Fulka Helena, Barnetova Irena, Mosko Tibor, Fulka Josef
Center for Cell Therapy and Tissue Repair, VUZV Laboratories, PO Box 1, CS-104 01 Prague 10, Czech Republic.
Hum Reprod. 2008 Mar;23(3):627-34. doi: 10.1093/humrep/dem406. Epub 2008 Jan 23.
The epigenetic status of human spermatozoa is difficult to analyse. The method of interspecies fertilization can be used for different purposes. The aim of our work was to adopt this approach for the detailed analysis of epigenetic status of human spermatozoa injected into mouse oocytes.
Human spermatozoa were injected into ovulated mouse oocytes. When both parental pronuclei were formed, the zygotes were fixed and labeled with antibodies against histones methylated or acetylated at different positions (residues).
Our results show that human spermatozoa injected into mouse oocytes fully respond to oocyte cytoplasmic factors and form analysable pronuclei. The labeling of zygotes showed that as in other species, the paternal chromatin is extensively epigenetically remodeled.
The interspecies ICSI may be a powerful tool for the analysis of sperm epigenetic status even with a very low number of spermatozoa available. This analysis could be used as an additional approach for the assessment of certain forms of human infertility, as well as for testing the normality of male gametes obtained from embryonic stem cells.
人类精子的表观遗传状态难以分析。种间受精方法可用于不同目的。我们工作的目的是采用这种方法来详细分析注入小鼠卵母细胞的人类精子的表观遗传状态。
将人类精子注入排卵的小鼠卵母细胞。当双亲原核形成后,将受精卵固定并用针对在不同位置(残基)甲基化或乙酰化的组蛋白的抗体进行标记。
我们的结果表明,注入小鼠卵母细胞的人类精子对卵母细胞胞质因子有充分反应并形成可分析的原核。受精卵的标记显示,与其他物种一样,父本染色质在表观遗传上被广泛重塑。
即使可用精子数量非常少,种间卵胞浆内单精子注射可能是分析精子表观遗传状态的有力工具。这种分析可作为评估某些形式人类不孕症的额外方法,以及用于测试从胚胎干细胞获得的雄配子的正常性。