Rago Vittoria, Siciliano Laura, Aquila Saveria, Carpino Amalia
Department of Cell Biology, Faculty of Pharmacy, University of Calabria, Cosenza, Italy.
Reprod Biol Endocrinol. 2006 Jul 17;4:36. doi: 10.1186/1477-7827-4-36.
A key role of estrogens in human sperm biology has been recently suggested by aromatase and estrogen receptor detection in human testicular germ cells and ejaculated spermatozoa. However, the involvement of these hormones in the sperm maturation process is still not defined. The aim of this work was to investigate the expression of estrogen receptors, ER-alpha and ER-beta, in human ejaculated immature spermatozoa with excess residual cytoplasm.
Immunofluorescence labelling has been used to localize ER-alpha and ER-beta proteins in immature spermatozoa isolated by Percoll gradient, while Western blot analysis was carried out on sperm protein extracts.
Both estrogen receptors were localized in excess residual cytoplasm of immature sperm, while sperm tails showed only ER-beta. Furthermore, in the same cells, immunoblots detected the presence of the full-length (approximately 67 kDa) ER-alpha and (approximately 59 kDa) ER-beta proteins, together with a approximately 50 kDa ER-beta species, lacking in mature sperm.
The present investigation demonstrated ER-alpha and ER-beta presence in excess residual cytoplasm of human abnormal sperm cells, suggesting the hypothesis that both the 'classical' ERs could be able to mediate estrogen action in spermatogenetic cells. Furthermore, the presence of the short ER-beta form in abnormal germ cells and its disappearance in mature sperm, support estrogen modulation via different ER forms during sperm maturation.
最近,在人类睾丸生殖细胞和射出的精子中检测到芳香化酶和雌激素受体,提示雌激素在人类精子生物学中起关键作用。然而,这些激素在精子成熟过程中的作用仍不明确。本研究旨在调查雌激素受体α(ER-α)和雌激素受体β(ER-β)在含有过量残留细胞质的人类射出未成熟精子中的表达情况。
采用免疫荧光标记法对通过Percoll梯度分离的未成熟精子中的ER-α和ER-β蛋白进行定位,同时对精子蛋白提取物进行蛋白质印迹分析。
两种雌激素受体均定位于未成熟精子的过量残留细胞质中,而精子尾部仅显示ER-β。此外,在同一细胞中进行的蛋白质印迹检测到全长(约67 kDa)的ER-α和(约59 kDa)的ER-β蛋白,以及成熟精子中缺乏的约50 kDa的ER-β蛋白。
本研究证明了人类异常精子细胞的过量残留细胞质中存在ER-α和ER-β,提示“经典”雌激素受体可能能够介导雌激素在生精细胞中的作用这一假说。此外,异常生殖细胞中短型ER-β的存在及其在成熟精子中的消失,支持了精子成熟过程中雌激素通过不同ER形式进行调节的观点。