Functional characterization of a glutamate/aspartate transporter from the mosquito Aedes aegypti.
Glutamate elicits a variety of effects in insects, including inhibitory and excitatory signals at both neuromuscular junctions and brain. Insect glutamatergic neurotransmission has been studied in great depth especially from the standpoint of the receptor-mediated effects, but the molecular mechanisms involved in the termination of the numerous glutamatergic signals have only recently begun to receive attention. In vertebrates, glutamatergic signals are terminated by Na(+)/K(+)-dependent high-affinity excitatory amino acid transporters (EAAT), which have been cloned and characterized extensively. Cloning and characterization of a few insect homologues have followed, but functional information for these homologues is still limited. Here we report a study conducted on a cloned mosquito EAAT homologue isolated from the vector of the dengue virus, Aedes aegypti. The deduced amino acid sequence of the protein, AeaEAAT, exhibits 40-50% identity with mammalian EAATs, and 45-50% identity to other insect EAATs characterized thus far. It transports L-glutamate as well as L- and D-aspartate with high affinity in the micromolar range, and demonstrates a substrate-elicited anion conductance when heterologously expressed in Xenopus laevis oocytes, as found with mammalian homologues. Analysis of the spatial distribution of the protein demonstrates high expression levels in the adult thorax, which is mostly observed in the thoracic ganglia. Together, the work presented here provides a thorough examination of the role played by glutamate transport in Ae. aegypti.
Umesh
Anita
A
Environmental Toxicology Graduate Program Department of Cell Biology and Neuroscience, University of California, Riverside, Riverside, CA 92521, USA.
Cohen
Bruce N
BN
Ross
Linda S
LS
Gill
Sarjeet S
SS
D000330:Aedes / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D027322:Amino Acid Transport System X-AG / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D016000:Cluster Analysis; D004594:Electrophysiology; D017952:Ganglia, Invertebrate / Q000378:metabolism; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D010802:Phylogeny
D027322:Amino Acid Transport System X-AG