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基于气相色谱-质谱联用代谢组学分析对与主观形态相关的山羊克隆胚胎培养基进行无创评估。

Non-invasive assessment of culture media from goat cloned embryos associated with subjective morphology by gas chromatography - mass spectroscopy-based metabolomic analysis.

作者信息

Zhang Yan-Li, Zhang Guo-Min, Jia Ruo-Xin, Wan Yong-Jie, Yang Hua, Sun Ling-Wei, Han Le, Wang Feng

机构信息

Jiangsu Livestock Embryo Engineering Laboratory, College of Animal Science, Nanjing Agricultural University, Nanjing, China.

出版信息

Anim Sci J. 2018 Jan;89(1):31-41. doi: 10.1111/asj.12885. Epub 2017 Aug 18.

Abstract

Pre-implantation embryo metabolism demonstrates distinctive characteristics associated with the development potential of embryos. We aim to determine if metabolic differences correlate with embryo morphology. In this study, gas chromatography - mass spectroscopy (GC-MS)-based metabolomics was used to assess the culture media of goat cloned embryos collected from high-quality (HQ) and low-quality (LQ) groups based on morphology. Expression levels of amino acid transport genes were further examined by quantitative real-time PCR. Results showed that the HQ group presented higher percentages of blastocysts compared with the LQ counterparts (P < 0.05). Metabolic differences were also present between HQ and LQ groups. The culture media of the HQ group showed lower levels of valin, lysine, glutamine, mannose and acetol, and higher levels of glucose, phytosphingosine and phosphate than those of the LQ group. Additionally, expression levels of amino acid transport genes SLC1A5 and SLC3A2 were significantly lower in the HQ group than the LQ group (P < 0.05, respectively). To our knowledge, this is the first report which uses GC-MS to detect metabolic differences in goat cloned embryo culture media. The biochemical profiles may help to select the most in vitro viable embryos.

摘要

植入前胚胎代谢表现出与胚胎发育潜力相关的独特特征。我们旨在确定代谢差异是否与胚胎形态相关。在本研究中,基于气相色谱 - 质谱联用(GC-MS)的代谢组学用于评估根据形态从高质量(HQ)和低质量(LQ)组收集的山羊克隆胚胎的培养基。通过定量实时PCR进一步检测氨基酸转运基因的表达水平。结果显示,与LQ组相比,HQ组的囊胚百分比更高(P <0.05)。HQ组和LQ组之间也存在代谢差异。HQ组的培养基中缬氨酸、赖氨酸、谷氨酰胺、甘露糖和丙酮醇的水平低于LQ组,而葡萄糖、植物鞘氨醇和磷酸盐的水平高于LQ组。此外,HQ组中氨基酸转运基因SLC1A5和SLC3A2的表达水平显著低于LQ组(分别为P <0.05)。据我们所知,这是第一份使用GC-MS检测山羊克隆胚胎培养基中代谢差异的报告。这些生化特征可能有助于选择体外活力最强的胚胎。

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