An electron microscopic study of sperm penetration into the rabbit egg after natural mating

The ultrastructure of fertilization has been studied in rabbit eggs recovered 11 to 15 hours after natural mating. Many sperm passing between the granulosa cells had undergone the acrosomal reaction, but this was not invariable, and, occasionally, intact sperm were present close to the zona pellucida. The cells of the corona radiata sometimes develop pseudopodial processes at the abovular surface and can ingest sperm after natural mating. The bulk of the content of the acrosome and the vesiculated elements formed during the acrosomal reaction, are lost before the sperm penetrates the zona pellucida, at which point the naked inner membrane of the acrosome is brought into intimate apposition with the zona. As the sperm cleaves a path through the zona pellucida, the posterior equatorial segment of the acrosome remains intact, and later persists as such in perivitelline sperm and quite possibly after incorporation of the sperm head into the vitellus. Sperm head entry into the vitellus is a two-fold process. The fertilizing sperm invariably fuses first with the vitelline membrane over the midposterior region of the head, whereas the rostral or acrosomal portion is drawn into the vitellus while encased by a flattened vesicle; this vesicle is comprised by the persistent inner membrane of the acrosome and externally by vitelline membrane sequestered from the egg surface. Soon after exposure to ooplasm, the sperm nucleus begins to decondense at a variable rate into a web of electron-dense strands; this process begins in the midposterior region, and then extends rostrally and caudally. At the same time the encasing membranes are reflected away from the anterior region of the nucleus, exposing subacrosomal material to the ooplasm. At this point the perforatorium remains, but this and the associated membranes are presumed to disintegrate eventually within the egg. After decondensation of the nucleus is complete, the faintly staining chromatin becomes enveloped by a series of compressed vesicles which together will form the porous limiting membrane of the male pronucleus. The last region to be incorporated is the sperm tail, the plasma membrane of which is lost as organelles of the tail pass into the ooplasm. During its incorporation, the midpiece engenders some reaction at the egg surface, and the mainpiece sometimes becomes fused with surface processes before it enters the body of the egg. The midpiece then commonly disintegrates, with dispersion of the mitochondrial sheath, whereas the mainpiece usually remains essentially intact until the time of syngamy or beyond.