Children With Idiopathic Generalized Epilepsy Susceptibility Genes Cacna1h And Gabrb3 Analysis

PART1 ANALYSIS OF CACNAIH GENE IN HAN ANCESTRY PATIENTS WITH IDIOPATHIC GENERALIZED EPILEPSYObjective:To explore the mutations of CACNA1H gene in GuangXi Han ancestry patients with idiopathie generalized epilepsy(IGE) and to analyze the relationship between the CACNA1H gene and IGE.Methods:100 GuangXi Han ancestry patients with IGE and 100 normal subjects of the same ethnical background were studied. Exons 9 to 11 and the intron-exon boundaries of CACNA1H were amplied by the polymerase chain reaction(PCR). The PCR products were directly sequenced with an ABI PRISM 3730 Automatic DNA sequencer. Using bioinformatics methods predicted whether CACNA1H gene point mutation affected its protein function.Results:1. Two novel heterozygous mutations were discovered in the CACNAIH gene, Arg735G1n and Arg766Trp both in exon 10 of CACNA1H, which were neither found in the control population nor never reported previously. 2. Alignment of CACNA1H gene amino acid sequences indicated Arginine residues at position 735 and 766 were partial conservation.3. One novel heterozygous CACNA1H mutation(Arg766Trp) was expected to affect its protein function by SIFT and PolyPhen programs.4. Fourteen polymorphisms were found in the CACNA1H gene and five variants were not reported in the NCBI SNP database. Single nucleotide polymorphisms(SNPs) were studied in both single locus and haplotype analyses.There were no difference between IGE patients and normal controls.Conclusions:Our findings suggest that the mutations of CACNA1H gene may be related to idiopathic generalized epilepsy in patienes of GuangXi China. T-type calcium channel gene-CACNA1H might be a susceptibility gene to idiopathic generalized epilepsy. PART2 ASSOCIATION OF POLYMORPHISMS IN GABRB3 GENE WITH IDIOPATHIC GENERALIZED EPILEPSY IN CHILDRENObjective:To investigate whether or not GABRB3 gene is associated with idiopathic generalized epilepsy(IGE) in GuangXi Han ancestry patients.Methods:Five single nucleotide polymorphisms(SNPs) rs4906902,rs981778,rs2059574,rs890317 and rs1426217100 were used as genetic markers. Genotypes of five SNPs were determined by using PCR/restriction enzymes and sequencing in 100 GuangXi Han ancestry patients with IGE and 200 normal subjects of the same ethnical background. Single nucleotide polymorphisms(SNPs) were studied in both single locus and haplotype analyses by spss17.0 and SHESIS software.Results:There were no significant difference in allele and genotype frequencies of five SNPs between IGE patients and normal controls. The frequency of haplotype A-A-T-A-G was significantly higher in IGE patients than those in controls(p＜0.0038).Conclusions:Our findings suggest that GAB A type-A receptor subunit gene GABRB3 may be associated with idiopathic generalized epilepsy in GuangXi patients. GABRB3 might be a susceptibility gene to idiopathic generalized epilepsy.