Hecht Nadine, Behr Rüdiger, Hild Anne, Bergmann Martin, Weidner Wolfgang, Steger Klaus
Department of Urology and Pediatric Urology, Justus Liebig University Giessen, Giessen, Germany.
Hum Reprod. 2009 Mar;24(3):536-45. doi: 10.1093/humrep/den390. Epub 2008 Dec 4.
Histone to protamine exchange, in man, is only 80% complete and spermatozoal histones are highly acetylated suggesting that their associated genes may be involved in early gene expression in the embryo.
Using in situ hybridization and immunocytochemistry, we analyzed expression of protamine-1 and protamine-2, and histone H4 specifically acetylated at lysine 5 (H4K5ac), 8 (H4K8ac), 12 (H4K12ac) and 16 (H4K16ac) in human (n = 22) and marmoset (n = 6) testes and ejaculates.
Protamine-1 and protamine-2 mRNA was present in round and elongating spermatids. All antibodies against acetylated histones revealed positive signals in these cells. Human spermatogonia showed positive signals for H4K8ac and H4K16ac, whereas marmoset spermatogonia were positive for H4K8ac and H4K12ac. In man, H4K16ac already displayed a positive immunoreaction with pachytene spermatocytes, starting at stage III of the seminiferous epithelial cycle. All antibodies showed positive immunostaining in ejaculated spermatozoa of both species.
The common marmoset represents a suitable animal model for studies on nuclear protein expression during human spermatogenesis. The two species exhibit a similar organization of seminiferous epithelium and an identical expression pattern of protamine-1 and protamine-2 mRNA in round and elongating spermatids. The presence of specifically acetylated histones H4 in testicular spermatids and ejaculated spermatozoa demonstrates an incomplete histone to protamine exchange in both species. As acetylated histones are known to be associated with genes involved in gene expression, the common marmoset may, in future, be used as a model for investigations on early embryo development which, in man, are not possible for ethical reasons.
在人类中,组蛋白向鱼精蛋白的交换仅完成80%,且精子组蛋白高度乙酰化,这表明其相关基因可能参与胚胎早期基因表达。
我们使用原位杂交和免疫细胞化学技术,分析了人类(n = 22)和狨猴(n = 6)睾丸及精液中鱼精蛋白-1、鱼精蛋白-2以及赖氨酸5(H4K5ac)、8(H4K8ac)、12(H4K12ac)和16(H4K16ac)特异性乙酰化的组蛋白H4的表达情况。
鱼精蛋白-1和鱼精蛋白-2 mRNA存在于圆形和伸长的精子细胞中。所有针对乙酰化组蛋白的抗体在这些细胞中均显示出阳性信号。人类精原细胞对H4K8ac和H4K16ac呈阳性信号,而狨猴精原细胞对H4K8ac和H4K12ac呈阳性信号。在人类中,从生精上皮周期的III期开始,H4K16ac就已在粗线期精母细胞中呈现阳性免疫反应。所有抗体在两个物种的射出精子中均显示出阳性免疫染色。
普通狨猴是研究人类精子发生过程中核蛋白表达的合适动物模型。这两个物种的生精上皮组织相似,圆形和伸长精子细胞中鱼精蛋白-1和鱼精蛋白-2 mRNA的表达模式相同。睾丸精子细胞和射出精子中特异性乙酰化组蛋白H4的存在表明两个物种中组蛋白向鱼精蛋白的交换均不完全。由于已知乙酰化组蛋白与参与基因表达的基因相关,未来普通狨猴可能会被用作研究早期胚胎发育的模型,而在人类中,出于伦理原因无法进行此类研究。