A high-affinity octopamine transporter cloned from the central nervous system of cabbage looper Trichoplusia ni.
A cDNA encoding a high-affinity Na(+)/anion(-)-dependent octopamine transporter (OAT) was isolated via an RT-PCR-based approach from caterpillars of the cabbage looper, Trichoplusia ni. The deduced amino acid sequence of the OAT cDNA predicts a 670 amino acid protein bearing strong homology to previously cloned monoamine transporters. The expression pattern of OAT mRNA in the central nervous system revealed by in situ hybridization closely resembles that of OA-ergic neurons identified by the presence of mRNA for tyramine beta-hydroxylase, a marker enzyme for OA-ergic neurons in invertebrates. In vitro, insect cells infected with OAT-expressing baculovirus accumulated both (3)H-OA and (3)H-dopamine with saturation kinetics typical of carrier-mediated processes. (3)H-dopamine uptake by OAT was most inhibited by tyramine, OA, dopamine and the tricyclic antidepressants desipramine and imipramine. Substitution studies for Na(+) and Cl(-) indicate that OAT has a strong requirement for Na(+) and a less stringent requirement for Cl(-). The pharmacological profile of OAT is distinct from those of other cloned monoamine transporters and makes OAT a potential target for neuro-active pest control agents.
Malutan
Tabita
T
Department of Zoology, University of Western Ontario, ON, N6A 5B7, London, Canada.
McLean
Heather
H
Caveney
Stan
S
Donly
Cam
C
D000316:Adrenergic alpha-Agonists / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002490:Central Nervous System / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D019476:Insect Proteins / Q000235:genetics*; D026901:Membrane Transport Proteins / Q000235:genetics*; D006899:Mixed Function Oxygenases / Q000235:genetics; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics* / Q000378:metabolism; D009655:Octopamine / Q000378:metabolism*; D012333:RNA, Messenger
D000316:Adrenergic alpha-Agonists; D018076:DNA, Complementary; D019476:Insect Proteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D009655:Octopamine; D006899:Mixed Function Oxygenases; C023205:tyramine beta-hydroxylase