Jiangsu Province Engineering Research Center for Aquatic Animals Breeding and Green Efficient Aquacultural Technology, College of Marine Science and Engineering, Nanjing Normal University, Nanjing, Jiangsu Province, 210023, China.
Jiangsu Province Engineering Research Center for Aquatic Animals Breeding and Green Efficient Aquacultural Technology, College of Marine Science and Engineering, Nanjing Normal University, Nanjing, Jiangsu Province, 210023, China.
Fish Shellfish Immunol. 2020 Jun;101:225-233. doi: 10.1016/j.fsi.2020.03.063. Epub 2020 Apr 1.
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.
Wnt 信号转导途径参与多种细胞过程，包括细胞增殖、分化、凋亡以及抵抗微生物感染的免疫反应。在本研究中，我们克隆并鉴定了日本沼虾（Macrobrachium nipponense）中的两个 Wnt 同源物（Mn-Wnt4 和 Mn-Wnt16）。Mn-Wnt4 的全长 cDNA 为 3144 bp，其中包含一个 1074 bp 的开放阅读框（ORF），编码一个含有 358 个氨基酸残基的蛋白质。Mn-Wnt16 转录本的全长 cDNA 为 2893 bp，其中包含一个 1281 bp 的 ORF，编码一个含有 427 个氨基酸的蛋白质。Mn-Wnt4 和 Mn-Wnt16 蛋白均含有高度保守的 WNT1 结构域。组织分布分析显示，Mn-Wnt4 和 Mn-Wnt16 在胃组织中高表达。在细菌（金黄色葡萄球菌和副溶血弧菌）和病毒（白斑综合征病毒）感染后，Mn-Wnt4 和 Mn-Wnt16 在胃组织中的转录水平在大多数测试时间点均上调。此外，在副溶血弧菌感染后，一些抗菌肽（AMPs）（包括抗脂多糖因子[ALF]和壳质[CRU]）的表达水平上调。我们进一步使用 dsRNA 介导的 RNA 干扰技术来探究这两个 Wnt 基因与副溶血弧菌感染期间 AMPs 表达水平之间的关系。Mn-Wnt4 敲低可显著抑制副溶血弧菌注射对虾胃中 ALF1 和 CRU4 的表达，而 Mn-Wnt16 沉默可导致副溶血弧菌感染对虾胃中 CRU3 和 CRU4 的表达水平受到抑制。这些发现表明，Wnt 基因家族可能通过调节多种 AMPs 的合成参与甲壳动物对弧菌感染的固有免疫反应。本研究将有助于了解 Wnt 信号通路在甲壳动物免疫反应中的作用。
