RRC ID 49260
著者 Kitaguchi T, Oya M, Wada Y, Tsuboi T, Miyawaki A.
タイトル Extracellular calcium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells.
ジャーナル Biochem J
Abstract Intracellular cAMP and Ca(2+) are important second messengers that regulate insulin secretion in pancreatic β-cells; however, the molecular mechanism underlying their mutual interaction for exocytosis is not fully understood. In the present study, we investigated the interplay between intracellular cAMP and Ca(2+) concentrations ([cAMP](i) and [Ca(2+)](i) respectively) in the pancreatic β-cell line MIN6 using total internal reflection fluorescence microscopy. For measuring [cAMP](i), we developed a genetically encoded yellow fluorescent biosensor for cAMP [Flamindo (fluorescent cAMP indicator)], which changes fluorescence intensity with cAMP binding. Application of high-KCl or glucose to MIN6 cells induced the elevation of [cAMP](i) and exocytosis. Furthermore, application of an L-type Ca(2+) channel agonist or ionomycin to induce extracellular Ca(2+) influx evoked the elevation of [cAMP](i), whereas application of carbachol or thapsigargin, which mobilize Ca(2+) from internal stores, did not evoke the elevation of [cAMP](i). We performed RT (reverse transcription)-PCR analysis and found that Ca(2+)-sensitive Adcy1 (adenylate cyclase 1) was expressed in MIN6 cells. Knockdown of endogenous ADCY1 by small interference RNA significantly suppressed glucose-induced exocytosis and the elevation of both [cAMP](i) and [Ca(2+)](i). Taken together, the findings of the present study demonstrate that ADCY1 plays an important role in the control of pancreatic β-cell cAMP homoeostasis and insulin secretion.
巻・号 450(2)
ページ 365-73
公開日 2013-3-1
DOI 10.1042/BJ20121022
PII BJ20121022
PMID 23282092
MeSH Adenylyl Cyclases / genetics Adenylyl Cyclases / metabolism* Animals COS Cells Calcium / metabolism* Calcium Signaling Cells, Cultured Chlorocebus aethiops Cyclic AMP / metabolism Glucose / metabolism Insulin / metabolism* Insulin Secretion Mice Transfection
IF 4.097
引用数 43
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
遺伝子材料 Flamindo/pcDNA3 (RDB15247) Flamindo (R279E)/pcDNA3 (RDB15248).