This research's long-term objective is to understand ion regulation in insects, particularly in mosquitoes. Our present grant has determined the structure of many key proteins involved in ion regulation in Aedes aegypti Malpighian tubules. Further we have preliminary data that show enhanced V-ATPase transcript levels following a blood meal. In the present proposal we plan to build on these findings. Our objective in this proposal in to molecularly characterize the proteins that are involved in Na+ and C1- transport in mosquitoes. In the first objective we plan to functionally characterize a Na+/K+/2C1- cotransporter that was isolated from A. aegypti Malpighian tubules. This cotransporter is significantly divergent from known mammalian and insect cotransporters. Further while the V-ATPase plays a crucial role in ion regulation in mosquitoes, other proteins facilitate actual cation transport. Hence in the second objective we will also characterize the cation/antiporter CDNA clone isolated from the midgut and/or Malpighian tubule of A. aegypti. In the third objective we will attempt to evaluate changes in Malpighian tubules function following a blood meal. Therefore we will follow up our preliminary studies on V-ATPase regulation. We will determine shether V1 and V0 dissociation- reassociation occurs in Malpighian tubules after blood feeding ans also whether these genes are transcriptionally regulated. Data from these experiments will help elucidate mechanisms of change in ion transport in mosquito Malpighian tubules that occurs following a blood meal. In addition we will evaluate whether the A. aegypti Na+/K+/2C1- cotransporter is regulated. In summary, this proposal is directed towards elucidating the molecular processes involved in ion secretion following a blood meal in mosquitoes. Characterization of these processes may aid future work that could focus on the disruption of these regulatory processes in an important vector of human disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI032572-06A2
Application #
6130394
Study Section
Special Emphasis Panel (ZRG1-TMP (01))
Program Officer
Aultman, Kathryn S
Project Start
1993-01-01
Project End
2005-03-28
Budget Start
2000-04-01
Budget End
2001-03-28
Support Year
6
Fiscal Year
2000
Total Cost
$217,332
Indirect Cost
Name
University of California Riverside
Department
Zoology
Type
Schools of Earth Sciences/Natur
DUNS #
City
Riverside
State
CA
Country
United States
Zip Code
92521
Kang'ethe, Wanyoike; Aimanova, Karlygash G; Pullikuth, Ashok K et al. (2007) NHE8 mediates amiloride-sensitive Na+/H+ exchange across mosquito Malpighian tubules and catalyzes Na+ and K+ transport in reconstituted proteoliposomes. Am J Physiol Renal Physiol 292:F1501-12
Pullikuth, Ashok K; Aimanova, Karlygash; Kang'ethe, Wanyoike et al. (2006) Molecular characterization of sodium/proton exchanger 3 (NHE3) from the yellow fever vector, Aedes aegypti. J Exp Biol 209:3529-44
Patrick, Marjorie L; Aimanova, Karlygash; Sanders, Heather R et al. (2006) P-type Na+/K+-ATPase and V-type H+-ATPase expression patterns in the osmoregulatory organs of larval and adult mosquito Aedes aegypti. J Exp Biol 209:4638-51
Sanders, Heather R; Foy, Brian D; Evans, Amy M et al. (2005) Sindbis virus induces transport processes and alters expression of innate immunity pathway genes in the midgut of the disease vector, Aedes aegypti. Insect Biochem Mol Biol 35:1293-307
Sanders, Heather R; Evans, Amy M; Ross, Linda S et al. (2003) Blood meal induces global changes in midgut gene expression in the disease vector, Aedes aegypti. Insect Biochem Mol Biol 33:1105-22
Filippov, V; Aimanova, K; Gill, S S (2003) Expression of an Aedes aegypti cation-chloride cotransporter and its Drosophila homologues. Insect Mol Biol 12:319-31
Jin, Xiangyang; Aimanova, Karlygash; Ross, Linda S et al. (2003) Identification, functional characterization and expression of a LAT type amino acid transporter from the mosquito Aedes aegypti. Insect Biochem Mol Biol 33:815-27
Pullikuth, Ashok K; Filippov, Valeri; Gill, Sarjeet S (2003) Phylogeny and cloning of ion transporters in mosquitoes. J Exp Biol 206:3857-68
Pullikuth, Ashok K; Gill, Sarjeet S (2002) In vivo membrane trafficking role for an insect N-ethylmaleimide-sensitive factor which is developmentally regulated in endocrine cells. J Exp Biol 205:911-26
Filippov, V; Filippov, M; Gill, S S (2001) Drosophila RNase H1 is essential for development but not for proliferation. Mol Genet Genomics 265:771-7

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