Dynamic status of lysosomal cathepsin in bovine oocytes and preimplantation embryos

Jianye Li, Mana Maeji, A. Balboula, Mansour Aboelenain, T. Fujii, S. Moriyasu, H. Bai, M. Kawahara, Masashi Takahashi
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引用次数: 7

Abstract

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.
溶酶体组织蛋白酶在牛卵母细胞和着床前胚胎中的动态变化
溶酶体组织蛋白酶,特别是组织蛋白酶B (CTSB),在着床、妊娠和胚胎发育中起重要作用。然而,对溶酶体组织蛋白酶在牛卵母细胞和着床前胚胎中动态状态的相关机制知之甚少。在本研究中,我们研究了CTSB在牛卵母细胞和着床前胚胎中的基因表达、活性、免疫定位以及溶酶体活性的动态变化。对几个组织蛋白酶相关基因进行基因表达分析后发现,CTSB、CTSD和CTSZ的转录水平在中期II (MII)卵母细胞中最高,从8细胞胚胎期开始显著降低。CTSB活性在1细胞期和桑葚胚中显著升高。溶酶体活性在1细胞期和桑葚胚期也显著升高,这与CTSB活性一致。然而,CTSB的免疫定位没有显示出类似的CTSB和溶酶体活性模式。我们还发现CTSB转录本在滋养外胚层(TE)中的表达水平明显高于内细胞团(ICM),而CTSB的活性和免疫定位表现出相反的模式,即在ICM中显著高于TE。使用分离的ICM和TE进行相同的分析,证实了这些模式。本研究结果表明,溶酶体CTSB在胚胎发育和分化,特别是受精和分化过程中起着关键作用。
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