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Aquaculture Breeding Enhancement: Maturation and Spawning in Sea Cucumbers Using a Recombinant Relaxin-Like Gonad-Stimulating Peptide

机译:水产养殖增强:使用重组松弛蛋白的Gonad刺激肽在海参中成熟和产卵

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摘要

Wild sea cucumber resources have been rapidly exhausted and therefore there is an urgent need to develop approaches that will help restocking. Currently, there is a lack of information regarding the genes involved in sea cucumber reproductive processes. The neurohormone relaxin-like gonad-stimulating peptide (RGP) has been identified as the active gonad-stimulating peptide in sea stars (Asteroidea), which could also be present in other echinoderm groups. In this study, a sea cucumber RGP was identified and confirmed by phylogenetic analysis. A recombinant Holothuria scabra RGP was produced in the yeast Pichia pastoris and confirmed by mass spectrometry. To assess bioactivity, four levels of purification were tested in an in vitro germinal vesicle breakdown (GVBD) bioassay. The most pure form induced 98.56 ± 1.19% GVBD in H. scabra and 89.57 ± 1.19% GVBD in Holothuria leucospilota. Cruder levels of purification still resulted in some GVBD. Upon single injection into female H. scabra, the recombinant RGP induced head waving behavior followed by spawning within 90–170 min. Spawned oocytes were fertilized successfully, larvae settled and developed into juveniles. Our results provide a key finding for the development of a break-through new artificial breeding approach in sea cucumber aquaculture.
机译:野生海洋黄瓜资源已经迅速疲惫,因此迫切需要开发有助于补充的方法。目前,缺乏有关涉及海参生殖过程的基因的信息。已经鉴定了神经期胃肠松弛素样刺激肽(RGP)作为海星(日龄儿藻)中的活性腺刺激肽,其也可以存在于其他棘皮胚层中。在该研究中,通过系统发育分析鉴定并确认了海参RGP。重组Holothuria scabra RGP在酵母Pichia Pastoris中产生并通过质谱法证实。为了评估生物活性,在体外发芽囊泡粒度(GVBD)生物测定中测试了四种纯化水平。最纯的形式诱导H.Scabra的98.56±1.19%GVBD和Holothuria Leucospilota的89.57±1.19%GVBD。粗糙的纯化水平仍然导致一些GVBD。单次注射到雌性H. scabra后,重组RGP诱导的头部挥动行为,然后在90-100分钟内产生。将生成的卵母细胞成功受精,幼虫沉淀并发展成青少年。我们的结果提供了在海参水产养殖中开发突破新的人工育种方法的关键。

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