Isolation and Characterization of the Corticotropin releasing factor-like diuretic hormone receptor in Rhodnius prolixus.

摘要

Rhodnius prolixus is a hemipteran insect which undergoes rapid post-feeding diuresis following ingestion of a large blood meal that can be up to 10 times initial body weight. Corticotropin releasing factor-like diuretic hormone (Rhopr-CRF/DH) is involved in the neurohormonal synergism that increases rates of fluid secretion by Malpighian tubules over 1000 fold during rapid post-feeding diuresis. In this study, the Rhopr-CRF/DH receptor was isolated and characterized from fifth instar R .prolixus. The receptor is a family B G-protein coupled receptor (GPCR) and was deorphaned using a Chinese hamster ovary (CHO) cell line. RT-qPCR demonstrated strong receptor expression in the digestive tissues, upper Malpighian tubules and the reproductive tissues. Finally, a modified Human embryonic kidney (HEK293) cell line was used to verify the initiation of the cyclic adenosine monophosphate (cAMP) pathway upon activation of the receptor. Characterization of the CRF/DH receptor will serve to elicit better understanding into the precise coordination of diuresis and other physiological processes.