| RRC ID |
49260
|
| Author |
Kitaguchi T, Oya M, Wada Y, Tsuboi T, Miyawaki A.
|
| Title |
Extracellular calcium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells.
|
| Journal |
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.
|
| Volume |
450(2)
|
| Pages |
365-73
|
| Published |
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
|
| Times Cited |
43
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| DNA material |
Flamindo/pcDNA3 (RDB15247)
Flamindo (R279E)/pcDNA3 (RDB15248). |
