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Role of Adenosine Triphosphate, Active Mitochondria, and Microtubules in the Acquisition of Developmental Competence of Parthenogenetically Activated Pig Oocytes¹

Department of Anatomy of Domestic Animals, Faculty of Veterinary Medicine, University of Milan, 10-20133 Milano, Italy

The purpose of this work was to determine the mechanisms regulating the acquisition of cytoplasmic maturation and embryonic developmental competence in pig oocytes. The presence or the absence of porcine follicular fluid (pff; 25% or 0%) in the maturation medium was used as a means to achieve complete nuclear maturation accompanied or not accompanied by cytoplasmic maturation. ATP content, active mitochondria relocation, and microtubule distribution were analyzed at different times during in vitro maturation (IVM). While nuclear maturation did not differ among the two groups, parthenogenetic embryonic development was significantly higher (41.5%) in the 25% pff group than in the 0% pff group (19.0%) with blastocysts that had a significantly higher number of blastomeres (76.1 ± 6.3, and 47.2 ± 6.5, respectively). Oocyte ATP content increased significantly during IVM, but at the end of maturation no significant differences were observed between high- and low-competence oocytes. An extensive relocation of mitochondria to the inner cytoplasm during IVM together with the formation of a well-developed mesh of cytoplasmic microtubules was observed only in the high-competence oocyte group. However, no differences in the formation of microtubules associated with the meiotic spindles were observed between high- and low-competence groups. We conclude that low developmental competence is associated with the lack of a microtubule cytoplasmic network, which prevents correct relocation of mitochondria and is likely to reflect a more generally altered compartmentalization of the ooplasm. This can be independent from the formation of the microtubule machinery required for the completion of chromosome disjunctions and does not affect the overall ATP content.

ATP, early development, gamete biology, meiosis, microtubule, mitochondria, ovum, parthenogenesis, pig oocyte

² Correspondence: Fulvio Gandolfi, Istituto di Anatomia degli Animali Domestici, via Celoria, 10-20133 Milano, Italy. FAX: 39 025 031 7980; fulvio.gandolfi{at}unimi.it