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KMID : 0811720110150010053
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Korean Journal of Physiology & Pharmacology
2011 Volume.15 No. 1 p.53 ~ p.60
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Ca2＋-induced Ca2＋ Release from Internal Stores in INS-1 Rat Insulinoma Cells
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Choi Kyung-Jin
Cho Dong-Su
Kim Ju-Young
Kim Byung-Joon
Lee Kyung-Moo
Kim Shin-Hye
Kim Dong-Kwan
Kim Se-Hoon
Park Hyung-Seo
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Abstract
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The secretion of insulin from pancreatic ?-cells is triggered by the influx of Ca2＋ through voltage-dependent Ca2＋ channels. The resulting elevation of intracellular calcium ([Ca2＋]i) triggers additional Ca2＋ release from internal stores. Less well understood are the mechanisms involved in Ca2＋ mobilization from internal stores after activation of Ca2＋ influx. The mobilization process is known as calcium-induced calcium release (CICR). In this study, our goal was to investigate the existence of and the role of caffeine-sensitive ryanodine receptors (RyRs) in a rat pancreatic ?-cell line, INS-1 cells. To measure cytosolic and stored Ca2＋, respectively, cultured INS-1 cells were loaded with fura-2/AM or furaptra/AM. [Ca2＋]i was repetitively increased by caffeine stimulation in normal Ca2＋ buffer. However, peak [Ca2＋]i was only observed after the first caffeine stimulation in Ca2＋ free buffer and this increase was markedly blocked by ruthenium red, a RyR blocker. KCl-induced elevations in [Ca2＋]i were reduced by pretreatment with ruthenium red, as well as by depletion of internal Ca2＋ stores using cyclopiazonic acid (CPA) or caffeine. Caffeine-induced Ca2＋ mobilization ceased after the internal stores were depleted by carbamylcholine (CCh) or CPA. In permeabilized INS-1 cells, Ca2＋ release from internal stores was activated by caffeine, Ca2＋, or ryanodine. Furthermore, ruthenium red completely blocked the CICR response in permeabilized cells. RyRs were widely distributed throughout the intracellular compartment of INS-1 cells. These results suggest that caffeine-sensitive RyRs exist and modulate the CICR response from internal stores in INS-1 pancreatic ?-cells.
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KEYWORD
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INS-1, Caffeine, Ryanodine, Calcium release, CICR
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