Lambard Sophie, Galeraud-Denis Isabelle, Bouraïma Hélène, Bourguiba Sonia, Chocat Anne, Carreau Serge
UPRES EA 2608-USC INRA, University-IRBA, Cedex, France.
Mol Hum Reprod. 2003 Mar;9(3):117-24. doi: 10.1093/molehr/gag020.
Cytochrome p450 aromatase (p450arom) is a key enzyme responsible for the irreversible transformation of androgens into estrogens. In the present study, we have analysed the ability of human ejaculated spermatozoa to produce estrogens and for that purpose we have looked for the expression of specific aromatase transcript and protein. We have confirmed the presence of p450arom transcript in all normospermic purified samples by nested PCR. The sequence of PCR products from purified spermatozoa shares 98% identity with published human p450arom sequence. Using a semi-quantitative approach, we have observed in immotile sperm a significant decrease (28%) of the aromatase/glyceraldehyde-3-phosphate dehydrogenase ratio compared with the motile sperm fraction. On Western blot with a monoclonal antibody directed against aromatase, we have detected two bands (53 and 49 kDa) in microsome preparations from purified spermatozoa. In total protein extracts of purified spermatozoa (with and without cytoplasmic droplets), we have only found the aromatase as a 49 kDa band with a stronger intensity when cytoplasmic droplets are present. Moreover, the band seems to be weaker in immotile spermatozoa (with and without cytoplasmic droplets). Our data demonstrate the expression of aromatase both in terms of mRNA and protein in each sample of human purified spermatozoa and in addition, our results suggest that aromatase could be concerned with the acquisition of sperm motility.
细胞色素P450芳香化酶(P450arom)是一种关键酶,负责将雄激素不可逆地转化为雌激素。在本研究中,我们分析了人类射出精子产生雌激素的能力,为此我们寻找了特定芳香化酶转录本和蛋白质的表达。我们通过巢式PCR在所有正常精子纯化样本中证实了P450arom转录本的存在。来自纯化精子的PCR产物序列与已发表的人类P450arom序列有98%的同一性。使用半定量方法,我们观察到与活动精子部分相比,不活动精子中芳香化酶/甘油醛-3-磷酸脱氢酶的比率显著降低(28%)。在用针对芳香化酶的单克隆抗体进行的蛋白质印迹分析中,我们在来自纯化精子的微粒体制剂中检测到两条带(53和49 kDa)。在纯化精子的总蛋白提取物中(有和没有细胞质滴),我们只发现芳香化酶为一条49 kDa的带,当存在细胞质滴时强度更强。此外,这条带在不活动精子(有和没有细胞质滴)中似乎较弱。我们的数据证明了在每个人类纯化精子样本中芳香化酶在mRNA和蛋白质水平上的表达,此外,我们的结果表明芳香化酶可能与精子运动能力的获得有关。