Selective adhesion of osteoblastic cells to different integrin ligands induces osteopontin gene expression.
Skeletal homeostasis is partly regulated by the mechanical environment and specific signals generated by a cell's adhesion to the matrix. Previous studies demonstrated that osteopontin (OPN) expression is stimulated in response to both cellular adhesion and mechanical stimulation. The present studies examine if specific integrin ligands mediate osteoblast selective adhesion and whether opn mRNA expression is induced in response to these same ligands. Embryonic chicken calvaria osteoblastic cells were plated on bacteriological dishes coated with fibronectin (FN), collagen type I (Col1), denatured collagen/gelatin (G), OPN, vitronectin (VN), laminin (LN) or albumin (BSA). Osteoblastic cells were shown to selectively adhere to FN, Col1, G and LN, yet not to VN, OPN or BSA. Opn mRNA expression was induced by adhesion to Col1, FN, LN and G, but neither OPN nor VN induced this expression. Examination of the activation of the protein kinases A and C second signaling systems showed that only adhesion to FN induced protein kinase A and protein kinase C (PKC) activity while adherence to Col1 induced PKC. Evaluation of the intracellular distribution of focal adhesion kinase (FAK) and p-tyrosine within cells after adherence to FN, VN or BSA demonstrated that adherence to FN stimulated FAK translocation from the nucleus to the cytoplasm and high levels of p-tyrosine localization at the cell surface. However, cell adherence to VN or BSA did not show these morphological changes. These data illustrate that osteoblast selective adhesion is mediated by specific integrin ligands, and induction of intracellular second signal kinase activity is related to the nature of the ligand.
Carvalho
R S
RS
Department of Orthopaedic Surgery, Orthopaedic Research Laboratory, Boston University School of Medicine, 715 Albany Street, R-205, Boston, MA 02118-2526, USA.
Kostenuik
P J
PJ
Salih
E
E
Bumann
A
A
Gerstenfeld
L C
LC
D000818:Animals; D002448:Cell Adhesion; D002642:Chick Embryo; D017868:Cyclic AMP-Dependent Protein Kinases / Q000096:biosynthesis; D004790:Enzyme Induction; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology; D005786:Gene Expression Regulation; D016023:Integrins / Q000378:metabolism*; D008024:Ligands; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D053495:Osteopontin; D019000:Phosphotyrosine / Q000378:metabolism; D011493:Protein Kinase C / Q000096:biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012795:Sialoglycoproteins / Q000235:genetics*
D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D016023:Integrins; D008024:Ligands; D012333:RNA, Messenger; D012795:Sialoglycoproteins; D053495:Osteopontin; D019000:Phosphotyrosine; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C