Bioorganic Research Institute, Suntory Foundation for Life Sciences, Kyoto, Japan.
Shimoda Marine Research Center, University of Tsukuba, Shimoda, Japan.
Front Endocrinol (Lausanne). 2021 May 6;12:668564. doi: 10.3389/fendo.2021.668564. eCollection 2021.
Oxytocin (OT) and vasopressin (VP) superfamily neuropeptides are distributed in not only vertebrates but also diverse invertebrates. However, no VPergic innervation of invertebrates has ever been documented. In the ascidian, Type A (), an OT/VP superfamily peptide was identified, and the vasopressin (CiVP) induces oocyte maturation and ovulation. In the present study, we characterize the innervation and phenotypes of genetically modified : promoter-Venus transgenic and mutants. promoter-Venus transgenic demonstrated that gene was highly expressed in the cerebral ganglion and several nerves. Fluorescence was also detected in the ovary of young promoter-Venus transgenic ascidians, suggesting that the gene is also expressed temporarily in the ovary of young ascidians. Furthermore, a marked decrease of post-vitellogenic (stage III) follicles was observed in the ovary of mutants, whereas pre-vitellogenic (stage I) and vitellogenic (stage II) follicles were increased in the mutant ovary, compared with that of wildtype . Gene expression profiles showed that the expression of various genes, including genes related to ovarian follicle growth, was altered in the ovary of mutants. Altogether, these results indicated that CiVP, mainly as a neuropeptide, plays pivotal roles in diverse biological functions, including growth of early-stage ovarian follicles regulation of the expression of a wide variety of genes. This is the first report describing a gene promoter-transgenic and gene-edited invertebrate and also on its gene expression profiles and phenotypes.
催产素（OT）和加压素（VP）神经肽超家族不仅分布在脊椎动物中，也分布在各种无脊椎动物中。然而，无脊椎动物中从未有过 VP 能神经支配的记录。在尾索动物中，鉴定出了一种 OT/VP 神经肽超家族肽，即血管加压素（CiVP），它能诱导卵母细胞成熟和排卵。在本研究中，我们对遗传修饰的：启动子-Venus 转基因和 突变体进行了神经支配和表型特征分析。启动子-Venus 转基因 显示， 基因在脑神经节和几条神经中高度表达。在年轻的 启动子-Venus 转基因尾索动物的卵巢中也检测到荧光，表明 基因也在年轻尾索动物的卵巢中暂时表达。此外，在 突变体的卵巢中，后卵黄生成期（阶段 III）滤泡明显减少，而在 突变体的卵巢中，前卵黄生成期（阶段 I）和卵黄生成期（阶段 II）滤泡增加，与野生型相比。基因表达谱显示，在 突变体的卵巢中，包括与卵巢滤泡生长相关的基因在内的各种基因的表达发生了改变。总之，这些结果表明，CiVP 主要作为一种神经肽，在包括早期卵巢滤泡生长在内的各种生物学功能中发挥着关键作用，并调节各种基因的表达。这是首次描述 CiVP 基因启动子转基因和 基因编辑的无脊椎动物及其基因表达谱和表型的报告。
