CHH binding protein (CHHBP): a newly identified receptor of crustacean hyperglycemic hormone (CHH)

Abstract

Crustacean hyperglycemic hormone (CHH) is a neurohormone found only in arthropods that plays a pivotal role in the regulation of hemolymph glucose levels, molting, and stress responses. Although it was determined that a membrane guanylyl cyclase (GC) acts as the CHH receptor in the Y-organ during ecdysteroidogenesis, the identity of the CHH receptor in the hepatopancreas has not been established. In this study, we identified a new molecular, CHH binding protein (CHHBP), as a potential receptor by screening the annotated unigenes from the transcriptome of Eriocheir sinensis, after removal of eyestalk. Analysis of the binding affinity between CHH and CHHBP provided direct evidence that CHH interacts with CHHBP in a specific binding mode. Subsequent analysis showed that CHHBP was expressed primarily in the hepatopancreas and localized on cell membrane. In addition, real-time PCR analysis showed that CHHBP transcript levels gradually increased in the hepatopancreas following eyestalk ablation. RNAi-mediated suppression of CHHBP expression resulted in decreased glucose levels. Furthermore, the reduction of blood glucose induced by CHHBP RNAi reached the same degree as that observed in the eyestalk ablation group, suggesting that CHHBP contributes to glucose metabolism regulated by CHH. Besides, compared to the control group, injection of CHH was unable to rescue the decreased glucose levels in CHHBP RNAi crabs. CHH induced transport of 2-NBDG to the outside of cells, with indispensable assist from CHHBP. Taken together, these findings imply that CHHBP probably acts as one type of the primary signal processor of CHH-mediated regulation of cellular glucose metabolism.