Functional ryanodine receptors in the plasma membrane of RINm5F pancreatic beta-cells.

The Journal of Biological Chemistry
Christian RoskerC W Taylor

Abstract

Ryanodine receptors (RyR) are Ca(2+) channels that mediate Ca(2+) release from intracellular stores in response to diverse intracellular signals. In RINm5F insulinoma cells, caffeine, and 4-chloro-m-cresol (4CmC), agonists of RyR, stimulated Ca(2+) entry that was independent of store-operated Ca(2+) entry, and blocked by prior incubation with a concentration of ryanodine that inactivates RyR. Patch-clamp recording identified small numbers of large-conductance (gamma(K) = 169 pS) cation channels that were activated by caffeine, 4CmC or low concentrations of ryanodine. Similar channels were detected in rat pancreatic beta-cells. In RINm5F cells, the channels were blocked by cytosolic, but not extracellular, ruthenium red. Subcellular fractionation showed that type 3 IP(3) receptors (IP(3)R3) were expressed predominantly in endoplasmic reticulum, whereas RyR2 were present also in plasma membrane fractions. Using RNAi selectively to reduce expression of RyR1, RyR2, or IP(3)R3, we showed that RyR2 mediates both the Ca(2+) entry and the plasma membrane currents evoked by agonists of RyR. We conclude that small numbers of RyR2 are selectively expressed in the plasma membrane of RINm5F pancreatic beta-cells, where they mediate Ca(2+) e...Continue Reading

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Citations

Jun 5, 2013·The International Journal of Biochemistry & Cell Biology·Gopinath Krishnan, Nivedita Chatterjee
Jan 17, 2019·American Journal of Physiology. Cell Physiology·Toshiaki SawataniTomohisa Ishikawa
Sep 14, 2016·The Biochemical Journal·Pia V RöderWeiping Han
Nov 26, 2009·Biochemistry·Colin W TaylorTaufiq Rahman

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Methods Mentioned

BETA
transfection
protein assay
PCR
transfections

Software Mentioned

pClamp

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