The Wnt signal transduction pathway is involved in a wide variety of cellular processes, including cell proliferation, differentiation, apoptosis, and immunity against microbial infection. In the current study, we cloned and characterized two Wnt homologues (Mn-Wnt4 and Mn-Wnt16) in Macrobrachium nipponense. The full length cDNA of Mn-Wnt4 was 3144 bp with a 1074 bp open reading frame (ORF) that encoded a protein containing 358 amino acid residues. The full length cDNA of Mn-Wnt16 transcript was 2893 bp with a 1281 bp ORF that encoded a 427 amino acid protein. Mn-Wnt4 and Mn-Wnt16 proteins contained a highly conserved WNT1 domain. Tissue distribution analysis showed that Mn-Wnt4 and Mn-Wnt16 were highly expressed in the stomach. The transcriptional levels of Mn-Wnt4 and Mn-Wnt16 in the stomach were upregulated at most tested time points after bacterial (Staphylococcus aureus and Vibrio parahaemolyticus) and viral (White spot syndrome virus) infection. Moreover, the expression levels of some antimicrobial peptides (AMPs) (including anti-lipopolysaccharide factor [ALF] and crustin [CRU]) were upregulated after V. parahaemolyticus infection. We further used dsRNA-mediated RNA interference technology to explore the relationship between these two Wnt genes and the expression levels of AMPs during V. parahaemolyticus infection. Mn-Wnt4 knockdown could significantly inhibit the expression of ALF1 and CRU4 in the stomach of V. parahaemolyticus-injected prawns, whereas Mn-Wnt16 silencing could result in the inhibition of the expression level of CRU3 and CRU4 in the stomach of V. parahaemolyticus-infected prawns. These findings indicated that the Wnt gene family might participate in the body's innate immune response to Vibrio infection by regulating the synthesis of a variety of AMPs. Our study will help to understand the role of the Wnt signaling pathway in the immune response of crustaceans.

中文翻译：

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两个Wnt基因调节日本沼虾弧菌感染过程中抗菌肽的表达水平。

Wnt信号转导途径涉及多种细胞过程，包括细胞增殖，分化，凋亡和抵抗微生物感染的免疫力。在当前的研究中，我们在日本沼虾中克隆并鉴定了两个Wnt同源物（Mn-Wnt4和Mn-Wnt16）。Mn-Wnt4的全长cDNA为3144 bp，具有1074 bp的开放阅读框（ORF），其编码包含358个氨基酸残基的蛋白质。Mn-Wnt16转录本的全长cDNA为2893 bp，ORF为1281 bp，编码427个氨基酸。Mn-Wnt4和Mn-Wnt16蛋白包含一个高度保守的WNT1域。组织分布分析表明，Mn-Wnt4和Mn-Wnt16在胃中高表达。在细菌（金黄色葡萄球菌和副溶血性弧菌）和病毒（白斑综合症病毒）感染后的大多数测试时间点，胃中Mn-Wnt4和Mn-Wnt16的转录水平上调。此外，副溶血性弧菌感染后某些抗菌肽（AMP）（包括抗脂多糖因子[ALF]和克鲁斯汀[CRU]）的表达水平上调。我们进一步使用dsRNA介导的RNA干扰技术来探索这两个Wnt基因与副溶血性弧菌感染过程中AMPs表达水平的关系。敲除Mn-Wnt4可以显着抑制副溶血弧菌对虾胃中ALF1和CRU4的表达，而Mn-Wnt16沉默可能导致抑制V.胃中CRU3和CRU4的表达水平。副溶血性对虾感染。这些发现表明，Wnt基因家族可能通过调节多种AMP的合成来参与机体对弧菌感染的先天免疫应答。我们的研究将有助于了解Wnt信号通路在甲壳类动物免疫应答中的作用。