aging; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D019556:COS Cells / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D002839:Chromans / Q000494:pharmacology; D020916:Conotoxins / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D018721:Muscarinic Agonists / Q000494:pharmacology*; D009543:Nifedipine / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D010095:Oxotremorine / Q000494:pharmacology*; D020033:Protein Isoforms; D017382:Reactive Oxygen Species; D011976:Receptors, Muscarinic / Q000145:classification / Q000187:drug effects / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014162:Transfection; D014882:Water-Electrolyte Balance / Q000187:drug effects
D000975:Antioxidants; D002121:Calcium Channel Blockers; D002839:Chromans; D020916:Conotoxins; D018721:Muscarinic Agonists; D020033:Protein Isoforms; D017382:Reactive Oxygen Species; D011976:Receptors, Muscarinic; D010095:Oxotremorine; D009543:Nifedipine; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D002118:Calcium; D004298:Dopamine