Mechanisms of a Human Skeletal Myotonia Produced by Mutation in the C-Terminus of NaV1.4: Is Ca2+ Regulation Defective?

Mutations in the cytoplasmic tail (CT) of voltage gated sodium channels cause a spectrum of inherited diseases of cellular excitability, yet to date only one mutation in the CT of the human skeletal muscle voltage gated sodium channel (hNa(V)1.4(F1705I)) has been linked to cold aggravated myotonia. The functional effects of altered regulation of hNa(V)1.4(F1705I) are incompletely understood. The location of the hNa(V)1.4(F1705I) in the CT prompted us to examine the role of Ca2+ and calmodulin (CaM) regulation in the manifestations of myotonia. To study Na channel related mechanisms of myotonia we exploited the differences in rat and human Na(V)1.4 channel regulation by Ca2+ and CaM. hNa(V)1.4(F1705I) inactivation gating is Ca2+-sensitive compared to wild type hNa(V)1.4 which is Ca2+ insensitive and the mutant channel exhibits a depolarizing shift of the V-1/2 of inactivation with CaM over expression. In contrast the same mutation in the rNa(V)1.4 channel background (rNa(V)1.4(F1698I)) eliminates Ca2+ sensitivity of gating without affecting the CaM over expression induced hyperpolarizing shift in steady-state inactivation. The differences in the Ca2+ sensitivity of gating between wild type and mutant human and rat Na(V)1.4 channels are in part mediated by a divergence in the amino acid sequence in the EF hand like (EFL) region of the CT. Thus the composition of the EFL region contributes to the species differences in Ca2+/CaM regulation of the mutant channels that produce myotonia. The myotonia mutation F1705I slows I-Na decay in a Ca2+-sensitive fashion. The combination of the altered voltage dependence and kinetics of I-Na decay contribute to the myotonic phenotype and may involve the Ca2+-sensing apparatus in the CT of Na(V)1.4.