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Dynamic status of lysosomal cathepsin in bovine oocytes and preimplantation embryos

机译:牛卵母细胞溶酶体组织蛋白酶素的动态状态及胚胎胚胎

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摘要

Lysosomal cathepsin, in particular cathepsin B (CTSB), plays an important role in implantation, pregnancy, and embryonic development. However, little is known about the mechanism related to the dynamic status of lysosomal cathepsins in bovine oocytes and preimplantation embryos. In the present study, we investigated the dynamics of gene expression, activity, and immunolocalization of CTSB, as well as the activities of lysosome, in bovine oocytes and preimplantation embryos. After gene expression analysis of several cathepsin-related genes, transcript levels of CTSB, CTSD and CTSZ were highest in Metaphase II (MII) oocytes followed by a significant decrease from the 8-cell embryo stage. Activity of CTSB showed a significant increase in 1-cell and morula stage embryos. Lysosomal activity was also significant higher in 1-cell and morula stages, which was consistent with CTSB activities. However, immunolocalization of CTSB did not show the similar pattern of CTSB and lysosomal activities. We also found significantly higher expression levels of CTSB transcript in the trophectoderm (TE) compared to inner cell mass (ICM), whereas activity and immunolocalization of CTSB showed an opposite pattern, i.e. significantly higher in ICM than TE. These patterns were confirmed by the same analysis using separated ICM and TE. Our results suggest that lysosomal CTSB has a pivotal role during embryonic development and differentiation, especially fertilization and the differentiation period.
机译:溶酶体组织蛋白酶,特别是组织蛋白酶B(CTSB),在植入,妊娠和胚胎发育中起重要作用。然而,关于与牛卵母细胞和预溶解胚胎的溶酶体组织蛋白酶的动态状态有关的机制很少。在本研究中,我们研究了CTSB的基因表达,活性和免疫悬垂性的动态,以及溶酶体,牛卵母细胞和预体胚胎的活性。在几个组织素相关基因的基因表达分析后,CTSB,CTSD和CTSZ的转录水平最高(MII)卵母细胞,然后从8细胞胚胎阶段显着降低。 CTSB的活性显示出1-细胞和割胶阶段胚胎的显着增加。溶酶体活性在1细胞和森拉阶段也显着较高,这与CTSB活性一致。然而,CTSB的免疫循环化并未显示出CTSB和溶酶体活性的类似模式。与内部电池质量(ICM)相比,我们还发现促杂细胞(TE)中CTSB转录物的显着更高的表达水平,而CTSB的活性和免疫循环化显示相反的图案,即ICM显着高于TE。通过使用分离的ICM和TE的相同的分析证实了这些模式。我们的研究结果表明,溶酶体CTSB在胚胎发育和分化期间具有关键作用,特别是施肥和分化期。

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