O-169 Conventional ovarian stimulation with gonadotropins depletes the developmental proteome of mouse oocytes, reducing their size and compromising their fetal yield
O-169 Conventional ovarian stimulation with gonadotropins depletes the developmental proteome of mouse oocytes, reducing their size and compromising their fetal yield
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O-169 使用促性腺激素进行常规卵巢刺激会消耗小鼠卵母细胞的发育蛋白质组，从而减小其大小并损害其胎儿产量
DOI：
10.1093/humrep/deab127.050
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发表时间：
2021
影响因子：
6.1
通讯作者：
Taher L
中科院分区：
文献类型：
--
作者：
Boiani M;Drexler HC;Fuellen G;Israel S;Makalowski W;Suzuki Y;Taher L
研究问题：促性腺激素常规促排卵是否会改变超数排卵小鼠卵母细胞的分子组成、大小和发育适应性？概要答案卵巢刺激干扰31%的发育蛋白质组和2%的转录组，产生较小的卵母细胞，形成囊胚，原始内胚层较少，胎儿产量减少。已知的是什么先前的小鼠研究旨在评估超排卵对卵母细胞和胚胎质量的影响，提供了不同的结果。在一些研究中观察到了异常，但在其他研究中没有观察到异常，并且与在取出进行体外培养之前在输卵管中花费的不同时间有关。虽然小鼠的自然卵巢周期持续4天，但常规刺激方案的时间跨度为2天。目前还没有全基因组的研究致力于超数排卵小鼠卵母细胞和衍生胚胎的整体基因表达，以确定基因产物是否积累到与自然周期相同的程度。研究设计，规模，持续时间大约。1100只雌性小鼠，一半注射连续马和人绒毛膜促性腺激素，eCG和hCG，以诱导超数排卵;另一半注射生理盐水作为对照。两组均与输精管切除或可育雄性交配，分别获得中期II和受精卵母细胞。将这些卵母细胞从输卵管中取出，并在体外随访至胚泡，或在手术转移至自然周期女性后在体内随访至足月。eCG+hCG）或自然排卵的胚胎测量直径，并且在受精后，将它们在KSOM（aa）中培养以收集植入前阶段用于分析（质谱法; RNA测序;免疫荧光用于滋养外胚层、上胚层和原始内胚层细胞的计数）。将4细胞期的胚胎移植到自然循环的雌性（8只/雌性，45只受体）。主要结果和机会的作用与自然排卵和间隔48小时（常规）或72小时的eCG-hCG刺激的超数排卵相比，超数排卵的卵母细胞的31%的蛋白质（893 / 2855）和2%的转录物（482 / 21784）受到影响（adj.P<0.05，Wilcoxon检验）。干扰蛋白质组的基因集富集分析返回最高术语“薄透明细胞”（ZP 1、ZP 2、ZP 3）和“异常内细胞团凋亡”（DAB 2、STAT 3）。显微镜测量证实，超数排卵卵母细胞的透明膜更薄（p = 0.077，Wilcoxon检验），沿着，直径更小（p < 0.0001，Wilcoxon检验），这导致囊胚缺乏原始内胚层（p < 0.013，Fisher精确检验）。由于893种不同表达的蛋白质中有529种被耗尽，我们认为卵巢刺激为蛋白质积累提供的时间不足。将eCG-hCG间隔从48 h（常规）增加到72 h恢复了卵母细胞的直径，并将其胎儿产量从25%提高到59%，而自然排卵为54%（每组15个胚胎移植）。相反，卵母细胞失去了他们的发育潜力的一部分，以显微操作辅助减少其体积。这项研究提供了证据的额外的新的影响，暴露于促性腺激素对小鼠卵母细胞的质量，其机制是介导的刺激生殖道，但通过时间依赖性的积累的蛋白质在卵ocytes.Limitations，原因谨慎这是一个动物模型研究的基础上一个小鼠品系。小鼠和..之间的卵巢刺激方案不同
Study questionDoes conventional ovarian stimulation with gonadotropins alter the molecular composition, size, and developmental fitness of superovulated mouse oocytes?Summary answerOvarian stimulation perturbs 31% of developmental proteome vs. 2% of transcriptome, yielding smaller oocytes that form blastocysts with less primitive endoderm and diminished fetal yield.What is known alreadyPrior mouse studies aimed to assess the impact of superovulation on oocyte and embryo quality provided variable results. Aberrations were observed in some studies but not in others, and were associated with a variable time spent in the oviduct until retrieval for in vitro culture. Although the natural ovarian cycle lasts 4 days in mice, the conventional stimulation protocol time spans 2 days. No genome-wide studies have been devoted yet to the global gene expression of superovulated mouse oocytes and derivative embryos, to determine if gene products accumulate to the same extent as in natural cycles.Study design, size, durationApprox. 1100 female mice were injected, half with serial equine and human chorionic gonadotropin, eCG and hCG, to induce superovulation; the other half were injected with saline as control. Both groups were mated to vasectomized or fertile males to obtain, respectively, metaphase II and fertilized oocytes. These were removed from the oviducts, and followed up in vitro to blastocyst, or in vivo to term after surgical transfer to naturally cycling females.Participants/materials, setting, methodsB6C3F1 oocytes (n= ∼16000) from superovulation (10 I.U. eCG+hCG) or natural ovulation were measured for diameter, and upon fertilization, they were cultured in KSOM(aa) to collect preimplantation stages for analyses (mass spectrometry; RNA sequencing; immunofluorescence for counting of trophectoderm, epiblast, and primitive endoderm cells). Embryos at the 4-cell stage were transplanted to naturally cycling females (8/female, 45 recipients). Results were compared between natural ovulation and superovulation with 48h (conventional) or 72h interval of eCG-hCG stimulation.Main results and the role of chancePreimplantation embryos of superovulated oocytes were affected in 31% of the proteins (893 / 2855) vs. 2% of the transcripts (482 / 21784), compared to natural counterparts (adj.P<0.05, Wilcoxon test). Gene set enrichment analysis of the perturbed proteome returned the top-terms ‘thin zona pellucida’ (ZP1, ZP2, ZP3) and ‘abnormal inner cell mass apoptosis’ (DAB2, STAT3). Microscope measurements verified a thinner zona pellucida (p = 0.077, Wilcoxon test) along with a smaller diameter (p < 0.0001, Wilcoxon test) of superovulated oocytes, which gave rise to blastocysts deficient in primitive endoderm (p < 0.013, Fisher’s exact test). Since 529 of the 893 differently expressed proteins were depleted, we considered that ovarian stimulation provided insufficient time for protein accumulation. Increasing the eCG-hCG interval from 48 h (conventional) to 72 h restored oocytes’ diameters, and improved their fetal output from 25% to 59%, compared to 54% of natural ovulation (15 embryo transfers per group). Conversely, oocytes lost part of their developmental potential to the micromanipulation-assisted reduction of their volume. This study provides evidence of an additional novel effect of exposure to gonadotropins on mouse oocyte quality, whose mechanism is mediated not by the stimulated genital tract, but by the time-dependent accumulation of proteins in oocytes.Limitations, reasons for cautionThis is an animal model study based on one mouse strain. Ovarian stimulation protocols differ between mice and …