Hypoxia increases intracellular calcium in glutamate-activated horizontal cells of goldfish retina via mitochondrial KATP channels and intracellular stores.

Central neurons of the common goldfish (Carassius auratus) are exceptional in their capacity to survive Ca2+-induced excitotoxicity and cell death during hypoxia. Horizontal cells (HCs) are inhibitory interneurons of the retina that are tonically depolarized by the neurotransmitter, glutamate, yet preserve intracellular Ca2+ homeostasis. In HCs isolated from goldfish, and in the absence of glutamatergic input, intracellular Ca2+ concentration ([Ca2+]i) is protected from prolonged exposure to hypoxia by mitochondrial ATP-dependent K+ (mKATP) channel activity. In the present study, we investigated the effects of hypoxia upon [Ca2+]i in isolated HCs during tonic activation by glutamate to better predict the effects of hypoxia in the active retina. Dynamic changes in [Ca2+]i were measured using the ratiometric Ca2+ indicator, Fura-2. Application of 100 μM glutamate during hypoxia (PO2 = 25 mmHg) produced a 1.3-fold greater rise in [Ca2+]i compared to the same glutamate stimulus during normoxia. The hypoxia-dependent increase in [Ca2+]i was abolished by application of 5-hydroxydecanoic acid, which renders mKATP channels inactive. Extracellular Ca2+ did not contribute to the elevated [Ca2+]i observed during hypoxia, as the effect persisted in Ca2+-free solution and during application of verapamil, an L-type Ca2+ channel blocker. By contrast, inhibition of the mitochondrial Ca2+ uniporter or ryanodine receptors (with ruthenium red or ryanodine, respectively) abolished the hypoxia-dependent rise in [Ca2+]i. This study reports an mKATP-dependent rise in [Ca2+]i during hypoxia in HCs activated by glutamate, and suggests roles for the mitochondria and intracellular Ca2+ stores in regulating this mechanism.