Cellular and developmental biological aspects of bovine meiotic maturation, fertilization, and preimplantation embryogenesis in vitro

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Abstract
Cellular aspects of reinitiated meiosis, fertilization, and early preimplantation embryogenesis in the bovine species were examined under in vitro conditions. An analysis of the cytoplasmic distribution of mitochondria, lipid droplets and vesicles in over 5,000 living GV-stage oocytes, with subsequent examination by electron microscopy, indicated that the organization of cytoplasm was pleomorphic and that five distinct cellular phenotypes could be identified. Inspection of oocytes during the resumption of arrested meiosis in vivo and in vitro demonstrated that the subcellular organization of the bovine oocyte cytoplasm remains unchanged during maturation to metaphase II. The influence of culture conditions and media on the frequency of maturation, cytoplasmic organization, fertilizability, and early preimplantation embryogenesis was also determined. The findings indicate that meiotic maturation and fertilization in the bovine species can occur at high frequency under comparatively simple and defined conditions. However, the acquisition of developmental competence for fertilization, and the ability of the egg to develop progressively after fertilization appears to be related to the organization of the cytoplasm at the GV stage. The relationship between cytoplasmic organization and conditions of maturation, fertilization, presence or absence of cumulus cells, and the acquisition of developmental competence is discussed with respect to (1) cell biological aspects of mammalian oocyte maturation, (2) the potential influence of extrinsic factors (e. g., differential intrafollicular biochemistry and morphophysiology) on sub-cellular organization of the GV-stage oocyte, and (3) the finding that morphologically equivalent bovine embryos derived from the in vitro fertilization of in vitro-matured oocytes may be developmentally heterogeneous. The studies also revealed that nuclear and cytoplasmic aberrations which could preclude normal embryogenesis can develop shortly after fertilization. The significance of this finding with respect to cytoplasmic phenotype of the oocyte and conditions of maturation, fertilization, and early embryo culture is discussed.