Nucleolar ultrastructural changes occurring in vivo in bovine oocytes during follicular growth were analyzed by electron microscopy. The rates of in vitro incorporation of 3H‐uridine by oocytes of the same size class were evaluated by autoradiography. One to two large fibrillogranular, vacuolated nucleoli were present in oocytes from small to medium antral follicles 0.5–3 mm in diameter. These oocytes showed intense hnRNA and rRNA synthesis. The homogeneous, agranular nucleoli in oocytes from follicles 3–4 mm in diameter were composed of a compact fibrillar material. This morphological change was accompanied by an impairment of nucleolar transcriptional activity as well as by a decrease in hnRNA synthesis.
The aim of the present study was to investigate the effect of size of from which goat oocytes originate on their subsequent ability to be fertilized and to undergo early embryonic development in vitro. Nonatretic follicles larger than 2 mm in diameter were dissected and distributed into three groups according to size (small: 2-3 mm; medium: 3.1-5 mm; large: > 5 mm). Cumulus-oocyte complexes were isolated from the follicles and only those with a compact multilayered cumulus were selected for in vitro maturation. After maturation, 70%, 83% and 97% of oocytes from small, medium and large follicles, respectively, were at metaphase II. After in vitro fertilization, no significant difference was observed in the cleavage rate 40 h after insemination between oocytes from small (46%) and medium (55%) follicles, and between oocytes from large follicles (69%) and ovulated oocytes (75%). After in vitro culture, significantly more embryos from small follicles arrested before or at the 8-16 cell stage (84% compared with 53%, 45% and 39% of embryos from medium and large follicles and ovulated oocytes, respectively). The proportion of morulae and blastocysts obtained was 10% and 6% from small follicles, 35% and 12% from medium follicles, 29% and 26% from large follicles and 20% and 41% from ovulated oocytes. Oocytes from small and medium follicles yielded a significantly lower proportion of hatched blastocysts (0% and 3%, respectively) than did those from large follicles and from ovulated oocytes (15% and 34%, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
Pig oocytes were isolated from early antral follicles of different sizes and their abilities to resume and complete meiotic maturation in vitro were compared. After 24 h of culture, more than 80% of the oocytes from follicles 0.3-0.7 mm in diameter remained at the germinal vesicle stage, while 66, 94.3 and 100% oocytes from follicles 0.8-1.6, 1.7-2.2 and 3-5 mm in diameter, respectively, completed germinal vesicle breakdown. After 48 h of culture, 35% of the oocytes in the smallest follicle class progressed to prometaphase and only 4% to metaphase I. Of the oocytes from follicles 0.8-1.6 mm in diameter, 23% reached metaphase I and 17.3% metaphase II. About 50 and 76% of the oocytes from follicles 1.8-2.2 mm and 3-5 mm in diameter, respectively, extruded the first polar body. The ability to resume meiosis (i.e. to undergo germinal vesicle breakdown) is reached by porcine oocytes when they approach their full size in antral follicles greater than 0.8 mm in diameter and before they are capable of completing it (i.e. reaching metaphase II). The ability to complete meiotic maturation acquired in antral follicles of about 2 mm in diameter coincided with a significant decrease in the nucleolar transcriptional activity of the oocytes.
Fertilization abnormalities (premature chromosome condensation of spermatozoa (PCC), triploidy, haploidy) were analysed in order to determine their origin. PCC occurs in 9% of unfertilized oocytes and seems to be the consequence of a failure of oocyte activation, leading to the continuing presence of cytoplasmic chromosome-condensing factors, causing the sperm nucleus to undergo chromosome condensation prematurely. This anomaly appears to be related to incomplete nuclear and/or cytoplasmic maturation. Triploid zygotes (6.5% of fertilized oocytes) display an original type of division: half of them divide into 3 and 6 cells, whereas at the same time diploid zygotes divide into 2 and then 4 cells. A cytological study, using both antitubulin antibodies and Hoechst dye, allowed us to demonstrate that they divide into 3 cells by means of a tripolar spindle. Triploidy seems to be correlated with four of 16 clinical or biological parameters examined: semen origin (fresh or frozen), type of stimulation treatment, number of oocytes recovered and embryo morphology. Haploid eggs (1.6% of inseminated oocytes) result from parthenogenetic activation. A correlation was found between a high number of recovered oocytes and triploid zygotes, and the occurrence of oocyte activation. These data show that increasing follicular recruitment decreases the overall oocyte quality and maturity leading to an overall 9% with impaired fertilization.
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