In the present study, we test the hypothesis that mouse skeletal muscle in culture retains the fundamental properties of excitation-sarcoplasmic reticulum Ca2 release coupling reported for young-adult (3–4 mo) and senescent (22–23) mice. Dissociated flexor digitorum brevis (FDB) muscles from young-adult and senescent mice were cultured for 7 d in a serum-free medium. During this period, the overall morphology of cultured fibers resembled that exhibited by acutely dissociated cells. In addition, survival analysis revealed that more than 70% of the fibers from both young and old mice remained suitable for electrophysiological studies during this same culture period. Charge movement and intracellular Ca2 recordings in FDB fibers, voltage clamped in the whole cell configuration of the patch-clamp technique, reproduced the maximal values, and voltage dependence similarly displayed by acutely dissociated cells for both parameters in young-adult and senescent mice. The analysis of the dihydropyridine receptor by immunoblots confirmed, in the culture system, the age-dependent decrease in the expression of this protein. In conclusion, FDB fibers from young-adult and old mice retain the excitation–contraction coupling phenotype during the course of a week in serum-free medium culture.
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22 August 2007
Muscle fibers from senescent mice retain excitation–contraction coupling properties in culture
Zhong-Min Wang,
Zhenlin Zheng,
María L. Messi,
Osvaldo Delbono
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In Vitro Cellular & Developmental Biology - Animal
Vol. 43 • No. 7
July 2007
Vol. 43 • No. 7
July 2007
aging
Dihydropyridine receptor
intracellular calcium
ryanodine receptor
Skeletal muscle