Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody
<p>Abstract</p> <p>Background</p> <p>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 contro...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-03-01
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| Series: | Molecular Neurodegeneration |
| Subjects: | |
| Online Access: | http://www.molecularneurodegeneration.com/content/7/1/8 |
| _version_ | 1818491456799637504 |
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| author | 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 Yu Chunjiang Hartley Dean M LaDu Mary Jo |
| author_facet | 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 Yu Chunjiang Hartley Dean M LaDu Mary Jo |
| author_sort | Youmans Katherine L |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>MOAB-2 (mouse IgG<sub>2b</sub>) 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<sub>Swe </sub>or HEK-APP<sub>Swe</sub>/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<sup>-/- </sup>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.</p> <p>Conclusions</p> <p>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.</p> |
| first_indexed | 2024-12-10T17:31:04Z |
| format | Article |
| id | doaj.art-ae3fd868e21a44f0b15e44c06b021fa6 |
| institution | Directory Open Access Journal |
| issn | 1750-1326 |
| language | English |
| last_indexed | 2024-12-10T17:31:04Z |
| publishDate | 2012-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Neurodegeneration |
| spelling | doaj.art-ae3fd868e21a44f0b15e44c06b021fa62022-12-22T01:39:42ZengBMCMolecular Neurodegeneration1750-13262012-03-0171810.1186/1750-1326-7-8Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibodyYoumans Katherine LTai Leon MKanekiyo TakahisaStine W BlaineMichon Sara-ClaudeNwabuisi-Heath EvelynManelli Arlene MFu YifanRiordan SeanEimer William ABinder LesterBu GuojunYu ChunjiangHartley Dean MLaDu Mary Jo<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>MOAB-2 (mouse IgG<sub>2b</sub>) 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<sub>Swe </sub>or HEK-APP<sub>Swe</sub>/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<sup>-/- </sup>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.</p> <p>Conclusions</p> <p>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.</p>http://www.molecularneurodegeneration.com/content/7/1/8IntraneuronalAβAPPMOAB-23xTg5xFADAntibodyAlzheimer's disease |
| spellingShingle | 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 Yu Chunjiang Hartley Dean M LaDu Mary Jo Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody Molecular Neurodegeneration Intraneuronal Aβ APP MOAB-2 3xTg 5xFAD Antibody Alzheimer's disease |
| title | Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody |
| title_full | Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody |
| title_fullStr | Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody |
| title_full_unstemmed | Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody |
| title_short | Intraneuronal Aβ detection in 5xFAD mice by a new Aβ-specific antibody |
| title_sort | intraneuronal aβ detection in 5xfad mice by a new aβ specific antibody |
| topic | Intraneuronal Aβ APP MOAB-2 3xTg 5xFAD Antibody Alzheimer's disease |
| url | http://www.molecularneurodegeneration.com/content/7/1/8 |
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