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

Katherine L. Youmans, Leon M. Tai, Takahisa Kanekiyo, W. Blaine Stine, Sara Claude Michon, Evelyn Nwabuisi-Heath, Arlene M. Manelli, Yifan Fu, Sean Riordan, William A. Eimer, Lester Binder, Guojun D Bu, Chunjiang Yu, Dean M. Hartley, Mary Jo Ladu

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

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 Sweor 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.

Original languageEnglish (US)
Article number8
JournalMolecular Neurodegeneration
Volume7
Issue number1
DOIs
StatePublished - 2012

Fingerprint

Antibodies
Brain
Transgenic Mice
Alzheimer Disease
Cathepsin D
Amyloid
Organelles
Culture Media
Anti-Idiotypic Antibodies
Cell Culture Techniques
Immunoglobulin G
Cell Membrane
Pathology
Neurons
Mutation

Keywords

  • 3xTg
  • 5xFAD
  • Alzheimer's disease
  • Antibody
  • APP
  • Intraneuronal
  • MOAB-2

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Clinical Neurology
  • Molecular Biology

Cite this

Youmans, K. L., Tai, L. M., Kanekiyo, T., Stine, W. B., Michon, S. C., Nwabuisi-Heath, E., ... Ladu, M. J. (2012). Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody. Molecular Neurodegeneration, 7(1), [8]. https://doi.org/10.1186/1750-1326-7-8

Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody. / Youmans, Katherine L.; Tai, Leon M.; Kanekiyo, Takahisa; Stine, W. Blaine; Michon, Sara Claude; Nwabuisi-Heath, Evelyn; Manelli, Arlene M.; Fu, Yifan; Riordan, Sean; Eimer, William A.; Binder, Lester; Bu, Guojun D; Yu, Chunjiang; Hartley, Dean M.; Ladu, Mary Jo.

In: Molecular Neurodegeneration, Vol. 7, No. 1, 8, 2012.

Research output: Contribution to journalArticle

Youmans, KL, Tai, LM, Kanekiyo, T, Stine, WB, Michon, SC, Nwabuisi-Heath, E, Manelli, AM, Fu, Y, Riordan, S, Eimer, WA, Binder, L, Bu, GD, Yu, C, Hartley, DM & Ladu, MJ 2012, 'Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody', Molecular Neurodegeneration, vol. 7, no. 1, 8. https://doi.org/10.1186/1750-1326-7-8
Youmans, Katherine L. ; Tai, Leon M. ; Kanekiyo, Takahisa ; Stine, W. Blaine ; Michon, Sara Claude ; Nwabuisi-Heath, Evelyn ; Manelli, Arlene M. ; Fu, Yifan ; Riordan, Sean ; Eimer, William A. ; Binder, Lester ; Bu, Guojun D ; Yu, Chunjiang ; Hartley, Dean M. ; Ladu, Mary Jo. / Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody. In: Molecular Neurodegeneration. 2012 ; Vol. 7, No. 1.
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AU - Youmans, Katherine L.

AU - Tai, Leon M.

AU - Kanekiyo, Takahisa

AU - Stine, W. Blaine

AU - Michon, Sara Claude

AU - Nwabuisi-Heath, Evelyn

AU - Manelli, Arlene M.

AU - Fu, Yifan

AU - Riordan, Sean

AU - Eimer, William A.

AU - Binder, Lester

AU - Bu, Guojun D

AU - Yu, Chunjiang

AU - Hartley, Dean M.

AU - Ladu, Mary Jo

PY - 2012

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N2 - 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 Sweor 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.

AB - 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 Sweor 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.

KW - 3xTg

KW - 5xFAD

KW - Aβ

KW - Alzheimer's disease

KW - Antibody

KW - APP

KW - Intraneuronal

KW - MOAB-2

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