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Expression and assessment of double genes of tyrosine hydroxylase gene and aromatic L-amino acid decarboxylase gene in vitro

Su Yue, Duan Chunli, Zhao Chunli, Zhao Huanying, Xu Qunyuan, Yang Hui

Beijing Institute for Neuroscience，Beijing Center for Neural Regeneration and Repair, Capital University of Medical Sciences. Bejing 100054,China

Abstract

The characteristic pathological changes of Parkinson's disease (PD) include a severe loss ofdopamine neurons in the substantia nigra and a severe decrease in dopamine in the striatum. Since the ex-pression of tyrosine hydroxylase (TH) and aromatic L-amino acid decarboxylase (AADC ) in the biosynthet-ic pathway for dopamine are low , a promising approach to the gene therapy of PD is to augment the gene ex-pression of the enzymes in the biosynthetic pathway for dopamine. In the present study, human TH andAADC genes were reconstructed into retrovirous vectors pLHCX and pLNCX_2 respectively. Then pLHCX/TH and pLNCX_2/AADC were transfected into packaging cell line PA317 with liposome. PA317/TH andPA317/AADC were selected by different antibiotics. Gene expression was examined by methods of immuno-histochemistry and in situ hybridization. The catalytic activity of two cloned gene enzymes was assessed invitro by HPLC-EC. Immunocytochemical staining showed that TH and AADC were expressed efficiently invitro.

Key words: Cell biology;Parkinson's disease;Tyrosine hydroxylase;Aromatic L-amino acid decarboxylase;Retrovirous vector;Gene therapy

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Citing This Article：

Su Yue, Duan Chunli, Zhao Chunli, Zhao Huanying, Xu Qunyuan, Yang Hui. Expression and assessment of double genes of tyrosine hydroxylase gene and aromatic L-amino acid decarboxylase gene in vitro. Acta Physiol Sin 2003; 55 (5): (in Chinese with English abstract).