mRNA expression of leukaemia inhibitory factor (LIF) and its receptor subunits glycoprotein 130 and LIF-receptor-β in bovine embryos derived in vitro or in vivo
mRNA expression of leukaemia inhibitory factor (LIF) and its receptor subunits glycoprotein 130 and LIF-receptor-β in bovine embryos derived in vitro or in vivo
复制标题
DOI：
10.1093/molehr/4.10.957
复制
发表时间：
1998-10-01
影响因子：
4
通讯作者：
Niemann, H
中科院分区：
文献类型：
--
作者：
Eckert, J;Niemann, H
白血病抑制因子（I，IF）在小鼠细胞分化和植入过程中发挥重要作用，通过两个二聚化受体亚基糖蛋白130（gp 130）和LIF-受体β（LR-β）发挥其作用。本研究探讨了牛LIF（bLIF），gp 130和LR-β的mRNA表达在合并或个别牛卵母细胞和体外或体内超排卵后产生的胚胎。gp 130和LR-β的转录本可在1/3卵母细胞当量和bLIF转录本的两个胚胎当量中检测到，采用简单，快速和稳健的一步逆转录-聚合酶链反应（RT-PCR）方案，在逆转录过程中使用随机六聚体引发，并在PCR中进行60个扩增循环。在卵丘细胞和各种牛组织中也发现了所有的mRNA片段。虽然gp 130和LR-β mRNA存在于汇集的材料在体外从未成熟的卵母细胞到孵化的胚泡的整个胚胎发育过程中，但bLIF-转录物在未成熟的卵母细胞中不存在，从成熟的卵母细胞到16细胞阶段以及在胚泡中不一致地表达，这取决于胚胎批次。在桑椹胚中没有发现，但在孵化的囊胚中也存在。相比之下，在体内衍生的胚胎中没有发现bLIF，LR-β在桑椹胚到胚泡的转变中没有检测到，而从桑椹胚到孵化的胚泡观察到gp 130转录物。在单个胚胎中，体外和体内衍生胚胎的mRNA表达模式与合并材料中发现的相似。这些结果表明，在体外产生的胚胎中，特异性LIF-LIF受体系统的mRNA表达模式的扰动可能导致形成胚泡的细胞隔室的异常分化。卵丘细胞，经常被用作在牛体外胚胎生产中的共培养物中的支持因子，是以旁分泌和/或自分泌方式起作用的LIF-LIF受体系统的丰富来源。在牛植入前发育过程中LIF-LIF受体系统的生物学功能值得进一步研究。
Leukaemia inhibitory factor (I,IF) plays an essential role during cell differentiation and implantation in the mouse exerting its effects via the two dimerizing receptor subunits glycoprotein 130 (gp130) and LIF-receptor p (LR-beta). This study investigated the mRNA expression of bovine LIF (bLIF), gp130 and LR-beta in pooled or individual bovine oocytes and embryos generated in vitro or in vivo after superovulation. Transcripts of gp130 and LR-beta were detectable in as little material as 1/3 oocyte equivalent and bLIF transcripts in two embryo equivalents employing a simple, rapid and robust one-step reverse transcription-polymerase chain reaction (RT-PCR) protocol using random hexamer priming during reverse transcription and 60 amplification cycles in the PCR. All mRNA fragments were also found in cumulus cells and various bovine tissues. While gp130 and LR-beta mRNA were present in pooled material throughout embryo development in vitro from the immature oocyte to the hatched blastocyst, bLIf-transcripts were absent in immature oocytes, inconsistently expressed from the matured oocyte up to the 16-cell stage as well as in blastocysts dependent on embryo batch. It was not found in morulae, but again present in hatched blastocysts. In contrast, in in-vivo derived embryos no bLIF was found, LR-beta was not detected at the morula to blastocyst transition while gp130 transcripts were observed from the morula to the hatched blastocyst. In individual embryos the mRNA expression pattern was similar for both in-vitro and in-vivo derived embryos as found in pooled material. These results indicate perturbation of the mRNA expression pattern of the specific LIF-LIF-receptor system in embryos generated in vitro that could lead to abnormal differentiation of the cell compartments forming the blastocyst. Cumulus cells, frequently used as supportive factor in co-culture in bovine in-vitro embryo production, are a rich source of the LIF-LIF-receptor system acting in a paracrine and/or autocrine manner. The biological function of the LIF-LIF-receptor system during bovine preimplantation development warrants further investigation.