Dell'Aquila M E, Masterson M, Maritato F, Hinrichs K
Department of Animal Production-Section of Reproduction, University of Bari, Valenzano, Bari, Italy.
Mol Reprod Dev. 2001 Sep;60(1):79-88. doi: 10.1002/mrd.1064.
There is a great variability in the success of horse oocyte maturation and fertilization among laboratories. This study was conducted to determine if the meiotic and developmental competence of horse oocytes could be dependent on the method of oocyte collection, i.e., aspiration of follicular fluid with a vacuum apparatus, or opening follicles and scraping the granulosa layer. Horse oocytes were recovered from abattoir ovaries by aspiration or scraping and classified as having compact (Cp), expanded (Ex), or partial (P) cumuli. In Experiment 1 (Part A in May and Part B in October), oocytes were fixed immediately after collection to assess whether the collection method influenced the initial chromatin configuration of oocytes. In Experiment 2, in vitro maturation rates of oocytes recovered by aspiration or scraping were compared. In Experiment 3, oocytes were matured in vitro and submitted to intracytoplasmic sperm injection (ICSI). Initial chromatin configuration differed according to collection method in that there was a significantly higher prevalence of diffuse chromatin within the germinal vesicle in oocytes recovered by scraping than in oocytes recovered by aspiration (29/87, 33% and 28/166, 17%, respectively; P < 0.01). Maturation of oocytes to metaphase II did not significantly differ between scraped and aspirated oocytes (56/101, 55.4 % vs. 65/106, 61.4%, respectively). The overall pronucleus formation rate after ICSI of oocytes recovered by scraping was not significantly different than that of oocytes recovered by aspiration (50/99, 52.6% vs. 50/85, 68.5 %, respectively); however, the rate of abnormal fertilization was significantly higher for oocytes collected by aspiration (14/73, 19% vs. 6/94, 6%, respectively; P <0.05). These results demonstrate that the collection method affects the population of recovered oocytes and may contribute to differences in results observed among laboratories working with horse oocytes.
不同实验室在马卵母细胞成熟和受精的成功率上存在很大差异。本研究旨在确定马卵母细胞的减数分裂和发育能力是否取决于卵母细胞采集方法，即使用真空装置抽吸卵泡液，还是打开卵泡并刮取颗粒层。通过抽吸或刮取从屠宰场的卵巢中回收马卵母细胞，并根据卵丘细胞的状态分为紧密型（Cp）、扩展型（Ex）或部分型（P）。在实验1（5月的A部分和10月的B部分）中，卵母细胞在采集后立即固定，以评估采集方法是否影响卵母细胞的初始染色质构型。在实验2中，比较了通过抽吸或刮取回收的卵母细胞的体外成熟率。在实验3中，卵母细胞在体外成熟后进行胞浆内单精子注射（ICSI）。初始染色质构型因采集方法而异，刮取回收的卵母细胞中，生发泡内弥散染色质的发生率显著高于抽吸回收的卵母细胞（分别为29/87，33%和28/166，17%；P<0.01）。刮取和抽吸的卵母细胞成熟至中期II的比例无显著差异（分别为56/101，55.4%和65/106，61.4%）。刮取回收的卵母细胞ICSI后总的原核形成率与抽吸回收的卵母细胞无显著差异（分别为50/99，52.6%和50/85，68.5%）；然而，抽吸采集的卵母细胞异常受精率显著更高（分别为14/73，19%和6/94，6%；P<0.05）。这些结果表明，采集方法会影响回收的卵母细胞群体，并可能导致在处理马卵母细胞的实验室之间观察到的结果差异。
