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MUSCLE CELL BIOLOGY AND CELL MOTILITY

Satellite cell activation in stretched skeletal muscle and the role of nitric oxide and hepatocyte growth factor

¹Department of Animal Science, Hokkaido University, Sapporo, Hokkaido, Japan; ²Department of Bioscience and Biotechnology, Kyushu University, Fukuoka, Japan; ³Muscle Biology Group, Department of Animal Sciences, and ⁴Veterinary Diagnostic Laboratory, College of Agriculture and Life Sciences, University of Arizona, Tucson, Arizona

Submitted 14 October 2005 ; accepted in final form 28 December 2005

In the present study, we examined the roles of hepatocyte growth factor (HGF) and nitric oxide (NO) in the activation of satellite cells in passively stretched rat skeletal muscle. A hindlimb suspension model was developed in which the vastus, adductor, and gracilis muscles were subjected to stretch for 1 h. Satellite cells were activated by stretch determined on the basis of 5-bromo-2'-deoxyuridine (BrdU) incorporation in vivo. Extracts from stretched muscles stimulated BrdU incorporation in freshly isolated control rat satellite cells in a concentration-dependent manner. Extracts from stretched muscles contained the active form of HGF, and the satellite cell-activating activity could be neutralized by incubation with anti-HGF antibody. The involvement of NO was investigated by administering nitro-L-arginine methyl ester (L-NAME) or the inactive enantiomer N^G-nitro-D-arginine methyl ester HCl (D-NAME) before stretch treatment. In vivo activation of satellite cells in stretched muscle was not inhibited by D-NAME but was inhibited by L-NAME. The activity of stretched muscle extract was abolished by L-NAME treatment but could be restored by the addition of HGF, indicating that the extract was not inhibitory. Finally, NO synthase activity in stretched and unstretched muscles was assayed in muscle extracts immediately after 2-h stretch treatment and was found to be elevated in stretched muscle but not in stretched muscle from L-NAME-treated rats. The results of these experiments demonstrate that stretching muscle liberates HGF in a NO-dependent manner, which can activate satellite cells.

muscle regeneration

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