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Comparison of the MicroRNA Expression Profiles of Male and Female Avian Primordial Germ Cell Lines

机译:男性和女性禽原始生殖细胞系MicroRNA表达谱的比较

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摘要

Primordial germ cells (PGCs) are the precursors of adult germ cells, and among the embryonic stem-like cells in the bird embryo, only they can transmit the genetic information to the next generation. Despite the wide range of applications, very little is known about the mechanism that governs primordial germ cell self-renewal and differentiation. As a first step, we compared 12 newly established chicken PGC lines derived from two different chicken breeds, performing CCK-8 proliferation assay. All of the lines were derived from individual embryos. A significant difference was found among the lines. As microRNAs have been proved to play a key role in the maintenance of pluripotency and the cell cycle regulation of stem cells, we continued with a complex miRNA analysis. We could discover miRNAs expressing differently in PGC lines with high proliferation rate, compared to PGC lines with low proliferation rate. We found that gga-miR-2127 expresses differently in female and male cell lines. The microarray analysis also revealed high expression level of the gga-miR-302b-3p strand (member of the miR-302/367 cluster) in slowly proliferating PGC lines compared to the gga-miR-302b-5p strand. We confirmed that the inhibition of miR-302b-5p significantly increases the doubling time of the examined PGC lines. In conclusion, we found that gga-miR-181-5p, gga-miR-2127, and members of the gga-miR-302/367 cluster have a dominant role in the regulation of avian primordial germ cell proliferation.
机译:原始生殖细胞(PGC)是成年生殖细胞的前体,在鸟类胚胎的胚胎干样细胞中,只有它们才能将遗传信息传递给下一代。尽管应用范围很广,但对于控制原始生殖细胞自我更新和分化的机制知之甚少。第一步,我们比较了来自两个不同鸡品种的12个新建立的鸡PGC品系,进行了CCK-8增殖测定。所有品系均来自单个胚胎。发现各行之间存在显着差异。由于已证明microRNA在维持多能性和干细胞的细胞周期调控中起关键作用,因此我们继续进行复杂的miRNA分析。我们可以发现,与低增殖率的PGC系相比,miRNA在高增殖率的PGC系中表达不同。我们发现gga-miR-2127在雌性和雄性细胞系中表达不同。微阵列分析还显示,与gga-miR-302b-5p链相比,在缓慢增殖的PGC系中gga-miR-302b-3p链(miR-302 / 367簇的成员)的高表达水平。我们证实,miR-302b-5p的抑制作用显着增加了所检查PGC品系的倍增时间。总之,我们发现gga-miR-181-5p,gga-miR-2127和gga-miR-302 / 367簇的成员在禽原始生殖细胞增殖的调节中起主导作用。

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