Nuclear and cytoplasmic dynamics of sperm penetration, pronuclear formation and microtubule organization during fertilization and early preimplantation development in the human.
This report describes spatial and temporal aspects of sperm penetration and intracytoplasmic migration, pronuclear evolution and the specificity of presyngamic opposition, stage-specific changes in cytoskeletal organization and the relative contribution of maternal and paternal components to mitotic spindle formation. These studies involved observations of living human oocytes during conventional insemination in vitro and after intracytoplasmic deposition of spermatozoa, analysis of chromatin organization and distribution during pronuclear evolution, and detection of actin and alpha-, beta- and gamma-tubulin by confocal immunofluorescence microscopy. Immature and mature oocytes, penetrated but unfertilized oocytes, fertilized but arrested eggs, and cleavage-stage embryos from normal and dispermic fertilizations were examined. The results demonstrate that sperm nuclear migration to the maternal perinuclear region is rapid and linear, occurs in the absence of a detectable cytoskeletal system and appears to be assisted by an unusual configuration of the sperm tail principal piece which results from either retained intracytoplasmic motility or the process by which the sperm tail is progressively incorporated into the oocyte. Our findings also show a specificity of pronuclear alignment that is associated with a polarized distribution of both maternal and paternal chromatin, and with the position of the sperm centrosome and the presence of microtubules nucleated from this structure. The results also indicate that a maternal microtubule nucleating capacity is present in the immature oocyte but is apparently inactive until spindle formation. The poles of the first mitotic spindle appear to be derived from the sperm centrosome, although some maternal contribution cannot be excluded. The sperm tail and centrosome persist in a single cell through the cleavage stages, and the latter serves as a prominent site of cytoplasmic microtubule nucleation. The results provide a detailed understanding of the cellular and nuclear morphodynamics of the human fertilization process and indicate subtle defects that may be responsible for early developmental failure.
Van Blerkom
J
J
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder 80309, USA.
Davis
P
P
Merriam
J
J
Sinclair
J
J
D002467:Cell Nucleus / Q000502:physiology; D016764:Cell Polarity; D002843:Chromatin; D003593:Cytoplasm / Q000502:physiology; D047108:Embryonic Development; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005307:Fertilization in Vitro; D006801:Humans; D007315:Insemination, Artificial / Q000379:methods; D008297:Male; D008870:Microtubules / Q000502:physiology; D008938:Mitosis / Q000502:physiology; D009865:Oocytes / Q000502:physiology; D011247:Pregnancy; D013084:Sperm-Ovum Interactions
D002843:Chromatin