Cerebrospinal Fluid Biomarkers for Kii Amyotrophic Lateral Sclerosis/Parkinsonism-Dementia Complex

Objective. Amyotrophic lateral sclerosis/parkinsonism-dementia complex is classified as one of the tauopathies. Methods. The total tau, phosphorylated tau, and amyloid β42 levels were assayed in cerebrospinal fluid from patients with Kii amyotrophic lateral sclerosis/parkinsonism-dementia complex (), Alzheimer’s disease (), Parkinson’s disease (), amyotrophic lateral sclerosis (), and controls () using specific enzyme-linked immunosorbent assay methods. Results. Total tau and phosphorylated tau did not increase and amyloid β42 was relatively reduced in Kii amyotrophic lateral sclerosis/parkinsonism-dementia complex. Relatively reduced amyloid β42 might discriminate Kii amyotrophic lateral sclerosis/parkinsonism-dementia complex from amyotrophic lateral sclerosis and Parkinson’s disease, and the ratios of phosphorylated-tau to amyloid β42 could discriminate Kii amyotrophic lateral sclerosis/parkinsonism-dementia complex from Alzheimer’s disease. Conclusions. Cerebrospinal fluid analysis may be useful to differentiate amyotrophic lateral sclerosis/parkinsonism-dementia complex from Alzheimer’s disease, amyotrophic lateral sclerosis, and Parkinson’s disease

Four mutations of the spastin gene in Japanese families with spastic paraplegia

Hereditary spastic paraplegia (HSP) is a group of genetically heterogeneous neurodegenerative disorders characterized by slowly progressive spasticity and weakness of the lower limbs. HSP is caused by failure of development or selective degeneration of the corticospinal tracts, which contain the longest axons in humans. The most common form of HSP is caused by mutations of the spastin gene (SPAST), located on chromosome 2p21-p22, which encodes spastin, one of the ATPases associated with diverse cellular activities (AAA). In this study, we detected four causative mutations of SPAST among 14 unrelated patients with spastic paraplegia. Two missense mutations (1447A-->G, 1207C-->G) and two deletion mutations (1465delT, 1475-1476delAA) were located in the AAA cassette region. Three of these four mutations were novel. Previous reports and our results suggest that the frequency of SPAST mutations is higher among Japanese patients with autosomal dominant HSP, although SPAST mutations are also observed in patients with sporadic spastic paraplegia

紀伊半島のAＬＳとパーキンソン痴呆複合におけるＳＮＰｓ解析による発病因子の検討

application/pdf紀伊半島に多発する筋萎縮性側索硬化症/パーキンソン認知症複合(紀伊ALS/PDC)における発症要因と病態解明を目的に、下記の点について検討を行った。1)遺伝子検索(1)候補遺伝子解析:パーキンソン病関連遺伝子(alpha-synuclein, LRRK2, parkin, DJ-1, PINK1, ATP13A2)、認知症関連遺伝子(APOE4, MAPT, PGRN, TARDBP, GSK3β)、運動ニューロン疾患関連遺伝子(SOD1, SOD2, SOD3,ALS2 alsin, SMN1, ANG, VEGF, VCP, VAPB, DCTN1 CHMP2B)について、既知の変異の有無を検討したが、すべての遺伝子について変異は認めなかった。また、平行して行ったGene dosage analyses(MAPT, alpha-synuclein, TARDBP, GSK3β,parkin)についても異常を認めなかった。さらに、若年性ALSの原因遺伝子として2009年春に同定されたFUS/TLS遺伝子の解析を行ったが、異常を認めなかった。(2)TRPM7遺伝子:細胞内カルシウムとマグネシウムの調節に関与し、グアム症例でミスセンス変異の認められたTRPM7遺伝子について検討したが、紀伊症例では同一の変異は認めなかった。2)TDP-43蛋白:2006年に発見されたALSと前頭側頭型認知症の神経細胞に特異的に出現するTDP-43蛋白にっいて生化学的、免疫組織化学的に検討した。紀伊ALS/PDC 5例での検討では、全例で海馬と脊髄の神経細胞内にTDP-43陽性封入体を認めた。ALS/PDCは、tau,α-synuclein, TDP-43が複合して蓄積する疾患であることがわかった。3)OPTN蛋白:2010年に遺伝性ALSの発症遺伝子として同定されたOPTN蛋白は、紀伊ALS/PDCの脊髄運動ニューロンにおいて主な蓄積蛋白としては認められず、また遺伝子変異もなかった。4)Ataxin遺伝子解析:現在、ALSのリスク遺伝子として報告されたataxin遺伝子のpoly Q延長について、解析を進めている。Objectives : To clarify the genetic background of amyotrophic lateral sclerosis/parkinsonism-dementia complex (ALS/PDC) of the Kii peninsula, Japan (KiiALS/PDC). Methods : We performed extended mutation analyses of three patients with pathologically diagnosed Kii ALS/PDC. Direct sequencing analyses were performed in 19 genes, including amyotrophic lateral sclerosis(ALS)/frontotemporal lobar degeneration (FTLD)-related genes(SOD2, SOD3, ALS2 alsin, SMN1, PGRN, ANG, VEGF, VCP, VAPB, DCTN1, CHMP2B, and TARDBP), tauopathy-related gene (GSK3β), and parkinsonism-related genes(alpha-synuclein, LRRK2, parkin, DJ-1, PINK1, and ATP13A2). Gene dosage analyses were conducted in screening of MAPT, alpha-synucleln, TARDBP,GSK3β, and parkin. Results : We found no mutation in the 19 genes. We found a homozygous non-synonymous SNP(ALS2 alsin V368M) shared in all the three patients. Gene dosage was normal in MAPT, alpha-synuclein, TARDBP, GSK3β, and pakin.Conclusions : The present findings, together with a previous negative study on MAPT and SOD1 mutation, further elucidated the lack of causative mutation in all exons, exon intron boundaries, or some rearrangements of the reported major causative or susceptible genes related to ALS, FTLD, parkinsonism, synucleinopathy, and tauopathy. However, the familial aggregation and lack of any environment factors suggest that Kii ALS/PDC is caused by other yet unidentified genetic factors.平成20~22年度科学研究費補助金(基盤研究(C))研究成果報告書2059099

Lifestyle Changes and Oxidative Stress in a High-incidence Area of Amyotrophic Lateral Sclerosis in the Southwestern Kii Peninsula, Japan

[Objective] Lifestyle changes may play an important role in the incidence reduction and delay of onset age of amyotrophic lateral sclerosis (ALS) in the Koza/Kozagawa/Kushimoto (K) area. The aim of this study was to evaluate recent lifestyle changes in the K area and to investigate the relationships between lifestyle and oxidative stress among the residents. [Methods] We conducted a medical checkup for elderly residents in the K area and the control area and evaluated the urinary 8-OHdG levels, cognitive function test scores and metal contents in serum and scalp hair, coupled with a lifestyle questionnaire survey between 2010 and 2015. [Results] Recent lifestyle changes among the K residents, including a decrease in the Japanese pickle consumption, increase in fresh vegetable consumption and decrease in farm work, were evaluated in this study. Low consumption of Japanese pickles, high consumption of fresh vegetables, rare farm work and low levels of 8-OHdG/creatinine were all associated with high scores in the cognitive function tests. Frequent farm work and consumption of Japanese pickles was associated with high contents of transition metals, such as Mn, Al and V, in the scalp hair. [Conclusion] These lifestyle changes among residents in the K area may be associated with their oxidative stress

Nitrative Stress and Tau Accumulation in Amyotrophic Lateral Sclerosis/Parkinsonism-Dementia Complex (ALS/PDC) in the Kii Peninsula, Japan

Objective: The Kii Peninsula of Japan is known to be a high incidence area of amyotrophic lateral sclerosis/parkinsonism-dementia complex (Kii ALS/PDC) with tauopathy. Nitrative stress and oxidative stress on ALS/PDC and their relationship to tau pathology were clarified.Methods: Seven patients with Kii ALS/PDC (3 males and 4 females, average age 70.7 years, 3 with ALS, 2 with ALS with dementia, and 2 with PDC) were analyzed in this study. Five patients with Alzheimer's disease and five normal aged subjects were used as controls. Immunohistochemical analysis was performed on formalin-fixed, paraffin-embedded temporal lobe sections (the hippocampal area including hippocampus, prosubiculum, subiculum, presubiculum, and parahippocampal gyri) using antibodies to detect phosphorylated tau (anti-AT-8), nitrated guanine (anti-8-NG), anti-iNOS, anti-NFκB, and oxidized guanine (anti-8-OHdG) antibodies.Results: Most hippocampal neurons of Kii ALS/PDC patients were stained with anti-8-NG, anti-iNOS, anti-NFκB, and anti-8-OHdG antibodies and some AT-8 positive neurons were co-stained with anti-8-NG antibody. The numbers of 8-NG positive neurons and 8-OHdG positive neurons were greater than AT-8 positive neurons and the number of 8-NG positive neurons was larger in patients with Kii ALS/PDC than in controls.Conclusion: Nitrative and oxidative stress may take priority over tau accumulation and lead to the neurodegeneration in Kii ALS/PDC