Lee Winchester

Hyperhomocysteinemia associated skeletal muscle weakness involves mitochondrial dysfunction and epigenetic modifications

HHcy has been implicated in elderly frailty, but the underlying mechanisms are poorly understood. Using C57 and CBS+/- mice and C2C12 cell line, we investigated mechanisms behind HHcy induced skeletal muscle weakness and fatigability. Possible alterations in metabolic capacity (levels of LDH, CS, MM-CK and COX-IV), in structural proteins (levels of dystrophin) and in mitochondrial function (ATP production) were examined. An exercise regimen was employed to reverse HHcy induced changes. CBS+/- mice exhibited more fatigability, and generated less contraction force. No significant changes in muscle morphology were observed. However, there is a corresponding reduction in large muscle fiber number in CBS+/- mice. Excess fatigability was not due to changes in key enzymes involved in metabolism, but was due to reduced ATP levels. A marginal reduction in dystrophin levels along with a decrease in mitochondrial transcription factor A (mtTFA) were observed. There was also an increase in the mir-31, and mir-494 quantities that were implicated in dystrophin and mtTFA regulation respectively. The molecular changes elevated during HHcy, with the exception of dystrophin levels, were reversed after exercise. In addition, the amount of NRF-1, one of the transcriptional regulators of mtTFA, was significantly decreased. Furthermore, there was enhancement in mir-494 levels and a concomitant decline in mtTFA protein quantity in homocysteine treated cells. These changes in C2C12 cells were also accompanied by an increase in DNMT3a and DNMT3b proteins and global DNA methylation levels. Together, these results suggest that HHcy plays a causal role in enhanced fatigability through mitochondrial dysfunction which involves epigenetic changes.

Sex Differences in Time to Fatigue at 100% VO2peak When Normalized for Fat Free Mass

The purpose of this investigation was to compare sexes for time to fatigue at 100% VO2peak in recreationally trained individuals. Ten men (age: 23.4 ± 1.8; height: 177 ± 6.7; body mass: 83.8 ± 11.3; ± SD) and nine women (age: 25.0 ± 2.5; height: 165.6 ± 5.5; body mass: 62.7 ± 6.7) participated in this investigation after providing written consent. One week after assessing VO2peak, subjects exercised on an electrically braked cycle ergometer at 100% of VO2peak until fatigue. The time taken to fatigue was 48.9% longer for men than women (274 ± 13s vs. 184 ± 14s; p < 0.001, for men and women, respectively). When normalized for fat free mass (ffm; s/kg ffm) no significant differences between men and women were observed (3.99 ± 0.21s/kg ffm vs. 3.72 ± 0.28s/kg ffm for men and women, respectively, p = 0.431). The difference in fatigability between the sexes at this exercise intensity is to a large degree related to the difference in fat free mass.