A novel application of the fluorescent dye bis-ANS for labeling neurons in acute brain slices

Mózes, Emese; Hunya, Ákos; Tóth, Anikó M.; Ayaydin, Ferhan; Penke, Botond; Datki, Zsolt

doi:10.1016/j.brainresbull.2011.07.004

Cited by 6 publications

(6 citation statements)

References 15 publications

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“…Fu et al (2005) [14] revealed a chaperone-like activity for BisANS in preventing protein aggregation and in partially attenuating the heat-inactivation of enzymes. In our previous study we described a novel application of BisANS, which is capable of labelling damaged live/degenerated neurons and neuroblastoma cells [15]. Additionally, we used in vivo detection of exogenic peptide aggregates (e.g.…”

Section: Introductionmentioning

confidence: 99%

“…This non-covalent dye binds to non-specifically at multiple sites of many proteins [19] through its hydrophobic and electrostatic interactions [18]. The main advantages of BisANS are the high fluorescent intensity and great sensitivity, since it lacks an aspecific background resulted by different wavelength ranges of excitation (380–410 nm) and emission (510–530 nm) [15]. All these characteristics of the dye enable the application of the BisANS in a greater variety of protein research.…”

Section: Introductionmentioning

confidence: 99%

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Application of BisANS fluorescent dye for developing a novel protein assay

et al. 2019

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In many biology- and chemistry-related research fields and experiments the quantification of the peptide and/or protein concentration in samples are essential. Every research environment has unique requirements, e.g. metal ions, incubation times, photostability, pH, protease inhibitors, chelators, detergents, etc. A new protein assay may be adequate in different experiments beyond or instead of the well-known standard protocols (e.g. Qubit, Bradford or bicinchoninic acid) in related conceptions. Based on our previous studies, we developed a novel protein assay applying the 4,4′-Dianilino-1,1′-binaphthyl-5,5′-disulfonic acid dipotassium salt (BisANS) fluorescent dye. This molecule has several advantageous properties related to protein detection: good solubility in water, high photostability at adequate pH, quick interaction kinetics (within seconds) with proteins and no exclusionary sensitivity to the chelator, detergent and inhibitor ingredients. The protocol described in this work is highly sensitive in a large spectrum to detect protein (100-fold diluted samples) concentrations (from 0.28 up to more than 100 μg/mL). The BisANS protein assay is valid and applicable for quantification of the amount of protein in different biological and/or chemical samples.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Application of BisANS fluorescent dye for developing a novel protein assay

et al. 2019

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“…To investigate (n=5, well) the amount of protein in the animals ( Figure 2G), BisANS was applied parallelly with detecting the total amount of nucleic acid, where PI was used (Mozes et al, 2011) after 10 min incubation and washing. The extinction/emission were 405/520 nm for BisANS and 530/620 nm for PI, measured by a microplate reader (NOVOstar, BMG, Germany).…”

Section: Treatment and Monitoringmentioning

confidence: 99%

Neurodegeneration-related beta-amyloid as autocatabolism-attenuator in amicro-in vivosystem

Balázs

Gálik-Oláh

Gálik

et al. 2020

Preprint

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StatementIn our current work we investigated the neurodegeneration-related and well-known neurotoxic beta-amyloid aggregate as an attenuator of autocatabolism-based organ shrinkage in an invertebrate micro-in vivo system. AbstractInvestigation of human neurodegeneration-related aggregates of beta-amyloid 1-42 (Aβ42) on bdelloid rotifers is a novel interdisciplinary approach in life sciences. We reapplied an organ size-based in vivo monitoring system, exploring the autocatabolism-related alterations evoked by Aβ42, in a glucosesupplemented starvation model. The experientially easy-to-follow size reduction of the bilateral reproductive organ (germovitellaria) in fasted rotifers was rescued by Aβ42, serving as a nutrient sourceand peptide sequence-specific attenuator of the organ shrinkage phase and enhancer of the regenerative one including egg reproduction. Recovery of the germovitellaria was significant in comparison with the greatly shrunken form. In contrast to the well-known neurotoxic Aβ42 (except the bdelloids) with specific regulatory roles, the artificially designed scrambled version (random order of amino acids) was inefficient in autocatabolism attenuation, behaving as negative control. This native Aβ42-related modulation of the 'functionally reversible organ shrinkage' can be a potential experiential and supramolecular marker of autocatabolism in vivo.

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“…In the following paragraphs, I will briefly describe the findings we made about the neuron labelling properties of bis-ANS, that has first been shown in our laboratory (Mozes et al 2011).…”

Section: A Novel Application Of the Fluorescent Dye Bis-ans Labelling Neurons In Acute Brain Tissuementioning

confidence: 97%

“…In vitro studies on Aβ oligomerization and on acute hippocampal slices executed in our laboratory lead us to the discovery and further exploration of the neuron labelling properties of the oligomer-specific fluorescent dye, 4,4-bis-1-anilinonaphtalene-8-sulfonate (bis-ANS) (Mozes et al, 2011) Due to their sensitivity and versatility, extrinsic, non-covalent fluorescent dyes such as 8-Anilinonaphthalene-1-sulfonic acid (ANS), bis-ANS, Nile Red and Thioflavin T find widespread application in various fields of protein analysis such as characterizing folding intermediates or to detect aggregation and fibrillation (Reviewed by Hawe et al, 2008).…”

Section: Ex Vivo Fluorescent Imaging Of Neuronal Structures: a Novel Application Of Bis-ansmentioning

confidence: 99%

Innovative approach for the application of MTT, LDH and bis-ANS

Jánosi-Mózes¹

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The acute hippocampus slice-based models can serve as a feasible tool to gain deeper understanding on how pathological events in AD, in particular Aβ oligomerization, influence hippocampal functionality and therefore paving the way to develop and test related hypothesis but also to screen potential protective agents or validate results seen in vivo or predicted in silico. To overcome the shortcomings of the in vitro and in vivo methods, we developed a cost effective and simple apparatus for maintaining the viability of acute tissue slices, named: ExViS (Ex Vivo System; Universal Mini-Chamber Tube System for Acute Tissue Slices). The system allowed us to further develop methods that use acute (ex vivo) hippocampal slices to model AD-related patomechanisms. Over the course of my PhD studies we have developed the following techniques and ex vivo models based on the features of acute hippocampal slices: 1. Rapid, reliable and quantitative determination of Aβ1-42 toxicity in ageing (OGD) acute hippocampal slice model using MTT and LDH assays. 2. Quantitative determination of zinc-induced Aβ1-42 oligomerization toxicity in acute hippocampal slice model using MTT assay. 3. Fluorescent imaging of neurite cross-sections and detailed imaging of neurite structures in acute hippocampal slices via a novel application of bis-ANS and its co-staining variations. 4. Modelling the effect of Zn2+ released in glutamergic synaptic cleft in the hippocampus on Aβ1-42 aggregation and its consequences on cell viability and synaptic functionality, learning and memory (LTP).

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A novel application of the fluorescent dye bis-ANS for labeling neurons in acute brain slices

Cited by 6 publications

References 15 publications

Application of BisANS fluorescent dye for developing a novel protein assay

Application of BisANS fluorescent dye for developing a novel protein assay

Neurodegeneration-related beta-amyloid as autocatabolism-attenuator in amicro-in vivosystem

Innovative approach for the application of MTT, LDH and bis-ANS

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