Functional characterization and neuronal modeling of the effects of childhood absence epilepsy variants of CACNA1H, a T-type calcium channel

J Neurosci. 2005 May 11;25(19):4844-55. doi: 10.1523/JNEUROSCI.0847-05.2005.

Abstract

Sequencing of the T-type Ca2+ channel gene CACNA1H revealed 12 nonsynonymous single nucleotide polymorphisms (SNPs) that were found only in childhood absence epilepsy (CAE) patients. One SNP, G773D, was found in two patients. The present study reports the finding of a third patient with this SNP, as well as analysis of their parents. Because of the role of T-channels in determining the intrinsic firing patterns of neurons involved in absence seizures, it was suggested that these SNPs might alter channel function. The goal of the present study was to test this hypothesis by introducing these polymorphisms into a human Ca(v)3.2a cDNA and then study alterations in channel behavior using whole-cell patch-clamp recording. Eleven SNPs altered some aspect of channel gating. Computer simulations predict that seven of the SNPs would increase firing of neurons, with three of them inducing oscillations at similar frequencies, as observed during absence seizures. Three SNPs were predicted to decrease firing. Some CAE-specific SNPs (e.g., G773D) coexist with SNPs also found in controls (R788C); therefore, the effect of these polymorphisms were studied. The R788C SNP altered activity in a manner that would also lead to enhanced burst firing of neurons. The G773D-R788C combination displayed different behavior than either single SNP. Therefore, common polymorphisms can alter the effect of CAE-specific SNPs, highlighting the importance of sequence background. These results suggest that CACNA1H is a susceptibility gene that contributes to the development of polygenic disorders characterized by thalamocortical dysrhythmia, such as CAE.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Calcium Channels, T-Type / genetics
  • Calcium Channels, T-Type / physiology*
  • Cell Line
  • Dose-Response Relationship, Radiation
  • Electric Stimulation / methods
  • Epilepsy, Absence / genetics*
  • Epilepsy, Absence / physiopathology*
  • Excitatory Postsynaptic Potentials / physiology
  • Excitatory Postsynaptic Potentials / radiation effects
  • Female
  • Humans
  • Kinetics
  • Male
  • Membrane Potentials / genetics
  • Membrane Potentials / physiology*
  • Membrane Potentials / radiation effects
  • Models, Neurological*
  • Mutagenesis, Site-Directed / methods
  • Neurons / physiology*
  • Patch-Clamp Techniques / methods
  • Polymorphism, Single Nucleotide / physiology
  • Time Factors
  • Transfection / methods

Substances

  • CACNA1H protein, human
  • Calcium Channels, T-Type