Reciprocal regulation of mitochondrial dynamics and calcium signaling in astrocyte processes

Abstract

We recently showed that inhibition of neuronal activity, glutamate uptake, or reversed-Na+/Ca2+-exchange with TTX, TFB-TBOA, or YM-244769, respectively, increases mitochondrial mobility in astrocytic processes. In the present study, we examined the interrelationships between mitochondrial mobility and Ca2+ signaling in astrocyte processes in organotypic cultures of rat hippocampus. All of the treatments that increase mitochondrial mobility decreased basal Ca2+.As recently reported, we observed spontaneous Ca2+ spikes with half-lives of ~ 1 s that spread ~6 μ mand are almost abolished by a TRPA1 channel antagonist. Virtually all of these Ca2+spikes overlap mitochondria (98%), and 62% of mitochondria are overlapped by these spikes. Although tetrodotoxin, TFB-TBOA, or YM-244769 increased Ca2+ signaling, the specific effects on peak, decay time, and/or frequency were different. To more specifically manipulate mitochondrial mobility, we explored the effects of Miro motor adaptor proteins. We show that Miro1 and Miro2 are both expressed in astrocytes and that exogenous expression of Ca2+-insensitive Miro mutants (KK) nearly doubles the percentage of mobile mitochondria. Expression of Miro1KK had a modest effect on the frequency of these Ca2+ spikes but nearly doubled the decay half-life. The mitochondrial proton ionophore, FCCP, caused a large, prolonged increase in cytosolic Ca2+ followed by an increase in the decay time and the spread of the spontaneous Ca2+ spikes. Photo-ablation of mitochondria in individual astrocyte processes has similar effects on Ca2+. Together, these studies show that Ca2+ regulates mitochondrial mobility, and mitochondria in turn regulate Ca2+ signals in astrocyte processes.