Calcium regulation in mouse mesencephalic neurons-Differential roles of Na(+)/Ca(2+) exchanger, mitochondria and endoplasmic reticulum

Abstract

Midbrain dopaminergic (DA) neurons are the key to finely tune the voluntary movement, habit and motivation. The progressive and selective degeneration of these neurons is a pathological hallmark of Parkinson's disease (PD). The susceptibility of DA neurons in the SNpc may result from differences in how Ca(2+) is handled. However, very little information is available about the mechanisms involved in the regulation of intracellular Ca(2+) concentration ([Ca(2+)]i) in DA neurons. In this study, the relative contributions of various Na(+)/Ca(2+) exchangers and their interplay with internal Ca(2+) stores, endoplasmic reticulum (ER) and the mitochondria, in the regulation of the [Ca(2+)]i of mouse mesencephalic neurons were characterized. Both the K(+)-dependent Na(+)/Ca(2+) exchanger (NCKX) and the K(+)-independent Na(+)/Ca(2+) exchanger (NCX) can be detected and are functional in DA and non-DA neurons. NCX accounts for the larger component of Na(+)/Ca(2+) exchange activity. Single-cell RT-PCR analysis showed each individual neuron expressed a distinct set of the Na(+)/Ca(2+) exchangers. Furthermore, the Na(+)/Ca(2+) exchangers play prominent roles in removing [Ca(2+)]i induced by glutamate but not [Ca(2+)]i induced by depolarization. The mitochondria serve as a major Ca(2+) sink and are functionally located close to NCX. In contrast, the ER is functionally located close to NCKX and acts primarily as a Ca(2+) source with marginal effects. This study reveals that the Na(+)/Ca(2+) exchangers, the ER and the mitochondria, which cooperate interactively, act similarly when regulating [Ca(2+)]i in mesencephalic DA and non-DA neurons. The heterogeneous expression of multiple types of Na(+)/Ca(2+) exchangers and the quantitative differences found in [Ca(2+)]i regulation, together with other risk factors specific to DA neurons such as dopamine oxidation resulting in oxidative stress, may drive these cells to undergo selective degeneration.

Keywords: Ca(2+); Dopaminergic neurons; Endoplasmic reticulum; Mitochondria; Na(+)/Ca(2+) exchanger; Parkinson’s disease.