title: Rac-mediated actin remodeling and myosin II are involved in KATP channel trafficking in pancreatic β-cells.
abstract: AMP-activated protein kinase (AMPK) is a metabolic sensor activated during metabolic stress and it regulates various enzymes and cellular processes to maintain metabolic homeostasis. We previously reported that activation of AMPK by glucose deprivation (GD) and leptin increases KATP currents by increasing the surface levels of KATP channel proteins in pancreatic β-cells. Here, we show that the signaling mechanisms that mediate actin cytoskeleton remodeling are closely associated with AMPK-induced KATP channel trafficking. Using F-actin staining with Alexa 633-conjugated phalloidin, we observed that dense cortical actin filaments present in INS-1 cells cultured in 11 mM glucose were disrupted by GD or leptin treatment. These changes were blocked by inhibiting AMPK using compound C or siAMPK and mimicked by activating AMPK using AICAR, indicating that cytoskeletal remodeling induced by GD or leptin was mediated by AMPK signaling. AMPK activation led to the activation of Rac GTPase and the phosphorylation of myosin regulatory light chain (MRLC). AMPK-dependent actin remodeling induced by GD or leptin was abolished by the inhibition of Rac with a Rac inhibitor (NSC23766), siRac1 or siRac2, and by inhibition of myosin II with a myosin ATPase inhibitor (blebbistatin). Immunocytochemistry, surface biotinylation and electrophysiological analyses of KATP channel activity and membrane potentials revealed that AMPK-dependent KATP channel trafficking to the plasma membrane was also inhibited by NSC23766 or blebbistatin. Taken together, these results indicate that AMPK/Rac-dependent cytoskeletal remodeling associated with myosin II motor function promotes the translocation of KATP channels to the plasma membrane in pancreatic β-cells.
authors.affiliation: Department of Physiology and Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Young-Eun
authors.initials: YE
authors.lastname: Han
authors.affiliation: Department of Physiology and Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Ajin
authors.initials: A
authors.lastname: Lim
authors.affiliation: Department of Physiology and Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Sun-Hyun
authors.initials: SH
authors.lastname: Park
authors.affiliation: Department of Physiology and Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Sunghoe
authors.initials: S
authors.lastname: Chang
authors.affiliation: Department of Physiology and Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Suk-Ho
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Physiology and Biomembrane Plasticity Research Center, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Won-Kyung
authors.initials: WK
authors.lastname: Ho
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D005947:Glucose / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D054086:KATP Channels / Q000378:metabolism*; D020738:Leptin / Q000378:metabolism; D024462:Myosin Type II / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D015398:Signal Transduction; D020744:rac GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D000199:Actins; D054086:KATP Channels; D020738:Leptin; D055372:AMP-Activated Protein Kinases; D024462:Myosin Type II; D020744:rac GTP-Binding Proteins; D005947:Glucose