Mutation Analysis Of The MYOC Gene In Families With Primary Open-angle Glaucoma

Purpose: The aim of this study was to indicate the disease causative role of the myocilin gene(MYOC) and cytochrome-P450 gene(CYP1B1) mutations in patients with Primary open-angle glaucoma(POAG) by screening the MYOC gene and CYP1B1 gene for mutations among POAG families in the southeast China.Patients:Family G-05,a three-generation POAG family,consisted of 2 patients and 2 suspects and 2 normal member.There were 5 members participating in this study.Family G-04 was a five-generation POAG family.There were 11 members suffering from POAG.In the 19 volunteers collected,6 members were POAG patients,3 were considered to be possible to suffer from POAG,and others were normal.Surgeries were underwent on all the patients in the two families.Methods: ? The participants experienced detailed clinical ophthalmic examinations and all patients' treatments were recorded;(2) 4ml Peripheral blood samples were collected from 24 family members from G-05 and G-04;(3)Genomic DNA was extracted and purified from peripheral leukocytes by using Wizard Genomic DNA Purification kit;(4)PCR(polymerase chain reaction) primers were designed according to the MYOC and CYP1B1 genes' DNA sequences;(5)The entire coding exons and splice junctions of the human MYOC and CYP1B1 gene were amplified by PCR from the genomic DNA of two patients come from the G-05 and G-04 respectively and then the PCR products was purified and directly sequenced using the forward and inverse primers;(6)When a mutation was found, all available related family members were tested. Sequence variations and numbering were assessed by comparison with GenBank reference sequences associated with MYOC and CYP1B1.Results:We found two different MYOC mutations in families G-05 and G-04,and both of them exhibited an autosomal dominant pattern of inheritance with incomplete penetrance.3 members from G-05 carry MYOC/R46 X mutation and 9 members from G-04 carry MYOC/P370 L mutation.Both of the two families didn't carry CYP1B1 mutation.Conclusion: The disease causing gene of the G-05 and G-04 is Arg46 Stop and Pro370 Leu,and the Pro370 Leu have close connection with serious POAG.Gene screening can be a positive method in the early diagnosis to POAG,meanwhile it benefits the POAG suspect and conduct the early treatment.