Dopamine induces apoptosis in young, but not in neonatal, neurons via Ca2+-dependent signal

Dopamine signaling plays a major role in regulation of neuronal apoptosis. During the postnatal period, dopamine signaling is known to be dramatically changed in the striatum. However, because it is difficult to culture neurons after birth, little is known about developmental changes in dopamine-mediated apoptosis. To examine such changes, we established the method of primary culture of striatal neurons from 2- to 3-wk-old ( young) mice. Dopamine, via D-1-like receptors, induced apoptosis in young, but not neonatal, striatal neurons, suggesting that the effect of dopamine on apoptosis changed with development. In contrast, although isoproterenol (Iso), a beta-adrenergic receptor agonist, increased cAMP production to a greater degree than dopamine, Iso did not increase apoptosis in striatal neurons from young and neonatal mice, suggesting a minor role of cAMP in dopamine-mediated apoptosis. Next, we examined the effect of dopamine on Ca2+ signaling. Dopamine, but not Iso, markedly increased intracellular Ca2+ in striatal neurons from young mice, and Ca2+-chelating agents abolished dopamine-induced apoptosis, suggesting that Ca2+ played a major role in the dopamine-mediated apoptosis pathway. In contrast, dopamine failed to increase intracellular Ca2+ in neonatal neurons, and the expression of PLC, which can increase intracellular Ca2+ via D-1-like receptor activation, was significantly greater in young than in neonatal striatal neurons. These data suggest that the developmental change in dopamine-mediated Ca2+ signaling was responsible for differences between young and neonatal striatum in induction of apoptosis. Furthermore, the culture of young striatal neurons is feasible and may provide a new tool for developmental studies.