An evaluation of common markers of muscle denervation in denervated young-adult and old rat gastrocnemius muscle

A large part of age-related muscle wasting is due to incomplete reinnervation of fibres that have become denervated following motoneuron loss. Neural cell adhesion molecule (NCAM) and sodium channel NaV1.5 are considered markers for denervation, but the time course of changes in their expression following denervation has never been systematically evaluated in young-adult and old muscle. To assess the time course of denervation-induced changes in their expression, the left gastrocnemius muscle in 15 young-adult (5-month) and 10 old (25-month) male Wistar rats was denervated for 1, 2 or 4 weeks, while the right muscle served as an internal control. Sections were stained for alpha-bungarotoxin, to visualise the neuromuscular junctions, combined with NCAM, polysialylated NCAM (PSA-NCAM) or NaV1.5. In young-adult animals, denervation induced a transient decrease in junctional and cytoplasmic NCAM expression, while in the old NCAM expression was increased after 2 weeks. Cytoplasmic PSA-NCAM was increased in both young-adult and old fibres after 2 weeks denervation with a further increase after 4 weeks in the young only. The junctional PSA-NCAM was transiently increased or decreased in the young and old muscles, respectively. NaV1.5 expression decreased after 1 and 2 weeks of denervation in NaV1.5 in young muscle fibres before returning to control levels, whereas old muscle fibres displayed a transient increase after 1 week followed by a decrease and a return to control levels after 2 and 4 weeks respectively. In conclusion, NCAM and NaV1.5 are not unequivocally elevated with denervation and consequently are not adequate markers of fibre denervation.