Oocyte-Specific Modulation of Female Pronuclear Development in Mice

Abstract

Cell fusion experiments were undertaken to determine both whether oocytes possess, in a stage-specific manner, suppressors of pronuclear development and whether these factors could be used to modulate female pronuclear development. The fusion of telophase II eggs obtained 2 hr after activation (A2 eggs) to germinal vesicle (GV)-staged oocytes (GV x A2) suppresses female pronuclear development. The cytoplasm of GV x A2 heterokaryons contains, at 10 hr postfusion, a micropronucleus and a GV which is significantly larger (approximately x1.6) than normal. Fusion of early pronucleate eggs (3 hr postactivation, A3) to GV-staged oocytes (GV x A3) results in the formation of a retarded half-sized pronucleus and a slightly enlarged (approximately x1.2) GV at 10 hr postactivation. Normal pronuclear formation occurs when eggs at 4 hr postactivation (A4) are fused to GV oocytes (GV x A4). Although pronuclear size is suppressed in heterokaryons formed when GV-staged oocytes are fused to eggs activated 2-3 hr earlier (A2 or A3 stage), entry into S-phase and DNA synthesis is not inhibited even in the micropronuclei. Meiotic progression in the oocyte germinal vesicle is arrested after fusion to activated eggs throughout the period during which the pronucleus is undergoing G1 and S-phase. However, at the end of S-phase in the pronucleate partner, germinal vesicle breakdown occurs, cell cycle progression is accelerated, and both nuclei reach M-phase by 12 hr postactivation. That suppressors of pronuclear development are found only in GV (G2)-staged oocytes, and not in mitotic cells at the same cell cycle stage, was demonstrated by fusing G2-staged blastomeres to A2-staged eggs (G2 mitotic x A2). By contrast to the micropronuclei formed in GV x A2 heterokaryons, normal pronuclear development occurred in G2 mitotic x A2 heterokaryons. Our results show that suppressor activity, present only in GV oocytes, restricts female pronuclear development, but only in the first 3 hr after activation. We propose to use the ability to modulate either female (this study) or male pronuclear formation in studies (i) on imprinting and (ii) on the developmental consequences of early asynchrony between male and female pronuclear development.

IVM Advances for Early Antral Follicle-Enclosed Oocytes Coupling Reproductive Tissue Engineering to Inductive Influences of Human Chorionic Gonadotropin and Ovarian Surface Epithelium Coculture

“…However, techniques aimed at improving pronuclear manipulation and quality, as well as at optimizing the culture environment to help maintain embryo developmental potential, are still under investigation [ 122 ]. Some indications for the amelioration of embryo development might be obtained from oocyte pronuclear manipulation in mouse models where preliminary data postulate that abnormal pronuclear development could determine some detrimental epigenetic effects [ 123 , 124 ].…”