The gut is a major route of entry in insects. However, our knowledge of insect gut function, most of which is primarily physiological, is inadequate. To enable us to better comprehend how insects adapt to and succeed in the environment, it is critical that we have a much better understanding of gut function, because the gut plays a critical role in maintaining insect homeostasis. The long-term objective of this research therefore is to establish the structure of proteins involved in ion regulation in insects. The specific objectives of this proposal are to deduce the primary structure of the vacuolar (V-type) ATPase subunits from the midgut and Malpighian tubules of the mosquito Aedes aegypti. The V-type ATPase(s) play an essential role in the maintenance of ion balance in insects, including mosquitoes. In addition these ATPase(s) also play an important role in excretion, and as a consequence, water balance. Once the subunits are characterized, their role in proton transport will be determined, and the spatial and temporal expression of the ATPase analyzed. Significant amino acid homology exists between V-type ATPase subunits from various sources. Utilizing this fact oligonucleotide primers will be made for each subunit, using the conserved domains as templates for primer design. The primers will then be used for the polymerase chain reaction (PCR) amplification of the putative subunit genes from cDNA prepared from midgut and Malpighian tubules of Ae. aegypti. These PCR products will be used for screening of cDNA libraries. However, if this strategy fails, alternative protocols, i.e., oligonucleotides based on the N-terminal sequence and/or antibodies raised against the subunits, will be used in screening the cDNA library for isolation of clones. Once the genes for these subunits are cloned, the spatial and temporal expression of the V-type ATPase will be determined using established methods. An understanding of mosquito gut function will facilitate our understanding of the mechanisms by which mosquitoes adapt to environmental conditions. Such knowledge may enable us to target the gut as a site for manipulating mosquito populations. The control of mosquitoes is critical for the disruption of many mosquito borne diseases of man.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI032572-05
Application #
2003779
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Project Start
1993-01-01
Project End
1998-12-31
Budget Start
1997-01-01
Budget End
1998-12-31
Support Year
5
Fiscal Year
1997
Total Cost
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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