RGD and YIGSR synthetic peptides facilitate cellular adhesion identical to that of laminin and fibronectin but alter the physiology of neonatal cardiac myocytes.
In the mammalian heart, the extracellular matrix plays an important role in regulating cell behavior and adaptation to mechanical stress. In cell culture, a significant number of cells detach in response to mechanical stimulation, limiting the scope of such studies. We describe a method to adhere the synthetic peptides RGD (fibronectin) and YIGSR (laminin) onto silicone for culturing primary cardiac cells and studying responses to mechanical stimulation. We first examined cardiac cells on stationary surfaces and observed the same degree of cellular adhesion to the synthetic peptides as their respective native proteins. However, the number of striated myocytes on the peptide surfaces was significantly reduced. Focal adhesion kinase (FAK) protein was reduced by 50% in cardiac cells cultured on YIGSR peptide compared with laminin, even though beta(1)-integrin was unchanged. Connexin43 phosphorylation increased in cells adhered to RGD and YIGSR peptides. We then subjected the cardiac cells to cyclic strain at 20% maximum strain (1 Hz) for 48 h. After this period, cell attachment on laminin was reduced to approximately 50% compared with the unstretched condition. However, in cells cultured on the synthetic peptides, there was no significant difference in cell adherence after stretch. On YIGSR peptide, myosin protein was decreased by 50% after mechanical stimulation. However, total myosin was unchanged in cells stretched on laminin. These results suggest that RGD and YIGSR peptides promote the same degree of cellular adhesion as their native proteins; however, they are unable to promote the signaling required for normal FAK expression and complete sarcomere formation in cardiac myocytes.
Boateng
Samuel Y
SY
Department of Physiology and Biophysics, University of Illinois at Chicago, 60612-7342, USA.
Lateef
Syed S
SS
Mosley
William
W
Hartman
Thomas J
TJ
Hanley
Luke
L
Russell
Brenda
B
D000818:Animals; D000831:Animals, Newborn; D002448:Cell Adhesion / Q000187:drug effects* / Q000502:physiology; D018031:Connexin 43 / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000494:pharmacology*; D010766:Phosphorylation; D010812:Physical Stimulation; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D012828:Silicones
D018031:Connexin 43; D016326:Extracellular Matrix Proteins; D009842:Oligopeptides; D012828:Silicones; C081571:tyrosyl-isoleucyl-glycyl-seryl-arginine; C047981:arginyl-glycyl-aspartic acid