Ca2+ leakage out of the sarcoplasmic reticulum is increased in type I skeletal muscle fibres in aged humans

Abstract

Key points: The amount of Ca stored in the sarcoplasmic reticulum (SR) of muscle fibres is decreased in aged individuals, and an important question is whether this results from increased Ca leakage out through the Ca release channels (ryanodine receptors; RyRs). The present study examined the effects of blocking the RyRs with Mg, or applying a strong reducing treatment, on net Ca accumulation by the SR in skinned muscle fibres from Old (∼70 years) and Young (∼24 years) adults. Raising cytoplasmic [Mg] and reducing treatment increased net SR Ca accumulation in type I fibres of Old subjects relative to that in Young. The densities of RyRs and dihydropyridine receptors were not significantly changed in the muscle of Old subjects. These findings indicate that oxidative modification of the RyRs causes increased Ca leakage from the SR in muscle fibres in Old subjects, which probably deleteriously affects normal muscle function both directly and indirectly. The present study examined whether the lower Ca storage levels in the sarcoplasmic reticulum (SR) in vastus lateralis muscle fibres in Old (70 ± 4 years) relative to Young (24 ± 4 years) human subjects is the result of increased leakage of Ca out of the SR through the Ca release channels/ryanodine receptors (RyRs) and due to oxidative modification of the RyRs. SR Ca accumulation in mechanically skinned muscle fibres was examined in the presence of 1, 3 or 10 mm cytoplasmic Mg because raising [Mg] strongly inhibits Ca efflux through the RyRs. In type I fibres of Old subjects, SR Ca accumulation in the presence of 1 mm Mg approached saturation at shorter loading times than in Young subjects, consistent with Caleakage limiting net uptake, and raising [Mg] to 10 mm in such fibres increased maximal SR Ca accumulation. No significant differences were seen in type II fibres. Treatment with dithiothreitol (10 mm for 5 min), a strong reducing agent, also increased maximal SR Ca accumulation at 1 mm Mg in type I fibres of Old subjects but not in other fibres. The densities of dihydropyridine receptors and RyRs were not significantly different in muscles of Old relative to Young subjects. These findings indicate that Ca leakage from the SR is increased in type I fibres in Old subjects by reversible oxidative modification of the RyRs; this increased SR Ca leak is expected to have both direct and indirect deleterious effects on Ca movements and muscle function