Introducing Amylo-Glo, a novel fluorescent amyloid specific histochemical tracer especially suited for multiple labeling and large scale quantification studies

Schmued, Larry C.; Raymick, James; Tolleson, William H.; Sarkar, Sumit; Zhang, Yihong; Bell-Cohn, Ashlee

doi:10.1016/j.jneumeth.2012.05.019

Cited by 36 publications

(30 citation statements)

References 8 publications

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“…7) and the number and size of β-amyloid aggregations increases with longer culture. Amylo-Glo 45 , a fluorescent amyloid dye can also be used to detect Aβ deposits, particularly after 8-weeks differentiation (Supplementary Fig. 2).…”

Section: Anticipated Resultsmentioning

confidence: 99%

A 3D human neural cell culture system for modeling Alzheimer's disease

et al. 2015

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Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer’s disease (AD) because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel three-dimensional (3D) culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of β-amyloid and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix, and the analysis of AD pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of β-amyloid is observed after 6-weeks of differentiation followed by robust tau pathology after 10–14 weeks.

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Section: Anticipated Resultsmentioning

confidence: 99%

A 3D human neural cell culture system for modeling Alzheimer's disease

et al. 2015

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“…Sections were incubated in primary antibodies overnight followed by detection with appropriate Alexa Fluor-conjugated secondary antibody (Life Technologies) and coverslipped using Fluoromount-G with or without DAPI (Southern Biotech). Fibrillar amyloid was visualized using Amylo-Glo (Biosensis) diluted 1:100 in 0.01 M PBS (61).…”

Section: Methodsmentioning

confidence: 99%

The adaptive immune system restrains Alzheimer’s disease pathogenesis by modulating microglial function

Marsh

Abud

Lakatos

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

348

287

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The innate immune system is strongly implicated in the pathogenesis of Alzheimer's disease (AD). In contrast, the role of adaptive immunity in AD remains largely unknown. However, numerous clinical trials are testing vaccination strategies for AD, suggesting that T and B cells play a pivotal role in this disease. To test the hypothesis that adaptive immunity influences AD pathogenesis, we generated an immune-deficient AD mouse model that lacks T, B, and natural killer (NK) cells. The resulting "Rag-5xfAD" mice exhibit a greater than twofold increase in β-amyloid (Aβ) pathology. Gene expression analysis of the brain implicates altered innate and adaptive immune pathways, including changes in cytokine/chemokine signaling and decreased Ig-mediated processes. Neuroinflammation is also greatly exacerbated in Rag-5xfAD mice as indicated by a shift in microglial phenotype, increased cytokine production, and reduced phagocytic capacity. In contrast, immune-intact 5xfAD mice exhibit elevated levels of nonamyloid reactive IgGs in association with microglia, and treatment of Rag-5xfAD mice or microglial cells with preimmune IgG enhances Aβ clearance. Last, we performed bone marrow transplantation studies in Rag-5xfAD mice, revealing that replacement of these missing adaptive immune populations can dramatically reduce AD pathology. Taken together, these data strongly suggest that adaptive immune cell populations play an important role in restraining AD pathology. In contrast, depletion of B cells and their appropriate activation by T cells leads to a loss of adaptiveinnate immunity cross talk and accelerated disease progression.is the leading cause of age-related neurodegeneration, affecting over 5.2 million people in the United States alone (1). Pathologically, AD is characterized by two hallmark protein aggregates, amyloid-β (Aβ) plaques and neurofibrillary tangles, that are accompanied by neuroinflammation, including microgliosis, elevated cytokine production, and activation of complement pathways (2-5). Initially, microglia respond to and surround plaques, degrading Aβ by phagocytosis (for review, see refs. 6-8). However, chronic activation of these cells shift microglia to a more proinflammatory and less phagocytic state (9, 10). Although much of the data implicating microglia in AD has come from neuropathological investigation, recent genome-wide association studies have provided the first genetic evidence (to our knowledge) linking microglia dysfunction to AD, with the discovery of risk polymorphisms in several immune system genes: CR1, TREM2, CD33, HLA-DRB5, MS4A6A, and ABCA7 (8,(11)(12)(13)(14)(15).In contrast to the field's increasing understanding of the role of innate immunity in AD, comparatively little is known about whether the adaptive immune system might also influence AD. Those studies that have examined these peripheral populations have largely focused on questions about their potential as biomarkers or their role in active Aβ immunization (3, 16). However, the adaptive and innate immune systems rarely fu...

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“…Our previous studies indicate it has twice the brightness of Congo Red or Thioflavin S, when image analysis was done at higher magnification (20×) and 5–6 times brighter when imaged at lower magnification (4×) [53]. It is important to note that at such low magnification neither Congo-Red nor Thioflavin S labeling was visible for image analysis.…”

Section: Amylo-glomentioning

confidence: 98%

“…While Congo Red is excited by green light and emits red light, Thioflavin S is excited by blue and UV light and emits green light, Amylo-Glo is excited by UV light and emits a bright yellow light. Amylo-Glo appears to have a relatively sharp excitation profile and broad emission profile [53]. Another important aspect of this tracer, its unusual brightness and contrast that attributed to the compounds 5.46-fold increase in fluorescent emission intensity upon binding A- beta aggregates.…”

Section: Amylo-glomentioning

confidence: 99%

The Use of Recently Developed Histochemical Markers for Localizing Neurotoxicant Induced Regional Brain Pathologies

Sarkar

Raymick

Schmued

2014

Toxins

Self Cite

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Neuronal and vascular brain components are interrelated morphologically, physiologically and developmentally. Due to this close interrelationship, it is often difficult to understand the cause and effect relationship between neuronal vs. vascular dysfunction and pathology. This review will discuss four of the more promising recent developments for detecting vascular pathology, and will compare them with the labeling pattern seen with markers of glial and neuronal pathology; following exposure to well characterized neurotoxicants. To detect the vascular dysfunction in the brain, we recently developed a Fluoro-Turquoise gelatin conjugate (FT-gel), a fluorescent probe that helps to delineate between healthy vs. sclerotic vessels. Similarly, we have investigated the potential for Fluoro-Gold to label in vivo all the endothelial cells in the brain as they co-localize with RECA, an endothelial cell marker. We have also developed Amylo-Glo, a fluorescent tracer that can detect neurotoxic A-beta aggregates in the brain. In this article, we will discuss the potential use of these novel histochemical markers to study the neurotoxicant induced brain. We will also discuss neurovascular strategies that may offer novel therapeutic opportunities for neurodegenerative disorders.

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Introducing Amylo-Glo, a novel fluorescent amyloid specific histochemical tracer especially suited for multiple labeling and large scale quantification studies

Cited by 36 publications

References 8 publications

A 3D human neural cell culture system for modeling Alzheimer's disease

A 3D human neural cell culture system for modeling Alzheimer's disease

The adaptive immune system restrains Alzheimer’s disease pathogenesis by modulating microglial function

The Use of Recently Developed Histochemical Markers for Localizing Neurotoxicant Induced Regional Brain Pathologies

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