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Effects of Newly Synthesized Recombinant Human Amyloid-β Complexes and Poly-Amyloid-β Fibers on Cell Apoptosis and Cognitive Decline

  • Park, Soojin (Graduate School of Medicine, Osaka University) ;
  • Huh, Jae-Won (National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Eom, Taekil (Subtropical/Tropical Organism Gene Bank, Jeju National University) ;
  • Park, Naeun (College of Pharmacy, Catholic University of Daegu) ;
  • Lee, Youngjeon (National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Ju-Sung (College of Applied Life Sciences, the Research Institute for Subtropical Agriculture and Biotechnology, Jeju National University) ;
  • Kim, Sun-Uk (National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Shim, Insop (Department of Science in Korean Medicine, Graduate School, College of Korean Medicine, Kyung Hee University) ;
  • Lee, Sang-Rae (National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Ekyune (College of Pharmacy, Catholic University of Daegu)
  • Received : 2017.07.05
  • Accepted : 2017.09.12
  • Published : 2017.11.28

Abstract

The main pathological hallmark of Alzheimer's disease is the deposition of amyloid-beta ($A{\beta}$) peptides in the brain. $A{\beta}$ has been widely used to mimic several aspects of Alzheimer's disease. However, several characteristics of amyloid-induced Alzheimer's disease pathology are not well established, especially in mice. The present study aimed to develop a new Alzheimer's disease model by investigating how $A{\beta}$ can be effectively aggregated using prokaryotes and eukaryotes. To express the $A{\beta}42$ complex in HEK293 cells, we cloned the $A{\beta}42$ region in a tandem repeat and incorporated the resulting construct into a eukaryotic expression vector. Following transfection into HEK293 cells via lipofection, cell viability assay and western blotting analysis revealed that exogenous $A{\beta}42$ can induce cell death and apoptosis. In addition, recombinant His-tagged $A{\beta}42$ was successfully expressed in Escherichia coli BL21 (DE3) and not only readily formed $A{\beta}$ complexes, but also inhibited the proliferation of SH-SY5Y cells and E. coli. For in vivo testing, recombinant His-tagged $A{\beta}42$ solution ($3{\mu}g/{\mu}l$ in $1{\times}PBS$ containing $1mM\;Ni^{2+}$) was injected stereotaxically into the left and right lateral ventricles of the brains of C57BL/6J mice (n = 8). Control mice were injected with $1{\times}PBS$ containing $1mM\;Ni^{2+}$ following the same procedure. Ten days after the sample injection, the Morris water maze test confirmed that exogenous $A{\beta}$ caused an increase in memory loss. These findings demonstrated that $Ni^{2+}$ is capable of complexing the 50-kDa amyloid and that intracerebroventricular injection of $A{\beta}42$ can lead to cognitive impairment, thereby providing improved Alzheimer's disease models.

Keywords

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