Ultrastructural analysis of artificially activated rabbit eggs

Abstract

Ultrastructural investigations have been carried out on parthenogenetic rabbit eggs in an effort to elucidate events occurring during artificial activation and their similarity to processes of fertilization and embryogenesis. Rabbit eggs were artificially activated by culturing at 10°C for 24 hours followed by incubation at 37°C for 2 to 24 hours. Examination of eggs immediately after incubation at 10°C for 24 hours indicates that activation is initiated when the chromosomes coalesce to form a reticulum which is either surrounded completely by two parallel membranes or incompletely by cisternae of smooth endoplasmic reticulum. Aggregation of the chromosomes occurs as a result of a reduction in the number of microtubules making up the meiotic spindle. When cold treated ova are subsequently incubated at 37°C a nucleus is formed which moves centrad where it may participate in the cleavage of the egg. Formation of a second polar body and release of the contents of the cortical granules as reported for inseminated eggs was not found to be a part of activation of the egg by cold treatment. Approximately 95% of the ova cultured at 10°C for 24 hours followed by 37°C for 12 hours were activated, i.e., they possessed a nucleus or they had cleaved. Many of the activated eggs cultured for short periods at 37°C were structurally similar to fertilized ova, with further incubation fragmented eggs and abnormal multicellular stages predominated.