tailieunhanh - Báo cáo khoa học: The use of recombinant protein and RNA interference approaches to study the reproductive functions of a gonad-stimulating hormone from the shrimp Metapenaeus ensis

Although the crustacean crustacean hyperglycemic hormone⁄molt-inhibi-ting hormone⁄gonad-inhibiting hormone neuropeptides have been studied extensively in the last two decades and several neuropeptides from the shrimpMetapenaeus ensishave been cloned, the functions of most of these neuropeptides remained putative. | ỊFEBS Journal The use of recombinant protein and RNA interference approaches to study the reproductive functions of a gonad-stimulating hormone from the shrimp Metapenaeus ensis Shirley Hiu-Kwan Tiu and Siu-Ming Chan Department of Zoology The University of Hong Kong China Keywords eyestalk neuropeptide hormone RNA interference shrimp vitellogenin gene Correspondence . Chan Department of Zoology The University of Hong Kong Pokfulam Road Hong Kong Fax 852 2857 4672 Tel 852 2299 0864 E-mail chansm@ Received 25 January 2007 revised 15 June 2007 accepted 2 July 2007 doi Although the crustacean crustacean hyperglycemic hormone molt-inhibi-ting hormone gonad-inhibiting hormone neuropeptides have been studied extensively in the last two decades and several neuropeptides from the shrimp Metapenaeus ensis have been cloned the functions of most of these neuropeptides remained putative. In this article we describe the use of recombinant protein and an RNA interference approach to study the reproductive function of the previously reported molt-inhibiting hormone MeMIH-B in M. ensis. When hepatopancreas and ovary explants were cultured in medium containing recombinant MeMIH-B the vitellogenin gene MeVgl expression level was upregulated in a dose-dependent manner reaching a maximum in explants treated with nM recombinant MeMIH-B. Shrimp injected with recombinant MeMIH-B showed an increase in vitellogenin gene expression in the hepatopancreas. Moreover a corresponding increase in the vitellogenin-like immunoreactive protein was detected in the hemolymph and ovary of these females. Injection of MeMIH-B dsRNA into the female shrimp caused a decrease in MeMIH-B transcript level in thoracic ganglion and eyestalk. These shrimp also showed reduction of vitellogenin gene expression in the hepatopancreas and ovary. Furthermore the hemolymph vitellogenin level was also reduced in these animals. In summary the results from recombinant .