Dopamine turnover and metabolism in the striatum of parkinsonian rats grafted with genetically-modified human astrocytes.
The potential of a novel therapeutic approach for treating Parkinson's disease, which involves the transplantation of a transfected human astrocyte cell line SVG-TH, that stably expresses the rate-limiting enzyme for dopamine production, tyrosine hydroxylase, was examined. SVG-TH and untransfected parent cells were grafted into the diseased striatum of rats in which Parkinson's disease had been induced by the administration of 6-hydroxydopamine. The in situ production and spillover of 3,4-dihydroxyphenylalanine (the precursor of dopamine), dopamine and their metabolites in the striatal extracellular fluid of the grafted rats was determined in conscious animals using the microdialysis technique and a high pressure liquid chromatography apparatus. Alleviation of symptoms of Parkinson's disease (abnormal movements) was evaluated by rotation tests. Upon transplantation of the SVG-TH cells into the striatum of the parkinsonian rats, the levels of dopamine in extracellular fluid of the striatum reached those of the normal rats, and correlated well with the improvement (74%) in their rotating behaviour (behavioural deficit). The levels of the two main dopamine metabolites, dihydroxyphenylacetic acid and homovanillic acid, were low in the lesioned rats, even after SVG-TH transplantation. An alternative route of metabolism of dopamine may occur in the transplanted striatum, since the dopamine metabolite, 3-O-methoxy-4-hydroxy-phenylethylamine, appeared, which indicates activity of catechol-O-methyl transferase. Upon blockade of L-aromatic-amino acid decarboxylase, 3,4-dihydroxyphenylalanine accumulated in extracellular fluid of the 6-hydroxydopamine-lesioned and SVG-TH-grafted rats, which indicated that these cells produced active tyrosine hydroxylase in vivo. These findings indicate the potential of treating Parkinson's disease by the intrabrain grafting of human astrocyte cells transfected with the rate limiting enzyme for dopamine production.
Fitoussi
N
N
Department of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
Sotnik-Barkai
I
I
Tornatore
C
C
Herzberg
U
U
Yadid
G
G
D000818:Animals; D001253:Astrocytes / Q000378:metabolism / Q000637:transplantation*; D002460:Cell Line; D003342:Corpus Striatum / Q000166:cytology / Q000378:metabolism*; D004298:Dopamine / Q000096:biosynthesis / Q000378:metabolism*; D015316:Genetic Therapy; D006801:Humans; D007980:Levodopa / Q000096:biosynthesis / Q000378:metabolism; D008297:Male; D017551:Microdialysis; D010302:Parkinson Disease, Secondary / Q000235:genetics* / Q000628:therapy*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D019076:Transgenes; D019737:Transplants; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
D007980:Levodopa; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine