Sheldon S Shen1, Richard A Steinhardt. 1. Department of Genetics, Development and Cellular Biology, Iowa State University, Ames, Iowa, USA.
Abstract
PURPOSE: To examine membrane repair mechanisms in rabbit corneal epithelial (RCE) cells. METHODS: Microneedle puncture and fluorescent dye loss were used to wound membranes and assay resealing, respectively. Different repair mechanisms were detected pharmacologically and with antisense oligonucleotides. RESULTS: The RCE cells rapidly reseal plasma membranes by calcium-dependent exocytotic mechanisms that exhibit both facilitated and potentiated responses to multiple wounding. The facilitated response was inhibited by specific inhibitors of protein kinase C (PKC) and brefeldin A, and the potentiated response was blocked by inhibitors of cAMP-dependent protein kinase (PKA). Reduction of myosin IIA inhibited the facilitated response, and reduction of IIB inhibited the initial resealing. CONCLUSIONS: RCE cells rapidly repair plasma membrane disruptions. At a second wound at the same site, PKC stimulated vesicle formation from the Golgi apparatus, resulting in more rapid membrane resealing for a facilitated response. The RCE cell also contains a PKA-dependent global potentiation of membrane resealing.
PURPOSE: To examine membrane repair mechanisms in rabbit corneal epithelial (RCE) cells. METHODS: Microneedle puncture and fluorescent dye loss were used to wound membranes and assay resealing, respectively. Different repair mechanisms were detected pharmacologically and with antisense oligonucleotides. RESULTS: The RCE cells rapidly reseal plasma membranes by calcium-dependent exocytotic mechanisms that exhibit both facilitated and potentiated responses to multiple wounding. The facilitated response was inhibited by specific inhibitors of protein kinase C (PKC) and brefeldin A, and the potentiated response was blocked by inhibitors of cAMP-dependent protein kinase (PKA). Reduction of myosin IIA inhibited the facilitated response, and reduction of IIB inhibited the initial resealing. CONCLUSIONS: RCE cells rapidly repair plasma membrane disruptions. At a second wound at the same site, PKC stimulated vesicle formation from the Golgi apparatus, resulting in more rapid membrane resealing for a facilitated response. The RCE cell also contains a PKA-dependent global potentiation of membrane resealing.
Authors: William M Bement; Hoi-Ying E Yu; Brian M Burkel; Emily M Vaughan; Andrew G Clark Journal: Curr Opin Cell Biol Date: 2006-12-14 Impact factor: 8.382
Authors: Heather L Chandler; Tao Tan; Chunlin Yang; Anne J Gemensky-Metzler; Rita F Wehrman; Qiwei Jiang; Cornelia M W Peterson; Bingchuan Geng; Xinyu Zhou; Qiang Wang; Denis Kaili; T M Ayodele Adesanya; Frank Yi; Hua Zhu; Jianjie Ma Journal: Commun Biol Date: 2019-02-20