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The subcellular location and cell biological function of small GTPase Rab40c in mammalian cells have not been investigated in detail. In this study, we demonstrated that the exogenously expressed GFP-Rab40c associates with lipid droplets marked by neutral lipid specific dye Oil red or Nile red, but not with the Golgi or endosomal markers. Further examination demonstrated that Rab40c is also associated with ERGIC-53 containing structures, especially under the serum starvation condition. Rab40c is increasingly recruited to the surface of lipid droplets during lipid droplets formation and maturation in HepG2 cells. Rab40c knockdown moderately decreases the size of lipid droplets, suggesting that Rab40c is involved in the biogenesis of lipid droplets. Stimulation for adipocyte differentiation increases the expression of Rab40c in 3T3-L1 cells. Rab40c interacts with TIP47, and is appositionally associated with TIP47-labeled lipid droplets. In addition, over-expression of Rab40c causes the clustering of lipid droplets independent of its GTPase activity, but completely dependent of the intact SOCS box domain of Rab40c. In addition, Rab40c displayed self-interaction as well as interaction with TIP47 and the SOCS box is essential for its ability to induce clustering of lipid droplets. Our results suggest that Rab40c is a novel Rab protein associated with lipid droplets, and is likely involved in modulating the biogenesis of lipid droplets. [ABSTRACT FROM AUTHOR]

Abstract: Glucagon-like peptide-1 (GLP-1)-enhanced insulin secretion is mainly mediated by cAMP-PKA and cAMP-Epac2 signaling pathways at physiological glucose concentrations. However the cellular mechanisms underlying the insulinotropic action of GLP-1 at glucotoxicity remain largely unknown. In the present study, we examined the effects of GLP-1 on glucotoxicity-diminished insulin secretion and explored the roles of these two cAMP-linked pathways in mediating the effects of GLP-1 under glucotoxic conditions. Consistent with the previous reports, exposure of INS-1E cells and mouse islets to 30mM glucose for 72h almost abolished glucose-stimulated insulin secretion. Addition of 10nM GLP-1 significantly increased glucose-stimulated insulin secretion. This was not due to a protective effect of GLP-1 against glucotoxicity-induced apoptosis but instead improvement of the secretory capacity of the insulin-secreting β-cells. It is of note that GLP-1 preferentially increased the expression and activity of PKA, whereas had no effects on Epac2 at high glucose. In correlation with the observations, treatment of INS-1E cells with the specific PKA inhibitor Rp-cAMPS completely abolished the insulinotropic action of GLP-1, whereas knock-down of Epac2 did not interfere the effects of GLP-1. Moreover, GLP-1 did not increase further insulin secretion in the presence of the PKA agonist 6-Bnz-cAMP-AM. By contrast, it produced additional enhancement of insulin secretion when Epac2 was maximally stimulated by its selective agonist 8-pCPT-2′-O-Me-cAMP-AM. Taken together, our results suggest that GLP-1 potentiates glucotoxicity-diminished insulin secretion mainly through cAMP-PKA signaling pathway. [Copyright &y& Elsevier]