Regulation of gene expression in the bovine blastocyst by colony stimulating factor 2.
Regulation of gene expression in the bovine blastocyst by colony stimulating factor 2.
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通过刺激因子2调节牛胚泡中基因表达。
DOI：
10.1186/s13104-016-2038-y
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发表时间：
2016-04-29
影响因子：
1.8
通讯作者：
Hansen PJ
中科院分区：
文献类型：
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作者：
Ozawa M;Sakatani M;Dobbs KB;Kannampuzha-Francis J;Hansen PJ
集落刺激因子 2 可对植入前胚胎的功能产生多种影响，包括增加发育至囊胚阶段的潜力、减少细胞凋亡以及增强内细胞团 (ICM) 在培养后保持多能性的能力。当前实验的目的是鉴定牛胚泡的 ICM 和滋养外胚层 (TE) 中受 CSF2 调节的基因，目的是确定 CSF2 提高生存发育能力的可能分子途径。胚胎在体外产生，并在含有 10 ng/ml 重组牛 CSF2 或载体的无血清培养基中培养第 6 天至第 8 天。第 8 天收获囊胚，并通过磁激活细胞分选将 ICM 与 TE 分离。纯化RNA并用于制备扩增的cDNA，然后使用SOLiD 4.0系统对其进行高通量测序。每个处理分析三个扩增的 cDNA 池。使用 P < 0.05 和 >1.5 倍差异作为截止值，表达受 CSF2 调节的基因数量在 ICM 中为 945 个（242 个被 CSF2 上调，703 个下调），在 TE 中为 886 个（401 个被 CSF2 上调，485 个下调）。在这两种细胞类型中，只有 49 个基因以类似的方式受到 CSF2 的调节。受 ICM 调控的基因被过度表达的三个重要注释簇与膜信号传导有关。 ICM 中 CSF2 下调的基因在包括 ERK 和 AKT 信号通路在内的多个通路中出现过多。 TE 中含有过多受 CSF2 调节的基因的唯一重要注释簇是分泌蛋白或细胞外蛋白。此外，TE 中下调的基因在 TGFβ 和 Nanog 通路中过多。囊胚的分化使得受精后第 8 天，ICM 和 TE 对 CSF2 的反应不同。对 ICM 和 TE 中 CSF2 调节的基因的分析表明，CSF2 增强了两种细胞谱系的发育命运和功能。本文的在线版本 (doi:10.1186/s13104-016-2038-y) 包含补充材料，可供授权用户使用。
Colony stimulating factor 2 can have multiple effects on the function of the preimplantation embryo that include increased potential to develop to the blastocyst stage, reduced apoptosis, and enhanced ability of inner cell mass (ICM) to remain pluripotent after culture. The objective of the current experiment was to identify genes regulated by CSF2 in the ICM and trophectoderm (TE) of the bovine blastocyst with the goal of identifying possible molecular pathways by which CSF2 increases developmental competence for survival. Embryos were produced in vitro and cultured from Day 6 to 8 in serum-free medium containing 10 ng/ml recombinant bovine CSF2 or vehicle. Blastocysts were harvested at Day 8 and ICM separated from TE by magnetic-activated cell sorting. RNA was purified and used to prepare amplified cDNA, which was then subjected to high-throughput sequencing using the SOLiD 4.0 system. Three pools of amplified cDNA were analyzed per treatment. The number of genes whose expression was regulated by CSF2, using P < 0.05 and >1.5-fold difference as cut-offs, was 945 in the ICM (242 upregulated by CSF2 and 703 downregulated) and 886 in the TE (401 upregulated by CSF2 and 485 downregulated). Only 49 genes were regulated in a similar manner by CSF2 in both cell types. The three significant annotation clusters in which genes regulated by ICM were overrepresented were related to membrane signaling. Genes downregulated by CSF2 in ICM were overrepresented in several pathways including those for ERK and AKT signaling. The only significant annotation cluster containing an overrepresentation of genes regulated by CSF2 in TE was for secreted or extracellular proteins. In addition, genes downregulated in TE were overrepresented in TGFβ and Nanog pathways. Differentiation of the blastocyst is such that, by Day 8 after fertilization, the ICM and TE respond differently to CSF2. Analysis of the genes regulated by CSF2 in ICM and TE are suggestive that CSF2 reinforces developmental fate and function of both cell lineages. The online version of this article (doi:10.1186/s13104-016-2038-y) contains supplementary material, which is available to authorized users.