Mechanisms of dopamine-induced cell death in cultured rat forebrain neurons: interactions with and differences from glutamate-induced cell death.
Injury to the brain, whether by ischemia or trauma, results in the uncontrolled release of many neurotransmitters, including glutamate and dopamine. Both of these neurotransmitters are neurotoxic in high concentrations, and the oxidative stress caused by reactive oxygen species generation has been implicated in the mechanism of neurotoxicity. In this study, we used cultured rat forebrain neurons to characterize cell death caused by exposure to dopamine and/or glutamate and to investigate potential acute mechanisms of toxicity. Dopamine exposure (250 microM for 2 h) reduced cell viability to 34. 3 +/- 5.5% of untreated control 20 h later and increased the number of neurons with apoptotic morphology. The antioxidant N-acetylcysteine (100 microM) inhibited dopamine-induced toxicity and prevented the covalent binding of dopamine quinones to protein. In contrast, glutamate toxicity lacked the hallmark characteristics of apoptosis. When neurons were exposed successively to sublethal concentrations of dopamine and glutamate, cell viability at 20 h was reduced to 62.3 +/- 5.2% of untreated control. Apoptosis was not evident, and N-acetylcysteine blocked the potentiating effect of dopamine on glutamate-induced toxicity. We used single-cell fluorescence assays to measure changes in intraneuronal glutathione, intraneuronal Ca2+, mitochondrial membrane potential, and DNA integrity as potential acute inducers of neuronal injury. While changes in these parameters could be demonstrated, none were identified as the sole acute inducer of cell death caused by dopamine. In summary, we have characterized a number of neuronal responses to lethal dopamine injury. Also, we have demonstrated that dopamine and glutamate can interact in vitro to potentiate cell death and that the potentiation appears to be induced by oxidative stress.
Hoyt
K R
KR
Department of Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, 15261, USA.
Reynolds
I J
IJ
Hastings
T G
TG
D000818:Animals; D016923:Cell Death / Q000187:drug effects*; D002478:Cells, Cultured; D004298:Dopamine / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D018698:Glutamic Acid / Q000494:pharmacology*; D016548:Prosencephalon / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley
D018698:Glutamic Acid; D004298:Dopamine