前列腺和睾丸表达（PATE）样蛋白在精卵相互作用中的参与。
Involvement of the prostate and testis expression (PATE)-like proteins in sperm-oocyte interaction.
机构信息
Institute for the Study of Fertility, Lis Maternity Hospital, Tel Aviv Sourasky Medical Center, Tel Aviv University, Tel Aviv, Israel.
出版信息
Hum Reprod. 2012 May;27(5):1238-48. doi: 10.1093/humrep/des064. Epub 2012 Mar 8.
BACKGROUND
The prostate and testis expression (PATE)-like family of proteins are expressed mainly in the male genital tract. They are localized in the sperm head and are homologous to SP-10, the acrosomal vesicle protein also named ACRV1. Our aim was to characterize the expression and functional role of three PATE-like proteins in the testis and ejaculated sperm.
METHODS
The expression and localization of PATE-like proteins in human testis biopsies (n= 95) and sperm cells were assessed by RT-PCR, immunohistochemistry and immunofluorescence staining (at least 600 sperm cells per specimen). The function of the PATE protein was tested by the hemizona assay and hamster egg penetration test (HEPT).
RESULTS
PATE and PATE-M genes and proteins were present almost exclusively in germ cells in the testis: immunoflourescence showed that the percentage of germ cells positive for PATE, PATE-M and PATE-B was 85, 50 and 2%, respectively. PATE and PATE-M proteins were localized in the equatorial segment of the sperm head, while PATE-B protein was localized in the post-acrosomal region. A polyclonal antibody (Ab, at 1:50 and 1:200 dilutions) against the PATE protein did not inhibit sperm-zona binding in the hemizona assay (hemizona index of 89.6 ± 10 and 87 ± 36%, respectively). However, there was inhibition of sperm-oolemma fusion and penetration in the HEPT (penetration index: without Ab 7 ± 3.9; Ab dilution of 1:100, 4 ± 3.5; Ab dilution of 1:20, 0.6 ± 1.2, P < 0.001).
CONCLUSIONS
Our data suggest that PATE protein is involved in sperm-oolemma fusion and penetration but not sperm-zona binding.
背景
前列腺和睾丸表达（PATE）样蛋白家族主要在男性生殖道中表达。它们定位于精子头部，与顶体囊泡蛋白 SP-10 同源，也称为 ACRV1。我们的目的是研究睾丸和射出精子中三种 PATE 样蛋白的表达和功能作用。
方法
通过 RT-PCR、免疫组织化学和免疫荧光染色（每个标本至少 600 个精子细胞）评估 PATE 样蛋白在人睾丸活检组织（n=95）和精子细胞中的表达和定位。通过半透明带试验和仓鼠卵穿透试验（HEPT）测试 PATE 蛋白的功能。
结果
PATE 和 PATE-M 基因和蛋白几乎仅存在于睾丸生殖细胞中：免疫荧光显示 PATE、PATE-M 和 PATE-B 阳性的生殖细胞百分比分别为 85%、50%和 2%。PATE 和 PATE-M 蛋白定位于精子头部赤道段，而 PATE-B 蛋白定位于顶体后区。针对 PATE 蛋白的多克隆抗体（Ab，1:50 和 1:200 稀释度）在半透明带试验中未抑制精子-透明带结合（半透明带指数分别为 89.6±10%和 87±36%）。然而，在 HEPT 中抑制了精子-卵母细胞膜融合和穿透（穿透指数：无 Ab 为 7±3.9%；Ab 稀释度为 1:100 时为 4±3.5%；Ab 稀释度为 1:20 时为 0.6±1.2%，P<0.001）。
结论
我们的数据表明 PATE 蛋白参与精子-卵母细胞膜融合和穿透，但不参与精子-透明带结合。
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