Mouse Spam1 (PH-20) is a multifunctional protein: evidence for its expression in the female reproductive tract.
Mouse Spam1 (PH-20) is a multifunctional protein: evidence for its expression in the female reproductive tract.
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Mouse Spam1 (PH-20) 是一种多功能蛋白：其在女性生殖道中表达的证据。
DOI：
10.1095/biolreprod.102.013854
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发表时间：
2003
影响因子：
3.6
通讯作者：
Martin-DeLeon,PatriciaA
中科院分区：
文献类型：
--
作者：
Zhang,Hong;Martin-DeLeon,PatriciaA
精子粘附分子 1 (Spam1) 是一种广泛保守的精子表面蛋白，在哺乳动物受精中发挥多种作用。尽管根据 Northern 印迹分析，该蛋白的基因被认为是睾丸特异性的，但当通过更灵敏的技术分析转录本时，有证据表明该蛋白具有非睾丸表达。在本研究中，逆转录聚合酶链式反应测定、RNase 保护测定 (RPA) 和原位转录物杂交测定的结果表明 murineSpam1 基因在女性生殖道中转录。 RPA显示Spam1转录本以区域依赖性方式合成，输卵管的转录本水平低于子宫和阴道。女性生殖道中的转录物水平比睾丸中低 3 至 10 倍。原位转录本杂交分析显示，生殖道所有三个区域的管腔上皮以及子宫肌层和输卵管间皮中均存在 RNA。蛋白质印迹分析和免疫组织化学表明，女性组织中的蛋白质浓度比精子中低 1.5 至 3 倍，并且定位与转录本相似。该蛋白在中性 pH 值下具有透明质酸酶活性，这是精子透明质酸酶所特有的，但在酸性 pH 值下则不具有透明质酸酶活性。在子宫中，Spam1 的表达在动情周期期间波动。它的定位表明，除了作为分泌蛋白发挥作用外，它还可能参与女性生殖道中的透明质酸代谢或周转。我们的结果进一步证明 Spam1 是一种多功能蛋白，并且其表达受到的限制比之前报道的要少。
Sperm adhesion molecule 1 (Spam1) is a widely conserved sperm surface protein with multiple roles in mammalian fertilization. Although the gene for this protein has been thought to be testis specific based on Northern blot analysis, there is evidence for nontesticular expression when transcripts are analyzed by more sensitive techniques. In the present investigation, results of a reverse transcription polymerase chain reaction assay, an RNase-protection assay (RPA), and an in situ transcript hybridization assay revealed that the murineSpam1gene is transcribed in the female genital tract. RPA revealed thatSpam1transcripts are synthesized in a region-dependent manner, with the oviduct having lower transcript levels than the uterus and vagina. The transcripts levels were 3- to 10-fold lower in the female genital tract than in the testis. In situ transcript hybridization assay revealed RNA in the luminal epithelium in all three regions of the genital tract and in the uterine myometrium and the oviductal mesothelium. Western blot analysis and immunohistochemistry demonstrated that the protein concentration is 1.5- to 3-fold lower in female tissues than in sperm, and localization is similar to that of the transcripts. The protein has hyaluronidase activity at neutral pH, which is unique for sperm hyaluronidase, but not at acidic pH. In the uterus, Spam1 expression fluctuated during the estrous cycle. Its localization suggests that in addition to functioning as a secretory protein, it may be involved in hyaluronic acid metabolism or turnover in the female genital tract. Our results provide further evidence that Spam1 is a multifunctional protein and that it is less restricted in its expression than previously reported.