Characterization of calcium channels of a glucose-responsive rat insulinoma

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Abstract

Voltage-dependent calcium channels are important in the control of calcium influx into excitable cells. With the use of cells isolated from a glucose-responsive rat insulinoma, the effect of the calcium-channel blocker nitrendipine on insulin release was examined. When 50 nM nitrendipine was added with a stimulatory glucose concentration (30 mM), first-phase insulin release was preserved, whereas the second phase was inhibited by 39.4 ± 5.3%. When 50 nM nitrendipine was also present during basal and stimulated insulin release, the insulin release from fresh cells was significantly less during both phases compared with cells not exposed to nitrendipine (P < 0.05). In cells cultured for 1 day in 30 mM glucose, a diminished insulin response was seen on stimulation with 30 mM glucose. Nitrendipine (50 nM) did not lead to a further decrease in insulin release. Saturable binding sites in homogenate, whole cells, and purified plasma membranes prepared from the insulinoma were characterized using [3H]nitrendipine. The K(d) (in pM) for homogenate from uncultured cells was 225 ± 34 (n = 11), whereas the B(max) (in fmol/mg protein) was 52 ± 5. The number of apparent binding sites for [3H]nitrendipine was reduced by ~ 50% in cultured cells (B(max) = 30 ± 5 fmol/mg protein). The reduction in insulin release from cultured cells correlated well with the reduction in nitrendipine binding. It is concluded that the decrease in the number of apparent binding sites for nitrendipine after culture represents a reduction in functional calcium channels associated with influx of calcium and insulin release.

Original languageEnglish (US)
Pages (from-to)19/4
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume256
Issue number4
StatePublished - Jan 1 1989
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Physiology (medical)

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