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Vol. 13, Issue 8, 2909-2918, August 2002
¶
*Department of Animal Science, Hokkaido University, Sapporo,
Hokkaido, Japan; Application of mechanical stretch to cultured adult rat muscle
satellite cells results in release of hepatocyte growth factor (HGF)
and accelerated entry into the cell cycle. Stretch activation of
cultured rat muscle satellite cells was observed only when medium pH
was between 7.1 and 7.5, even though activation of satellite cells was
accelerated by exogenous HGF over a pH range from 6.9 to 7.8. Furthermore, HGF was only released in stretched cultures when the pH of
the medium was between 7.1 and 7.4. Conditioned medium from
stretched satellite cell cultures stimulated activation of unstretched
satellite cells, and the addition of anti-HGF neutralizing antibodies
to stretch-conditioned medium inhibited the stretch activation
response. Conditioned medium from satellite cells that were stretched
in the presence of nitric-oxide synthase (NOS) inhibitor
NDepartment of Bioscience and
Biotechnology, Kyushu University, Fukuoka, Japan;
§Department of Human Anatomy and Cell Science, University
of Manitoba, Winnipeg, Canada; and Muscle Biology Group,
Department of Animal Sciences, University of Arizona, Tucson, Arizona
35721
-nitro-L-arginine methyl
ester hydrochloride did not accelerate activation of unstretched
control satellite cells, and HGF was not released into the medium.
Conditioned medium from unstretched cells that were treated with a
nitric oxide donor, sodium nitroprusside dihydrate, was able to
accelerate the activation of satellite cells in vitro, and HGF was
found in the conditioned medium. Immunoblot analysis
indicated that both neuronal and endothelial NOS isoforms were present
in satellite cell cultures. Furthermore, assays of NOS activity in
stretched satellite cell cultures demonstrated that NOS is stimulated
when satellite cells are stretched in vitro. These experiments indicate
that stretch triggers an intracellular cascade of events, including
nitric oxide synthesis, which results in HGF release and satellite cell activation.
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