Abstract
We report that both Na + and Ca 2+ currents are involved in the action potentials and in the hormone release from rat somatotrophs in primary culture. Single somatotrophs were identified by reverse hemolytic plaque assay (RHPA) and transmembrane voltage and curents were recorded using the whole-cell mode of the patch-clamp technique. Somatotrophs displayed a mean resting potential of −80mV and an average input resistance of 5.7GΩ. Most of the cells showed spontaneous or evoked action potentials. Single action potentials or the initial spike in a burst were characterized by their high amplitude and short duration. Tetrodotoxin (TTX, 1μM) blocked single action potentials and the initial spikes in a burst, whereas action potentials of long duration and low amplitude persisted. Cobalt (2mM) plus TTX (1μM) blocked all the action potentials. Voltage-clamp experiments confirmed the presence of both a TTX-sensitive Na + current and Co 2+ -sensitive Ca 2+ currents. TTX or Na +-free medium slightly decreased the basal release of GH but did not markedly modify hGRF-stimulated GH release. However, Co 2+ (2mM), which partially decreased the basal release, totally blocked hGRF-stimulated release. We conclude that (1) Na + currents which initiate rapid action potentials may participate in spontaneous GH release; (2) Ca 2+ currents, which give rise to long duration action potentials and membrane voltage fluctuation, are probably involved in both basal and hGRF-stimulated GH releases.
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