Vascular endothelial growth factor expression in regenerating skeletal muscle ofthe rats after putted hindlimb in a cast :

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Reduction of muscular loading and paucity of neuronal im ulses to muscle fibers induces substantial atrophy of skeletal muscle fibers. Although these myofibers are known to recover to normal size and activity after various neuronal stimulations, it is still not clear whether the angiogenic response is involved in the recovery processes of atrophied skeletal muscle fibers because the atrophied muscles exihibit poor and pale vascularity. It has been shown that vascular endothelial growth factor (VEGF) plays an important role in the processes leading to angiogenesis. In addition, basic fibroblast growth factor (bFGF) synthesized within endothelial cells under several physiological and/or experimental conditions shows a direct mitogenic activity in endothelial cells. Therefore, in the present study, we have undertaken to examine by means of immunohistochemical methods whether VEGF and bFGF are induced in the recovering skeletal muscle after sustained hindlimb fixation. Wistar rats were used as the subjects. One leg was laced in a cast, and the animals were allowed to survive for a week, a er which they were housed normally for 3, 5 or 7 days, and then were anesthetized and fixed by vascular perfusion with 4% paraformaldehyde solution. The soleus muscle (SOL) was dissected out from the animals. 10 μm cryostat cross sections ofthe specimens were cut and reacted with anti-VEGF(polyclonal, Santa Cruze Bio. Inc. , USA) or anti-FGF abtibody (polyclonal, Chemicon, USA). Specimens were incubated in biotinilated JgG and HRP-conjugated streptavin, and were reacted with DAB. The immunohistochemical study revealed VEGF and bFGF-immunoreactivies in the cytoplasm of endothelial cells in skeletal muscles of rats 3 to 7 days after recovery from the hindlimb immobilization. With regard to the mode of expression, the immunoreactivities of both these cytokines contained to be expressed for 3 to 7 days after the release of hindlimb fixation. These results su ggested that expression of VEGF and bFGF may be involved during angiogenesis an remodeling of the vascular system during skeletal muscle recovery after forced atrophy.

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