Spermaurin, an La1-like peptide from the venom of the scorpion Scorpio maurus palmatus, improves sperm motility and fertilization in different mammalian species
Spermaurin, an La1-like peptide from the venom of the scorpion Scorpio maurus palmatus, improves sperm motility and fertilization in different mammalian species
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DOI：
10.1093/molehr/gaw075
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发表时间：
2017-02-01
影响因子：
4
通讯作者：
Arnoult, Christophe
中科院分区：
文献类型：
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作者：
Martinez, Guillaume;Hograindleur, Jean-Pascal;Arnoult, Christophe
研究问题：有没有可能从蝎子的毒液中鉴定出能够增强精子活力的原始化合物？总结回答：我们发现了一种有效的富含二硫键的肽（DRP）的73个氨基酸，显着提高了活力的新鲜和冻融精子在不同的哺乳动物物种，包括人类，并改善受精结果在小鼠IVF experiments.What是已知的：精子活力的任何障碍有很大的影响受精，并可能导致不孕不育。因此，已经做出了重大努力来确定可能改善精子活力的药理学药物。这类化合物在无精子症中特别有用，以改善睾丸精子提取，并且在冷冻保存领域中特别有用，因为冷冻-解冻精子的运动性降低。研究设计、规模、持续时间：这是一项基础科学/医学研究，旨在从能够增强哺乳动物精子运动性的毒液库中鉴定原始化合物，包括人类。我们首先在一种蝎子的毒液中发现了S.M.掌肌是一个能够有效激活精子活力的部分。我们接下来通过液相色谱、质谱和肽合成来纯化和表征该化合物。最后，使用不同的体外试验在不同的哺乳动物species.PARTICIPANTS/MATERIALS，SETTING，METHODS：对于人类精子，生物样品收集normozospermic捐助者和生育能力低下的患者参加生殖科诊断精液分析的效力和毒性的鉴定化合物。从出于特定授权研究项目的需要而实施安乐死的食蟹猴（Macaca fascicularis）中采集睾丸精子。该肽还在牛和小鼠精子中进行了测试。我们测量了不同的精子活力参数与计算机辅助精子分析系统中存在或不存在的peptide.Main结果和作用的机会：尺寸排阻色谱法使我们能够分离出一部分的毒液的S. m。掌骨能够增加精子活力。通过液相色谱和质谱分析，鉴定了一种由73个氨基酸组成的具有4个二硫键的肽，该肽具有生物活性，称为“精尿蛋白”。通过化学合成证实了精子蛋白的身份。我们发现，肽增加了新鲜和冻融人类精子的活力。我们观察到，肽的效力更高的新鲜射精精子与低活力，实现了100%的曲线速度增加表现不佳的精子。我们还证明了肽对牛和小鼠新鲜附睾、牛冻融射精和新鲜非人灵长类动物睾丸精子有效。最后，在小鼠体外受精，2-细胞胚胎的生产增加了24%，当精子用peptide.Limitations处理时，预防的原因：这项工作是在体外评价spermaurin提高精子活力参数的能力。本研究的另一个局限性是使用天然（n = 36）和合成（n = 12）肽检测的人类精子样本数量较少。此外，肽对IVF结果的影响仅在小鼠中进行了测试，需要进一步测试人类和牛的配子，以确认和扩展这一结果在其他哺乳动物species.WIDER影响的发现：这项工作证实了我们的初步研究表明，毒液代表一个有趣的来源，能够修改精子生理学的分子。此外，这项工作提出了第一个证明的毒肽的生物作用，从蝎子S。M.大规模数据：不适用研究资助/竞争兴趣：这项工作是项目“LAB COM-14 LAB 7 0004 01-LIPAV”的一部分，由法国研究机构（ANR）的LabCom 2014计划资助。Arnoult博士报告说，在研究进行期间，IMV技术公司提供了赠款。此外，Arnoult、Martinez、Ray和施密特博士的专利EP 16305642.7正在申请中，其中包含本手稿中的一些信息。
STUDY QUESTION: Is it possible to identify original compounds that are able to enhance sperm motility from the venom of the scorpion Scorpio maurus palmatus?SUMMARY ANSWER: We identified a potent disulfide-rich peptide (DRP) of 73 amino acids that significantly improved the motility of fresh and frozen-thawed sperm in different mammalian species, including human, and improved fertilization outcome in mouse IVF experiments.WHAT IS KNOWN ALREADY: Any disturbance of sperm motility has a strong impact on fertilization and can lead to subfertility or infertility. Significant efforts have, therefore, been made to identify pharmacological drugs that might improve sperm motility. Such compounds are particularly useful in azoospermia to improve testicular sperm extraction and in the domain of cryopreservation because the motility of frozen-thawed sperm is reduced.STUDY DESIGN, SIZE, DURATION: This was a basic science/medical research study aimed at identifying original compounds from a library of venoms able to enhance mammalian sperm motility, including human. We first identified in the venom of a scorpion S.m. palmatus a fraction able to potently activate sperm motility. We next purified and characterized the compound by liquid chromatography, mass spectrometry and peptide synthesis. Finally, the potency and toxicity of both purified and synthetic versions of the identified compound on sperm motility were assessed using different in vitro tests in different mammalian species.PARTICIPANTS/MATERIALS, SETTING, METHODS: For human sperm, biological samples were collected from normozoospermic donors and subfertile patients attending a reproduction department for diagnostic semen analysis. Testicular sperm was collected from cynomolgus monkeys (Macaca fascicularis) euthanized for the needs of specific authorized research projects. The peptide was also tested on bovine and mouse epidydimal sperm. We measured different sperm motility parameters with a computer-assisted sperm analysis system in the presence or absence of the peptide.MAIN RESULTS AND THE ROLE OF CHANCE: Size exclusion chromatography enabled us to isolate a fraction of the venom of S.m. palmatus able to increase sperm motility. By liquid chromatography and mass spectrometry, a peptide comprising 73 amino acids with 4 disulfide bridges was identified as responsible for the biological activity and called 'spermaurin'. The identity of spermaurin was confirmed by chemical synthesis. We showed that the peptide increased the motility of fresh and frozen-thawed human sperm. We observed that the potency of the peptide was higher on fresh ejaculated spermatozoa with a low motility, achieving a 100% increase of curvilinear velocity in poorly performing sperm. We also demonstrated that peptide is effective on bovine and mouse fresh epididymal, bovine frozen-thawed ejaculated and fresh non-human primate testicular sperm. Finally, in mouse IVF, the production of 2-cell embryos was increased by 24% when sperm were treated with the peptide.LIMITATIONS, REASONS FOR CAUTION: This work is an in vitro evaluation of the ability of spermaurin to improve sperm motility parameters. Another limitation of this study is the small number of human sperm samples tested with the natural (n = 36) and synthetic (n = 12) peptides. Moreover, the effect of the peptide on IVF outcome was only tested in mouse and further tests with human and bovine gametes are required to confirm and extend this result in other mammalian species.WIDER IMPLICATIONS OF THE FINDINGS: This work confirms our initial study showing that venoms represent an interesting source of molecules that are able to modify sperm physiology. Moreover, this work presents the first demonstrated biological action of a venom peptide from the scorpion S. m. palmatus with sequence similarities to La1 peptide from Liocheles australasiae (Wood scorpion), a widespread family of DRPs.LARGE SCALE DATA: Not applicable.STUDY FUNDING/COMPETING INTEREST(S): This work is part of the project 'LAB COM-14 LAB7 0004 01-LIPAV', funded by the program LabCom 2014 from the French Research Agency (ANR). Dr Arnoult reports grants from IMV Technologies during the conduct of the study. In addition, Drs Arnoult, Martinez, Ray and Schmitt have a patent EP16305642.7 pending containing some of the information presented in this manuscript.