Ba Huazhong, Yao Feng, Yang Lei, Qin Tong, Luan Hong, Li Zhengmin, Zou Xiangyang, Hou Lin
College of Life Sciences, Liaoning Normal University, 1, Liushu South Street, Ganjingzi District, Dalian 116081, People's Republic of China.
Department of Biology, Dalian Medical University, Dalian 116044, People's Republic of China.
Gene. 2015 Jul 1;565(1):96-105. doi: 10.1016/j.gene.2015.03.071. Epub 2015 Apr 2.
Sea cucumbers have a strong regenerative capacity. Many important genes involved in the molecular mechanism of regeneration and associated with intercellular signaling pathways of regeneration have been identified. The product of the fibropellin-ia gene forms a layer known as the apical lamina that surrounds the sea cucumber embryo throughout development. Meanwhile, the tenascin gene displays highly restricted and dynamic patterns of expression in the embryo and is expressed in the adult during normal processes such as wound healing, nerve regeneration and tissue involution. In this study, we cloned for the first time full-length cDNAs of fibropellin-ia (1390 bp, encoding a 199 amino acid protein) and tenascin (1366 bp, encoding a 179 amino acid protein) from Apostichopus japonicus (designated Aj-fnia and Aj-tenascin, respectively) using rapid amplification of cDNA ends. The structures and characteristics of these two genes were analyzed bioinformatically, and their expression patterns associated with extracellular matrix remodeling in regeneration of A. japonicus were investigated by real-time PCR and in situ hybridization (ISH). Expression levels of Aj-fnia and Aj-tenascin in the regeneration tissues were higher than those in normal tissues. The highest expression levels of Aj-fnia and Aj-tenascin were shown in the intestine and respiratory tree on the 15th and 20th days after sea cucumbers were eviscerated. In the body wall, the highest expression levels of Aj-fnia and Aj-tenascin occurred at 35 and 45 min during early regeneration and then emerged between 5 and 7 days again during late regeneration after the body wall was injured. ISH analysis revealed expression of these genes in the body wall, longitudinal muscle, intestine and respiratory tree. These findings suggest that Aj-fnia and Aj-tenascin are crucial genes that play important roles in the regeneration of the sea cucumber.
海参具有很强的再生能力。许多参与再生分子机制并与再生细胞间信号通路相关的重要基因已被鉴定出来。纤维膜蛋白-ia基因的产物形成一层称为顶端板的结构,在整个发育过程中围绕海参胚胎。同时,腱生蛋白基因在胚胎中表现出高度受限且动态的表达模式,并在成体的正常过程中表达,如伤口愈合、神经再生和组织退化。在本研究中,我们首次使用cDNA末端快速扩增技术从仿刺参中克隆了纤维膜蛋白-ia(1390 bp,编码一个199个氨基酸的蛋白质)和腱生蛋白(1366 bp,编码一个179个氨基酸的蛋白质)的全长cDNA(分别命名为Aj-fnia和Aj-tenascin)。对这两个基因的结构和特征进行了生物信息学分析,并通过实时PCR和原位杂交(ISH)研究了它们在仿刺参再生过程中与细胞外基质重塑相关的表达模式。Aj-fnia和Aj-tenascin在再生组织中的表达水平高于正常组织。在海参去内脏后的第15天和第20天,Aj-fnia和Aj-tenascin在肠道和呼吸树中的表达水平最高。在体壁中,Aj-fnia和Aj-tenascin在早期再生的35和45分钟时表达水平最高,然后在体壁受伤后的后期再生的5至7天再次出现高表达。ISH分析显示这些基因在体壁、纵肌、肠道和呼吸树中表达。这些发现表明Aj-fnia和Aj-tenascin是在海参再生中起重要作用的关键基因。