Meeting Abstract
P2.133 Sunday, Jan. 5 15:30 Tissue and developmental expression of three putative Na-coupled cation-chloride cotransporters in Aedes aegypti AKUMA, D.C.*; PIERMARINI, P.M.; GILLEN, C.M.; Kenyon College, Gambier OH; Ohio Agricultural Research and Development Center, Wooster; Kenyon College, Gambier OH gillenc@kenyon.edu
We evaluated the expression of three genes encoding putative Na-coupled cation-chloride cotransporters (CCCs) in the mosquito Aedes aegypti. Phylogenetic analysis reveals that the protein predicted by VectorBase gene AAEL006180 (tentatively named aeCCC1) groups closely with Drosophila melanogaster bumetanide-sensitive Na-K-Cl cotransporter dmNCC69 (CG4357) and shares strong similarity to mammalian Na-coupled CCCs. In contrast, proteins predicted by AAEL009888 (aeCCC2) and AAEL009886 (aeCCC3) group closely with Manduca sexta msBSC and Drosophila CG31457 and not with aeCCC1. All 3 genes cluster distinctly from mosquito and mammalian K-Cl cotransporters. We quantified expression levels of aeCCC1, aeCCC2, and aeCCC3 using quantitative PCR. Expression of aeCCC1 was similar among whole adults, pupae, and larvae, whereas aeCCC2 was expressed about 35-fold higher in larvae and adults than in pupae. aeCCC3 was predominately found in larvae, with about 500-fold greater expression than in pupae and adults. In adult female mosquitoes, aeCCC1 was expressed most highly in head, with 2.5-fold greater expression than in Malpighian tubules, 4.5-fold greater expression than in hindgut, and 20-fold greater expression than in midgut. In contrast, aeCCC2 was predominantly found in the hindgut of adult females, with more than 200-fold greater expression than in other tissues. In larvae, aeCCC3 was expressed 6,000-fold more highly in anal papillae than in Malpighian tubules. On the other hand, aeCCC1 and aeCCC2 were more highly expressed in larval Malpighian tubules compared to anal papillae. The different tissue and developmental expression patterns of aeCCC1, aeCCC2, and aeCCC3 suggest diverse functional roles for these transporters.
