体外马卵泡卵母细胞在纯卵泡液加白细胞介素-1中的成熟及卵胞浆内单精子注射后的受精情况

In vitro equine oocyte maturation in pure follicular fluid plus interleukin-1 and fertilization following ICSI.

作者信息

Caillaud M, Dell'aquila M E, De Santis T, Nicassio M, Lacalandra G M, Goudet G, Gérard N

机构信息

INRA, CNRS, Université de Tours, Haras Nationaux, IFR 135, 37380 Nouzilly, France.

出版信息

Anim Reprod Sci. 2008 Jul;106(3-4):431-9. doi: 10.1016/j.anireprosci.2007.06.005. Epub 2007 Jun 13.

DOI:10.1016/j.anireprosci.2007.06.005

PMID:17628364

Abstract

The interleukin-1 (IL-1) system is thought to be involved in periovulatory events in the mare. Previous in vivo studies have demonstrated that IL-1beta induces oocyte maturation, but depresses the pregnancy rate 14 days after ovulation. To better understand the role of IL-1 in oocyte maturation and fertilization, the effects of IL-1 on the in vitro maturation rate of equine oocytes in pure follicular fluid were evaluated and fertilization rate assessed following intracytoplasmic sperm injection (ICSI). Oocytes collected from slaughterhouse ovaries were cultured in four different media for 30 h prior to fertilization. Two experiments were performed, each using three maturation media as the experimental treatments. Medium 1 was pure follicular fluid from subordinate follicles. Medium 2 was medium 1 plus 50 ng/ml recombinant human IL-1beta. Medium 3 was pure follicular fluid collected from mares administered crude equine gonadotropin (CEG). Medium 4 was medium 2 plus 50 ng/ml of recombinant human IL-1 receptor antagonist. Media 1, 2 and 3 were compared in experiment 1. In experiment 2, media 1, 2 and 4 were compared. After maturation, metaphase II oocytes were submitted to microinjection and assessed for signs of fertilization. In experiment 1, 101 oocytes were evaluated. The rate of polar body extrusion was 66, 51 and 68% and the proportions of normally fertilized oocytes after ICSI were 40, 18 and 38% for media 1, 2 and 3, respectively. In experiment 2, 122 oocytes were evaluated. The rate of polar body extrusion was 55, 48 and 42% and the proportions showing normal fertilization after ICSI were 14, 25 and 29% for media 1, 2 and 4, respectively. There was no positive effect of IL-1beta on maturation in both experiments, but the fertilization rate and percentage of embryos reaching four-cell were low in the presence of IL-1beta, indicating that this cytokine may interfere with fertilization and early embryo development.

摘要

白细胞介素 -1（IL -1）系统被认为参与母马排卵前后的一系列事件。此前的体内研究表明，IL -1β可诱导卵母细胞成熟，但会降低排卵后14天的妊娠率。为了更好地理解IL -1在卵母细胞成熟和受精过程中的作用，研究人员评估了IL -1对纯卵泡液中马属动物卵母细胞体外成熟率的影响，并在卵胞浆内单精子注射（ICSI）后评估了受精率。从屠宰场卵巢采集的卵母细胞在受精前于四种不同培养基中培养30小时。进行了两项实验，每项实验都使用三种成熟培养基作为实验处理。培养基1是来自次级卵泡的纯卵泡液。培养基2是培养基1加50 ng/ml重组人IL -1β。培养基3是从接受粗制马促性腺激素（CEG）的母马采集的纯卵泡液。培养基4是培养基2加50 ng/ml重组人IL -1受体拮抗剂。在实验1中比较了培养基1、2和3。在实验2中比较了培养基1、2和4。成熟后，将处于中期II的卵母细胞进行显微注射，并评估受精迹象。在实验1中，评估了101个卵母细胞。培养基1、2和3的极体排出率分别为66%、51%和68%，ICSI后正常受精的卵母细胞比例分别为40%、18%和38%。在实验2中，评估了122个卵母细胞。培养基1、2和4的极体排出率分别为55%、48%和42%，ICSI后显示正常受精的比例分别为14%、25%和29%。在两项实验中，IL -1β对成熟均无积极作用，但在存在IL -1β的情况下，受精率和发育到四细胞阶段的胚胎百分比都很低，这表明这种细胞因子可能会干扰受精和早期胚胎发育。