Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody

Molecular Neurodegeneration

Mary Jo LaDu⁴  Dean M Hartley¹  Chunjiang Yu⁴  Guojun Bu²  Lester Binder³  William A Eimer³  Sean Riordan³  Yifan Fu³  Arlene M Manelli⁵  Evelyn Nwabuisi-Heath⁴  Sara-Claude Michon¹  W Blaine Stine⁶  Takahisa Kanekiyo²  Leon M Tai⁴  Katherine L Youmans⁴ 

[1] Department of Neurological Sciences, Rush University Medical Center, Chicago IL 60612, USA;Department of Neuroscience, Mayo Clinic, Jacksonville FL 32224, USA;Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago IL 60611, USA;Department of Anatomy and Cell Biology, University of Illinois at Chicago, Chicago IL 60612, USA;Neuroscience Research, Global Pharmaceutical Research and Development, Abbott, 100 Abbott Park Road, Abbott Park IL 60064-6123, USA;CMC Program Management, Global Pharmaceutical Operations, Abbott Bioresearch Center, 100 Research Drive, Worcester MA 01605, USA

关键词: Alzheimer's disease;    Antibody;    5xFAD;    3xTg;    MOAB-2;    APP;    Aβ;    Intraneuronal;   

Others  :  863926
DOI  :  10.1186/1750-1326-7-8

 received in 2011-12-04, accepted in 2012-03-16,  发布年份 2012

【 摘 要 】

Background

The form(s) of amyloid-β peptide (Aβ) associated with the pathology characteristic of Alzheimer's disease (AD) remains unclear. In particular, the neurotoxicity of intraneuronal Aβ accumulation is an issue of considerable controversy; even the existence of Aβ deposits within neurons has recently been challenged by Winton and co-workers. These authors purport that it is actually intraneuronal APP that is being detected by antibodies thought to be specific for Aβ. To further address this issue, an anti-Aβ antibody was developed (MOAB-2) that specifically detects Aβ, but not APP. This antibody allows for the further evaluation of the early accumulation of intraneuronal Aβ in transgenic mice with increased levels of human Aβ in 5xFAD and 3xTg mice.

Results

MOAB-2 (mouse IgG_2b) is a pan-specific, high-titer antibody to Aβ residues 1-4 as demonstrated by biochemical and immunohistochemical analyses (IHC), particularly compared to 6E10 (a commonly used commercial antibody to Aβ residues 3-8). MOAB-2 did not detect APP or APP-CTFs in cell culture media/lysates (HEK-APP_Swe or HEK-APP_Swe/BACE1) or in brain homogenates from transgenic mice expressing 5 familial AD (FAD) mutation (5xFAD mice). Using IHC on 5xFAD brain tissue, MOAB-2 immunoreactivity co-localized with C-terminal antibodies specific for Aβ40 and Aβ42. MOAB-2 did not co-localize with either N- or C-terminal antibodies to APP. In addition, no MOAB-2-immunreactivity was observed in the brains of 5xFAD/BACE^-/- mice, although significant amounts of APP were detected by N- and C-terminal antibodies to APP, as well as by 6E10. In both 5xFAD and 3xTg mouse brain tissue, MOAB-2 co-localized with cathepsin-D, a marker for acidic organelles, further evidence for intraneuronal Aβ, distinct from Aβ associated with the cell membrane. MOAB-2 demonstrated strong intraneuronal and extra-cellular immunoreactivity in 5xFAD and 3xTg mouse brain tissues.

Conclusions

Both intraneuronal Aβ accumulation and extracellular Aβ deposition was demonstrated in 5xFAD mice and 3xTg mice with MOAB-2, an antibody that will help differentiate intracellular Aβ from APP. However, further investigation is required to determine whether a molecular mechanism links the presence of intraneuronal Aβ with neurotoxicity. As well, understanding the relevance of these observations to human AD patients is critical.

【 授权许可】

   
2012 Youmans et al; licensee BioMed Central Ltd.

【 图 表 】

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