Hormone-stimulated cellular Ca2+ mobilization in the isolated perfused rat pancreas was investigated by analyzing the efflux profiles of 45Ca2+ from 45Ca(2+)-loaded pancreata following agonist stimulation. The increased 45Ca2+ efflux reflects the enhanced exchange of Ca2+ across the plasma membrane as a result of increased [Ca2+]i. Both high and low concentrations of the cholecystokinin analog, cerulein, applied to the isolated perfused pancreas gave rise to an increased release of 45Ca3+. The patterns of the increase in 45Ca2+ release were consistently different for high and low concentrations of the agonist. Cerulein infused at a concentration of 10(-11) M induced a release of a small but significant amount of 45Ca2+ which could be abolished by 8-(N,N-diethylamine)octyl-3,4,5-trimethoxy-benzoate (TMB-8), but was not affected by ethyleneglycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA). Cerulein stimulation at 10(-9) M elicited a marked increase in 45Ca2+ release which was minimized by EGTA, but not by TMB-8. Also, infusion of cerulein stimulated a concentration-dependent amylase secretion response which displayed the same TMB-8- and EGTA-sensitivity pattern as the 45Ca2+ release response. The present study suggests (i) that cellular Ca2+ influx is a prominent feature of the increased 45Ca2+ efflux (i.e., increased [Ca2+]i) induced by pharmacological concentrations of cerulein while physiological concentrations of cerulein cause an increase in [Ca2+]i which is due predominantly to a release of internal Ca2+; and (ii) [Ca2+]i changes are essential for pancreatic enzyme secretion. Although isolated pancreatic acini or cells may lose their sensitivity and physiological responses to various agonists during isolation and preparation, the isolated perfused pancreas is a suitable and very sensitive model in which to study the physiology of Ca2+ mobilization and enzyme secretion.
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