Protamine Withdrawal from Human Sperm Nuclei Following Heterologous ICSI into Hamster Oocytes

Estella      L. Jones; Andrei      O. Zalensky; Irina      A. Zalenskaya

doi:10.2174/092986611795713925

Abstract

During late stages of spermatogenesis in mammals, most histones bound to DNA are replaced by protamines (PRM), which results in formation of supercondensed and genetically inert sperm chromatin. At fertilization, mature spermatozoon penetrates oocyte and chromatin is remodeled “back” from nucleoprotamine to nucleohistone state. While being crucial for activation of male genome and ultimately for initiation of embryonic development, this process is poorly studied, especially in humans. Data on model animals concerning PRM to histones exchange post fertilization are few and contradictory. As direct experimentation with human embryos is impossible due to ethical, legal and technical reasons, we evaluate the timing and mode of PRM removal in a heterologous ICSI system using hamster ova injected with human sperm. Localization of human PRM 1 and 2 in hybrid zygotes was established using immunofluorescence. We observed a marked zygote to zygote variability in male pronuclei size for any time point post ICSI and demonstrated that PRM removal correlates with the developing pronuclei area rather than time after injection. Overall, the disappearance of protamines from sperm is rather rapid and most likely completed within 1 hr. We propose that the critical characteristic influencing PRM removal after heterologous fertilization is the intrinsic heterogeneity of the human sperm population. The same yet unexplored variance may be one of the reasons for canceled, delayed or aberrant early embryonic development during natural or artificial fertilization in humans.

Keywords: Fertilization, sperm, protamine, heterologous ICSI, male pronuclei, zygote, Spermatozoa, spermatogenesis, germ cells, nucleoprotamine, oocytes, Histones, Spermiogenesis, Mitosis, Immunolocalization, ooplasm, immunofluorescence, homogeneous PRM, Non-decondensed sperm, AntiserumFertilization, sperm, protamine, heterologous ICSI, male pronuclei, zygote, Spermatozoa, spermatogenesis, germ cells, nucleoprotamine, oocytes, Histones, Spermiogenesis, Mitosis, Immunolocalization, ooplasm, immunofluorescence, homogeneous PRM, Non-decondensed sperm, Antiserum