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Am J Physiol Gastrointest Liver Physiol 276: G227-G237, 1999;
0193-1857/99 $5.00
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Vol. 276, Issue 1, G227-G237, January 1999

Bombesin-evoked gastrin release and calcium signaling in human antral G cells in culture

Paul E. Squires, R. Mark Meloche, and Alison M. J. Buchan

Department of Physiology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3

Amplification of mRNA from a human antral cell culture preparation demonstrated the presence of two receptors of the bombesin and gastrin-releasing peptide family, GRPR-1 and BRS-3. Single cell microfluorometry demonstrated that most cells that exhibited bombesin-evoked changes in intracellular Ca2+ concentration were gastrin immunoreactive, indicating that antral G cells express the GRPR subtype. There were two components to the intracellular Ca2+ response: an initial nitrendipine-insensitive mobilization followed by a sustained phase that was inhibited by removal of extracellular Ca2+ and 20 mM caffeine and was partially inhibited by 10 µM nitrendipine. Preexposure of cells to thapsigargin and caffeine prevented the response to bombesin, indicating activation of inositol 1,4,5-trisphosphate (IP3)-sensitive stores. Gastrin release could be partially reversed by removal of extracellular Ca2+ and blockade of L-type voltage-dependent Ca2+ channels, indicating that a component of the secretory response to bombesin was dependent on Ca2+ influx. These data demonstrated that bombesin-stimulated gastrin release from human antral G cells resulted from activation of GRPRs and involved both release of intracellular Ca2+ and influx of extracellular Ca2+ through a combination of L-type voltage-gated and IP3-gated Ca2+ channels.

gastrin-releasing peptide; calcium imaging; nitrendipine; calcium-sensing receptor


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