Kang H M, Kim K, Kwon H B, Cho W K
Department of Zoology, College of Natural Sciences, Seoul National University, Korea.
Mol Reprod Dev. 1990 Nov;27(3):191-9. doi: 10.1002/mrd.1080270303.
This study aims to elucidate gene expression of laminin and its role in expansion of the blastocyst during mouse early embryo-genesis. The gene expression of laminin, in particular the B1 subunit and the synthesis of laminin polypeptides, was examined during the expansion of blastocyst by a RNA-blot hybridization with 32P-labeled laminin B1 cDNA and immunoprecipitation followed by a SDS-PAGE, respectively. Laminin B1 transcript was actively expressed in the blastocyst stage of embryos. The gene expression of laminin B1 and the synthesis of laminin protein were also increased when blastocyst was expanded. Treatments of cAMP analogue, isobutylmethylxanthine, forskolin, and cholera toxin, which are known to stimulate the blastocyst expansion, increased laminin B1 transcript levels and synthesis of laminin polypeptides. Treatment with retinoic acid, a known regulator of laminin gene expression, not only increased the gene expression of laminin but stimulated the blastocoel expansion without a significant increase in intracellular cAMP levels. These results indicate that laminin gene expression may play an important role in the process of blastocyst expansion in the mouse preimplantation embryos.
本研究旨在阐明层粘连蛋白的基因表达及其在小鼠早期胚胎发育过程中对囊胚扩张的作用。在囊胚扩张过程中,分别通过用32P标记的层粘连蛋白B1 cDNA进行RNA印迹杂交以及免疫沉淀后进行SDS-PAGE,检测层粘连蛋白的基因表达,特别是B1亚基以及层粘连蛋白多肽的合成。层粘连蛋白B1转录本在胚胎的囊胚阶段活跃表达。当囊胚扩张时,层粘连蛋白B1的基因表达和层粘连蛋白的合成也增加。已知能刺激囊胚扩张的环磷酸腺苷类似物、异丁基甲基黄嘌呤、福斯可林和霍乱毒素处理,增加了层粘连蛋白B1转录本水平和层粘连蛋白多肽的合成。用视黄酸(一种已知的层粘连蛋白基因表达调节剂)处理,不仅增加了层粘连蛋白的基因表达,还刺激了囊胚腔扩张,而细胞内环磷酸腺苷水平没有显著增加。这些结果表明,层粘连蛋白基因表达可能在小鼠植入前胚胎的囊胚扩张过程中起重要作用。