Effects of different magnitudes of cyclic stretch on Na+-K+-ATPase in skeletal muscle cells in vitro.
The Na(+)-K(+)-ATPase, which plays a major role in modulation of skeletal muscle excitability and contractility, is one of the marker enzymes that senses the mechanical strain and adapts to the stimuli. Although many papers had been published on the effects of mechanical stress on Na(+)-K(+)-ATPase in aortic smooth muscle cells, little was known about the effects of different magnitudes of mechanical stretch on Na(+)-K(+)-ATPase in skeletal muscle cells. In the present study, we determined the effect of different magnitudes(6%, 12%, or 25% elongation) of cyclic stretch on the activity of the Na(+)-K(+)-ATPase and investigated possible mechanisms that might be involved in the action of stretch. The results showed the application of different magnitudes of cyclic stretch induced a magnitude-dependent increase of Na(+)-K(+)-ATPase activity in cultured skeletal muscle cells. Furthermore, inhibition of ionic fluxes through SACs prevented the action of stretch on Na(+)-K(+)-ATPase activity. The stretch-induced increase in Na(+)-K(+)-ATPase activity was not blocked by Actinomycin D. No significant changes in mRNA and total cell protein levels of Na(+)-K(+)-ATPase were detected after stretched continuous for 24 h. However, cyclic stretch increased cell surface expression of Na(+)-K(+)-ATPase alpha(1)- and alpha(2)-subunit proteins by 1.3- and 1.75-fold, respectively, and the increases in Na(+)-K(+)-ATPase activity and cell surface expression were abolished by LY-294002. These data indicated that cyclic stretch induced a "magnitude-dependent" increase of Na(+)-K(+)-ATPase activity in cultured skeletal muscle cells in vitro. The upregulation involved translocation of Na(+)-K(+)-ATPase alpha(1)- and alpha(2)-subunits to plasma membrane, not increased gene transcription. These results suggested a novel nontranscriptional mechanism for regulation of Na(+)-K(+)-ATPase in skeletal muscle cells by cyclic stretch.
Yuan
Xiao
X
Department of Orthodontics, 4th Military Medical University, Xi'an, Shanxi Province, People's Republic of China.
Lin
Zhu
Z
Luo
Songjiao
S
Ji
Guoping
G
Yuan
Changqing
C
Wu
Yong
Y
D062565:Acid Sensing Ion Channels; D000818:Animals; D000831:Animals, Newborn; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D015640:Ion Channel Gating; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D009025:Morpholines / Q000494:pharmacology; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D009470:Muscle Spindles / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015222:Sodium Channels / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*; D013314:Stress, Mechanical; D015854:Up-Regulation
D062565:Acid Sensing Ion Channels; D002867:Chromones; D003609:Dactinomycin; D007770:L-Lactate Dehydrogenase; D008565:Membrane Proteins; D009025:Morpholines; D009419:Nerve Tissue Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D015222:Sodium Channels; D000254:Sodium-Potassium-Exchanging ATPase; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C499075:Atp1a1 protein, rat; C498532:Atp1a2 protein, rat