【摘要】 为了解中华蛸(Octopus sinensis)受精卵的孵化及幼体发育机制,本研究对其章鱼胺受体基因(OsOARβ2R)进行克隆和生物信息学分析,同时研究了OsOARβ2R在不同组织/器官、刚孵出的幼体不同饥饿时间及不同的胚胎发育时期表达水平的变化。结果为:OsOARβ2R开放阅读框全长为1 158 bp,编码385个氨基酸,有7个跨膜结构域,具有G蛋白偶联受体的共性,其中TM3、TM5、TM6、TM7 4个跨膜结构相对保守。经氨基酸同源对比及构建系统进化树分析,其与加州双斑蛸(O. bimaculoides)的OAR的一致性最高。荧光定量PCR结果显示,在成体10个组织/器官中,OsOARβ2R在后唾液腺中表达水平最高,其次是脑和小肠;在幼体饥饿实验中,随着饥饿时间的推移,OsOARβ2R的表达水平在饥饿2 d后显著降低,在饥饿3 d时表达水平显著升高,且表达水平达到最高,之后表达水平开始回落;OsOARβ2R在中华蛸整个胚胎发育周期均可检测到,且在多细胞期表达水平最高,后显著下降,黑珠期显著高于红珠期及初孵幼体。这些结果为研究中华蛸受精卵的孵化及幼体发育机制、提高人工育苗效率提供了基础资料。更多还原

【Abstract】 To understand the mechanism of hatching and larval development in Octopus sinensis, we cloned its OsOARβ2R gene and performed a bioinformatics analysis. Meanwhile, the expression of OsOARβ2R in different tissues/organs after different periods of starvation in newly hatched larvae and at different stages of embryonic development was analyzed. The results revealed that the open reading frame of OsOARβ2R is 1158 bp long, encodes 385 amino acids, and contains 7 transmembrane domains(TMs), which have commonality with G protein-coupled receptors. Among the TMs, TM3, TM5, TM6, and TM7 are relatively conserved. According to the amino acid homology comparison and phylogenetic tree analysis, OsOARβ2R shares the highest identity with the OAR of O. bimaculoides. The results of quantitative polymerase chain reaction revealed that the expression of OsOARβ2R was widespread in the 10 examined tissues/organs, with the highest expression level in the posterior salivary gland, followed by the brain and intestine. In the larval starvation experiment, the expression of OsOARβ2R decreased significantly after 2 days of starvation; increased significantly, reaching a peak at 3 days of starvation; and then declined to normal levels. OsOARβ2R could be detected throughout the embryonic development cycle, and its expression level was the highest at the multicellular stage, decreasing significantly thereafter. OsOARβ2R expression was also significantly higher in the black-bead stage than in the red-bead stage and in newly hatched larvae. These results provide basic data for studying the hatching and larval development mechanisms of fertilized O. sinensis eggs and for improving the efficiency of artificial seeding with O. sinensis.更多还原