PDF(Pubmed)
Abstract：
BACKGROUND: Sea urchins have contributed greatly to knowledge of fertilization, embryogenesis, and cell biology. However, until now, they have not been genetic model organisms because of their long generation times and lack of tools for husbandry and gene manipulation. We recently established the sea urchin Lytechinus pictus, as a multigenerational model Echinoderm, because of its relatively short generation time of 4-6 months and ease of laboratory culture. To take full advantage of this new multigenerational species, methods are needed to biobank and share genetically modified L. pictus sperm.
RESULTS: Here, we describe a method, based on sperm ion physiology that maintains L. pictus and Strongylocentrotus purpuratus sperm fertilizable for at least 5-10 weeks when stored at 0°C. We also describe a new method to cryopreserve sperm of both species. Sperm of both species can be frozen and thawed at least twice and still give rise to larvae that undergo metamorphosis.
CONCLUSIONS: The simple methods we describe work well for both species, achieving >90% embryo development and producing larvae that undergo metamorphosis to juvenile adults. We hope that these methods will be useful to others working on marine invertebrate sperm.
RESULTS: Here, we describe a method, based on sperm ion physiology that maintains L. pictus and Strongylocentrotus purpuratus sperm fertilizable for at least 5-10 weeks when stored at 0°C. We also describe a new method to cryopreserve sperm of both species. Sperm of both species can be frozen and thawed at least twice and still give rise to larvae that undergo metamorphosis.
CONCLUSIONS: The simple methods we describe work well for both species, achieving >90% embryo development and producing larvae that undergo metamorphosis to juvenile adults. We hope that these methods will be useful to others working on marine invertebrate sperm.
摘要：
背景：海胆对受精知识有很大的贡献，胚胎发生,和细胞生物学。然而,直到现在,它们不是遗传模式生物，因为它们的世代时间长，缺乏饲养和基因操纵的工具。我们最近建立了海胆Lytechinuspictus，作为一个多代模型，由于其4-6个月的世代时间相对较短，并且易于实验室培养。为了充分利用这种新的多代物种，需要方法来进行生物样本库和共享转基因的皮氏乳杆菌精子。
结果：这里，我们描述了一种方法，基于精子离子生理学，当储存在0°C时，可保持皮氏L.pictus和Strongylocentrotuspuricatus精子受精至少5-10周。我们还描述了一种冷冻保存这两个物种精子的新方法。两种物种的精子都可以冷冻和解冻至少两次，并且仍然会产生经历变态的幼虫。
结论：我们描述的简单方法对两个物种都有效，实现>90％的胚胎发育和产生的幼虫经历变态成虫。我们希望这些方法对其他研究海洋无脊椎动物精子的人有用。
结果：这里，我们描述了一种方法，基于精子离子生理学，当储存在0°C时，可保持皮氏L.pictus和Strongylocentrotuspuricatus精子受精至少5-10周。我们还描述了一种冷冻保存这两个物种精子的新方法。两种物种的精子都可以冷冻和解冻至少两次，并且仍然会产生经历变态的幼虫。
结论：我们描述的简单方法对两个物种都有效，实现>90％的胚胎发育和产生的幼虫经历变态成虫。我们希望这些方法对其他研究海洋无脊椎动物精子的人有用。
