Expression and function of WNT4 involved in larvae development and limb regeneration in <i>Portunus</i> <i>trituberculatus</i> -- Journal of Oceanology and Limnology,2021,39(1):306-316

	Cite this paper:
	Zhiming REN, Yuanyuan FU, Lei LIU, Xiao LIU, Chunlin WANG. Expression and function of WNT4 involved in larvae development and limb regeneration in Portunus trituberculatus[J]. Journal of Oceanology and Limnology, 2021, 39(1): 306-316

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

Zhiming REN^1,2, Yuanyuan FU^1,2, Lei LIU^1,2, Xiao LIU^1,2, Chunlin WANG^1,2

1 Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo 315832, China;
2 Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture, Ningbo University, Ningbo 315832, China

Abstract:

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 different larvae development stages of the swimming crab (Portunus trituberculatus) were characterized using immunofluorescence, real-time PCR, and Western blotting. Significant 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 significantly 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 specific stages.

Received: 2019-12-12 Revised: 2020-04-13

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Articles by Zhiming REN

Articles by Yuanyuan FU

Articles by Lei LIU

Articles by Xiao LIU

Articles by Chunlin WANG

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