Abstract
Effects of cooling and calmodulin antagonist on ionomycin-induced increase in oxidative metabolism (or formation of reactive oxygen species) of rat cerebellar neurons was examined using a flow cytometer and 2′,7′-dichlorofluorescin diacetate, a fluorescent dye for intracellular hydrogen peroxide. Cooling neurons to temperatures below 16°C greatly attenuated ionomycin-induced augmentation of oxidative metabolism without affecting the Ca 2+ influx produced by ionomycin. Rewarming neurons to 36°C in peesence of ionomycin increased the oxidative metabolism, indicating a temperature-sensitive metabolic process. Substitution of Ca 2+ with Ba 2+ or Sr 2+ completely abolished an ionomycin-induced increase in the oxidative metabolism. Pretreatment with W-7, a calmodulin antagonist, at concentrations of 10 μM or higher (up to 100 μM) produced a dose-dependent attenuation of ionomycin-induced increase in oxidative metabolism. Results suggest that calmodulin is involved in the ionomycin-induced increase in oxidative metabolism of dissociated cerebellar neurons.
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