924 results on '"T Nagatsu"'
Search Results
2. Continuous measurement in a greenhouse reveals high COP of air-source heat pump in winter
- Author
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T. Nagatsu, F. Goto, K. Shoji, and T. Ito
- Subjects
Continuous measurement ,Meteorology ,Air source heat pumps ,Environmental science ,Greenhouse ,Horticulture - Published
- 2015
- Full Text
- View/download PDF
3. A New Assay of Monoamine Oxidase Activity for Selective Substrates and Inhibitors
- Author
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S. Kawai, T. Nagatsu, and T. Nakano
- Subjects
Biochemistry ,Monoamine oxidase ,Chemistry ,Monoamine oxidase B - Published
- 2015
- Full Text
- View/download PDF
4. A branch site mutation leading to aberrant splicing of the human tyrosine hydroxylase gene in a child with a severe extrapyramidal movement disorder
- Author
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Jeroen Luyten, Ron A. Wevers, G.C.H. Steenbergen-Spanjers, L.P.W.J. van den Heuvel, Rolf J.R.J. Janssen, Martin Häussler, Georg F. Hoffmann, and T. Nagatsu
- Subjects
Heterozygote ,Tyrosine 3-Monooxygenase ,RNA Splicing ,DNA Mutational Analysis ,Molecular Sequence Data ,Restriction Mapping ,Biology ,Polymerase Chain Reaction ,Exon ,Basal Ganglia Diseases ,Transcription (biology) ,Genetics ,Humans ,Child ,Gene ,Genetics (clinical) ,Polymorphism, Single-Stranded Conformational ,DNA Primers ,chemistry.chemical_classification ,Inherited neurotransmitten diseases ,Messenger RNA ,Tyrosine hydroxylase ,Base Sequence ,Alternative splicing ,Intron ,Exons ,Erfelijke ziekten in de neurotransmissie ,Amino acid ,chemistry ,Chronic Disease ,Mutation ,Female - Abstract
We report a branch site mutation in the gene of the enzyme tyrosine hydroxylase (TH): a −24t > a substitution two bases upstream of the adenosine in the branchpoint sequence (BPS) of intron 11. As normal lariat formation is therefore prevented, alternative splicing takes place: use of the BPS of intron 12 results in skipping of exon 12, whereas the use of a cryptic branch site in intron 11 leads to partial retention of this intron in the mRNA. This leads in both cases to an aberrant protein product. In the one case, skipping of exon 12 results in the absence of 32 amino acids. In the other, retention of 36 nucleotides of intron 11 in the mRNA results in the incorporation of twelve additional amino acids. The functional consequences of this mutation for the patient, who is also heterozygous for another previously identified mutation, become apparent in a severe clinical phenotype.
- Published
- 2000
5. Neopterin and Cytokines in Hereditary Dystonia and Parkinson's Disease
- Author
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Makio Mogi, Akifumi Togari, Hiroshi Ichinose, and T Nagatsu
- Subjects
Parkinson's disease ,Crystallography ,business.industry ,striatum ,Clinical Biochemistry ,Neopterin ,Striatum ,medicine.disease ,Biochemistry ,cytokines ,cerebrospinal fluid ,nervous system diseases ,chemistry.chemical_compound ,Cerebrospinal fluid ,chemistry ,neopterin ,immune system diseases ,QD901-999 ,Immunology ,parkinson's disease ,medicine ,Molecular Medicine ,Hereditary Dystonia ,business - Abstract
β Both neopterin and biopterin concentrations in cerebrospinal fluid from patients with Parkinson's disease, in which the nigrostriatal dopamine neurons degenerate, were lower than those from age-matched older control subjects. However, the decrease in biopterin was more marked than that in neopterin, resulting in the increase in the neopterin/ biopterin ratio in Parkinson's disease. These results suggests that neopterin in cerebrospinal fluid in Parkinson's disease may partly be derived from immunoactivated glial cells, besides catecholamine or serotonin n eurons including nigrostriatal dopamine neurons. In accordance to this hypothesis, cytokines (TNF-α, IL-1, IL-2 , IL-6, EGF, TGF-α, TGF-β1) were found to be increased in the striatum and/or in cerebrospinal fluid. The increment of cytokines in the brain in Parkinson's disease may be related to the mechanism of neurodegeneration of dopaminergic neurons in Parkinson's disease . In contrast to Parkinson's disease, in hereditary progressive dystonia/ dopa-responsive dystonia, which is a dopamine deficiency caused by mutations in GTP cyclohydrolase I without neuronal cell death (Segawa's disease), neopterin and biopterin in cerebrospinal fluid decreases in parallel owing to the decreased activity in GTP cyclohydrolase I .
- Published
- 1999
6. Quantification of tyrosine hydroxylase mRNA
- Author
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H, Ichinose, T, Ohye, T, Suzuki, H, Inagaki, and T, Nagatsu
- Abstract
The main biochemical characteristic of Parkinson's disease (PD) is reduction of the neurotransmitter dopamine and the dopamine-synthesizing enzyme system, including tyrosine hydroxylase (TH, tyrosine 3-monooxygenase, EC 1.14.16.2) and tetrahydrobiopterin (BH(4) co-factor, in nigrostriatal neurons (1). The deficiency in dopamine-synthesizing enzymes is accompanied by cell loss, which is thought to be caused by unknown exogenous environmental factors as well as endogenous genetic factors.
- Published
- 2011
7. The relationship between depression and regional cerebral blood flow in Parkinson's disease and the effect of selegiline treatment
- Author
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K, Imamura, N, Okayasu, and T, Nagatsu
- Subjects
Aged, 80 and over ,Male ,Psychiatric Status Rating Scales ,Depressive Disorder ,Brain ,Parkinson Disease ,Middle Aged ,Neuropsychological Tests ,Antiparkinson Agents ,Blood Circulation Time ,Treatment Outcome ,Cerebrovascular Circulation ,Selegiline ,Humans ,Female ,Radionuclide Imaging ,Aged - Abstract
We examined the relationship between severity of depression in Parkinson's disease (PD) and regional cerebral blood flow (rCBF) using single photon emission computed tomography (SPECT) and the reaction to levodopa-selegiline combination therapy.We evaluated 52 patients with PD and nine age-matched controls with SPECT and the Unified Parkinson's Disease Rating Scale (UPDRS) part III, Mini-Mental State Examination (MMSE), and Beck Depression Inventory (BDI) to evaluate depression severity and its connection with rCBF. Furthermore, we examined rCBF in patients with PD treated with levodopa with or without selegiline.A significant fall in rCBF was observed in the bilateral posterior cingulate, hippocampus, and cuneus and the superior parietal and primary visual areas in PD patients with minor depression and in all regions in those with major depression. Elevations in UPDRS part III and BDI scores and falls in MMSE scores were of significantly lower magnitude in the levodopa-selegiline group than in the levodopa group. Whole brain rCBF fell significantly less in the levodopa-selegiline group than in the levodopa group.These results indicate that selegiline controlled not only worsening of motor function and cognitive function in PD but also aggravation of minor depression, and restrained a fall in whole brain rCBF.
- Published
- 2010
8. Biochemistry of postmortem brains in Parkinson's disease: historical overview and future prospects
- Author
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T, Nagatsu and M, Sawada
- Subjects
Epinephrine ,Tyrosine 3-Monooxygenase ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Dopamine ,Parkinson Disease ,Dopamine beta-Hydroxylase ,Biopterin ,Corpus Striatum ,Substantia Nigra ,Norepinephrine ,Humans ,RNA, Messenger ,GTP Cyclohydrolase ,Microdissection ,Forecasting - Abstract
Biochemical studies on postmortem brains of patients with Parkinson's disease (PD) have greatly contributed to our understanding of the molecular pathogenesis of this disease. The discovery by 1960 of a dopamine deficiency in the nigro-striatal dopamine region of the PD brain was a landmark in research on PD. At that time we collaborated with Hirotaro Narabayashi and his colleagues in Japan and with Peter Riederer in Germany on the biochemistry of PD by using postmortem brain samples in their brain banks. We found that the activity, mRNA level, and protein content of tyrosine hydroxylase (TH), as well as the levels of the tetrahydrobiopterin (BH4) cofactor of TH and the activity of the BH4-synthesizing enzyme, GTP cyclohydrolase I (GCHI), were markedly decreased in the substantia nigra and striatum in the PD brain. In contrast, the molecular activity (enzyme activity/enzyme protein) of TH was increased, suggesting a compensatory increase in the enzyme activity. The mRNA levels of all four isoforms of human TH (hTH1-hTH4), produced by alternative mRNA splicing, were also markedly decreased. This finding is in contrast to a completely parallel decrease in the activity and protein content of dopamine beta-hydroxylase (DBH) without changes in its molecular activity in cerebrospinal fluid (CSF) in PD. We also found that the activities and/or the levels of the mRNA and protein of aromatic L-amino acid decarboxylase (AADC, DOPA decarboxylase), DBH, phenylethanolamine N-methyltransferase (PNMT), which synthesize dopamine, noradrenaline, and adrenaline, respectively, were also decreased in PD brains, indicating that all catecholamine systems were widely impaired in PD brains. Programmed cell death of the nigro-striatal dopamine neurons in PD has been suggested from the following findings on postmortem brains: (1) increased levels of pro-inflammatory cytokines such as TNF-alpha and IL-6; (2) increased levels of apoptosis-related factors such as TNF-alpha receptor R1 (p 55), soluble Fas and bcl-2, and increased activities of caspases 1 and 3; and (3) decreased levels of neurotrophins such as brain-derived nerve growth factor (BDNF). Immunohistochemical data and the mRNA levels of the above molecules in PD brains supported these biochemical data. We confirmed by double immunofluorescence staining the production of TNF-alpha and IL-6 in activated microglia in the putamen of PD patients. Owing to the recent development of highly sensitive and wide-range analytical methods for quantifying mRNAs and proteins, future assays of the levels of various mRNAs and proteins not only in micro-dissected brain tissues containing neurons and glial cells, but also in single cells from frozen brain slices isolated by laser capture micro-dissection, coupled with toluidine blue, Nissl staining or immunohistochemical staining, should further contribute to the elucidation of the molecular pathogenesis of PD and other neurodegenerative or neuropsychiatric diseases.
- Published
- 2007
9. Molecular mechanism of the relation of monoamine oxidase B and its inhibitors to Parkinson's disease: possible implications of glial cells
- Author
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T, Nagatsu and M, Sawada
- Subjects
Monoamine Oxidase Inhibitors ,Animals ,Humans ,Parkinson Disease ,Monoamine Oxidase - Abstract
Monoamine oxidases A and B (MAO A and MAO B) are the major enzymes that catalyze the oxidative deamination of monoamine neurotaransmitters such as dopamine (DA), noradrenaline, and serotonin in the central and peripheral nervous systems. MAO B is mainly localized in glial cells. MAO B also oxidizes the xenobiotic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to a parkinsonism-producing neurotoxin, 1-methyl-4-phenyl-pyridinium (MPP+). MAO B may be closely related to the pathogenesis of Parkinson's disease (PD), in which neuromelanin-containing DA neurons in the substantia nigra projecting to the striatum in the brain selectively degenerate. MAO B degrades the neurotransmitter DA that is deficient in the nigro-striatal region in PD, and forms H2O2 and toxic aldehyde metabolites of DA. H2O2 produces highly toxic reactive oxygen species (ROS) by Fenton reaction that is catalyzed by iron and neuromelanin. MAO B inhibitors such as L-(-)-deprenyl (selegiline) and rasagiline are effective for the treatment of PD. Concerning the mechanism of the clinical efficacy of MAO B inhibitors in PD, the inhibition of DA degradation (a symptomatic effect) and also the prevention of the formation of neurotoxic DA metabolites, i.e., ROS and dopamine derived aldehydes have been speculated. As another mechanism of clinical efficacy, MAO B inhibitors such as selegiline are speculated to have neuroprotective effects to prevent progress of PD. The possible mechanism of neuroprotection of MAO B inhibitors may be related not only to MAO B inhibition but also to induction and activation of multiple factors for anti-oxidative stress and anti-apoptosis: i.e., catalase, superoxide dismutase 1 and 2, thioredoxin, Bcl-2, the cellular poly(ADP-ribosyl)ation, and binding to glyceraldehydes-3-phosphate dehydrogenase (GAPDH). Furthermore, it should be noted that selegiline increases production of neurotrophins such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and glial cell line-derived neurotrphic factor (GDNF), possibly from glial cells, to protect neurons from inflammatory process.
- Published
- 2007
10. Role of cytokines in inflammatory process in Parkinson's disease
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M, Sawada, K, Imamura, and T, Nagatsu
- Subjects
Inflammation ,Neostriatum ,Substantia Nigra ,Putamen ,Animals ,Cytokines ,Humans ,Tetradecanoylphorbol Acetate ,Parkinson Disease ,Microglia ,Hippocampus - Abstract
We investigated whether the cytokines produced in activated microglia in the substantia nigra (SN) and putamen in sporadic Parkinson's disease (PD) are neuroprotective or neurotoxic. In autopsy brains of PD, the number of MHC class II (CR3/43)-positive activated microglia, which were also ICAM-1 (CD 54)-, LFA-1 (CD 11a)-, TNF-alpha-, and IL-6-positive, increased in the SN and putamen during progress of PD. At the early stage activated microglia were mainly associated with tyrosine hydroxylase (TH)-positive neurites in the putamen, and at the advanced stage with damaged TH-positive neurons in the SN. The activated microglia in PD were observed not only in the nigro-striatal region, but also in various brain regions such as the hippocampus and cerebral cortex. We examined the distribution of activated microglia and the expression of cytokines and neurotrophins in the hippocampus of PD and Lewy body disease (LBD). The levels of IL-6 and TNF-alpha mRNAs increased both in PD and LBD, but those of BDNF mRNA and protein drastically decreased specifically in LBD, in which neuronal loss was observed not only in the nigro-striatum but also in the hippocampus. The results suggest activated microglia in the hippocampus to be probably neuroprotective in PD, but those to be neurotoxic in LBD. As an evidence supporting this hypothesis, two subsets of microglia were isolated from mouse brain by cell sorting: one subset with high production of reactive oxygen species (ROS) and the other with no production of ROS. When co-cultured with neuronal cells, one microglia clone with high ROS production was neurotoxic, but another clone with no ROS production neuroprotective. On the other hand, Sawada with coworkers found that a neuroprotective microglial clone in a culture experiment converted to a toxic microglial clone by transduction of the HIV-1 Nef protein with increasing NADPH oxidase activity. Taken together, all these results suggest that activated microglia may change in vivo from neuroprotective to neurotoxic subtsets as degeneration of dopamine neurons in the SN progresses in PD. We conclude that the cytokines from activated microglia in the SN and putamen may be initially neuroprotective, but may later become neurotoxic during the progress of PD. Toxic change of activated microglia may also occur in Alzheimer's disease and other neurodegenerative diseases in which inflammatory process is found.
- Published
- 2006
11. [Huntington's disease model mouse and neuronal cell death]
- Author
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H, Ishiguro, H, Sawada, K, Nishii, K, Yamada, and T, Nagatsu
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Mice, Knockout ,Neurons ,Disease Models, Animal ,Huntingtin Protein ,Mice ,Huntington Disease ,Cell Death ,Animals ,Brain ,Nuclear Proteins ,Mice, Transgenic ,Nerve Tissue Proteins - Published
- 2001
12. DOPA-responsive dystonia. From causative gene to molecular mechanism
- Author
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H, Ichinose, T, Suzuki, H, Inagaki, T, Ohye, and T, Nagatsu
- Subjects
Family Health ,Dystonia ,Dopamine Agents ,Humans ,Dihydroxyphenylalanine ,Pedigree - Published
- 2001
13. Neural degeneration in Parkinson's disease
- Author
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P, Riederer, H, Reichmann, B, Janetzky, J, Sian, K P, Lesch, K W, Lange, K L, Double, T, Nagatsu, and M, Gerlach
- Subjects
Nerve Degeneration ,Animals ,Humans ,Parkinson Disease - Published
- 2001
14. Catecholamines and serotonin are differently regulated by tetrahydrobiopterin. A study from 6-pyruvoyltetrahydropterin synthase knockout mice
- Author
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C, Sumi-Ichinose, F, Urano, R, Kuroda, T, Ohye, M, Kojima, M, Tazawa, H, Shiraishi, Y, Hagino, T, Nagatsu, T, Nomura, and H, Ichinose
- Subjects
Mice, Inbred C57BL ,Mice, Knockout ,Mice ,Serotonin ,Catecholamines ,Gene Expression Regulation ,Animals ,Phosphorus-Oxygen Lyases ,Biopterin - Abstract
(6R)-L-erythro-5,6,7,8-Tetrahydrobiopterin (BH4) is an essential cofactor for tyrosine hydroxylase (TH), tryptophan hydroxylase, phenylalanine hydroxylase, and nitric-oxide synthase. These enzymes synthesize neurotransmitters, e.g. catecholamines, serotonin, and nitric oxide (NO). We established mice unable to synthesize BH4 by disruption of the 6-pyruvoyltetrahydropterin synthase gene, the encoded protein of which catalyzes the second step of BH4 biosynthesis. Homozygous mice were born at the almost expected Mendelian ratio, but died within 48 h after birth. In the brain of homozygous mutant neonates, levels of biopterin, catecholamines, and serotonin were extremely low. The number of TH molecules was highly dependent on the intracellular concentration of BH4 at nerve terminals. Alteration of the TH protein level by modulation of the BH4 content is a novel regulatory mechanism. Our data showing that catecholaminergic, serotonergic, and NO systems were differently affected by BH4 starvation suggest the possible involvement of BH4 synthesis in the etiology of monoamine-based neurological and neuropsychiatric disorders.
- Published
- 2001
15. Cotransduction of tyrosine hydroxylase and aromatic L-amino acid decarboxylase genes into cultured striatal cells using adeno-associated virus vectors
- Author
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D, Fan, D, Kang, M, Ogawa, I, Nakano, T, Nagatsu, G J, Kurtzman, and K, Ozawa
- Subjects
Tyrosine 3-Monooxygenase ,Genetic Vectors ,Gene Expression ,Dependovirus ,Embryo, Mammalian ,Kidney ,Corpus Striatum ,Rats ,Aromatic-L-Amino-Acid Decarboxylases ,Transduction, Genetic ,Animals ,Humans ,Rats, Wistar ,Cells, Cultured - Abstract
To examine whether tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC) genes can be cotransduced into the same target striatal cells using adeno-associated virus (AAV) vectors, and to determine whether the cotransduction would result in better biochemical change than the TH gene alone.TH and AADC genes were cotransduced into cultured striatal cells with separate AAV vectors. Expressions of TH and AADC were detected by immunocytochemistry; intracellular catecholamine levels were assayed by high-performance liquid chromatography (HPLC).TH and AADC genes were efficiently cotransduced into the striatal cells. Specifically, the coexpression of TH and AADC resulted in more effective dopamine production compared with the TH gene alone.Using AAV vectors, coexpression of TH and AADC in the striatal cells might be a useful approach to gene therapy for Parkinson's disease.
- Published
- 2001
16. Changes in cytokines and neurotrophins in Parkinson's disease
- Author
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T, Nagatsu, M, Mogi, H, Ichinose, and A, Togari
- Subjects
Major Histocompatibility Complex ,Parkinsonian Disorders ,Proto-Oncogene Proteins c-bcl-2 ,Caspases ,Neurotoxins ,Animals ,Brain ,Cytokines ,Humans ,Apoptosis ,Parkinson Disease ,Nerve Growth Factors ,fas Receptor - Abstract
Degeneration of the dopamine (DA) neurons of the substantia nigra pars compacta and the resulting loss of nerve terminals accompanied by DA deficiency in the striatum are responsible for most of the movement disturbances called parkinsonism, observed in Parkinson's disease (PD). One hypothesis of the cause of degeneration of the nigrostriatal DA neurons is that PD is caused by programmed cell death (apoptosis) due to increased levels of cytokines and/or decreased ones of neurotrophins. We and other workers found markedly increased levels of cytokines, such as tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-2, IL-4, IL-6, transforming growth factor (TFG)-alpha, TGF-beta1, and TGF-beta2, and decreased ones of neurotrophins, such as brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF), in the nigrostriatal DA regions and ventricular and lumbar cerebrospinal fluid of PD patients. Furthermore, the levels of TNF-alpha receptor R1 (TNF-R1, p55), bcl-2, soluble Fas (sFas), and the activities of caspase-1 and caspase-3 were also elevated in the nigrostriatal DA regions in PD. In experimental animal models of PD, IL-1beta level was increased and NGF one decreased in the striatum of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonian mice, and TNF-alpha level was increased in the substantia nigra and striatum of the 6-hydroxydopamine (6OHDA)-injected side of hemiparkinsonian rats. L-DOPA alone or together with 6OHDA does not increase the level of TNF-alpha in the brain in vivo. Increased levels of proinflammatory cytokines, cytokine receptors and caspase activities, and reduced levels of neurotrophins in the nigrostriatal region in PD patients, and in MPTP- and 6OHDA-produced parkinsonian animals suggest increased immune reactivity and programmed cell death (apoptosis) of neuronal and/or glial cells. These data indicate the presence of such proapoptotic environment in the substantia nigra in PD that may induce increased vulnerability of neuronal or glial cells towards a variety of neurotoxic factors. The probable causative linkage among the increased levels of proinflammatory cytokines and the decreased levels of neurotrophins, candidate parkinsonism-producing neurotoxins such as isoquinoline neurotoxins (Review; Nagatsu, 1997), and the genetic susceptibility to toxic factors, remains for further investigation in the molecular mechanism of PD. The increased cytokine levels, decreased neurotrophin ones, and the possible immune response in the nigrostriatal region in PD indicate new neuroprotective therapy including nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, immunosuppressive or immunophilin-binding drugs such as FK-506, and drugs increasing neurotrophins.
- Published
- 2001
17. Cytokines in Parkinson's disease
- Author
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T, Nagatsu, M, Mogi, H, Ichinose, and A, Togari
- Subjects
Postmortem Changes ,Animals ,Brain ,Cytokines ,Humans ,Apoptosis ,Nerve Tissue Proteins ,Parkinson Disease - Abstract
We found that in Parkinson's disease (PD) the levels of various cytokines [tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-2, IL-4, IL-6, epidermal growth factor (EGF), transforming growth factor (TGF)-alpha, TGF-beta1] were significantly increased in the striatum (caudate and putamen) of the postmortem brain and in ventricular or spinal cerebrospinal fluid (VCSF, LCSF). Furthermore, the levels of the apoptosis-related proteins such as bcl-2 and soluble Fas (sFas) in the striatum were also elevated in PD. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated parkinsonism mice, the levels of IL-1beta in the striatum were significantly increased, but those of nerve growth factor (NGF) were significantly decreased, compared with control mice. In hemiparkinsonism rats produced by injection of 6-hydroxydopamine (6-OHDA) into one side of the median forebrain bundle, the levels of TNF-alpha in the 6-OHDA-treated side were increased in the striatum and substantia nigra, but not in the cerebral cortex, compared with those in the control side. Repeated administration of L-DOPA in the 6-OHDA-treated rats did not change the TNF-alpha levels in the control side and in the 6-OHDA-treated side in the substantia nigra, striatum, and cerebral cortex. Our results suggest that the changes in the levels of cytokines, neurotrophins, and apoptosis-related proteins in the nigrostriatal regions of PD may be involved in apoptosis and degeneration of the nigrostriatal DA neurons.
- Published
- 2000
18. Direct imaging of phosphorylation-dependent conformational change and DNA binding of CREB by electron microscopy
- Author
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J, Usukura, Y, Nishizawa, A, Shimomura, K, Kobayashi, T, Nagatsu, and M, Hagiwara
- Subjects
Tyrosine 3-Monooxygenase ,Protein Conformation ,Nuclear Proteins ,DNA ,CREB-Binding Protein ,Recombinant Proteins ,Rats ,Microscopy, Electron ,Trans-Activators ,Animals ,Nucleic Acid Conformation ,Phosphorylation ,Cyclic AMP Response Element-Binding Protein ,Promoter Regions, Genetic ,Somatostatin ,Dimerization ,Protein Binding - Abstract
The second messenger cAMP stimulates the expression of numerous genes through the PKA-dependent phosphorylation of CREB. The cAMP-regulated transcription factor CREB undergoes conformational change in response to phosphorylation by PKA at Ser 133. The phosphorylation enables interaction between the kinase-inducible domain (KID) of CREB and KIX domain of CREB binding protein (CBP).To understand the activation mechanism of CREB-mediated gene expression, we performed the electron-microscope imaging of the transcription machinery. We improved the metal shadowing techniques to achieve higher resolution to detect phosphorylation-induced conformation change of the protein. Homodimer formation of CREB and the complex formation of phosphorylated CREB with CBP were observed under the electron microscope. The binding of the CREB dimer to CREs on the somatostatin and tyrosine hydroxylase promoters were also visualized directly and stereoscopically.Greatly improved resolution achieved by our modified metal shadowing techniques makes it possible to visualize that the shape of CREB homodimer was changed in phosphorylation-dependent manner and that the promoter DNA strands containing CREs appeared to be bent and twisted slightly by the holding in the crevice of the CREB homodimer. This method may be applicable to visualize transcriptional activation process of nuclear receptors or general transcription machinery.
- Published
- 2000
19. Expression of human tyrosine hydroxylase type I in Escherichia coli as a protease-cleavable fusion protein. Short communication
- Author
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A, Nakashima, K, Mori, T, Nagatsu, and A, Ota
- Subjects
Enteropeptidase ,Bacterial Proteins ,Monosaccharide Transport Proteins ,Tyrosine 3-Monooxygenase ,Escherichia coli Proteins ,Recombinant Fusion Proteins ,Escherichia coli ,Humans ,ATP-Binding Cassette Transporters ,Carrier Proteins ,Maltose-Binding Proteins - Abstract
Wild-type and N-terminal 35-, 38-, and 44-amino acid-deleted mutants of human tyrosine hydroxylase type 1 (hTH1) fused to maltose-binding protein via the target sequence for a restriction protease were expressed in Escherichia coli and purified. The fused protein was treated with the restriction protease factor Xa or enterokinase to isolate hTH1 from the fused form. The treatment of fused wild-type and 35-amino acid-deleted mutant with factor Xa and enterokinase caused non-specific cleavages in the vicinity of the phosphorylation sites, Ser19 and Ser40, due to the flexible conformation of the N-terminus of hTH1.
- Published
- 1999
20. [The relation between metabolism of biopterin and dystonia-parkinsonism]
- Author
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H, Ichinose, T, Ohye, T, Suzuki, H, Inagaki, and T, Nagatsu
- Subjects
Chromosomes, Human, Pair 14 ,Substantia Nigra ,Dystonia ,Heterozygote ,Dopamine ,Animals ,Humans ,Parkinson Disease ,GTP Cyclohydrolase ,Biopterin - Abstract
Tetrahydrobiopterin (BH4) is an essential cofactor for tyrosine hydroxylase. BH4 can be synthesized from GTP through three enzymatic reactions. The rate-limiting step of the BH4 synthesis is catalyzed by GTP cyclohydrolase I (GCH). Recently, we found that GCH is a causative gene for hereditary progressive dystonia/dopa-responsive dystonia (HPD/DRD). However, several problems still remain to be solved. The first concern is the presence of asymptomatic carriers in the disease. The difference between symptomatic and asymptomatic carriers is unknown. Second, we cannot find any mutation in the coding region of the GCH gene in about 40% of the patients. What kind of mutation would be present in these patients. The last concern is the molecular mechanism how the enzymatic activity is decreased to less than 20% of normal values. Further studies are required to solve the questions.
- Published
- 1999
21. Neurotrophins and cytokines in Parkinson's disease
- Author
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M, Mogi and T, Nagatsu
- Subjects
Proto-Oncogene Proteins c-bcl-2 ,Solubility ,Reference Values ,Brain ,Cytokines ,Humans ,Apoptosis ,Parkinson Disease ,Nerve Growth Factors ,fas Receptor - Published
- 1999
22. Molecular genetics of DOPA-responsive dystonia
- Author
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H, Ichinose and T, Nagatsu
- Subjects
Dystonia ,Mutation ,Humans ,Genes, Recessive ,Parkinson Disease ,Child ,Molecular Biology ,Dihydroxyphenylalanine ,Genes, Dominant - Published
- 1999
23. Molecular biology of catecholamine-related enzymes in relation to Parkinson's disease
- Author
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T, Nagatsu and H, Ichinose
- Subjects
Catecholamines ,Animals ,Humans ,Parkinson Disease - Abstract
1. Catecholamine (dopamine, norepinephrine, and epinephrine) biosynthesis is regulated by tyrosine hydroxylase (TH). TH activity is regulated by the concentration of the cofactor tetrahydrobiopterin (BH4), whose level is regulated by GTP cyclohydrolase I (GCH) activity. Thus, GCH activity indirectly regulates TH activity and catecholamine levels. 2. TH activity in the nigrostriatal dopaminergic neurons is most sensitive to the decrease in BH4. 3. Mutations of GCH result in reductions in GCH activity, BH4, TH activity, and dopamine, causing either recessively inherited GCH deficiency or dominantly inherited hereditary progressive dystonia [HPD; Segawa's disease; also called dopa-responsive dystonia (DRD)]. 4. In juvenile parkinsonism and Parkinson's disease, which have dopamine deficiency in the basal ganglia as HPD/DRD, the GCH gene may be normal, and the molecular mechanism of the dopamine deficiency in the basal ganglia is different from that in HPD/DRD.
- Published
- 1999
24. [Structure and function of a novel ErbB ligand, NTAK]
- Author
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H, Ishiguro, S, Higashiyama, K, Yamada, N, Ichino, N, Taniguchi, and T, Nagatsu
- Subjects
Animals ,Humans ,Nerve Growth Factors ,Cloning, Molecular ,Ligands ,Rats - Abstract
A novel member of the epidermal growth factor (EGF) family, the neural and thymus-derived activator for ErbB kinase (NTAK) has been cloned from the cDNA library of a rat pheochromocytoma cell line, PC12 cells and human neuroblastoma cell line, SK-N-SH cells. Four alternative spliced isoforms from rat cDNA have been detected by the methods of RT-PCR. The rat NTAK alpha 2a isoform exhibits 94% identity in its sequence with the human NTAK alpha isoform. Three characteristic Ig-like, EGF-like and hydrophobic domains have been identified in rat and human NTAK molecules. Recombinant NTAK, the soluble 46 kDa form, binds directly to ErbB3 and ErbB4, but not ErbB1 and B2. NTAK, however, transactivates with heterodimer such as ErbB1/B3, B1/B4, B2/B3, B2/B4, and B3/B4. NTAK stimulates the differentiation of MDA-MB-453 cells, derived from blast carcinoma. NTAK competitively inhibits the binding of [125I] NRG-1 to these cells. Thus, NTAK is a new member of the EGF family displaying NRG-1 properties.
- Published
- 1998
25. [Autosomal-dominant DOPA-responsive dystonia, caused by mutations in the GTP-cyclohydrolase I gene]
- Author
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T, Nagatsu and H, Ichinose
- Subjects
Dystonia ,Mutation ,Humans ,Genes, Recessive ,Parkinson Disease ,GTP Cyclohydrolase ,Dihydroxyphenylalanine ,Genes, Dominant - Abstract
The development of autosomal dominant DOPA-responsive dystonia (AD-DRD) is stipulated by mutation in GTP-cyclohydrolase I gene. GTP-cyclohydrolase I is the first and key enzyme of tetrahydrobiopterin biosynthesis. Its deficiency in nigrostriatal dopaminergic neurons cause a decrease in tyrosine hydroxylase activity and therefore dopamine deficiency. However, administration of low doses of dopamine can control the development of AD-DRD. Determination of GTP-cyclohydrolase I activity in mononuclear blood cells is convenient diagnostic method.
- Published
- 1998
26. [Molecular biology of hereditary dystonia]
- Author
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T, Nagatsu and H, Ichinose
- Subjects
Diagnosis, Differential ,Dystonia ,Mice ,Tyrosine 3-Monooxygenase ,Molecular Sequence Data ,Mutation ,Animals ,Humans ,Parkinson Disease ,Amino Acid Sequence ,GTP Cyclohydrolase ,Genes, Dominant - Abstract
The causative genes of hereditary dystonia (hereditary progressive dystonia, HPD; dopa-responsive dystonia, DRD) were discovered in 1994-1995. HPD/DRD is caused by the deficiency of dopamine to less than 20% of the normal level in the nigro-striatum of the brain owing to the mutations of the dopamine synthesizing enzymes. Autosomal dominant dystonia (Segawa's disease) was found to be caused by mutations of GTP cyclohydrolase I which synthesizes tetrahydrobiopterin, the cofactor of tyrosine hydroxylase, by Ichinose et al. (Nature Genetics, 1994) in Japan. Autosomal recessive dystonia was reported to be caused by mutations of tyrosine hydroxylase by Lüdecke et al. (Human Genetics, 1995) in Germany. Hereditary dystonia, especially autosomal dominant Segawa's disease can be completely controlled by L-dopa administration. Measurement of the activity of GTP cyclohydrolase I in mononuclear blood cells is useful for the diagnosis of Segawa's disease.
- Published
- 1998
27. Autonomic neuropathy in transgenic mice caused by immunotoxin targeting of the peripheral nervous system
- Author
-
H, Sawada, K, Nishii, T, Suzuki, K, Hasegawa, T, Hata, I, Nagatsu, R J, Kreitman, I, Pastan, T, Nagatsu, and K, Kobayashi
- Subjects
Cytotoxicity, Immunologic ,Chromatography, Gas ,Sympathetic Nervous System ,Immunotoxins ,Mice, Transgenic ,Dopamine beta-Hydroxylase ,Mice ,Catecholamines ,Phenotype ,Autonomic Nervous System Diseases ,Nerve Degeneration ,Peripheral Nervous System ,Animals ,Humans ,Interleukin-2 - Abstract
Autonomic neuropathy in several neurodegenerative disorders results from disturbance in physiological functions of different cell types in the central and peripheral nervous systems. For a clearer understanding of the etiology and pathogenesis of the autonomic disorders it is necessary to create animal models in which degeneration of the causative neuronal types can be induced. Immunotoxin-mediated cell targeting (IMCT) is a novel transgenic mouse technology for eliminating selective cell types with the cytotoxic activity of a recombinant immunotoxin anti-Tac(Fv)-PE40. In this study we conditionally disrupted peripheral catecholaminergic cells with IMCT to generate a mouse model developing autonomic failure based on primary defects of the sympathetic nervous system. Transgenic mice expressing human interleukin-2 receptor alpha subunit under the control of the dopamine beta-hydroxylase gene promoter were intravenously treated with a proper dose of anti-Tac(Fv)-PE40. The immunotoxin induced a selective loss of the target cells in peripheral tissues of the transgenic mice and an impairment of catecholamine metabolism in the tissues. Targeting of the peripheral catecholaminergic cells resulted in severe and progressive phenotypic abnormalities mainly characterized by cardiac dysfunction, hypoactivity, and hypothermia, which explain development of autonomic neuropathy. Our IMCT strategy is useful for elucidating the involvement of different neuronal types and their interactions in the development and symptom of autonomic disorders.
- Published
- 1998
28. Genetic basis of dominant dystonia
- Author
-
T, Nagatsu and H, Ichinose
- Subjects
Chromosomes, Human, Pair 14 ,Male ,Dystonia ,Dopamine ,Chromosome Mapping ,Humans ,Point Mutation ,Female ,Frameshift Mutation ,GTP Cyclohydrolase ,Corpus Striatum ,Sequence Deletion - Published
- 1997
29. Immunocytochemical evidence of novel catecholamine- or biopterin-related neurons of mammalian brain
- Author
-
I, Nagatsu, M, Sakai, N, Karasawa, T, Takeuchi, R, Arai, K, Yamada, and T, Nagatsu
- Subjects
Mammals ,Neurons ,Serotonin ,Tyrosine 3-Monooxygenase ,Dopamine ,Brain ,Mice, Transgenic ,Biopterin ,Immunohistochemistry ,Dihydroxyphenylalanine ,Rats ,Mice ,Catecholamines ,Animals ,Humans ,GTP Cyclohydrolase - Published
- 1997
30. Catecholamine synthesis and release. Overview
- Author
-
T, Nagatsu and L, Stjärne
- Subjects
Mice, Knockout ,Mice ,Catecholamines ,Tyrosine 3-Monooxygenase ,Phenylethanolamine N-Methyltransferase ,Animals ,Humans ,Mice, Transgenic ,Dopamine beta-Hydroxylase ,Exocytosis ,Gene Expression Regulation, Enzymologic - Published
- 1997
31. Population genetics of a functional variant of the dopamine beta-hydroxylase gene (DBH)
- Author
-
J F, Cubells, K, Kobayashi, T, Nagatsu, K K, Kidd, J R, Kidd, F, Calafell, H R, Kranzler, H, Ichinose, and J, Gelernter
- Subjects
DNA, Complementary ,Genotype ,Substance-Related Disorders ,Genetic Variation ,Dopamine beta-Hydroxylase ,Blood Protein Electrophoresis ,Polymerase Chain Reaction ,Phenotype ,Gene Frequency ,Ethnicity ,Schizophrenia ,Humans ,Point Mutation ,Disease Susceptibility ,Alleles ,Polymorphism, Restriction Fragment Length - Abstract
Dopamine beta-hydroxylase (E.C. 1.14.17.1; protein abbreviation: DbetaH) catalyzes conversion of dopamine to norepinephrine. Previous work identified two expressed alleles of the gene encoding DbetaH (locus symbol DBH), containing either G or T at nucleotide position 910, resulting in specification by codon 304 of alanine (DBH*304A) or serine (DBH*304S), respectively. The current study employed denaturing gradient gel electrophoresis to identify these alleles, and after developing a PCR RFLP for rapid genotyping, estimated the frequencies of the alleles in African-Americans, European-Americans, and in several geographically dispersed populations (Mbuti, Danes, Adygei, Chinese, Japanese, Surui, Maya, and Nasioi). DBH*304A was the most common allele in all populations tested, with allele frequencies greater than 0.80 in each case. There was significant heterogeneity in allele frequency across population groups. The DBH*304S allele was most common in subjects of African descent, and least common in East Asians and individuals from indigenous populations of North and South America. The frequency of DBH*304S was significantly higher in African-Americans (0.16) than in European-Americans (0.06; P0.004). Of the four DBH*304S homozygotes observed, all were Europeans and three of the four were Danes. Based on empirical P-values generated by computer simulation, the observed proportions of DBH*304S homozygotes did not differ significantly from Hardy-Weinberg expectations in any of the populations after Bonferroni correction for multiple comparisons. The observation of significant heterogeneity in DBH*304S allele frequency across different population samples demonstrates the importance of controlling for population stratification in future studies testing for associations between DBH*304S and clinical phenotypes.
- Published
- 1997
32. The role of L-aromatic amino acid decarboxylase in serotonin-stimulated aldosterone secretion in response to salt intake
- Author
-
T Nagatsu, Bryan Williams, L Brett, H J Olverman, M R Lee, and N. Burns
- Subjects
medicine.medical_specialty ,Serotonin ,Sodium ,chemistry.chemical_element ,Biology ,5-Hydroxytryptophan ,chemistry.chemical_compound ,Endocrinology ,Zona fasciculata ,Internal medicine ,medicine ,Animals ,Salt intake ,Rats, Wistar ,Aldosterone ,Aromatic L-amino acid decarboxylase ,Adrenal cortex ,Sodium, Dietary ,General Medicine ,Rats ,medicine.anatomical_structure ,chemistry ,Zona glomerulosa ,Aromatic-L-Amino-Acid Decarboxylases ,Zona Glomerulosa ,Adrenal medulla - Abstract
In this study we tested a new hypothesis namely that serotonin (5-hydroxytryptamine, 5-HT) could be synthesised within the zona glomerulosa of the rat adrenal gland from exogenous 5-hydroxytryptophan (5-HTP) by the enzyme L-aromatic amino acid decarboxylase (L-AAAD). A specific monoclonal antibody against L-AAAD showed that the enzyme was present predominantly in the adrenal medulla but also in the zona glomerulosa and zona fasciculata. Wistar rats, maintained on a normal (NS), low (LS) or high (HS) salt diet for one week, were sacrificed by decapitation, blood samples taken and the adrenal glands removed. Plasma aldosterone concentrations were significantly higher in the LS diet group (2.91 +/- 0.35 nM) and significantly lower in the HS diet group (0.261 +/- 0.55 nM) compared with the NS diet group (1.025 +/- 0.133 nM) (p < 0.001). Capsules from the LS diet group synthesised significantly higher maximal levels of 5-HT (2615.463 +/- 480.88 nM/mg protein) than capsules from the NS (1219.117 +/- 150.259 nM/mg protein) and the HS (968.477 +/- 214.485 nM/mg protein) salt diet groups (p < 0.05). Maximal aldosterone secretion in adrenal capsules obtained from rats on the LS diet (73.428 +/- 4.053 nM/mg protein) was significantly higher than in those obtained from rats on the NS diet (41.658 +/- 1.87 nM/mg protein) (p < 0.05). Maximal aldosterone secretion in adrenal capsules from the HS diet group (30.624 +/- 2.114 nM/mg protein) was significantly lower than in the capsules from both the LS and NS groups (p < 0.05). Carbidopa (10(-4) M), a specific inhibitor of L-AAAD, markedly attenuated the secretion of aldosterone when adrenal capsules from all three salt diet groups were incubated with 10(-4) M 5-HTP (p < 0.05), but had no significant effect on basal aldosterone secretion. These results clearly demonstrate that L-AAAD is not only present in the medulla, but also in the zona glomerulosa and zona fasciculata of the rat adrenal gland. In addition, 5-HT can be synthesised in the zona glomerulosa/capsular region of the rat adrenal gland and both its biosynthesis and its ability to stimulate aldosterone secretion is increased by sodium depletion and attenuated by sodium loading. This raises the interesting possibility that L-AAAD could play a role in the regulation of aldosterone secretion during sodium deficiency in the rat by converting circulating 5-HTP (which is present in blood at concentrations exceeding 1 micromolar) into 5-HT within the adrenal cortex.
- Published
- 1996
33. Transient appearance of GTP cyclohydrolase I--positive non-monoaminergic neurons in the ventral lateral geniculate nucleus of postnatal mice
- Author
-
I, Nagatsu, T, Takeuchi, M, Sakai, R, Arai, N, Karasawa, and T, Nagatsu
- Subjects
Cerebral Cortex ,Male ,Neurons ,Neurotransmitter Agents ,Serotonin ,Superior Colliculi ,Tyrosine 3-Monooxygenase ,Age Factors ,NADPH Dehydrogenase ,Geniculate Bodies ,Hippocampus ,Immunohistochemistry ,Mice, Inbred C57BL ,Mice ,Antibody Specificity ,Aromatic-L-Amino-Acid Decarboxylases ,Cerebellum ,Animals ,Nitric Oxide Synthase ,GTP Cyclohydrolase - Abstract
The transient appearance of GTP cyclohydrolase I (GCH)-immunoreactive (ir) cells in the ventral lateral geniculate nuclear region of mice was detected by use of an avidin-biotin peroxidase complex method with an antibody specific for an oligopeptide of rat GCH (residues from 12 to 23, GFPERELPRPGA). In this brain region, we found for the first time novel GCH-ir cells already at postnatal day 1 (P1). The numbers reached maximum at P14 and decreased until P29, and they had mostly disappeared by P56. These cells were tyrosine hydroxylase negative and aromatic L-amino acid decarboxylase negative, indicating a lack of dopamine or serotonin production, and thus do not belong to the monoaminergic neuron system.
- Published
- 1996
34. [Amines and pteridines]
- Author
-
K, Shimpo, T, Chihara, M, Hibiya, S, Ito, and T, Nagatsu
- Subjects
Cysteinyldopa ,Neoplasms ,Biomarkers, Tumor ,Polyamines ,Humans ,Biopterin ,Melanoma ,Neopterin - Abstract
Polyamines (putrescine, spermidine and spermine) play important roles in cell proliferation and differentiation, and have been established as tumors markers. We and other workers have confirmed that N1-acetylspermidine in tumor tissues, spermidine and spermine in erythrocytes, and N1,N12-diacetylspermine in urine might be the most sensitive indicators for various forms of tumors. Neopterin is a marker of cell-mediated immunostimulation, and may be a helpful marker in monitoring cancer patients. HPLC and immunological assays of neopterin, biopterin, and N2-(3-aminopropyl)biopterin(oncopterin) in urine might be useful in the clinical study of pteridines, as cancer markers. Serum 5-S-cysteinyldopa level is a useful and specific biochemical marker for malignant melanoma.
- Published
- 1996
35. [Molecular genetics of hereditary progressive dystonia (HPD/Segawa's disease)]
- Author
-
H, Ichinose and T, Nagatsu
- Subjects
Dystonia ,Cell Death ,Tyrosine 3-Monooxygenase ,Dopamine ,Humans ,Point Mutation ,GTP Cyclohydrolase ,Corpus Striatum - Abstract
Hereditary progressive dystonia with marked diurnal fluctuation (HPD, Segawa's disease), also known as DOPA-responsive dystonia (DRD), was found to be caused by mutation of GTP cyclohydrolase I (GCH) gene. GCH activity in mononuclear blood cells was decreased to less than 20% of the normal values. The decrease in GCH activity causes the decrease in tetrahydrobiopterin (BH4) levels, resulting in decreased tyrosine hydroxylase (TH) activity and finally in decreased dopamine levels in the nigrostriatal dopamine neurons. In contrast, GCH activity in mononuclear blood cells in juvenile parkinsonism was normal. Recessive dystonia was shown to have a point mutation in TH gene. Thus, HPD (Segawa's disease) is distinct from recessive dystonia and juvenile parkinsonism. Patients with Parkinson's disease had decreased GCH activity in parallel with the decreases in TH activity and dopamine in the striatum, probably as the results of cell death.
- Published
- 1996
36. Molecular biology of catecholamine neurons in relation to Parkinson's disease
- Author
-
T, Nagatsu and H, Ichinose
- Subjects
Neurons ,Catecholamines ,Base Sequence ,Species Specificity ,Tyrosine 3-Monooxygenase ,Molecular Sequence Data ,Animals ,Humans ,Parkinson Disease ,Amino Acid Sequence - Published
- 1996
37. Inhibition of hepatocellular carcinoma development and erythrocyte polyamine levels in ODS rats fed on 3'-methyl-4-dimethylaminoazobenzene by hemicalcium ascorbate, 2-O-octadecylascorbic acid, and ascorbyl palmitate
- Author
-
K, Shimpo, H, Takahashi, H, Tsuda, T, Hibino, K, Kawai, C, Kimura, T, Nagatsu, and K, Fujita
- Subjects
Male ,Methyldimethylaminoazobenzene ,Erythrocytes ,Spermidine ,Body Weight ,Antimutagenic Agents ,Ascorbic Acid ,Free Radical Scavengers ,Organ Size ,Rats ,Liver Neoplasms, Experimental ,Liver ,Carcinogens ,Putrescine ,Animals ,Spermine - Abstract
We examined the modifying effect of hemicalcium ascorbate (Ca-Asc), and its lipophilic derivatives, 2-O-octadecylascorbic acid (CV-3611) and ascorbyl palmitate (AscP), on hepatocarcinogenesis by 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) in ODS rats (a mutant unable to synthesize ascorbic acid). Male 14-week-old ODS rats were given a modified AIN-A diet or the diet containing 0.06% 3'-Me-DAB, and drinking water containing 0.1% ascorbic acid. Rats were divided into the following eight groups: Group 1, no treatment (basal diet alone); Group 2, Ca-Asc; Group 3, CV-3611; Group 4, AscP;Group 5, 3'-Me-DAB; Group 6, 3'-Me-DAB + Ca-Asc; Group 7, 3'-Me-DAB + CV-3611; and Group 8, 3'-Me-DAB + AscP. Ca-Asc (2 g/kg), CV-3611 (0.2 g/kg), and AscP (0.6 g/kg) was administered once every day by gavage. 3'-Me-DAB was given in the basal diet. After 17 weeks, animals were killed by exsanguination, and the liver was weighed and processed for histological examination. Treatment by CV-3611 exerted a marked inhibitory effect on the development of 3'-Me-DAB-induced hepatocellular carcinomas (HCC) as measured by multiplicity. Although less effective than CV-3611, Ca-Asc and AscP also showed inhibitory effect. We have also studied the correlation of erythrocyte (RBC) polyamine levels and HCC development. RBC polyamine levels were inhibited by Ca-Asc and its derivatives, indicating it may be a marker of hepatocarcinogenesis.
- Published
- 1996
38. Interleukin-2 but not basic fibroblast growth factor is elevated in parkinsonian brain. Short communication
- Author
-
M, Mogi, M, Harada, T, Kondo, P, Riederer, and T, Nagatsu
- Subjects
Aged, 80 and over ,Male ,Substantia Nigra ,Case-Control Studies ,Humans ,Interleukin-2 ,Female ,Fibroblast Growth Factor 2 ,Parkinson Disease ,Middle Aged ,Corpus Striatum ,Aged - Abstract
The contents of interleukin (IL)-2 and basic fibroblast growth factor (bFGF) were measured in the brain (caudate nucleus, putamen, and cerebral cortex) from control and parkinsonian patients by highly sensitive enzyme-linked immunosorbent assay (ELISA). The concentrations of IL-2 in the brain were in the order of pg/mg protein, and the values were significantly higher in the caudate and putamen from parkinsonian patients than those from control patients. However, the levels of IL-2 in the cerebral cortex showed no significant difference between parkinsonian and control patients. In contrast to IL-2, the bFGF levels in the brain were high and in the order of ng/mg protein, and there was no significant difference in the caudate and putamen between parkinsonian and control patients. Although both IL-2 and bFGF may play important roles in dopaminergic neurons as neurotrophic factors, IL-2 but not bFGF may relate to the compensatory response in the nigrostriatal dopaminergic regions in parkinsonian brain during progress of neurodegeneration.
- Published
- 1996
39. Characterization of mouse and human GTP cyclohydrolase I genes. Mutations in patients with GTP cyclohydrolase I deficiency
- Author
-
H, Ichinose, T, Ohye, Y, Matsuda, T, Hori, N, Blau, A, Burlina, B, Rouse, R, Matalon, K, Fujita, and T, Nagatsu
- Subjects
Base Sequence ,Transcription, Genetic ,Molecular Sequence Data ,Chromosome Mapping ,Nucleic Acid Hybridization ,DNA ,Dystonia ,Mice ,Sequence Homology, Nucleic Acid ,Mutation ,Escherichia coli ,Animals ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,GTP Cyclohydrolase - Abstract
GTP cyclohydrolase I is the first and rate-limiting enzyme for the biosynthesis of tetrahydrobiopterin in mammals. Previously, we reported three species of human GTP cyclohydrolase I cDNA in a human liver cDNA library (Togari, A., Ichinose, H., Matsumoto, S., Fujita, K., and Nagatsu, T. (1992) Biochem. Biophys. Res. Commun. 187, 359-365). Furthermore, very recently, we found that the GTP cyclohydrolase I gene is causative for hereditary progressive dystonia with marked diurnal fluctuation, also known as DOPA-responsive dystonia (Ichinose, H., Ohye, T., Takahashi, E., Seki, N., Hori, T., Segawa, M., Nomura, Y., Endo, K., Tanaka, H., Tsuji, S., Fujita, K., and Nagatsu, T. (1994) Nature Genetics 8, 236-242). To clarify the mechanisms that regulate transcription of the GTP cyclohydrolase I gene and to generate multiple species of mRNA, we isolated genomic DNA clones for the human and mouse GTP cyclohydrolase I genes. Structural analysis of the isolated clones revealed that the GTP cyclohydrolase I gene is encoded by a single copy gene and is composed of six exons spanning approximately 30 kilobases. We sequenced all exon/intron boundaries of the human and mouse genes. Structural analysis also demonstrated that the heterogeneity of GTP cyclohydrolase I mRNA is caused by an alternative usage of the splicing acceptor site at the sixth exon. The transcription start site of the mouse GTP cyclohydrolase I gene and the 5'-flanking sequences of the mouse and human genes were determined. We performed regional mapping of the mouse gene by fluorescence in situ hybridization, and the mouse GTP cyclohydrolase I gene was assigned to region C2-3 of mouse chromosome 14. We identified missense mutations in patients with GTP cyclohydrolase I deficiency and expressed mutated enzymes in Escherichia coli to confirm alterations in the enzyme activity.
- Published
- 1995
40. Brain beta 2-microglobulin levels are elevated in the striatum in Parkinson's disease
- Author
-
M, Mogi, M, Harada, T, Kondo, P, Riederer, and T, Nagatsu
- Subjects
Cerebral Cortex ,Immunoenzyme Techniques ,Male ,Tumor Necrosis Factor-alpha ,Putamen ,Humans ,Female ,Parkinson Disease ,Autopsy ,Middle Aged ,beta 2-Microglobulin ,Corpus Striatum ,Aged - Abstract
beta 2-Microglobulin (B2-MG) content was measured for the first time in the brain (caudate nucleus, putamen, and cerebral cortex) from control and parkinsonian patients by a highly sensitive sandwich enzyme immunoassay. The concentrations of B2-MG in dopaminergic striatal regions were significantly higher in parkinsonian patients than those in controls, whereas those in the cerebral cortex showed no significant difference between parkinsonian and control subjects. Tumor necrosis factor-alpha (TNF-alpha) concentrations were also increased in the striatum, confirming our previous findings, but not in the cerebral cortex. Since TNF-alpha may induce B2-MG expression, these results suggest that an immunological response may occur in the nigrostriatal dopaminergic regions in Parkinson's disease.
- Published
- 1995
41. Tyrosine hydroxylase: human isoforms, structure and regulation in physiology and pathology
- Author
-
T, Nagatsu
- Subjects
Isoenzymes ,Mice ,Base Sequence ,Tyrosine 3-Monooxygenase ,Cricetinae ,Pteridines ,Molecular Sequence Data ,Schizophrenia ,Animals ,Humans ,Parkinson Disease ,Amino Acid Sequence ,Gene Expression Regulation, Enzymologic - Abstract
TH is a tetrahydrobiopterin-requiring, iron-containing monooxygenase. It catalyses the conversion of L-tyrosine to L-dopa, which is the first, rate-limiting step in the biosynthesis of catecholamines (dopamine, noradrenaline and adrenaline), the central and sympathetic neurotransmitters and adrenomedullary hormones. The cofactor of TH is tetrahydrobiopterin, which is synthesized from GTP in three steps. The TH gene consists of 14 exons only in humans and 13 exons in animals. Human TH exists in four isoforms (hTH1-4) that are produced by alternative mRNA splicing from a single gene. A single mRNA and protein corresponding to hTH1 exists in non-primates. Monkey TH exists in two isoforms, corresponding to hTH1 and hTH2. TH activity is regulated in the short term by feedback inhibition of catecholamines in competition with tetrahydrobiopterin, and by activation and deactivation due to phosphorylation and dephosphorylation, mainly at Ser-19 and Ser-40 of hTH1. The multiple TH isoforms in humans and monkeys have additional phosphorylation, resulting in more subtle regulation. In long-term regulation under stress conditions, TH protein is induced. CRE and AP1 in the 5' flanking region of the TH gene may be the main functional elements for TH gene expression. TH may be closely related to the pathogenesis of neurological diseases, such as dystonia and Parkinson's disease, psychiatric diseases, such as affective disorders and schizophrenia, as well as cardiovascular diseases. The TH gene may prove useful in gene therapy to compensate for decreased levels of catecholamines in neurological diseases, for example, for supplementation of dopamine in Parkinson's disease.
- Published
- 1995
42. Recombinant human tyrosine hydroxylase types 1-4 show regulatory kinetic properties for the natural (6R)-tetrahydrobiopterin cofactor
- Author
-
S, Nasrin, H, Ichinose, H, Hidaka, and T, Nagatsu
- Subjects
Isoenzymes ,Kinetics ,DNA, Complementary ,Base Sequence ,Tyrosine 3-Monooxygenase ,Enzyme Stability ,Molecular Sequence Data ,Escherichia coli ,Humans ,Cloning, Molecular ,Hydrogen-Ion Concentration ,Biopterin ,Recombinant Proteins - Abstract
Human tyrosine hydroxylase (hTH) exists in four isoforms. The four recombinant hTH isoenzymes types 1-4 (hTH1-4) produced in and purified from Escherichia coli showed regulatory kinetic properties for the natural (6R)-L-erythro-tetrahydrobiopterin (RBPH4) as a cofactor. In contrast, the unnatural cofactor (6S)-L-erythro-tetrahydrobiopterin (SBPH4) and a synthetic cofactor (6RS)-methyl-tetrahydropterin (6MPH4) showed usual kinetic characteristics with each of hTH1-4. Substrate inhibition by tyrosine was observed for each of hTH1-4 with natural RBPH4. Two different Km values for pterin cofactor were observed at a high concentration (200 microM) of L-tyrosine only with natural RBPH4, in contrast to a single Km value for unnatural SBPH4 or synthetic 6MPH4. The present results suggest that in the presence of relatively high concentrations (approximately 100 microM) of tyrosine in vivo, RBPH4 cofactor may have a regulatory role for the activity of all four human isoenzymes in vivo. We also found that recombinant hTH1 and 3 were more unstable than hTH2 and 4, suggesting that the 4-amino-acid insertion in hTH2 and 4 may be responsible for the relative stability of hTH2 and 4 isoenzymes.
- Published
- 1994
43. Novel toxins and Parkinson's disease: N-methylation and oxidation as metabolic bioactivation of neurotoxin
- Author
-
M, Naoi, W, Maruyama, T, Niwa, and T, Nagatsu
- Subjects
Microdialysis ,Neurotoxins ,Brain ,Cell Count ,Parkinson Disease ,Isoquinolines ,Methylation ,PC12 Cells ,Rats ,Adenosine Triphosphate ,Animals ,Humans ,Tissue Distribution ,Oxidation-Reduction - Abstract
In human brains, a series of monoamine-derived 1,2,3,4-tetrahydroisoquinolines and the 6,7-dihydroxy derivatives has been identified. A tetrahydroisoquinoline was found to cause parkinsonism in monkey, but its toxicity was not so potent as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Two metabolic steps were found to increase cytotoxicity of isoquinolines. N-Methylation by a non-specific N-methyltransferase was proved by in vivo and in vitro experiments. The N-methylated compound was oxidized into N-methylisoquinolinium ion by monoamine oxidase from human brain mitochondria. The oxidation was proved by microdialysis in the rat brain. The isoquinolinium ion was more cytotoxic than the two metabolic precursors. N-Methylation of dopamine-derived 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines was detected by in vivo microdialysis in the rat striatum, and their presence in the human brain was confirmed by GC-MS. The metabolic bioactivation may be a general pathway to produce neurotoxins as the pathogenic agents of Parkinson's disease.
- Published
- 1994
44. Food-derived heterocyclic amines, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole and related amines, as inhibitors of monoamine metabolism
- Author
-
W, Maruyama, A, Ota, A, Takahashi, T, Nagatsu, and M, Naoi
- Subjects
Mice ,Monoamine Oxidase Inhibitors ,Tyrosine 3-Monooxygenase ,Food ,Animals ,Amines ,Carbolines ,Rats - Abstract
The effects of heterocyclic amines, pyrolysis products of tryptophan, on monoamine metabolism were examined. Among these amines, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) are potent inhibitors of the enzymes related to amine metabolism. They inhibited type A monoamine oxidase more markedly than type B. After culture of a dopamine cell model, clonal pheochromocytoma PC12h cells, with Trp-P-1 activity of tyrosine hydroxylase was decreased by reduction of its affinity to the biopterin cofactor. Trp-P-1 and Trp-P-2 inhibited tryptophan hydroxylase competitively with the substrate and non-competitively with biopterin. These results suggest that food-derived heterocyclic amines may perturb the monoamine levels in the brain through the inhibition of the biosynthesis and metabolism of biogenic amines.
- Published
- 1994
45. [Molecular genetics of aromatic L-amino acid decarboxylase]
- Author
-
H, Ichinose and T, Nagatsu
- Subjects
Base Sequence ,Liver ,Aromatic-L-Amino-Acid Decarboxylases ,Genome, Human ,Molecular Sequence Data ,Adrenal Gland Neoplasms ,Humans ,Amino Acid Sequence ,DNA ,Pheochromocytoma ,Cloning, Molecular - Abstract
Aromatic L-amino acid decarboxylase (AADC) decarboxylates both L-5-hydroxytryptophan to serotonin in serotonergic neurons and pineal cells, and L-dopa to dopamine in catecholaminergic neurons and adrenal medullary cells. Thus AADC produces two major mammalian neurotransmitters and hormones. We isolated and sequenced a full-length, neuronal-type, cDNA encoding human AADC. It consisted of 1932 bases containing an open reading frame encoding 480 amino acids residues with a molecular weight of 53,891. We expressed a recombinant human AADC in COS cells and proved that the expressed enzyme decarboxylated both L-5-hydroxytryptophan to serotonin and L-dopa to dopamine. We have cloned genomic DNA of human AADC and determined the structure. The genomic DNA of human AADC consists of 15 exons spanning about 100 kilobases and exists as a single copy in the hapoloid genome. We have mapped the gene to chromosome band 7p12.1-p12.3 by fluorescence in situ hybridization. We cloned the nonneuronal type cDNA from human liver and identified another first exon different from the neuronal type cDNA. This showed that an alternative usage of the first exon produced two types of mRNAs in AADC and suggested that alternative splicing would regulate the tissue-specific expression of AADC.
- Published
- 1993
46. Cloning and sequencing of cDNA encoding mouse GTP cyclohydrolase I
- Author
-
T, Nomura, H, Ichinose, C, Sumi-Ichinose, H, Nomura, Y, Hagino, K, Fujita, and T, Nagatsu
- Subjects
Mice ,Base Sequence ,Sequence Homology, Amino Acid ,Molecular Sequence Data ,Animals ,Brain ,Humans ,Amino Acid Sequence ,DNA ,Cloning, Molecular ,GTP Cyclohydrolase - Abstract
A full-length cDNA clone for GTP cyclohydrolase I (EC 3.5.4.16) was isolated from a mouse brain cDNA library by plaque hybridization. The nucleotide sequence determination revealed that the length of the cDNA insert was 994 base pairs. The coding region encoded a protein of 241 amino acid residues with a calculated molecular mass of 27,014 daltons. The deduced amino acid sequence of mouse GTP cyclohydrolase I was found to be highly homologous to rat (96%) and human type 1 (89%) enzymes.
- Published
- 1993
47. [Genetic conversion of neurotransmitter phenotype in transgenic mice]
- Author
-
K, Kobayashi and T, Nagatsu
- Subjects
Mice ,Catecholamines ,Phenotype ,Phenylethanolamine N-Methyltransferase ,Animals ,Humans ,Mice, Transgenic ,DNA ,Dopamine beta-Hydroxylase ,Cloning, Molecular - Abstract
The mammalian central and sympathetic nervous systems contain three kinds of catecholaminergic neuron (dopamine, norepinephrine and epinephrine neurons). Adrenal medulla also consists of cells producing either norepinephrine or epinephrine as hormones. To switch of catecholamine phenotype in the nervous and endocrine systems, we generated a line of transgenic mice carrying a chimeric gene containing human phenylethanolamine N-methyltransferase (PNMT) cDNA fused to the 4-kb fragment of the human dopamine beta-hydroxylase (DBH) gene promoter. Analysis of transgenic mice indicated that the additional expression of human PNMT in norepinephrine-producing cells can convert these cells to the epinephrine phenotype, and suggested that norepinephrine-producing cells normally possess the fundamental machinery required for the synthesis of epinephrine except for the PNMT expression.
- Published
- 1993
48. Cytotoxicity of dopamine-derived 6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines
- Author
-
W, Maruyama, T, Takahashi, M, Minami, A, Takahashi, P, Dostert, T, Nagatsu, and M, Naoi
- Subjects
Structure-Activity Relationship ,Salsoline Alkaloids ,Tyrosine 3-Monooxygenase ,Cell Survival ,Dopamine ,Quinolinium Compounds ,Tetrahydroisoquinolines ,Animals ,Isoquinolines ,PC12 Cells ,Cell Division ,Rats - Abstract
The in vivo effects of dopamine-derived alkaloids, 6,7-dihydroxy-1,2,3,4-tetrahydroisoquinolines, salsolinols, and their N-methylated derivatives on a dopaminergic cell model, clonal rat pheochromocytoma PC12h cells, were examined by culture in the presence of various concentrations of the agents. The effects were evaluated in comparison with those by 1,2,3,4-tetrahydroisoquinoline and its N-methylated derivatives. Among 1,2,3,4-tetrahydroisoquinolines, only N-methylisoquinolinium ion had cytotoxic effect on PC12h cells. In general, 6,7-dihydroxyisoquinolines had more potent cytotoxic effect than N-methylisoquinolinium ion, and they reduced protein amounts of PC12h cells at 100 microM and 1 mM concentration. The specific activity of tyrosine hydroxylase, the rate-limiting enzyme in dopamine biosynthesis, decreased with these isoquinolines at concentrations lower than those required to reduce the protein amount. The toxicity of N-methylated derivatives seems to be more potent than non-methylated isoquinolines. Salsolinols were proved to be accumulated in the mitochondrial fraction of the cells after 3 days in culture. N-methyl-1,2,3,4-tetrahydroisoquinoline depleted ATP from PC12h cells and it was prevented by preincubation with an inhibitor of type-A monoamine oxidase, clorgyline. These results indicate that N-methylated and oxidized derivatives of dopamine-derived alkaloids may be potent dopaminergic neurotoxins similar to 1-methyl-4-phenylpyridinium ion in the human brain and may induce Parkinson's disease after long years of accumulation.
- Published
- 1993
49. Elevated levels of oncopterin, N2-(3-aminopropyl)biopterin, a new pterin compound, in urine from patients with solid and blood cancers
- Author
-
S, Ogiwara, H, Hidaka, T, Sugimoto, R, Teradaira, K, Fujita, and T, Nagatsu
- Subjects
Adult ,Male ,Ovarian Neoplasms ,Carcinoma, Hepatocellular ,Lung Neoplasms ,Lymphoma ,Liver Neoplasms ,Prostatic Neoplasms ,Middle Aged ,Biopterin ,Neopterin ,Leukemia, Myeloid, Acute ,Urinary Bladder Neoplasms ,Neoplasms ,Biomarkers, Tumor ,Humans ,Female ,Multiple Myeloma ,Aged - Abstract
The concentrations of oncopterin, N2-(3-aminopropyl)biopterin, a new pterin compound, were determined in urine from various cancer patients by HPLC on a reverse-phase or ion-exchange column. The concentration of oncopterin increased after acid hydrolysis, indicating that it exists as an amide in urine. The oncopterin concentrations were very low in the urine of healthy controls. Among the urine samples examined, those from cases of solid cancers, e.g., hepatomas, prostatic cancer and bladder cancer, exhibited very high levels; and those from cases of blood cancers, e.g., myelomas, acute myelocytic leukemia, and lymphomas, showed moderate increases. Oncopterin may thus be a new biochemical marker of some types of cancer.
- Published
- 1993
50. Presence of tetrahydroisoquinoline-related compounds, possible MPTP-like neurotoxins, in parkinsonian brain
- Author
-
T, Niwa, N, Takeda, H, Yoshizumi, A, Tatematsu, M, Yoshida, P, Dostert, M, Naoi, and T, Nagatsu
- Subjects
Structure-Activity Relationship ,1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ,Culture Techniques ,Dopamine ,Tetrahydroisoquinolines ,Neurotoxins ,Humans ,Parkinson Disease ,Isoquinolines ,Biotransformation ,Gas Chromatography-Mass Spectrometry ,Frontal Lobe - Published
- 1993
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