Rho-ROCK signaling differentially regulates chondrocyte spreading on fibronectin and bone sialoprotein.
The mammalian growth plate is a dynamic structure rich in extracellular matrix (ECM). Interactions of growth plate chondrocytes with ECM proteins regulate cell behavior. In this study, we compared chondrocyte adhesion and spreading dynamics on fibronectin (FN) and bone sialoprotein (BSP). Chondrocyte adhesion and spreading were also compared with fibroblasts to analyze potential cell-type-specific effects. Chondrocyte adhesion to BSP is independent of posttranslational modifications but is dependent on the RGD sequence in BSP. Whereas chondrocytes and fibroblasts adhered at similar levels on FN and BSP, cells displayed more actin-dependent spread on FN despite a 16x molar excess of BSP adsorbed to plastic. To identify intracellular mediators responsible for this difference in spreading, we investigated focal adhesion kinase (FAK)-Src and Rho-Rho kinase (ROCK) signaling. Although activated FAK localized to the vertices of adhered chondrocytes, levels of FAK activation did not correlate with the extent of spreading. Furthermore, Src inhibition reduced chondrocyte spreading on both FN and BSP, suggesting that FAK-Src signaling is not responsible for less cell spreading on BSP. In contrast, inhibition of Rho and ROCK in chondrocytes increased cell spreading on BSP and membrane protrusiveness on FN but did not affect cell adhesion. In fibroblasts, Rho inhibition increased fibroblast spreading on BSP while ROCK inhibition changed membrane protrusiveness of FN and BSP. In summary, we identify a novel role for Rho-ROCK signaling in regulating chondrocyte spreading and demonstrate both cell- and matrix molecule-specific mechanisms controlling cell spreading.
Gill
Kamal S
KS
CIHR Group in Skeletal Development and Remodeling, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.
Beier
Frank
F
Goldberg
Harvey A
HA
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D048430:Cell Shape / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000502:physiology*; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology; D005353:Fibronectins / Q000378:metabolism*; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D058574:Integrin-Binding Sialoprotein; D051379:Mice; D009842:Oligopeptides / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D012795:Sialoglycoproteins / Q000378:metabolism*; D015398:Signal Transduction; D020741:rho GTP-Binding Proteins / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism
D005353:Fibronectins; C546872:Ibsp protein, mouse; D058574:Integrin-Binding Sialoprotein; D009842:Oligopeptides; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; C047981:arginyl-glycyl-aspartic acid; D051417:Focal Adhesion Kinase 1; C496051:Ptk2 protein, mouse; D019061:src-Family Kinases; D054460:rho-Associated Kinases; D020741:rho GTP-Binding Proteins