Polyspermically penetrated human zona-free eggs prepared from oocytes that had failed to be fertilized in an in-vitro fertilization programme were used. The pronuclear synthetic activity was evaluated by high-resolution autoradiography and correlated with the development of pronuclear structure. Incorporation of [3H]-thymidine, signalling the occurrence of a DNA synthetic phase, was only detected in structurally fully developed pronuclei previously shown to appear no sooner than 12 h after gamete union. However, [3H]adenosine was incorporated into very early pronuclei which had not yet completed the development of their nuclear envelopes and which first appeared about 4 h after sperm-egg fusion. In the absence of DNA synthesis (shown by the lack of thymidine incorporation), this early adenosine incorporation apparently reflects an early pronuclear RNA synthesis. Taken together, these results indicate that nucleic acid synthesis in human male pronuclei is tightly bound to the development of a corresponding pronuclear structure and that DNA synthesis, beginning about 12 h after fertilization, is preceded by a slight but evident RNA synthesis taking place during an early stage of human male pronuclear formation.

Centre for Reproductive Medicine Purkyne University Medical Faculty Brno Czechoslovakia

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