Regulation of fibronectin, integrin and cytoskeleton expression in differentiating adipocytes: inhibition by extracellular matrix and polylysine

The differentiation of 3T3 preadipocytes into adipocytes is characterized by major changes in cell morphology from a fibroblastic to a rounded shape and by the induction of gene expression related to lipid metabolism. We have studied the synthesis and mRNA levels of proteins involved in the formation of cell-matrix contacts and in defining cell shape to determine the role and molecular basis of these morphological changes during adipose conversion. When confluent preadipocyte cultures were stimulated with adipogenic medium there was a gradual decrease in the expression of fibronectin, beta-integrin, actin and in the microfilament-associated proteins vinculin, alpha-actinin and tropomyosin. The changes in extracellular matrix and cytoskeletal mRNA levels were apparent before the accumulation of glycerophosphate dehydrogenase (GPD) mRNA and continued during the massive increase in GPD mRNA level. The culturing of preadipocytes on an extracellular matrix deposited on the dish by corneal endothelial cells, or on substrata coated with polylysine, prevented the morphological changes, the decrease in the level of assembled actin, the accumulation of lipid and the shifts in the expression of integrin, cytoskeletal proteins and GPD. In cells cultured on malleable hydrated collagen gels, adipocyte differentiation proceeded at normal rates. The results suggest that the regulated expression of proteins involved in the formation of the transmembrane linkage between the extracellular matrix and the microfilaments are programmed regulatory events that affect cell adhesion and thereby cell shape during adipocyte differentiation.