Impaired sperm fertilizing ability in mice lacking Cysteine-RIch Secretory Protein 1 (CRISP1)
Impaired sperm fertilizing ability in mice lacking Cysteine-RIch Secretory Protein 1 (CRISP1)
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DOI：
10.1016/j.ydbio.2008.03.015
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发表时间：
2008-08-01
影响因子：
2.7
通讯作者：
Cuasnicu, Patricia S.
中科院分区：
文献类型：
--
作者：
Da Ros, Vanina G.;Maldera, Julieta A.;Cuasnicu, Patricia S.
哺乳动物受精是一个复杂的多步骤过程，由两个配子上存在的不同分子介导。附睾蛋白CRISP 1是本实验室鉴定的半胱氨酸-RICH分泌蛋白（CRISP）家族的一员，推测其通过与卵子互补位点结合参与精子-透明带（ZP）相互作用和配子融合。为了阐明CRISP 1在体内的功能作用，我们破坏了Crisp 1基因，并评估了对动物生育力和几个精子参数的影响。与对照组相比，雄性和雌性Crisp 1（-/-）动物的生育力无差异。Crisp 1（-/-）小鼠的精子活力和自发或孕酮诱导的顶体反应的能力均未受到影响。然而，在获能过程中的蛋白质酪氨酸磷酸化水平明显低于突变体精子比对照组。体外受精试验表明，Crisp 1（-/-）精子穿透ZP完整和ZP游离卵子的能力也显着降低。此外，当在竞争试验中将不含ZP的卵子与Crisp 1（+/+）和Crisp 1（-/-）精子同时授精时，突变精子在其融合能力方面表现出更大的劣势。最后，在配子共孵育过程中，CRISP 1或CRISP 2的存在进一步抑制了Crisp 1（-/-）精子的融合能力，这一发现支持另一种CRISP在受精过程中与CRISP 1合作，并可能弥补其在突变小鼠中的缺乏。总之，这些结果表明CRISP蛋白是哺乳动物受精过程中的参与者。据我们所知，这是针对CRISP蛋白产生的第一个敲除小鼠。所获得的信息可能对广泛分布和进化保守的CRISP家族的其他成员具有重要的功能意义。(C)2008年爱思唯尔公司All rights reserved.
Mammalian fertilization is a complex multi-step process mediated by different molecules present on both gametes. Epididymal Protein CRISP1, a member of the Cysteine-RIch Secretory Protein (CRISP) family, was identified by our laboratory and postulated to participate in both sperm-zona pellucida (ZP) interaction and gamete fusion by binding to egg-complementary sites. To elucidate the functional role of CRISP1 in vivo, we disrupted the Crisp1 gene and evaluated the effect on animal fertility and several sperm parameters. Male and female Crisp1(-/-) animals exhibited no differences in fertility compared to controls. Sperm motility and the ability to undergo a spontaneous or progesterone-induced acrosome reaction were neither affected in Crisp1(-/-) mice. However, the level of protein tyrosine phosphorylation during capacitation was clearly lower in mutant sperm than in controls. In vitro fertilization assays showed that Crisp1(-/-) sperm also exhibited a significantly reduced ability to penetrate both ZP-intact and ZP-free eggs. Moreover, when ZP-free eggs were simultaneously inseminated with Crisp1(+/+) and Crisp1(-/-) sperm in a competition assay, the mutant sperm exhibited a greater disadvantage in their fusion ability. Finally, the finding that the fusion ability of Crisp1(-/-) sperm was further inhibited by the presence of CRISP1 OF CRISP2 during gamete co-incubation, supports that another CRISP cooperates with CRISP1 during fertilization and might compensate for its lack in the mutant mice. Together, these results indicate that CRISP proteins are players in the mammalian fertilization process. To our knowledge this is the first knockout mice generated for a CRISP protein. The information obtained might have important functional implications for other members of the widely distributed and evolutionarily conserved CRISP family. (C) 2008 Elsevier Inc. All rights reserved.