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初产兔非超排卵促性腺激素刺激后的卵巢反应和胚胎基因表达模式。

Ovarian response and embryo gene expression patterns after nonsuperovulatory gonadotropin stimulation in primiparous rabbits does.

机构信息

Department Producción Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain.

出版信息

Theriogenology. 2013 Jan 15;79(2):323-30. doi: 10.1016/j.theriogenology.2012.09.019. Epub 2012 Nov 13.

Abstract

Ovarian stimulation with equine chorionic gonadotropin (eCG) is largely used in animal reproductive technologies to provide a larger number of oocytes and embryos and to improve the reproductive outcome. However, the consequences of maternal treatment with eCG on embryo gene expression patterns are not widely studied. The aim of this work was to assess the ovarian response (preovulatory follicular population, oocyte maturation, ovulation rate, and serum steroid concentrations), the early embryo survival and gene expression patterns of a panel of quality-genes involved in glucose intake, oxidative stress, apoptosis, proliferation, implantation, and fetal growth in embryos of lactating rabbits treated with eCG. A total of 34 primiparous rabbit does (Oryctolagus cuniculus) were randomly distributed at Day 23 postpartum into a treatment group receiving a unique nonsuperovulatory dose (25 IU) of eCG (eCG group; N = 17 does); or a control group without eCG treatment previously to artificial insemination (control group; N = 17 does). After 48 hours, 8 does of each group were euthanized and their ovarian response was studied. The rest of animals were artificially inseminated and their ovulation was induced with a GnRH analogue. Embryos were recovered 3.5 days later. The oocytes retrieved for in vitro maturation showed no differences in metaphase II rate in both experimental groups, although oocyte cytoplasmic maturation, in terms of cortical granule migration rate, was improved in eCG-treated does (P < 0.05). The mean number of preovulatory follicles was similar between groups but the ovulation rate was significantly higher in eCG-treated does compared with does not stimulated (P < 0.05). No differences were found in serum estradiol and progesterone concentrations of does the day of oocyte and embryo recovery, respectively. However, progesterone:estradiol ratio was slightly increased in eCG group on embryo recovery day (P = 0.1). The percentage of embryos recovered at the blastocyst stage was also increased in eCG-treated does (P < 0.05), nevertheless, there were no differences in the gene expression patterns of candidate genes SLC2A4, IGF1R, IGF2R, SHC1-SHC, TP53, PTGS2, and PLAC8; except for the transcripts of SOD1 mRNA which were downregulated in eCG-derived embryos (P < 0.05). In conclusion, the administration of eCG improves ovulation rate, oocyte cytoplasmic maturation, and blastocyst formation in primiparous rabbit does inseminated on Day 25 postpartum. Although it seems not to influence the gene expression patterns studied, a lower antioxidant defense of embryos developed after the maternal eCG treatment is suggested.

摘要

用马绒毛膜促性腺激素(eCG)进行卵巢刺激在动物生殖技术中被广泛用于提供更多的卵母细胞和胚胎,并提高生殖结果。然而,母体用 eCG 治疗对胚胎基因表达模式的后果并没有得到广泛研究。本研究的目的是评估卵巢反应(排卵前卵泡群体、卵母细胞成熟、排卵率和血清类固醇浓度)、哺乳期母兔接受 eCG 处理后的早期胚胎存活率和一组与葡萄糖摄取、氧化应激、凋亡、增殖、植入和胎儿生长有关的质量基因的表达模式。总共 34 只初产母兔(Oryctolagus cuniculus)在产后第 23 天随机分为治疗组(n = 17 只),接受单次非超排卵剂量(25 IU)的 eCG;或对照组(n = 17 只),在人工授精前不接受 eCG 治疗。48 小时后,每组 8 只母兔被安乐死,研究其卵巢反应。其余动物接受人工授精,并使用 GnRH 类似物诱导排卵。3.5 天后回收胚胎。用于体外成熟的卵母细胞在两个实验组中的中期 II 率没有差异,尽管 eCG 处理的母兔的卵母细胞细胞质成熟度(以皮质颗粒迁移率表示)有所提高(P <0.05)。排卵前卵泡的平均数量在两组之间相似,但 eCG 处理的母兔的排卵率明显高于未刺激的母兔(P <0.05)。卵母细胞和胚胎回收当天母兔的血清雌二醇和孕酮浓度没有差异。然而,在胚胎回收日,eCG 组的孕酮:雌二醇比值略有升高(P = 0.1)。eCG 处理的母兔回收的囊胚阶段胚胎的百分比也增加(P <0.05),但候选基因 SLC2A4、IGF1R、IGF2R、SHC1-SHC、TP53、PTGS2 和 PLAC8 的基因表达模式没有差异;除了 SOD1 mRNA 的转录物在 eCG 衍生的胚胎中下调(P <0.05)。总之,在产后第 25 天接受人工授精的初产母兔中,eCG 的给药可提高排卵率、卵母细胞细胞质成熟度和囊胚形成。尽管它似乎不影响所研究的基因表达模式,但提示母体 eCG 处理后胚胎的抗氧化防御能力较低。

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