TETRAHYDROBIOPTERIN-DEPENDENT PRODUCTION OF L-DOPA IN NRK FIBROBLASTS TRANSFECTED WITH TYROSINE-HYDROXYLASE CDNA - FUTURE USE FOR INTRACEREBRAL GRAFTING

被引:24
作者
UCHIDA, K
ISHII, A
KANEDA, N
TOYA, S
NAGATSU, T
KOHSAKA, S
机构
[1] NATL INST NEUROSCI,DEPT NEUROCHEM,4-1-1 OGAWA HIGASHI,KODAIRA,TOKYO 187,JAPAN
[2] KEIO UNIV,SCH MED,DEPT NEUROSURG,TOKYO 108,JAPAN
[3] NAGOYA UNIV,SCH MED,DEPT BIOCHEM,NAGOYA,AICHI 466,JAPAN
关键词
(6R)-l-Erythro-tetrahydrobiopterin; Intracerebral grafting; l-3,4-Dihydroxyphenylalanine; NRK fibroblast; Tyrosine hydroxylase; Tyrosine hydroxylase cDNA;
D O I
10.1016/0304-3940(90)90008-W
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
In the present study, tyrosine hydroxylase (TH; EC 1.14.16.2) cDNA was transfected into cultured fibroblasts and the production of l-3,4-dihydroxyphenylalanine (l-DOPA) was determined. Type 2 TH cDNA was transfected into fibroblasts (NRK-49F) derived from the normal rat kidney, and the expression of the TH minigene was screened by immunocytochemical staining and immunoblotting analysis with TH antiserum. Several clones of the NRK transfectants that produce TH molecules were obtained. The expressed TH molecules showed high enzyme activity in a complete assay system in vitro. However, l-DOPA was not detected in the cultured cells due to the possible absence of de novo synthesis of (6R)-l-erythro-tetrahydrobiopterin (BH4) in these cells. When BH4 was added to the medium, a large amount of l-DOPA was detected not only in the cells but also in the medium. These findings may aid in regulating the amount of l-DOPA secretion from cells after they are transplanted into the brain. © 1990.
引用
收藏
页码:282 / 286
页数:5
相关论文
共 12 条
[1]   NEURAL GRAFTING IN ANIMAL-MODELS OF NEURODEGENERATIVE DISEASES [J].
BJORKLUND, A ;
GAGE, FH .
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1985, 457 :53-81
[2]   EPITHELIOID AND FIBROBLASTIC RAT-KIDNEY CELL CLONES - EPIDERMAL GROWTH-FACTOR (EGF) RECEPTORS AND EFFECT OF MOUSE SARCOMA-VIRUS TRANSFORMATION [J].
DELARCO, JE ;
TODARO, GJ .
JOURNAL OF CELLULAR PHYSIOLOGY, 1978, 94 (03) :335-342
[3]   ON-OFF PHENOMENON IN PARKINSONS-DISEASE - RELATIONSHIP BETWEEN DOPA AND OTHER LARGE NEUTRAL AMINO-ACIDS IN PLASMA [J].
ERIKSSON, T ;
GRANERUS, AK ;
LINDE, A ;
CARLSSON, A .
NEUROLOGY, 1988, 38 (08) :1245-1248
[4]   AUTO-TRANSPLANTATION OF THE SUPERIOR CERVICAL-GANGLION INTO THE BRAIN - A POSSIBLE THERAPY FOR PARKINSONS-DISEASE [J].
ITAKURA, T ;
KAMEI, I ;
NAKAI, K ;
NAKA, Y ;
NAKAKITA, K ;
IMAI, H ;
KOMAI, N .
JOURNAL OF NEUROSURGERY, 1988, 68 (06) :955-959
[5]   ISOLATION OF A NOVEL CDNA CLONE FOR HUMAN TYROSINE-HYDROXYLASE - ALTERNATIVE RNA SPLICING PRODUCES 4 KINDS OF MESSENGER-RNA FROM A SINGLE GENE [J].
KANEDA, N ;
KOBAYASHI, K ;
ICHINOSE, H ;
KISHI, F ;
NAKAZAWA, A ;
KUROSAWA, Y ;
FUJITA, K ;
NAGATSU, T .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1987, 146 (03) :971-975
[6]   DRAMATIC RESULTS WITH BRAIN GRAFTS [J].
LEWIN, R .
SCIENCE, 1987, 237 (4812) :245-247
[7]   EXPRESSION OF FUNCTIONAL ACETYLCHOLINE-RECEPTOR FROM CLONED CDNAS [J].
MISHINA, M ;
KUROSAKI, T ;
TOBIMATSU, T ;
MORIMOTO, Y ;
NODA, M ;
YAMAMOTO, T ;
TERAO, M ;
LINDSTROM, J ;
TAKAHASHI, T ;
KUNO, M ;
NUMA, S .
NATURE, 1984, 307 (5952) :604-608
[8]   BIOPTERIN COFACTOR AND MONOAMINE-SYNTHESIZING MONOOXYGENASES [J].
NAGATSU, T .
NEUROCHEMISTRY INTERNATIONAL, 1983, 5 (01) :27-38
[9]   NERVE-CELL GRAFTING IN PARKINSONS-DISEASE [J].
SLADEK, JR ;
GASH, DM .
JOURNAL OF NEUROSURGERY, 1988, 68 (03) :337-351
[10]  
SOUTHERN P J, 1982, Journal of Molecular and Applied Genetics, V1, P327