TAILIEUCHUNG - Báo cáo khoa học: Functionally distinct dopamine and octopamine transporters in the CNS of the cabbage looper moth*

A cDNA was cloned from the cabbage looperTricho-plusia ni based on similarity to other cloned dopamine transporters (DATs). The total nucleotide sequence is in length and contains an open reading frame for a protein of 612 amino acids. The predicted moth DAT protein (TrnDAT) has greatest amino acid sequence identity with Drosophila melanogasterDAT (73%) andCaenorhabdi-tis elegans DAT (51%). TrnDAT shares only 45% amino acid sequence identity with an octopamine transporter (TrnOAT) cloned recently from this moth. . | Eur. J. Biochem. 270 664-674 2003 FEBS 2003 doi Functionally distinct dopamine and octopamine transporters in the CNS of the cabbage looper moth Pamela Gallant1 2 Tabita Malutan1 2 Heather McLean2 LouAnn Verellen1 Stanley Caveney2 and Cam Donly1 1 Southern Crop Protection and Food Research Centre Agriculture and Agri-Food Canada and department of Biology The University of Western Ontario London Ontario Canada A cDNA was cloned from the cabbage looper Tricho-plusia ni based on similarity to other cloned dopamine transporters DATs . The total nucleotide sequence is kb in length and contains an open reading frame for a protein of 612 amino acids. The predicted moth DAT protein TrnDAT has greatest amino acid sequence identity with Drosophila melanogaster DAT 73 and Caenorhabdi-tis elegans DAT 51 . TrnDAT shares only 45 amino acid sequence identity with an octopamine transporter TrnOAT cloned recently from this moth. The functional properties of TrnDAT and TrnOAT were compared through transient heterologous expression in Sf9 cells. Both transporters have similar transport affinities for DA Km and M respectively . However the competitive substrates octopamine and tyramine are more potent blockers of 3H dopamine DA uptake by TrnOAT than by TrnDAT. D-Amphetamine is a strong inhibitor and L-nor-epinephrine a weak inhibitor of both transporters. TrnDAT- mediated DA uptake is approximately 100-fold more sensitive to selective blockers of vertebrate transporters of dopamine and norepinephrine such as nisoxetine nomifensine and dibenzazepine antidepressants than TrnOAT-mediated DA uptake. TrnOAT is 10-fold less sensitive to cocaine than TrnDAT. None of the 15 monoamine uptake blockers tested was TrnOAT-selective. In situ hybridization shows that TrnDAT and TrnOAT transcripts are expressed by different sets of neurons in caterpillar brain and ventral nerve cord. These results show that the caterpillar CNS contains both a phenolamine

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