The distribution of Alz-50 immunoreactivity in the normal human brain

Rye, David B.; Leverenz, James B.; Greenberg, Sharon G.; Davies, Peter; Saper, Clifford B.

doi:10.1016/0306-4522(93)90567-y

Cited by 31 publications

(13 citation statements)

References 36 publications

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“…The Alz50 antibody recognizes an early stage conformational tau epitope that labels neurons undergoing early degenerative events associated with NFT formation but has also been shown to label neurons in normal brains [33–37]. Similar to 6E10 staining, the number of Alz50-ir neurons increased with age in all regions examined (Figure 4).…”

Section: Resultsmentioning

confidence: 75%

Staging of Alzheimer's Pathology in Triple Transgenic Mice: A Light and Electron Microscopic Analysis

Perez

Lagalwar

et al. 2010

International Journal of Alzheimer's Disease

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The age-related pathological cascade underlying intraneuronal tau formation in 3xTg-AD mice, which harbor the human APPSwe, PS1M126V , and TauP301L gene mutations, remains unclear. At 3 weeks of age, AT180, Alz50, MC1, AT8, and PHF-1 intraneuronal immunoreactivity appeared in the amygdala and hippocampus and at later ages in the cortex of 3xTg-AD mice. AT8 and PHF-1 staining was fixation dependent in young mutant mice. 6E10 staining was seen at all ages. Fluorescent immunomicroscopy revealed CA1 neurons dual stained for 6E10 and Alz50 and single Alz50 immunoreactive neurons in the subiculum at 3 weeks and continuing to 20 months. Although electron microscopy confirmed intraneuronal cytoplasmic Alz50, AT8, and 6E10 reaction product in younger 3xTg-AD mice, straight filaments appeared at 23 months of age in female mice. The present data suggest that other age-related biochemical mechanisms in addition to early intraneuronal accumulation of 6E10 and tau underlie the formation of tau filaments in 3xTg-AD mice.

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Section: Resultsmentioning

confidence: 75%

Staging of Alzheimer's Pathology in Triple Transgenic Mice: A Light and Electron Microscopic Analysis

Perez

Lagalwar

et al. 2010

International Journal of Alzheimer's Disease

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“…Any discrepancy may at least in part be related to the artificial antibody epitopes that are generated in the blots. It is well established that both the MC1 and PHF1 antibodies show greater specificity toward pathological tau on histological sections than in Western blots (Greenberg et al, 1992;Rye et al, 1993;Jicha et al, 1997;Weaver et al, 2000). For example, the MC1 antibody binds to recombinant tau on Western blots (Jicha et al, 1997), indicating its lack of specificity under those conditions as opposed to on fixed brain sections, on which it specifically binds to abnormal tau conformation (Weaver et al, 2000).…”

Section: Tau Pathologymentioning

confidence: 99%

mentioning

confidence: 99%

Immunotherapy Targeting Pathological Tau Conformers in a Tangle Mouse Model Reduces Brain Pathology with Associated Functional Improvements

Asuni

Boutajangout²,

Quartermain³

et al. 2007

J. Neurosci.

464

524

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Immunotherapies for various neurodegenerative diseases have recently emerged as a promising approach for clearing pathological protein conformers in these disorders. This type of treatment has not been assessed in models that develop neuronal tau aggregates as observed in frontotemporal dementia and Alzheimer's disease. Here, we present that active immunization with a phosphorylated tau epitope, in P301L tangle model mice, reduces aggregated tau in the brain and slows progression of the tangle-related behavioral phenotype. Females had more tau pathology than males but were also more receptive to the immunotherapy. The tau antibodies generated in these animals recognized pathological tau on brain sections. Performance on behavioral assays that require extensive motor coordination correlated with tau pathology in corresponding brain areas, and antibody levels against the immunogen correlated inversely with tau pathology. Interestingly, age-dependent autoantibodies that recognized recombinant tau protein but not the immunogen were detected in the P301L mice. To confirm that anti-tau antibodies could enter the brain and bind to pathological tau, FITC-tagged antibodies purified from a P301L mouse, with a high antibody titer against the immunogen, were injected into the carotid artery of P301L mice. These antibodies were subsequently detected within the brain and colocalized with PHF1 and MC1 antibodies that recognize pathological tau. Currently, no treatment is available for clearing tau aggregates. Our present findings may lead to a novel therapy targeting one of the major hallmarks of Alzheimer's disease and frontotemporal dementia.

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“…Indeed, Alz50 has been shown to react with tau proteins isolated from normal brain (15), recombinant sources (12), and PHFs (8) by Western analysis. Nonetheless, the ability of Alz50 to label distinct populations of neurons in normal human brain (16), fetal brain (17), and early stage neurofibrillary degeneration (4,18) suggests that Alz50 selectively recognizes a distinct subset of tau proteins.…”

mentioning

confidence: 99%

The Structural Basis of Monoclonal Antibody Alz50's Selectivity for Alzheimer's Disease Pathology

Carmel¹,

Mager

Binder

et al. 1996

Journal of Biological Chemistry

393

414

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The epitope on tau protein recognized by the monoclonal antibody Alz50 was defined through internal deletion mutagenesis and quantified by affinity measurements. The epitope is discontinuous and requires both a previously identified N-terminal segment and the microtubule binding region for efficient binding of Alz50. The interaction between these regions is consistent with an intramolecular reaction mechanism, suggesting that Alz50 binding depends on the conformation of individual tau monomers. The results suggest that tau adopts a distinct conformation when polymerized into filaments and that this conformation is recognized selectively by Alz50.Alz50 is an IgM-class monoclonal antibody that stains the fibrillar pathology (dystrophic neurites, neurofibrillary tangles, and neuropil threads) commonly observed in postmortem histological analysis of Alzheimer's disease (AD) 1 brain (1, 2). Because of these properties, it has emerged as an important tool for gauging the temporal and spatial severity of Alzheimer's disease pathology (3, 4). The major components of the fibrillar pathology are straight and paired helical filaments (PHF) (5), which themselves are comprised largely of hyperphosphorylated forms of the microtubule-associated protein tau (6 -10). Previous studies have shown that Alz50 reacts with tau and that its epitope is located at the N terminus (11-13) in a region conserved in all known splice variants of human tau (14). Indeed, Alz50 has been shown to react with tau proteins isolated from normal brain (15), recombinant sources (12), and PHFs (8) by Western analysis. Nonetheless, the ability of Alz50 to label distinct populations of neurons in normal human brain (16), fetal brain (17), and early stage neurofibrillary degeneration (4, 18) suggests that Alz50 selectively recognizes a distinct subset of tau proteins.To place the many observations on Alz50 immunocytochemistry into a structural context, we reinvestigated its epitope selectivity in vitro. The results suggest that individual tau monomers adopt a specific conformation preceding or during filament formation that is selectively recognized by Alz50. EXPERIMENTAL PROCEDURESMaterials-All monoclonal antibodies were prepared from supernatants of hybridoma cells grown in serum-free medium. Supernatants containing Alz50 were pooled, precipitated with 45% ammonium sulfate, resuspended in TBS (50 mM Tris HCl, pH 7.5, 50 mM NaCl, and 1 mM EGTA), and dialyzed twice against 100 volumes of TBS. The dialysate was clarified by centrifugation and redialyzed against 5 mM sodium phosphate, pH 7.5, to precipitate the IgM. The resultant fine precipitate was resuspended in S300 buffer (50 mM Tris HCl, pH 7.5, 700 mM NaCl, and 1 mM EGTA), dispersed with 30 strokes of a glass-Teflon homogenizer, and loaded onto a 400-ml (2.6 ϫ 100-cm) S300HR gel filtration column equilibrated and run in S300 buffer. The IgM fraction emerging in the void volume was pooled, dialyzed against storage buffer (10 mM HEPES, pH 7.4, 50% glycerol, and 150 mM NaCl), and stored at Ϫ20°C until use...

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The distribution of Alz-50 immunoreactivity in the normal human brain

Cited by 31 publications

References 36 publications

Staging of Alzheimer's Pathology in Triple Transgenic Mice: A Light and Electron Microscopic Analysis

Staging of Alzheimer's Pathology in Triple Transgenic Mice: A Light and Electron Microscopic Analysis

Immunotherapy Targeting Pathological Tau Conformers in a Tangle Mouse Model Reduces Brain Pathology with Associated Functional Improvements

The Structural Basis of Monoclonal Antibody Alz50's Selectivity for Alzheimer's Disease Pathology

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