Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai, 201306, China; National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, 201306, China; Shanghai Engineering Research Center of Aquaculture, Shanghai, 201306, China.
Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, Shanghai Ocean University, Shanghai, 201306, China; National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai, 201306, China; Shanghai Engineering Research Center of Aquaculture, Shanghai, 201306, China.
Fish Shellfish Immunol. 2019 Aug;91:99-107. doi: 10.1016/j.fsi.2019.05.009. Epub 2019 May 7.
β-catenin is a multifunctional protein that participates in a variety of physiological activities, including immune regulation, sex determination, nervous system development and, cell differentiation. However, the function of β-catenin in freshwater mussel Hyriopsis cumingii remains unclear. Herein, the gene encoding β-catenin from H. cumingii (Hc-β-catenin) was cloned and characterised. The full-length 5544 bp gene includes an open reading frame (ORF) of 2463 bp encoding a putative protein of 820 amino acids residues containing 12 armadillo (ARM) repeats. After injecting H. cumingii with Aeromonas hydrophila or lipopolysaccharides, Hc-β-catenin transcription was induced in hemocytes and gills, and the greatest responses occurred at 24 h after bacterial challenge, confirming an important role in immune responses. Quantitative real-time PCR analysis showed that Hc-β-catenin mRNA was distributed in the gill, foot, liver, kidney, mantle, adductor muscle and gonad of male and female mussels. In gonad, Hc-β-catenin expression was markedly higher in females than males. During the embryonic period, Hc-β-catenin expression was highest at 3 day. In 1-, 2- and 3-year-old mature mussels, Hc-β-catenin expression in female gonad tissue was notably higher than in males. In situ hybridisation revealed a significant hybridisation signal in female gonads, indicating that Hc-β-catenin is a pro-ovarian, anti-testis gene. Our findings demonstrate that Hc-β-catenin is important in immune regulation and sex determination in freshwater mussel.
β-连环蛋白是一种多功能蛋白，参与多种生理活动，包括免疫调节、性别决定、神经系统发育和细胞分化。然而，β-连环蛋白在淡水贻贝 Hyriopsis cumingii 中的功能尚不清楚。本文从 H. cumingii 中克隆并鉴定了编码β-连环蛋白的基因（Hc-β-catenin）。全长 5544 bp 的基因包含一个 2463 bp 的开放阅读框（ORF），编码一个含有 12 个角蛋白重复序列的推定蛋白，由 820 个氨基酸残基组成。在向 H. cumingii 注射嗜水气单胞菌或脂多糖后，在血细胞和鳃中诱导了 Hc-β-catenin 的转录，在细菌攻击后 24 小时达到最大反应，证实其在免疫反应中具有重要作用。定量实时 PCR 分析显示，Hc-β-catenin mRNA 分布在雌雄贻贝的鳃、足、肝、肾、套膜、闭壳肌和性腺中。在性腺中，Hc-β-catenin 在雌性中的表达明显高于雄性。在胚胎期，Hc-β-catenin 在第 3 天表达最高。在 1 年、2 年和 3 年成熟贻贝中，雌性性腺组织中 Hc-β-catenin 的表达明显高于雄性。原位杂交显示在雌性性腺中有显著的杂交信号，表明 Hc-β-catenin 是一种促卵巢、抗睾丸基因。我们的研究结果表明，Hc-β-catenin 在淡水贻贝的免疫调节和性别决定中具有重要作用。
