Abstract:
:The steroid hormone 1 alpha,25-dihydroxyvitamin D3 [1,25-(OH)2D3] can elicit biological responses via a nongenomic pathway that involves rapid opening of the plasma membrane Ca2+ channels. There is also evidence that 1,25-(OH)2D3 influences insulin secretion in the pancreatic beta-cell, which is primarily mediated by a rapid rise in the concentration of intracellular free Ca2+ ([Ca2+]i). We employed fluorescent digital ratiometric video imaging at the single cell level to study the effects of 1,25-(OH)2D3 on [Ca2+]i in a pancreatic beta-cell line, RINr1046-38. In RIN cells equilibrated at a steady state glucose concentration (5.5 mM), 1,25-(OH)2D3 (2-20 nM) rapidly, within 5-10 sec, increased [Ca2+]i and evoked sinusoidal [Ca2+]i oscillations with a frequency of 1.87 +/- 0.13 min-1 and an amplitude of 236 +/- 3 nM (from the initial basal level of 110 +/- 2 nM). The [Ca2+]i oscillations were acutely dependent on extracellular Ca2+, but not on extracellular glucose. Further, we investigated the mechanisms of activation by 1,25-(OH)2D3 of the Ca2+ entry pathway in the plasma membrane and analyzed the relationship between 1,25-(OH)2D3-stimulated Ca2+ entry and Ca2+ release from intracellular stores. The 1,25-(OH)2D3-evoked [Ca2+]i oscillations were mediated by nonselective Ca2+ channels, which are permeable to Mn2+ and suppressed by extracellular La3+. Blockage of voltage-dependent Ca2+ channels by nifedipine significantly decreased the amplitude of the oscillations. Depletion of intracellular Ca2+ stores with thapsigargin did not affect the 1,25-(OH)2D3-stimulated Ca2+ entry estimated by the Mn2+ entry and fura-2 fluorescence quench, which implies that the hormone directly activates nonselective Ca2+ channels. The 1,25-(OH)2D3-evoked increase in the background Ca2+ influx appears to generate [Ca2+]i oscillations by triggering Ca2+ release through the ryanodine receptor/Ca2+ release channel, but not through activation of the inositol 1,4,5-triphosphate receptor. Our findings are consistent with a role of the plasmalemmal vitamin D receptor coupled to the plasma membrane Ca2+ channels in mediating rapid effects of the hormone. We propose that the 1,25-(OH)2D3-mediated Ca2+ signaling pathway may be involved in the regulation of insulin secretion from the pancreatic beta-cell.
journal_name
Endocrinologyjournal_title
Endocrinologyauthors
Sergeev IN,Rhoten WBdoi
10.1210/endo.136.7.7789310subject
Has Abstractpub_date
1995-07-01 00:00:00pages
2852-61issue
7eissn
0013-7227issn
1945-7170journal_volume
136pub_type
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