Intrastriatal grafting of Cos cells stably expressing human aromatic L- amino acid decarboxylase

Neurochemical effects

Farida G. Kaddis, Edward D. Clarkson, Michel J. Weber, David J. Vandenbergh, David M. Donovan, Jacques Mallet, Philippe Horellou, George R. Uhl, Curt R. Freed

Research output: Contribution to journalArticle

Abstract

To study the possibility that increasing striatal activity of aromatic L-amino acid decarboxylase (AADC; EC 4.1.1.28) can increase dopamine production in dopamine denervated striatum in response to L-3,4- dihydroxyphenylalanine (L-DOPA) administration, we grafted Cos cells stably expressing the human AADC gene (Cos-haadc cells) into 6-hydroxydopamine denervated rat striatum. Before grafting, the catalytic activity of the enzyme was assessed in vitro via the generation of 14CO2 from L[14C]DOPA. The K(m) value for L-DOPA in intact and disrupted cells was 0.60 and 0.56 mM, respectively. The cofactor, pyridoxal 5-phosphate, enhanced enzymatic activity with maximal effect at 0.1 mM. The pH optimum for enzyme activity was 6.8. Grafting Cos-haadc cells into denervated rat striatum enhanced striatal dopamine levels measured after systemic administration of L-DOPA. When measured 2 h after L-DOPA administration, the mean dopamine level in the striata of Cos-haadc-grafted animals was 2 μg/g of tissue, representing 31% of normal striatal dopamine concentration. The mean dopamine concentration in the striata grafted with untransfected Cos cells (Cos-ut cells) was 1 μg/9. At 6-8 h after L-DOPA administration, striatal dopamine content in the Cos-haadc-grafted animals was 0.67 μg/g of tissue weight, representing 9% of intact striatum dopamine content. By contrast, the average dopamine content in the Cos-ut-grafted animals was undetectable. These findings demonstrate that enhancing striatal AADC activity can improve dopamine bioformation in response to systemically administered L-DOPA.

Original languageEnglish (US)
Pages (from-to)1520-1526
Number of pages7
JournalJournal of Neurochemistry
Volume68
Issue number4
StatePublished - Apr 1997

Fingerprint

Aromatic-L-Amino-Acid Decarboxylases
Dopamine
Corpus Striatum
Levodopa
Animals
Rats
Tissue
Pyridoxal Phosphate
Oxidopamine
Enzyme activity
Enzymes
Catalyst activity
Genes

Keywords

  • Aromatic L-amino acid decarboxylase
  • Cos cells
  • L-DOPA
  • Parkinson's disease
  • Stable expression
  • Transfected cells
  • Transplantation

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Kaddis, F. G., Clarkson, E. D., Weber, M. J., Vandenbergh, D. J., Donovan, D. M., Mallet, J., ... Freed, C. R. (1997). Intrastriatal grafting of Cos cells stably expressing human aromatic L- amino acid decarboxylase: Neurochemical effects. Journal of Neurochemistry, 68(4), 1520-1526.

Intrastriatal grafting of Cos cells stably expressing human aromatic L- amino acid decarboxylase : Neurochemical effects. / Kaddis, Farida G.; Clarkson, Edward D.; Weber, Michel J.; Vandenbergh, David J.; Donovan, David M.; Mallet, Jacques; Horellou, Philippe; Uhl, George R.; Freed, Curt R.

In: Journal of Neurochemistry, Vol. 68, No. 4, 04.1997, p. 1520-1526.

Research output: Contribution to journalArticle

Kaddis, FG, Clarkson, ED, Weber, MJ, Vandenbergh, DJ, Donovan, DM, Mallet, J, Horellou, P, Uhl, GR & Freed, CR 1997, 'Intrastriatal grafting of Cos cells stably expressing human aromatic L- amino acid decarboxylase: Neurochemical effects', Journal of Neurochemistry, vol. 68, no. 4, pp. 1520-1526.
Kaddis FG, Clarkson ED, Weber MJ, Vandenbergh DJ, Donovan DM, Mallet J et al. Intrastriatal grafting of Cos cells stably expressing human aromatic L- amino acid decarboxylase: Neurochemical effects. Journal of Neurochemistry. 1997 Apr;68(4):1520-1526.
Kaddis, Farida G. ; Clarkson, Edward D. ; Weber, Michel J. ; Vandenbergh, David J. ; Donovan, David M. ; Mallet, Jacques ; Horellou, Philippe ; Uhl, George R. ; Freed, Curt R. / Intrastriatal grafting of Cos cells stably expressing human aromatic L- amino acid decarboxylase : Neurochemical effects. In: Journal of Neurochemistry. 1997 ; Vol. 68, No. 4. pp. 1520-1526.
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abstract = "To study the possibility that increasing striatal activity of aromatic L-amino acid decarboxylase (AADC; EC 4.1.1.28) can increase dopamine production in dopamine denervated striatum in response to L-3,4- dihydroxyphenylalanine (L-DOPA) administration, we grafted Cos cells stably expressing the human AADC gene (Cos-haadc cells) into 6-hydroxydopamine denervated rat striatum. Before grafting, the catalytic activity of the enzyme was assessed in vitro via the generation of 14CO2 from L[14C]DOPA. The K(m) value for L-DOPA in intact and disrupted cells was 0.60 and 0.56 mM, respectively. The cofactor, pyridoxal 5-phosphate, enhanced enzymatic activity with maximal effect at 0.1 mM. The pH optimum for enzyme activity was 6.8. Grafting Cos-haadc cells into denervated rat striatum enhanced striatal dopamine levels measured after systemic administration of L-DOPA. When measured 2 h after L-DOPA administration, the mean dopamine level in the striata of Cos-haadc-grafted animals was 2 μg/g of tissue, representing 31{\%} of normal striatal dopamine concentration. The mean dopamine concentration in the striata grafted with untransfected Cos cells (Cos-ut cells) was 1 μg/9. At 6-8 h after L-DOPA administration, striatal dopamine content in the Cos-haadc-grafted animals was 0.67 μg/g of tissue weight, representing 9{\%} of intact striatum dopamine content. By contrast, the average dopamine content in the Cos-ut-grafted animals was undetectable. These findings demonstrate that enhancing striatal AADC activity can improve dopamine bioformation in response to systemically administered L-DOPA.",
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