Essential role of Epac2/Rap1 signaling in regulation of insulin granule dynamics by cAMP.
cAMP is well known to regulate exocytosis in various secretory cells, but the precise mechanism of its action remains unknown. Here, we examine the role of cAMP signaling in the exocytotic process of insulin granules in pancreatic beta cells. Although activation of cAMP signaling alone does not cause fusion of the granules to the plasma membrane, it clearly potentiates both the first phase (a prompt, marked, and transient increase) and the second phase (a moderate and sustained increase) of glucose-induced fusion events. Interestingly, all granules responsible for this potentiation are newly recruited and immediately fused to the plasma membrane without docking (restless newcomer). Importantly, cAMP-potentiated fusion events in the first phase of glucose-induced exocytosis are markedly reduced in mice lacking the cAMP-binding protein Epac2 (Epac2(ko/ko)). In addition, the small GTPase Rap1, which is activated by cAMP specifically through Epac2 in pancreatic beta cells, mediates cAMP-induced insulin secretion in a protein kinase A-independent manner. We also have developed a simulation model of insulin granule movement in which potentiation of the first phase is associated with an increase in the insulin granule density near the plasma membrane. Taken together, these data indicate that Epac2/Rap1 signaling is essential in regulation of insulin granule dynamics by cAMP, most likely by controlling granule density near the plasma membrane.
Division of Cellular and Molecular Medicine, Department of Physiology and Cell Biology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe, Hyogo 650-0017, Japan.
Tadao
T
Shibasaki
Harumi
H
Takahashi
Takashi
T
Miki
Yasuhiro
Y
Sunaga
Kimio
K
Matsumura
Mami
M
Yamanaka
Changliang
C
Zhang
Atsuko
A
Tamamoto
Takaya
T
Satoh
Jun-Ichi
J
Miyazaki
Susumu
S
Seino
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D000242:Cyclic AMP / Q000502:physiology*; D003594:Cytoplasmic Granules / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D005089:Exocytosis / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007328:Insulin / Q000032:analysis / Q000378:metabolism*; D000078790:Insulin Secretion; D051379:Mice; D018345:Mice, Knockout; D011188:Potassium / Q000494:pharmacology; D015398:Signal Transduction; D020827:rap1 GTP-Binding Proteins / Q000032:analysis / Q000378:metabolism*
D002352:Carrier Proteins; D020662:Guanine Nucleotide Exchange Factors; D007328:Insulin; C486707:Rapgef4 protein, mouse; D000242:Cyclic AMP; D020827:rap1 GTP-Binding Proteins; D005947:Glucose; D011188:Potassium