disrupted membrane structure and intracellular ca2+ signaling in adult skeletal muscle with acute knockdown of bin1破坏膜结构和细胞内钙离子信号在成人骨骼肌急性bin1击倒.pdfVIP

Disrupted Membrane Structure and Intracellular Ca2+ Signaling in Adult Skeletal Muscle with Acute Knockdown of Bin1 1 1 1 2 1 Andoria Tjondrokoesoemo , Ki Ho Park , Christopher Ferrante , Shinji Komazaki , Sebastian Lesniak , 3 1 4 1 1 Marco Brotto , Jae-Kyun Ko , Jingsong Zhou , Noah Weisleder *, Jianjie Ma 1 Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey (UMDNJ), Piscataway, New Jersey, United States of America, 2 Department of Anatomy, Saitama Medical School, Saitama, Japan, 3 Muscle Biology Research Group, University of Missouri, Kansas City, Missouri, United States of America, 4 Department of Molecular Biophysics and Physiology, Rush University, Chicago, Illinois, United States of America Abstract Efficient intracellular Ca2+ ([Ca2+]i) homeostasis in skeletal muscle requires intact triad junctional complexes comprised of t- tubule invaginations of plasma membrane and terminal cisternae of sarcoplasmic reticulum. Bin1 consists of a specialized BAR domain that is associated with t-tubule development in skeletal muscle and involved in tethering the dihydropyridine receptors (DHPR) to the t-tubule. Here, we show that Bin1 is important for Ca2+ homeostasis in adult skeletal muscle. Since systemic ablation of Bin1 in mice results in postnatal lethality, in vivo electroporation mediated transfection method was used to deliver RFP-tagged plasmid that produced short –hairpin (sh)RNA targeting Bin1 (shRNA-Bin1) to study the effect of Bin1 knockdown in adult mouse FDB skeletal muscle. Upon confirming the reduction of endogenous Bin1 expression, we