Dynamic status of lysosomal cathepsin in bovine oocytes and preimplantation embryos

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  • LI Jianye
    Laboratory of Animal Genetics and Reproduction, Graduate School of Agriculture, Hokkaido University, Hokkaido 060-8589, Japan
  • MAEJI Mana
    Laboratory of Animal Genetics and Reproduction, Graduate School of Agriculture, Hokkaido University, Hokkaido 060-8589, Japan
  • BALBOULA Ahmed Zaky
    Animal Sciences Research Center, College of Agriculture, Food & Natural Resources, University of Missouri, Columbia, MO 65211, USA
  • ABOELENAIN Mansour
    Department of Theriogenology, Faculty of Veterinary Medicine, Mansoura University, Mansoura 35516, Egypt
  • FUJII Takashi
    Animal Research Center, Hokkaido Research Organization, Hokkaido 081-0038, Japan
  • MORIYASU Satoru
    Animal Research Center, Hokkaido Research Organization, Hokkaido 081-0038, Japan
  • BAI Hanako
    Laboratory of Animal Genetics and Reproduction, Graduate School of Agriculture, Hokkaido University, Hokkaido 060-8589, Japan
  • KAWAHARA Manabu
    Laboratory of Animal Genetics and Reproduction, Graduate School of Agriculture, Hokkaido University, Hokkaido 060-8589, Japan
  • TAKAHASHI Masashi
    Graduate School of Global Food Resources(GSF), Hokkaido University, Hokkaido 060-0809, Japan Global Station for Food, Land and Water Resources, Global Institution for Collaborative Research and Education(GI-CoRE), Hokkaido University, Hokkaido, 060-0815, Japan

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<p> 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.</p>

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