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Apparent Reduction of ADAM10 in Scrapie-Infected Cultured Cells and in the Brains of Scrapie-Infected Rodents

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Abstract

It has been described that A disintegrin and metalloproteinase (ADAM10) may involve in the physiopathology of prion diseases, but the direct molecular basis still remains unsolved. In this study, we confirmed that ADAM10 was able to cleave recombinant human prion protein in vitro. Using immunoprecipitation tests (IP) and immunofluorescent assays (IFA), reliable molecular interaction between the native cellular form of PrP (PrPC) and ADAM10 was observed not only in various cultured neuronal cell lines but also in brain homogenates of healthy hamsters and mice. Only mature ADAM10 (after removal of its prodomain) molecules showed the binding activity with the native PrPC. Remarkably more prion protein (PrP)-ADAM10 complexes were detected in the membrane fraction of cultured cells. In the scrapie-infected SMB cell model, the endogenous ADAM10 levels, especially the mature ADAM10, were significantly decreased in the fraction of cell membrane. IP and IFA tests of prion-infected SMB-S15 cells confirmed no detectable PrP-ADAM10 complex in the cellular lysates and PrP-ADAM10 co-localization on the cell surface. Furthermore, we demonstrated that the levels of ADAM10 in the brain homogenates of scrapie agent 263K-infected hamsters and agent ME7-infected mice were also almost diminished at the terminal stage, showing time-dependent decreases during the incubation period. Our data here provide the solid molecular basis for the endoproteolysis of ADAM10 on PrP molecules and interaction between ADAM10 and PrPC. Obvious loss of ADAM10 during prion infection in vitro and in vivo highlights that ADAM10 may play essential pathophysiological roles in prion replication and accumulation.

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Acknowledgement

This work was supported by the China Mega-Project for Infectious Disease (2011ZX10004-101, 2012ZX10004215), Chinese National Natural Science Foundation Grants (81100980), Young Scholar Scientific Research Foundation of China CDC (2012A102), and the SKLID Development Grant (2012SKLID102, 2011SKLID104 and 2011SKLID211).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changbai Road 155, Beijing 102206, People’s Republic of China

    Cao Chen, Yan Lv, Bao-Yun Zhang, Jin Zhang, Qi Shi, Jing Wang, Chan Tian, Chen Gao, Kang Xiao, Ke Ren, Wei Zhou & Xiao-Ping Dong

  2. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University, Hangzhou 310003, People’s Republic of China

    Cao Chen, Yan Lv, Bao-Yun Zhang, Jin Zhang, Qi Shi, Jing Wang, Chan Tian, Chen Gao, Kang Xiao, Ke Ren, Wei Zhou & Xiao-Ping Dong

  3. Chinese Academy of Sciences Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, People’s Republic of China

    Xiao-Ping Dong

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  1. Cao Chen
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Chen, C., Lv, Y., Zhang, BY. et al. Apparent Reduction of ADAM10 in Scrapie-Infected Cultured Cells and in the Brains of Scrapie-Infected Rodents. Mol Neurobiol 50, 875–887 (2014). https://doi.org/10.1007/s12035-014-8708-7

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  • DOI: https://doi.org/10.1007/s12035-014-8708-7

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