Cunha, L

Grazina, M

Januário, C

Oliveira, CR

Oliveira, M

Silva, F

Name not available

Parkinson’s Disease and Mitochondrial DNA NADH Dehydrogenase Subunit 1 Nucleotides 3337–3340: Study in a Population from the Central Region of Portugal (Coimbra)

Repositório Institucional dos Hospitais da Universidade de Coimbra

60 Short ReportsReferences1 Bogousslavsky J, Pierre P: Ischemic stroke in patients under age 45. NeurolClin 1992;10:113–124.2 Hirst AE, John VJ Jr, Kime SW Jr: Dissecting aneurysm of the aorta: Areview of 505 cases. Medicine 1958;37:217–219.3 Burkland CW: Spontaneous dissecting aneurysm of the cervical carotidartery. Johns Hopkins Med J 1970;126:154–159.4 Graham JM, Miller T, Stinnett DM: Spontaneous dissection of the com-mon carotid artery: Case report and review of the literature. J Vasc Surg1988;6:811–813.5 Tzeng SS, Hu HH, Kao KP, Chu FL: Common carotid artery dissectiondiagnosed by ultrasonic image: Report of a case. J Formos Med Assoc1990;89:1093–1095.6 Early TF, Gregory RT, Wheeler JR, Snyder SO, Gayle RG, Parent FN 3rd,Sorrell K: Spontaneous carotid dissection: Duplex scanning in diagnosisand management. J Vasc Surg 1991;14:391–397.7 Kawajiri K, Kiyama M, Hayazaki K: Spontaneous dissection in the com-mon carotid artery – Case report. Neurol Med Chir (Tokyo) 1995;35:373–376.8 Lubin J, Capparella J, Vecchion M: Acute monocular blindness associatedwith spontaneous common carotid artery dissection. Ann Emerg Med2001;38:332–335.9 O’Dwyer JA, Moscow N, Trevor R, Ehrenfeld WK, Newton TH: Sponta-neous dissection of the carotid artery. Radiology 1980;137:379–385.10 Zurbrugg HR, Leupi F, Schupbach P, Althaus U: Duplex scanner study ofcarotid artery dissection following surgical treatment of aortic dissectiontype A. Stroke 1988;19:970–976.11 Hart RG, Easton JD: Dissections of cervical and cerebral arteries. NeurolClin 1983;1:155–182.12 Schievink WI: Spontaneous dissection of the carotid and vertebral arteries.N Engl J Med 2001;344:898–906.Sien-Tsong Chen, MDDepartment of Neurology, Chang Gung Memorial Hospital199 Tung Hwa North Road, Taipei 10591 (Taiwan)Tel. +886 3 3281200, ext. 8340, Fax +886 3288849E-Mail asd108@adm.cgmh.org.twEur Neurol 2003;50:60–61DOI: 10.1159/000070863Parkinson’s Disease and Mitochondrial DNANADH Dehydrogenase Subunit 1 Nucleotides3337–3340: Study in a Population from theCentral Region of Portugal (Coimbra)Manuela Grazina a, b, Filipe Silvab, c, Cristina Janua´rio c,Miguel Oliveira b, Luı´s Cunha a–c, Catarina Oliveira a–caFaculty of Medicine, University of Coimbra, bCentre forNeurosciences of Coimbra and c Neurological Unit of theUniversity Hospital of Coimbra, Coimbra, PortugalParkinson’s disease (PD) is an idiopathic, chronic neurodegener-ative disorder characterized by degeneration of the dopaminergicneurons of the substantia nigra pars compacta. Although the etiologyof this disease is still unknown, it has been associated with mitochon-drial respiratory chain dysfunction, particularly with complex I defi-ciency [1]. The first clue indicating an involvement of the mitochon-drial respiratory chain in the etiology of PD came from the observa-tion that contamination of a recreational drug by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine caused irreversible PD symptoms in hu-mans and nonhuman primates [2]. Additionally, Kosel et al. [3]found, in the substantia nigra of neuropathologically confirmed idio-pathic PD patients, a known missense mutation at nucleotide 3338of the NADH dehydrogenase subunit 1 (ND1) mitochondrial DNA(mtDNA) gene that changes the amino acid valine to alanine,described previously by Chalmers et al. [4] in a control subject bloodDNA. In order to determine if the mtDNA ND1 is related with PDand to find a peripheral tissue genetic risk factor of the disease, weperformed a polymerase chain reaction (PCR)-restriction fragmentlength polymorphism analysis specific to mtDNA ND1 nucleotides3337–3340 in 53 PD patients and 61 healthy age-matched controls.Fifty-three patients, 29 female and 24 male (age range 37–81years; mean 63.81 B 11.04 years), were followed in the MovementDisorders Consultation at the Neurological Unit of the UniversityHospital of Coimbra with the diagnosis of PD, as defined by the cri-teria of the United Kingdom Parkinson’s Disease Society BrainBank. Sixty-one healthy age-matched control subjects free of progres-sive neurological disorders, 32 female and 29 male (age range 37–84years; mean 61.23 B 11.90 years), were recruited at the NeurologicalUnit of the University Hospital of Coimbra, sharing a similar socio-economic status as the parkinsonian patients. Total cellular DNAwas isolated from peripheral leukocytes of the subjects by standardprocedures, and then a fragment with 289 base pairs (bp) was ampli-fied by using a T Gradient thermocycler (Biometra), for 30 cycles at95°C for 30 s, 57°C for 30 s and 72°C for 30 s. The mastermixincluded 0.25–0.5 Ìg of total cellular DNA, 200 ÌM dNTP (Amers-ham Biosciences), 1 unit of Taq polymerase (Amersham Bio-sciences), 1! Taq polymerase buffer (Amersham Biosciences) and1 ÌM each of the primers 3397-5) (5)-AAAGGACAAGAGAAATA-AGGCC-3)) and 3397-SEL (5)-GGGCCTTTGCGTAGTTGTCT-3),specially designed with 1 mismatch for allele-specific PCR for detec-tion of the wild-type allele at mtDNA ND1 nucleotide 3397) for areaction volume of 25 Ìl. The fragment obtained was digested withthe restriction endonuclease Csp6I (MBI-Fermentas) producing, ifno mutation was present, 2 fragments with 210 and 79 bp andremaining uncut if a mutation was present. In order to verify if thePCR product was an amplification of mtDNA genes or nuclear pseu-dogenes, we performed the PCR reaction using 250 ng of total cellu-lar DNA isolated from Ú0 cells, since these cells are devoid of mtDNA[5].The main finding of the present study was that all the PCR prod-ucts of the 53 parkinsonian patients and the 61 healthy age-matchedcontrols were completely cut by the restriction endonuclease Csp6I,meaning that we have not found any mutant in the 2 groups at themtDNA ND1 nucleotides 3337–3340. Another important fact is thatthere was no amplification of the total cellular DNA isolated from Ú0cells, confirming that we had amplified mtDNA genes and notnuclear pseudogenes from parkinsonian patients and controls.In conclusion, the mtDNA ND1 nucleotides 3337–3340 seemunlikely to contribute to susceptibility to PD in the population understudy (central region of Portugal), and our results seem to rule outthat mutations in this sequence are a primary cause of sporadic PD.However, this finding does not exclude involvement of the ND1 orany other mtDNA complex I gene in the pathogenesis of PD. In fact,somatic brain mitochondrial mutations may occur in subjects with-out mutations in blood and our results relate to peripheral blood,whereas Kosel et al. [3] report on brain tissue. Additionally, cautionis necessary when working with mtDNA because it has a high muta-tion rate due to lack of a histone coat and an inadequate repair sys-tem [6]. Given this vulnerability, free radical mtDNA damage mayShort Reports 61occur, particularly in aging and neurodegenerative disorders, causingadditional mtDNA injury, possibly initiating a vicious circle. Webelieve that the study of peripheral tissues may be a step forward inidentifying possible genetic risk factors in earlier stages of the disease,allowing earlier therapeutic approaches.AcknowledgementsWe would like to acknowledge Prof. A. Münnich, Dr. P. Rustinand Dr. A. Rötig for providing Ú0 cell DNA. The authors are alsograteful to Novartis for their partial contribution in acquiring the TGradient thermocycler (Biometra). This work was partially support-ed by a fellowship from the Portuguese Foundation for Science andTechnology (Praxis XXI/BD/4049/94).References1 Schapira AHV, Cooper JM, Dexter D, Jenner P, Clark JB, Marsden CD:Mitochondrial complex I deficiency in Parkinson’s disease. Lancet 1989;i:1269.2 Langston JW, Ballard P: Chronic parkinsonism in humans due to a productof meperidine-analog synthesis. Science 1983;219:979–980.3 Kosel S, Grasbon-Frodl EM, Mautsch U, Egensperger R, von Eitzen U,Frishman D, Hofmann S, Gerbitz KD, Mehraein P, Graeber MB: Novelmutations of mitochondrial complex I in pathologically proven Parkinsondisease. Neurogenetics 1998;1:197–204.4 Chalmers RM, Robertson N, Kellar-Wood H, Compston DAS, HardingAE: Sequence of the human homologue of a mitochondrially encodedmurine transplantation antigen in patients with multiple sclerosis. J Neurol1995;242:332–334.5 Parfait B, Rustin P, Münnich A, Rötig A: Coamplification of nuclear pseu-dogenes and assessment of heteroplasmy of mitochondrial DNA muta-tions. Biochem Biophys Res Commun 1998;217:57–59.6 Schapira AHV: Evidence for mitochondrial dysfunction in Parkinson’s dis-ease – A critical appraisal. Mov Disord 1994;9:125–138.Catarina Oliveira, Biochemistry Institute, Faculty of MedicineUniversity of Coimbra, Rua Larga, P–3004 504 Coimbra (Portugal)Tel. +351 239 820190, Fax +351 239 822776E-Mail catarina@cnc.cj.uc.ptCopyright: S. Karger AG, Basel 2003. Reproduced with the permission of S. Karger AG, Basel. Furtherreproduction or distribution (electronic or otherwise) is prohibited without permission from the copyrightholder.

http://rihuc.huc.min-saude.pt/bitstream/10400.4/1563/1/Eur%20Neurol.pdf

Parkinson’s Disease and Mitochondrial DNA NADH Dehydrogenase Subunit 1 Nucleotides 3337–3340: Study in a Population from the Central Region of Portugal (Coimbra)

Abstract

Similar works

Full text

Available Versions

Name not available

Repositório Institucional dos Hospitais da Universidade de Coimbra