Mechanical stimulus is a regulator of tenocyte metabolism. The present study investigated temporal regulation of the expression of selected genes by tenocytes in isolated fascicles subjected to tensile strain in vitro. Cyclic tensile strain with a 3% amplitude superimposed on a 2% static strain was provided for 10 min, followed by either an unstrained period or continuous cyclic strain until the end of a 24 hrs incubation period. mRNA expression of selected anabolic and catabolic genes were evaluated with qPCR at 10min, 1hr, 6hr and 24hrs. The application of 6 hours cyclic strain significantly upregulated type III collagen mRNA expression in strained fascicles compared to unstrained controls, but no alterations were observed in mRNA expression of type I collagen and biglycan. Significant downregulation in the expression of the decorin core protein was observed in fascicles subjected to 24 hours cyclic strain. MMP3 and MMP13 expression levels were upregulated by the application of 10 minutes of cyclic strain, followed by a progressive downregulation until the end of incubation period in both the absence and presence of the continuing cyclic strain. Accordingly, alterations in the expression of anabolic genes were limited to the upregulation of collagen III by prolonged exposure to cyclic strain, whereas catabolic genes were upregulated by a small number of strain cycles and downregulated by a prolonged cyclic strain. These findings demonstrate distinctive patterns of mechano-regulation for anabolic and catabolic genes, and help our understanding of tenocyte response to mechanical stimulation.