The Deletion Analysis of Mitochondrial Multicomplexes in Parkinson's Disease

Lee, Uhn; Yoo, Young Mi; Yoo, Chan Jong; Kim, Yong Jung; Lee, Chang Joong

Abstract

Arkinson's disease(PD) is a neurodegenerative disease involving mainly the loss of dopaminergic neurons in substantia nigra by several factors. The cause of dopaminergic cell death is unknown. Recently, it has been focused on that Parkinson's disease resulting from mitochondrial dysfunction. In the previous studies, it was found that a 5 kilobase(kb) deletion derived from mtDNA dysfunction. And this result leads to a reduction of ATP production, which ultimately causes result in cell death. Blood samples were collected from 6 positive control(PC) and 9 PD patients. Total DNA was extracted twice with phenol followed by chloroform:isoamylalcohol(24: 1). For the analysis of mtDNA, polymerase chain reaction(PCR) and long and accurate polymerase chain reaction(LA PCR) were performed by mitochondrial specific primers. As a result, a deletions of large quantity was detected within several regions of mtDNA in PD patients. The analysis of the partial sequence of the mitochondrial D-loop gene and restriction fragment length polymorphism(RFLP) technique were performed to investigate the point mutation and nucleotide sequence variations between PC and PD patients. Fragment variations between PC and PD were seen in the fragment digested by Hin d III, Eco R V. These variations are attributed to the presence or absence of recognition site by base substitution. Point mutation was observed in the D-loop region. Patients 1 and 2 had one point mutation. Patient 1 had a transition from T to C at 195, and patient 2 had a transversion from A to T. In addition to point mutation, the deletion of mtDNA occurred complexI, III, IV and V subunits in PD patients.