title: Impaired NaV1.2 function and reduced cell surface expression in benign familial neonatal-infantile seizures.
abstract: OBJECTIVE
  Mutations in SCN2A, the gene encoding the brain voltage-gated sodium channel alpha-subunit Na(V)1.2, are associated with inherited epilepsies including benign familial neonatal-infantile seizures (BFNIS). Functional characterization of three BFNIS mutations was performed to identify defects in channel function that underlie this disease.
  
  
  METHODS
  We examined three BFNIS mutations (R1319Q, L1330F, and L1563V) using whole-cell patch-clamp recording of heterologously expressed human Na(V)1.2. Membrane biotinylation was employed to examine the cell surface protein expression of the four Na(V)1.2 alleles.
  
  
  RESULTS
  R1319Q displayed mixed effects on activation and fast inactivation gating, consistent with a net loss of channel function. L1563V exhibited impaired fast inactivation predicting a net gain of channel function. The L1330F mutation significantly decreased overall channel availability during repetitive stimulation. Patch-clamp analysis also revealed that cells expressing BFNIS mutants exhibited lower levels of sodium current compared to wild type (WT) Na(V)1.2. Biochemical experiments demonstrated that all three BFNIS mutations exhibited a significant reduction in cell surface expression compared to WT.
  
  
  CONCLUSIONS
  Our findings indicate that BFNIS is associated with a range of biophysical defects accompanied by reduced levels of channel protein at the plasma membrane.
authors.affiliation: Department of Pharmacology, Division of GeneticMedicine, Vanderbilt University, Nashville, Tennessee 37232-0275, USA.
authors.forename: Sunita N
authors.initials: SN
authors.lastname: Misra
authors.forename: Kristopher M
authors.initials: KM
authors.lastname: Kahlig
authors.forename: Alfred L
authors.initials: AL
authors.lastname: George
mesh_terms: D019921:Biotinylation; D002462:Cell Membrane / Q000502:physiology; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D004594:Electrophysiology / Q000295:instrumentation; D020936:Epilepsy, Benign Neonatal / Q000235:genetics* / Q000503:physiopathology*; D053503:Epithelial Sodium Channels / Q000235:genetics*; D006801:Humans; D007231:Infant, Newborn; D018408:Patch-Clamp Techniques; D017354:Point Mutation / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D053503:Epithelial Sodium Channels