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给予猪促黄体素的奶牛排卵前卵泡中颗粒细胞骨形态发生蛋白15、生长分化因子9及基因表达的变化

Alterations in bone morphogenetic protein 15, growth differentiation factor 9, and gene expression in granulosa cells in preovulatory follicles of dairy cows given porcine LH.

作者信息

Behrouzi Amir, Colazo Marcos Germán, Ambrose Divakar Justus

机构信息

Livestock Research Branch, Alberta Agriculture and Forestry, Edmonton, AB, T6H 5T6, Canada; Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, T6G 2P5, Canada.

Livestock Research Branch, Alberta Agriculture and Forestry, Edmonton, AB, T6H 5T6, Canada.

出版信息

Theriogenology. 2016 Apr 15;85(7):1249-57. doi: 10.1016/j.theriogenology.2015.12.007. Epub 2015 Dec 20.

Abstract

In a previous work, using porcine LH (pLH) in lieu of GnRH for synchronizing ovulation in dairy cows improved pregnancy rates without increasing plasma progesterone concentrations after ovulation. The LH profile is known to remain elevated above basal concentrations (≥1 ng/mL) for up to 20 hours in pLH-treated cows compared to less than 6 hours in GnRH-treated cows. Because LH triggers a cascade of signaling networks in the preovulatory follicle to promote final maturation and support oocyte competence, we hypothesized that dissimilar LH profiles will differentially regulate the intrafollicular factors and expression of downstream genes associated with improved oocyte competence. Specific objectives were to determine differences in the abundance of oocyte-secreted factors in the preovulatory follicular fluid and target genes in granulosa cells associated with oocyte competence, in response to exogenous porcine LH or GnRH-induced endogenous bovine LH exposure, in dairy cows. Follicular contents were aspirated by a transvaginal ultrasound-guided procedure from the preovulatory follicle of cyclic, nonlactating Holstein cows 21 ± 1 hour after administration of either pLH (25-mg) or GnRH (100-μg). Mature forms of bone morphogenetic protein 15, growth differentiation factor 9, and transforming growth factorβ1 were approximately 2-fold more abundant in pLH-treated cows which were exposed to an extended, low LH profile, than in GnRH-treated cows that had a short, high LH profile. The relative abundance of messenger RNA for cyclooxygenase-2, LH receptor, and progesterone receptor in granulosa cells, was about two-, eight-, and two-fold higher, respectively, in cows subjected to pLH than GnRH treatment. We infer that the improved pregnancy rate after pLH-induced ovulation reported previously, occurred through greater activation of intrafollicular transforming growth factor-β1 superfamily members, as these proteins promote cumulus expansion and oocyte competence.

摘要

在之前的一项研究中,使用猪促黄体素(pLH)代替促性腺激素释放激素(GnRH)来同步奶牛排卵,提高了妊娠率,且排卵后血浆孕酮浓度并未增加。已知在接受pLH处理的奶牛中,促黄体素水平在基础浓度(≥1 ng/mL)之上保持升高状态长达20小时,而在接受GnRH处理的奶牛中,这一水平维持升高的时间不到6小时。由于促黄体素会触发排卵前卵泡中的一系列信号网络,以促进最终成熟并支持卵母细胞的能力,我们推测不同的促黄体素水平会对卵泡内因子以及与卵母细胞能力改善相关的下游基因表达产生不同的调节作用。具体目标是确定在奶牛中,外源性猪促黄体素或GnRH诱导的内源性牛促黄体素暴露后,排卵前卵泡液中卵母细胞分泌因子的丰度以及颗粒细胞中与卵母细胞能力相关的靶基因的差异。在给予pLH(25 mg)或GnRH(100 μg)后21±1小时,通过经阴道超声引导程序从周期性、非泌乳荷斯坦奶牛的排卵前卵泡中抽吸卵泡内容物。在接受pLH处理、促黄体素水平呈延长的低水平状态的奶牛中,骨形态发生蛋白15、生长分化因子9和转化生长因子β1的成熟形式的丰度比接受GnRH处理、促黄体素水平呈短暂的高水平状态的奶牛高出约2倍。与接受GnRH处理的奶牛相比,接受pLH处理的奶牛颗粒细胞中环氧合酶-2、促黄体素受体和孕酮受体的信使核糖核酸相对丰度分别高出约2倍、8倍和2倍。我们推断,先前报道的pLH诱导排卵后妊娠率提高,是通过卵泡内转化生长因子-β1超家族成员的更大激活实现的,因为这些蛋白质可促进卵丘扩展和卵母细胞能力。

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