Fouchécourt S, Dacheux F, Dacheux J L
Station de Physiologie de la Reproduction des Mammifères Domestiques, URA INRA-CNRS 1291, 37380 Nouzilly, France.
Biol Reprod. 1999 Mar;60(3):558-66. doi: 10.1095/biolreprod60.3.558.
Microsequencing after two-dimensional electrophoresis revealed a major protein, glutathione-independent prostaglandin D2 synthase (PGDS) in the anterior epididymal region fluid of the ram and stallion. In this epididymal region, PGDS was a polymorphic compound with a molecular mass around 30 kDa and a range of pI from 4 to 7. PGDS represented 15% and 8% of the total luminal proteins present in this region in the ram and stallion, respectively. The secretion of the protein as judged by in vitro biosynthesis, and the presence of its mRNA as studied by Northern blot analysis, were limited to the proximal caput epididymidis. Using a specific polyclonal antibody raised against a synthetic peptide, PGDS was found throughout the epididymis, decreasing in concentration toward the cauda region. PGDS was also detected in the testicular fluid and seminal plasma by Western blotting. Castration and efferent duct ligation in the ram led to a decrease in PGDS mRNA and secretion. PGDS mRNA was not detected in the stallion 1 mo after castration, and it was restored by testosterone supplementation. This study showed that PGDS is present in the environment of spermatozoa throughout the male genital tract. Its function in the maturation and/or protection of spermatozoa is unknown.
二维电泳后的微量测序显示，在公羊和种马附睾头区域的液体中，一种主要蛋白质——谷胱甘肽非依赖性前列腺素D2合成酶（PGDS）被发现。在这个附睾区域，PGDS是一种多态性化合物，分子量约为30 kDa，等电点范围为4至7。在公羊和种马中，PGDS分别占该区域腔内总蛋白质的15%和8%。通过体外生物合成判断该蛋白质的分泌情况，以及通过Northern印迹分析研究其mRNA的存在情况，结果显示其分泌和mRNA仅局限于附睾头近端。使用针对合成肽产生的特异性多克隆抗体，发现PGDS存在于整个附睾中，但其浓度朝着附睾尾区域逐渐降低。通过蛋白质免疫印迹法还在睾丸液和精浆中检测到了PGDS。公羊去势和结扎输出小管导致PGDS mRNA和分泌减少。种马去势1个月后未检测到PGDS mRNA，补充睾酮后其得以恢复。这项研究表明，PGDS存在于整个雄性生殖道的精子所处环境中。其在精子成熟和/或保护中的功能尚不清楚。
