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PMID:19116207

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Citation

Rosker, C, Meur, G, Taylor, EJ and Taylor, CW (2009) Functional ryanodine receptors in the plasma membrane of RINm5F pancreatic beta-cells. J. Biol. Chem. 284:5186-94

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+) entry.

Links

PubMed PMC2643496 Online version:10.1074/jbc.M805587200

Keywords

Animals; Caffeine/pharmacokinetics; Calcium/metabolism; Calcium Channel Agonists/pharmacology; Cell Line, Tumor; Cell Membrane/metabolism; Central Nervous System Stimulants/pharmacology; Cresols/pharmacology; Dose-Response Relationship, Drug; Fungicides, Industrial/pharmacology; Inositol 1,4,5-Trisphosphate Receptors/agonists; Inositol 1,4,5-Trisphosphate Receptors/metabolism; Insulin-Secreting Cells/metabolism; Male; Rats; Rats, Wistar; Ryanodine/pharmacology; Ryanodine Receptor Calcium Release Channel/biosynthesis

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

RAT:RYR2

involved_in

GO:0034220: ion transmembrane transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:RYR2

located_in

GO:0005886: plasma membrane

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:RYR2

involved_in

GO:0071421: manganese ion transmembrane transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:RYR2

involved_in

GO:0070588: calcium ion transmembrane transport

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:ITPR3

located_in

GO:0005783: endoplasmic reticulum

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:Q99M98

enables

GO:0005219: ryanodine-sensitive calcium-release channel activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:Q99M98

involved_in

GO:0031000: response to caffeine

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:Q99M98

involved_in

GO:0006816: calcium ion transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:Q99M98

located_in

GO:0005886: plasma membrane

ECO:0000315: mutant phenotype evidence used in manual assertion

C

Seeded From UniProt

complete

RAT:Q99M98

involved_in

GO:0051480: regulation of cytosolic calcium ion concentration

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:Q99M98

involved_in

GO:0051209: release of sequestered calcium ion into cytosol

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:Q9R273

enables

GO:0005219: ryanodine-sensitive calcium-release channel activity

ECO:0000304: author statement supported by traceable reference used in manual assertion

F

Seeded From UniProt

complete

RAT:Q9R273

involved_in

GO:0031000: response to caffeine

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

RAT:Q9R273

involved_in

GO:0006816: calcium ion transport

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

RAT:Q9R273

involved_in

GO:0051480: regulation of cytosolic calcium ion concentration

ECO:0000304: author statement supported by traceable reference used in manual assertion

P

Seeded From UniProt

complete

RAT:RYR1

enables

GO:0005219: ryanodine-sensitive calcium-release channel activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

RAT:RYR1

involved_in

GO:0006816: calcium ion transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:RYR1

involved_in

GO:0031000: response to caffeine

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:RYR1

involved_in

GO:0051209: release of sequestered calcium ion into cytosol

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

RAT:RYR1

involved_in

GO:0051480: regulation of cytosolic calcium ion concentration

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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