The role of type V collagen fibril as an ECM that induces the motility of glomerular endothelial cells.
Although type V collagen (Col V) is present in developing and mature connective tissues of glomeruli, its primary function has not been elucidated yet. The purpose of this study was to elucidate the role of Col V fibrils in glomerular cells. We isolated primary cells from porcine kidney and cultured them on Col V fibrils reconstructed from purified Col V molecules extracted from porcine cornea. Time-lapse observation showed that Col V fibrils induce dynamic movement of glomerular endothelial cells (GEC) by stimulating them to extend long filopodial protrusions and wide lamellipodia. Col V signaling mediated through beta1 integrin activated phosphorylation of paxillin at tyrosine 118 (paxillin-pY118) and of focal adhesion kinase at tyrosine 861 (FAKpY861) at the cell periphery; a second Col V signal was mediated through neuroglycan 2 and activated FAKpY397. FAKpY861 was present in loose attachment points between Col V fibrils and GEC, allowing the cells to migrate easily. Activation of FAKpY397 induced incomplete focal adhesion at the centers of cells and caused cell movement. Therefore both signaling pathways facilitated cell motility, which was inhibited by the addition of antibodies to beta1 integrin, NG2, and Col V. We suggest that Col V fibrils activate 'outside-in' signaling in GEC and induce their dynamic motility.
Murasawa
Yusuke
Y
Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba city, Ibaraki 305-8572, Japan. ore_mura@hotmail.com
Hayashi
Toshihiko
T
Wang
Pi-Chao
PC
D000818:Animals; D002465:Cell Movement; D002478:Cells, Cultured; D024062:Collagen Type V / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D042783:Endothelial Cells / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D024022:Fibrillar Collagens / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D007668:Kidney / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D008954:Models, Biological; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology; D013552:Swine
D024062:Collagen Type V; D024022:Fibrillar Collagens; D011956:Receptors, Cell Surface; C052131:extracellular matrix receptor; D011494:Protein Kinases