Generalized epilepsy with febrile seizures plus-associated sodium channel β1 subunit mutations severely reduce beta subunit-mediated modulation of sodium channel function

Two novel mutations (R85C and R85H) on the extracellular immunoglobulin-like domain of the sodium channel beta 1 subunit have been identified in individuals from two families with generalized epilepsy with febrile seizures plus (GEFS+). The functional consequences of these two mutations were determined by co-expression of the human brain NaV1.2 alpha subunit with wild type or mutant beta 1 subunits in human embryonic kidney (HEK)-293T cells. Patch clamp studies confirmed the regulatory role of beta 1 in that relative to NaV1.2 alone the NaV1.2+beta 1 currents had right-shifted voltage dependence of activation, fast and slow inactivation and reduced use dependence. In addition, the NaV1.2+beta 1 current entered fast inactivation slightly faster than NaV1.2 channels alone. The beta 1(R85C) subunit appears to be a complete loss of function in that none of the modulating effects of the wild type beta 1 were observed when it was co-expressed with NaV1.2. Interestingly, the beta 1(R85H) subunit also failed to modulate fast kinetics, however, it shifted the voltage dependence of steady state slow inactivation in the same way as the wild type beta 1 subunit. Immunohistochemical studies revealed cell surface expression of the wild type beta 1 subunit and undetectable levels of cell surface expression for both mutants. The functional studies suggest association of the beta 1(R85H) subunit with the a subunit where its influence is limited to modulating steady state slow inactivation. In summary, the mutant beta 1 subunits essentially fail to modulate a subunits which could increase neuronal excitability and underlie GEFS+ pathogenesis. (c) 2007 IBRO. Published by Elsevier Ltd. All rights reserved.