AIM： To identify a causative mutation in a three-generation family with autosomal dominant congenital total cataract and dissect the molecular consequence of the identified mutation.METHODS： Clinical and ophthalmolo...AIM： To identify a causative mutation in a three-generation family with autosomal dominant congenital total cataract and dissect the molecular consequence of the identified mutation.METHODS： Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by polymerase chain reaction（PCR） of the two reported genes（CRYAA and GJA8） which were linked to human total cataracts and direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected. The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for connexin 50（Cx50）, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy.RESULTS： Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by PCR of the two reported genes（CRYAA and GJA8） which were linked to human total cataractsand direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected.The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for Cx50, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy. CONCLUSION： This study has identified a novel cataract mutation in GJA8, which adds a novel mutation to the existing spectrum of Cx50 mutations with cataract.The molecular consequences of p.F32 I mutation in GJA8 exclude instability and the mislocalization of mutant Cx50 protein.展开更多
基金Supported by Natural Science Foundation of China(No.81270999No.81201181)+3 种基金Professor Academic Development Fund of Fujian Medical University(No.JS14019)Zhejiang Provincial&Ministry of Health Research Fund for Medical Sciences(No.2016KYA145No.2016KYA146)Wenzhou City Grant(No.Y20140663)
文摘AIM： To identify a causative mutation in a three-generation family with autosomal dominant congenital total cataract and dissect the molecular consequence of the identified mutation.METHODS： Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by polymerase chain reaction（PCR） of the two reported genes（CRYAA and GJA8） which were linked to human total cataracts and direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected. The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for connexin 50（Cx50）, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy.RESULTS： Clinical and ophthalmological examinations were performed on the affected and unaffected family members. Mutation were screened in recruited family members by PCR of the two reported genes（CRYAA and GJA8） which were linked to human total cataractsand direct sequencing of the PCR product. The molecular consequences of the identified mutation was dissected.The plasmids carrying wild-type and mutant mouse ORF of Gja8, coding for Cx50, were generated and ectopic expressed in 293 cells. Recombinant protein expression and cellular localization of recombinated Cx50 were assessed by confocal microscopy. CONCLUSION： This study has identified a novel cataract mutation in GJA8, which adds a novel mutation to the existing spectrum of Cx50 mutations with cataract.The molecular consequences of p.F32 I mutation in GJA8 exclude instability and the mislocalization of mutant Cx50 protein.
