Hyeogsun Kwon
"Cloning and Expression Analysis of the Diuretic Hormone 31 Receptor cDNA from the Malpighian Tubules of Aedes aegypti Females."
The Malpighian tubules of the yellow fever mosquito, Aedes aegypti (L.) are composed of two cell types, primary and stellate cells. Malpighian tubules are known to control hemolymph filtration and primary urine production in dipterans. Synergism between diuretic hormones, such as the corticotropin releasing factor-like peptide DH44 and the calcitonin-like peptide DH31 I have been shown to mediate osmotic balance in Drosophila melanogaster, Anopheles gambiae, and Ae. aegypti. In D. melanogaster, the receptor encoded by CG8422 has been identified as a functional diuretic hormone (DH44) receptor, while the receptor encoded by GC174·15 responds to neuropeptide DH31 . However, the identity of diuretic hormone receptors controlling water and salt homeostasis in Ae. aegypti has not been experimentally investigated. Thus, my research focuses on the characterization of the likely DH31 receptor in mosquito. Putative DH31 receptor cDNA from the Malpighian tubules of Ae. aegypti females was cloned using BLAST alignment similarity with GC17415 of Drosophila melanogaster. To determine if the transcript levels of receptor mRNA vary during' the life cycle of adnlt females, RT-PCR was used. Future experiments will investigate knock down of the mRNA transcript by RNA Interference. The receptor transcript and protein will be localized using in situ hybridization with an RNA probe and immunocytochemistry, respectively. The receptor will be expressed and changes in cAMP elicited by receptor activation will be measured.