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Publication : Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis.

First Author  Ohara-Imaizumi M Year  2007
Journal  J Cell Biol Volume  177
Issue  4 Pages  695-705
PubMed ID  17502420 Mgi Jnum  J:134722
Mgi Id  MGI:3789566 Doi  10.1083/jcb.200608132
Citation  Ohara-Imaizumi M, et al. (2007) Imaging analysis reveals mechanistic differences between first- and second-phase insulin exocytosis. J Cell Biol 177(4):695-705
abstractText  The mechanism of glucose-induced biphasic insulin release is unknown. We used total internal reflection fluorescence (TIRF) imaging analysis to reveal the process of first- and second-phase insulin exocytosis in pancreatic beta cells. This analysis showed that previously docked insulin granules fused at the site of syntaxin (Synt)1A clusters during the first phase; however, the newcomers fused during the second phase external to the Synt1A clusters. To reveal the function of Synt1A in phasic insulin exocytosis, we generated Synt1A-knockout (Synt1A(-/-)) mice. Synt1A(-/-) beta cells showed fewer previously docked granules with no fusion during the first phase; second-phase fusion from newcomers was preserved. Rescue experiments restoring Synt1A expression demonstrated restoration of granule docking status and fusion events. Inhibition of other syntaxins, Synt3 and Synt4, did not affect second-phase insulin exocytosis. We conclude that the first phase is Synt1A dependent but the second phase is not. This indicates that the two phases of insulin exocytosis differ spatially and mechanistically.
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