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Histamine-induced calcium entry in rat cerebellar astrocytes: evidence for capacitative and non-capacitative mechanisms

Silke Jung, Fatima Pfeiffer and Joachim W. Deitmer

1. We have investigated the effects of histamine on the intracellular calcium concentration ([Ca²⁺]_i) of cultured rat cerebellar astrocytes using fura-2-based Ca²⁺ imaging microscopy.

2. Most of the cells responded to the application of histamine with an increase in [Ca²⁺]_i which was antagonized by the H₁ receptor blocker mepyramine. When histamine was applied for several minutes, the majority of the cells displayed a biphasic Ca²⁺ response consisting of an initial transient peak and a sustained component. In contrast to the initial transient [Ca²⁺]_i response, the sustained, receptor-activated increase in [Ca²⁺]_i was rapidly abolished by chelation of extracellular Ca²⁺ or addition of Ni²⁺, Mn²⁺, Co²⁺ and Zn²⁺, but was unaffected by nifedipine, an antagonist of L-type voltage-activated Ca²⁺ channels. These data indicate that the sustained increase in [Ca²⁺]_i was dependent on Ca²⁺ influx.

3. When intracellular Ca²⁺ stores were emptied by prolonged application of histamine in Ca²⁺-free conditions, Ca²⁺ re-addition after removal of the agonist did not lead to an 'overshoot' of [Ca²⁺]_i indicative of store-operated Ca²⁺ influx. However, Ca²⁺ stores were refilled despite the absence of any substantial change in the fura-2 signal.

4. Depletion of intracellular Ca²⁺ stores using cyclopiazonic acid in Ca²⁺-free saline and subsequent re-addition of Ca²⁺ to the saline resulted in an increase in [Ca²⁺]_i that was significantly enhanced in the presence of histamine.

5. The results suggest that besides capacitative mechanisms, a non-capacitative, voltage-independent pathway is involved in histamine-induced Ca²⁺ entry into cultured rat cerebellar astrocytes.

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