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Fertilization is a precisely orchestrated cascade of events that results in the union of paternal and maternal genomes and in the establishment of mitotic potential of the zygote. To initiate embryonic development, the structures of the fertilizing sperm have to be disassembled and transformed into zygotic components by interactions with the cytoplasm of the egg. These interactions include the decondensation of the sperm nucleus into male pronucleus, the assembly of the zygotic centrosome, and the gathering of centrosomal proteins and sperm aster microtubules around the sperm centriole. Both the formation of the male pronucleus and the assembly of the zygotic centrosome are crucial steps required for pronuclear apposition and genomic union. The discovery of previously undetected fertilization failures that are due to defects in the assembly of the zygotic centrosome, abnormal pronuclear development, and compromised cytoskeletal dynamics enforces the development of new diagnostic strategies. Moreover, the introduction of new methods of infertility treatments, such as intracytoplasmic sperm injection and round spermatid nucleus injection into assisted human reproductive technology programs, emphasizes our lack of understanding of the cellular and molecular basis of human fertilization and evokes the need for additional experimentation. These efforts, however, are compromised by the sensitive nature of human embryo research and thus are severely restricted. Animal models that are reliable and cost-effective and that feature the characteristics of human fertilization have therefore been sought. Rodents such as the rat, mouse, and hamster are poor models owing to their maternal inheritance of the zygotic centrosome that is in strong contrast with the biparentally contributed assembly of the human zygotic centrosome during fertilization. Although rabbits are similar to humans from the standpoint of mitotic potential inheritance, information on postfertilization events in rabbits are lacking. Nonhuman primates represented by the rhesus monkey proved to be a reliable model for human in vitro fertilization and intracytoplasmic sperm injection, an advantage that is further emphasized by phyllogenetic similarity. In situations in which the high cost of primate research does not allow for large-scale experimentation (i.e., when large numbers of oocytes and embryos are needed), ruminants would be an ideal solution. Represented by the cow and sheep, domestic ruminants feature a fertilization strategy similar to that of the human. In addition, large numbers of gametes can be obtained wherever farms and slaughterhouses are accessible. Moreover, the detailed information on ruminant fertilization is strengthened by years of research and well-defined reproductive technology aimed at increasing the productivity of farm animals. Ruminants and rhesus monkeys have been extensively studied, and the data from these studies have been extrapolated in order to propose new strategies for the diagnosis and treatment of human infertility.

Fertilization in humans follows a complex series of events including binding of the sperm to the oocyte plasma membrane, oocyte activation, the completion of meiotic maturation of the oocyte with the extrusion of the second polar body, the decondensation of the sperm nucleus and the maternal chromosomes into male and female pronuclei and the restoration of the sperm centrosome. This duplicates and separates, forming two mitotic spindle poles upon which the parental genomes can intermix to complete fertilization. The use of intracytoplasmic sperm injection (ICSI) has been highly effective as a treatment for severe male infertility and thousands of ICSI babies have been born world-wide. Working with rhesus monkey gametes, we have developed a preclinical animal model for understanding the cell biological basis of ICSI. Typically, ICSI results in abnormal nuclear remodeling during sperm decondensation due to the presence of the sperm acrosome and perinuclear structures normally removed at the oolemma during in vitro fertilization. These unusual modifications raise concerns that the ICSI procedure itself might lead to the observed increase in chromosome anomalies reported for

During fertilization in humans, the sperm introduces the centrosome, the oocyte's microtubule organizing centre (MTOC), restoring centrosome function. The newly activated oocyte initiates extrusion of the second polar body and begins pronuclear formation. Apposition of the male and female pronuclei requires microtubule-mediated motility in the form of an aster of microtubules emanating from the sperm centrosome. The centrosome then duplicates and separates, forming the two poles of the mitotic spindle, upon which the parental genomes intermix, completing fertilization. The restoration and function of the centrosome is critical for successful fertilization suggesting that a defective sperm centrosome will lead to fertilization arrest and may be a new cause of male infertility. Several types of fertilization failure, associated with the sperm centrosome, are documented. These include: i) failure of the sperm to nucleate microtubules after sperm incorporation; ii) detachment of the sperm centrosome from the sperm head; and iii) failure in microtubule elongation after successful sperm aster formation. Although some types of severe male infertility can be overcome with the use of intracytoplasmic sperm injection (ICSI), men with defective sperm centrosomes are unlikely to benefit. The rate of sperm aster formation, size and organization during bovine fertilization has been used as a measurement of bovine sperm quality. Sperm from bulls which developed large highly organized sperm asters resulted in a higher rate of fertilization in vitro. The development of an assay using Xenopus laevis oocyte extracts has also been used to test centrosome function and sperm aster formation using sperm from men with questionable fertility. In general, these sperm were incapable of forming sperm asters and when used for in vitro fertilization, resulted typically in fertilization failure. These discoveries on the inheritance and function of the sperm centrosome have revealed a new cause of fertilization failure linked to male infertility which may not be circumvented using ICSI.