Enhancement of human sperm motility by trophinin binding peptide
Enhancement of human sperm motility by trophinin binding peptide
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DOI：
10.1016/j.juro.2008.03.185
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发表时间：
2008-08-01
影响因子：
6.6
通讯作者：
Fukuda, Michiko N.
中科院分区：
文献类型：
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作者：
Hatakeyama, Shingo;Sugihara, Kazuhiro;Fukuda, Michiko N.
目的：以前我们发现，滋养蛋白结合肽GWRQ（甘氨酸，色氨酸，精氨酸，谷氨酸）激活人滋养层细胞。虽然营养素在人类精子中表达，但据我们所知，这种蛋白质的功能尚不清楚。在这项研究中，我们测试了GWRQ对人类精子的影响，以评估是否肽增强人类精子motility.Materials和方法：免疫组织化学进行使用特异性的trophinin，bystin或tastin的单克隆抗体。化学合成GWRQ-MAPS（多价8分支GWRQ肽）。将来自平均+/- SD年龄为35.75 +/- 3.4岁的4名志愿者的人精子悬浮在含有GWRQ或对照肽的缓冲液中。在平均年龄为25.5 ± 2.5岁的23名志愿者中，计数不活动精子的数量，并从精子总数中减去以确定活动精子的数量。使用Transwell测定来测量游泳运动。用荧光细胞活力测定法和Fluo-4钙离子测定法分别测定了GWRQ或对照肽孵育的精子细胞中三磷酸腺苷和细胞内钙的水平。结果：免疫组化证实了人精子中存在营养素和营养素相关蛋白bystin和tastin。人精子与GWRQ-MAPS孵育后，在精子计数试验和向下游动Transwell试验中显示出增强的活动性。与GWRQ-MAPS孵育的精子细胞显示三磷酸腺苷减少和细胞内Ca 2+增加。结论：结果表明，GWRQ-MAPS可能有利于优化体外受精的结果，并有助于避免需要在严重低精子活力的情况下进行卵胞浆内单精子注射。滋养蛋白可能在调节人类精子中的腺苷三磷酸酶中起作用。
Purpose: Previously we found that the trophinin-binding peptide GWRQ (glycine, tryptophane, arginine, glutamic acid) activated human trophoblastic cells. Although trophinin is expressed in human sperm, to our knowledge the function of this protein is not known. In this study we tested the effect of GWRQ on human sperm to evaluate whether the peptide enhances human sperm motility.Materials and Methods: Immunohistochemistry was performed using monoclonal antibodies specific to trophinin, bystin or tastin. GWRQ-MAPS (multivalent 8-branched GWRQ peptide) was chemically synthesized. Human sperm from 4 volunteers with a mean +/- SD age of 35.75 +/- 3.4 years was suspended in buffer with GWRQ or control peptides. In 23 volunteers with a mean age of 25.5 +/- 2.5 years the number of immotile sperm was counted and subtracted from the total number of sperm to determine the number of motile sperm. A Transwell assay was used to measure swim-down motility. Levels of adenosine triphosphate and intracellular calcium in sperm cells incubated with GWRQ or control peptide were measured using a luminescent cell viability assay and a fluo-4 calcium assay, respectively.Results: The presence of trophinin and the trophinin associated proteins bystin and tastin in human sperm was confirmed by immunohistochemistry. Human sperm incubated with GWRQ-MAPS showed enhanced motility on sperm count assay and swim-down Transwell assay. Sperm cells incubated with GWRQ-MAPS showed decreased adenosine triphosphate and increased intracellular calcium.Conclusions: Results suggest that GWRQ-MAPS may facilitate optimized in vitro fertilization outcomes and help avert the need for intracytoplasmic sperm injection in cases of severely low sperm motility. Trophinin may have a role in regulating adenosine triphosphatase in human sperm.