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 

机构地区：[1]Key Laboratory of Applied Marine Biotechnology,Ministry of Education,Ningbo University,Ningbo 315832,China [2]Collaborative Innovation Center for Zhejiang Marine High-effi ciency and Healthy Aquaculture,Ningbo University,Ningbo 315832,China

出　　处：《Journal of Oceanology and Limnology》2021年第1期306-316,共11页海洋湖沼学报（英文）

基　　金：Supported by the National Natural Science Foundation of China(No.31602152);the Major Agriculture Program of Ningbo(No.2017C110007);the K.C.Wong Magana Fund in Ningbo University.The funding body had no role in the study design,experimental implementation,interpretation of data,or writing of the manuscript。

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.

关 键 词：wingless-type MMTV integration site family member-4(WNT4) limb regeneration larvae development Portunus trituberculatus expression pattern RNAi 

分 类 号：S917.4[农业科学—水产科学]