Expression and function of WNT4 involved in larvae development and limb regeneration in Portunus trituberculatus

作者：Zhiming REN,Yuanyuan FU,Lei LIU,Xiao LIU,Chunlin WANG

摘要：The wingless-related integration site(WNT)proteins are a family of secreted glycoproteins that are evolutionarily conserved and are believed to be involved in evolution in vertebrates and invertebrates.WNT signaling pathways may be associated with limb regeneration and development in crustaceans.However,the detail mechanisms remain unclear.Therefore,the distribution of WNT4 in the hepatopancreas,muscle,hemocyte,ganglion,heart,eyestalk,gill tissue,and diff erent larvae development stages of the swimming crab(Portunus trituberculatus)were characterized using immunofl uorescence,real-time PCR,and Western blotting.Signifi cant PtWNT4 expression was detected in heart and eyestalk.In addition,PtWNT4 was expressed in all larval stages of P.trituberculatus with a dynamic expression pattern,especially in the eyestalk and other organs in the carapace area.The injection of WNT4 dsRNA into regenerative limbs signifi cantly decreased PtWNT4 mRNA levels in the eyestalk,heart,and muscle,resulting in 1.9-fold,2.2-fold,and 2.7-fold decreases compared with those detected in the group injected with crab saline(P<0.05),respectively,indicating successful gene silencing.Overall,expression analysis on the WNT4 using RNAi provides an insight to its functional mechanism during limb regeneration in P.trituberculatus.The results not only demonstrated the requirement for WNT4 in limb regeneration of Crustaceans,but also suggested its ability to promote larval development at specifi c stages.

发文机构：Key Laboratory of Applied Marine Biotechnology Collaborative Innovation Center for Zhejiang Marine High-effi ciency and Healthy Aquaculture

关键词：wingless-typeMMTVintegrationsitefamilymember-4(WNT4)limbregenerationlarvaedevelopmentPortunustrituberculatusexpressionpatternRNAi

分类号： S91[农业科学—水产科学]