Improved skeletal muscle Ca2+ regulation in vivo following contractions in mice overexpressing PGC-1α

In skeletal muscle, resting intracellular Ca2+ concentration ([ Ca2+]i) homeostasis is exquisitely regulated by Ca2+ transport across the sarcolemmal, mitochondrial, and sarcoplasmic reticulum (SR) membranes. Of these three systems, the relative importance of the mitochondria in [ Ca2+] i regulation remains poorly understood in in vivo skeletal muscle. We tested the hypothesis that the capacity for Ca2+ uptake by mitochondria is a primary factor in determining [ Ca2+] i regulation in muscle at rest and following contractions. Tibialis anterior muscle of anesthetized peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1 alpha)-overexpressing (OE, increased mitochondria model) and wild-type (WT) littermate mice was exteriorized in vivo and loaded with the fluorescent probe fura 2-AM, and Rhod 2-AM Ca2+ buffering and mitochondrial [ Ca2+] were evaluated at rest and during recovery from fatiguing tetanic contractions induced by electrical stimulation (120 s, 100 Hz). In addition, the effects of pharmacological inhibition of SR (thapsigargin) and mitochondrial [ carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP)] function were examined at rest. [ Ca2+] i in WT remained elevated for the entire postcontraction recovery period (+6 +/- 1% at 450 s), but in PGC-1 alpha OE [ Ca2+] i returned to resting baseline within 150 s. Thapsigargin immediately and substantially increased resting [ Ca2+] i in WT, whereas in PGC-1 alpha OE this effect was delayed and markedly diminished (WT, +12 +/- 3; PGC-1 alpha OE, + 1 +/- 2% at 600 s after thapsigargin treatment, P < 0.05). FCCP abolished this improvement of [ Ca2+] i regulation in PGC-1 alpha OE. Mitochondrial [ Ca2+] accumulation was observed in PGC-1 alpha OE following contractions and thapsigargin treatment. In the SR, PGC-1 alpha OE downregulated SR Ca2+ ATPase 1 (Ca2+ uptake) and parvalbumin (Ca2+ buffering) protein levels, whereas mitochondrial Ca2+ uptake-related proteins (Mfn1, Mfn2, and mitochondrial Ca2+ uniporter) were upregulated. These data demonstrate a heretofore unappreciated role for skeletal muscle mitochondria in [ Ca2+] i regulation in vivo following fatiguing tetanic contractions and at rest.