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首页> 外文期刊>The Journal of Reproduction and Development >The gene expression pattern induced by high plating density in cultured bovine and buffalo granulosa cells might be regulated by specific miRNA species
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The gene expression pattern induced by high plating density in cultured bovine and buffalo granulosa cells might be regulated by specific miRNA species

机译：高培养密度在培养的牛和水牛颗粒细胞中诱导的基因表达模式可能受特定miRNA种类的调控

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Precise regulation of cell type-specific gene expression profiles precedes the profound morphological reorganization of somatic cell layers during folliculogenesis, ovulation and luteinization. Cell culture models are essential to the study of corresponding molecular mechanisms of gene regulation. In a recent study, it was shown that an increased cell plating density can largely change gene expression profiles of cultured bovine granulosa cells. In our present study, we comparatively analyzed cell plating density effects on cultured bovine and buffalo granulosa cells. Cells were isolated from small- to medium-sized follicles (2–6 mm) and cultured under serum-free conditions at different plating densities. The abundance of selected marker transcripts and associated miRNA candidates was determined by quantitative real-time RT-PCR. We found in both species that the abundance of CYP19A1 , CCNE1 and PCNA transcripts was remarkably lower at a high plating density, whereas VNN2 and RGS2 transcripts significantly increased. In contrast, putative regulators of CYP19A1 , miR-378, miR-106a and let-7f were significantly higher in both species or only in buffalo, respectively. Also miR-15a, a regulator of CCNE1 , was upregulated in both species. Thus, increased plating density induced similar changes of mRNA and miRNA expression in granulosa cells from buffalo and cattle. From these data, we conclude that specific miRNA species might be involved in the observed density-induced gene regulation.
机译：细胞类型特异性基因表达谱的精确调节先于卵泡形成，排卵和黄体化过程中体细胞层的深刻形态重组。细胞培养模型对于研究相应的基因调控分子机制至关重要。在最近的研究中，表明增加的细胞平板密度可以大大改变培养的牛颗粒细胞的基因表达谱。在我们目前的研究中，我们比较分析了细胞培养密度对培养的牛和水牛颗粒细胞的影响。从小至中等大小的卵泡（2–6 mm）中分离细胞，并在无血清条件下以不同的接种密度进行培养。通过定量实时RT-PCR确定所选标记转录物和相关miRNA候选物的丰度。我们发现在这两个物种中，CYP19A1，CCNE1和PCNA转录本的含量在高接种密度下均显着降低，而VNN2和RGS2转录本则显着增加。相反，推定的CYP19A1，miR-378，miR-106a和let-7f的调节剂在两个物种中或仅在水牛中显着更高。在两个物种中，CCR1的调节子miR-15a也被上调。因此，增加的接种密度诱导了来自水牛和牛的颗粒细胞中mRNA和miRNA表达的类似变化。从这些数据，我们得出结论，特定的miRNA物种可能与观察到的密度诱导的基因调控有关。

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