Otoi T, Fujii M, Tanaka M, Ooka A, Suzuki T
Department of Veterinary Sciences Yamaguchi University, Japan.
Theriogenology. 2000 Sep 1;54(4):535-42. doi: 10.1016/s0093-691x(00)00368-x.
This study was conducted to determine the diameter of canine oocytes that are able to attain full meiotic competence and sperm penetration. Oocytes were collected from ovaries of bitches at various stages of the estrous cycle. Only healthy-looking cumulus-oocyte complexes were used for in vitro maturation, and were divided into four groups based on diameter: <100, 100 to <110, 110 to <120 and >120 microm. Following in vitro maturation or fertilization, oocytes were stained to assess nuclear maturation and penetration rates. The mean oocyte diameter was 108.5 +/- 0.4 microm. The oocytes displayed size-related ability to undergo meiotic maturation. After culture for 72 h, the rates of oocytes that remained at the germinal vesicle stage in the <110 microm groups were significantly higher (P<0.01) than in the > or = 110 microm groups. None of the oocytes <110 microm reached metaphase II (MU), but 4.9 and 21.5% of the oocytes that were greater than 110 and 120 microm, respectively, progressed to MII. After in vitro fertilization for 20 h, 10 to 25% of oocytes were penetrated by spermatozoa, but there were no clear relationships between oocyte diameter and penetration rates of the oocyte by sperm. In the <120 microm groups, sperm penetration was mostly found in oocytes arrested at the germinal vesicle stage. However, a total of eight oocytes > or = 120 microm in diameter were penetrated by spermatozoa, of which five oocytes reached MII. These results suggest that there is a clear relationship between oocyte diameter and meiotic competence, but no relationship between oocyte diameter and sperm penetration. Canine oocytes may have acquired meiotic competence once they reach at a diameter of 120 microm, but the oocytes may allow the entry of spermatozoa into the ooplasm irrespective of oocyte diameter.
本研究旨在确定能够获得完全减数分裂能力和精子穿透能力的犬类卵母细胞的直径。在发情周期的各个阶段从母犬卵巢中收集卵母细胞。仅使用外观健康的卵丘-卵母细胞复合体进行体外成熟,并根据直径分为四组:<100、100至<110、110至<120和>120微米。体外成熟或受精后,对卵母细胞进行染色以评估核成熟和穿透率。卵母细胞的平均直径为108.5±0.4微米。卵母细胞表现出与大小相关的减数分裂成熟能力。培养72小时后,<110微米组中处于生发泡期的卵母细胞比例显著高于(P<0.01)≥110微米组。<110微米的卵母细胞均未达到减数第二次分裂中期(MII),但分别有4.9%和21.5%的大于110微米和120微米的卵母细胞发育至MII期。体外受精20小时后,10%至25%的卵母细胞被精子穿透,但卵母细胞直径与精子对卵母细胞的穿透率之间没有明确关系。在<120微米组中,精子穿透主要发生在停滞于生发泡期的卵母细胞中。然而,共有8个直径≥120微米的卵母细胞被精子穿透,其中5个卵母细胞达到MII期。这些结果表明,卵母细胞直径与减数分裂能力之间存在明确关系,但卵母细胞直径与精子穿透之间没有关系。犬类卵母细胞一旦直径达到120微米可能已获得减数分裂能力,但无论卵母细胞直径如何,卵母细胞都可能允许精子进入卵质。