Caillaud Maud, Duchamp Guy, Gérard Nadine
Physiologie de la Reproduction et des Comportements, INRA-CNRS-Université de Tours-Haras Nationaux, IFR135, 37380 Nouzilly, France.
Reprod Biol Endocrinol. 2005 Jun 22;3:26. doi: 10.1186/1477-7827-3-26.
A growing body of evidence suggests that the interleukin-1 system is involved in periovulatory events. Previous work from our lab demonstrated that in the mare, interleukin-1beta (IL-1beta) increases the ovulatory rate of metaphase II oocytes. The present study was conducted to analyze in vivo the effect of IL-1 on oocyte cytoplasmic maturation, ovulation and pregnancy rate. In the present work, IL-1beta (experiment 1, n = 13; experiment 2, n = 25) and interleukin-1RA (IL-1RA; experiment 1, n = 25) were injected intrafollicularly by using the transvaginal ultrasound-guided injection method. Injections were performed on cyclic mares when the diameter of the growing dominant follicle reached 30-34 mm. In experiment 1, mares were inseminated the day of the treatment and all the other day until ovulation. The time of ovulation was determined and a pregnancy diagnosis was performed 14 days after ovulation of the injected follicle. In experiment 2, the cumulus-oocyte complex from each injected follicle was collected by transvaginal ultrasound-guided aspiration 38 h after the intrafollicular injection. Oocyte nuclear stage and oocyte cytoplasmic maturation were assessed by analyzing chromatin configuration, cortical granules migration and mitochondria distribution under a confocal microscope. The results from experiment 1 confirm that an intrafollicular injection of 1 microgram IL-1beta induces ovulation in the mare whereas IL-1RA has no effect at the dose used in the present study. Furthemore, we demonstrated, that in our experimental conditions, IL-1beta and IL-1RA induced a decrease in embryo development. Experiment 2 leads us to observe that IL-1beta is unable to induce cortical granules migration and remodelling of mitochondria, that commonly occurs during oocyte maturation, whereas it acts on nuclear maturation. This result may explain the decrease in embryo development we observed after IL-1beta intrafollicular injection. In conclusion, the present study tends to demonstrate that IL-1beta plays a role in the ovulatory process and may acts on oocyte maturation in the mare, but additional factors are required to complete equine oocyte cytoplasmic maturation to allow embryo development.
越来越多的证据表明,白细胞介素-1系统参与排卵前后的一系列事件。我们实验室之前的研究表明,在母马中,白细胞介素-1β(IL-1β)可提高中期II期卵母细胞的排卵率。本研究旨在分析IL-1对卵母细胞细胞质成熟、排卵和妊娠率的体内影响。在本研究中,采用经阴道超声引导注射法,将IL-1β(实验1,n = 13;实验2,n = 25)和白细胞介素-1受体拮抗剂(IL-1RA;实验1,n = 25)注射到卵泡内。当优势生长卵泡直径达到30 - 34毫米时,对处于发情周期的母马进行注射。在实验1中,母马在治疗当天及之后每天进行授精,直至排卵。确定排卵时间,并在注射卵泡排卵后14天进行妊娠诊断。在实验2中,卵泡内注射38小时后,通过经阴道超声引导抽吸收集每个注射卵泡的卵丘-卵母细胞复合体。通过共聚焦显微镜分析染色质构型、皮质颗粒迁移和线粒体分布,评估卵母细胞核阶段和卵母细胞细胞质成熟情况。实验1的结果证实,卵泡内注射1微克IL-1β可诱导母马排卵,而在本研究中所用剂量下,IL-1RA没有作用。此外,我们证明,在我们的实验条件下,IL-1β和IL-1RA会导致胚胎发育减少。实验2使我们观察到,IL-1β无法诱导皮质颗粒迁移和线粒体重塑,而这通常发生在卵母细胞成熟过程中,但其作用于核成熟。这一结果可能解释了我们在卵泡内注射IL-1β后观察到的胚胎发育减少现象。总之,本研究倾向于证明IL-1β在排卵过程中起作用,可能作用于母马的卵母细胞成熟,但需要其他因素来完成马卵母细胞的细胞质成熟以促进胚胎发育。