Suppression of programmed neuronal death by a thapsigargin-induced Ca2+ influx.
Rat sympathetic neurons undergo programmed cell death (PCD) in vitro and in vivo when they are deprived of nerve growth factor (NGF). Chronic depolarization of these neurons in cell culture with elevated concentrations of extracellular potassium ([K+]o) prevents this death. The effect of prolonged depolarization on neuronal survival is thought to be mediated by a rise of intracellular calcium concentration ([Ca2+]i) caused by Ca2+ influx through voltage-gated channels. In this report we investigate the effects of chronic treatment of rat sympathetic neurons with thapsigargin, an inhibitor of intracellular Ca2+ sequestration. In medium containing a normal concentration of extracellular Ca2+ ([Ca2+]o), thapsigargin caused a sustained rise of intracellular Ca2+ concentration and partially blocked death of NGF-deprived cells. Elevating [Ca2+]o in the presence of thapsigargin further increased [Ca2+]i, suggesting that the sustained rise of [Ca2+]i was caused by a thapsigargin-induced Ca2+ influx. This treatment potentiated the effect of thapsigargin on survival. The dihydropyridine Ca2+ channel antagonist, nifedipine, blocked both a sustained elevation of [Ca2+]i and enhanced survival caused by depolarization with elevated [K+]o, suggesting that these effects are mediated by Ca2+ influx through L-type channels. Nifedipine did not block the sustained rise of [Ca2+]i or enhanced survival caused by thapsigargin treatment, indicating that these effects were not mediated by influx of Ca2+ through L-type channels. These results provide additional evidence that increased [Ca2+]i can suppress neuronal PCD and identify a novel method for chronically raising neuronal [Ca2+]i for investigation of this and other Ca(2+)-dependent phenomena.
Lampe
P A
PA
Department of Molecular Biology and Pharmacology, Washington University School of Medicine, Saint Louis, Missouri 63110, USA.
Cornbrooks
E B
EB
Juhasz
A
A
Johnson
E M
EM
Franklin
J L
JL
D000320:Adrenergic Fibers / Q000473:pathology / Q000648:ultrastructure; D000818:Animals; D002118:Calcium; D016923:Cell Death / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D009414:Nerve Growth Factors / Q000502:physiology; D009474:Neurons; D011188:Potassium; D051381:Rats; D017207:Rats, Sprague-Dawley; D013729:Terpenes; D019284:Thapsigargin
D009414:Nerve Growth Factors; D013729:Terpenes; D019284:Thapsigargin; D011188:Potassium; D002118:Calcium