A study of hetero‐specific sperm‐egg interactions in the rat, mouse, and deer mouse using in vitro fertilization and sperm injection

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Abstract
Hetero-Specific fertilization of zona-free eggs is used in these experiments as a tool to analyze the barriers to hybridization and to gain insight into the mechanisms of normal fertilization. When the zonae of rat eggs, which are a barrier to hetero-specific fertilization, are removed with pronase, the eggs can be fertilized by mouse sperm and the zygotes start to develop normally. A rat egg fertilized with mouse sperm completes meiosis and forms both male and female pronuclei. Chromosomes from both parents are found on the spindle at the metaphase stage of the first cleavage division. Under present culture conditions, embryos develop only to the two-cell stage, but this initial development of the hybrid is apparently normal. The question of whether sperm and egg membrane fusion is requisite for normal development is addressed by injecting sperm directly into the cytoplasm of unfertilized eggs. The injection of mouse sperm into rat eggs frequently leads to activation and formation of male and female pronuclei. The first cleavage division is indistinguishable from that following hetero-specific fetilization. Capacitated and uncapacitated sperm react alike when injected into eggs. Egg activation, however, is necessary for male pronucleus formation. Sperm from the deer mouse Peromyscus maniculatus bairdii, which are incapable of fertilizing even zona-free rat eggs, respond like mouse sperm when injected into rat eggs. These data indicate that sperm interactions with the egg cytoplasm are less species-specific than interactions at the egg surface. Furthermore, the normal surface interactions of sperm and eggs are not essential for the start of development.