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Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease

BACKGROUND: Alzheimer’s disease (AD) is characterized by the pathological deposition of amyloid-β (Aβ) protein-containing plaques. Microglia and astrocytes are commonly attracted to the plaques by an unknown mechanism that may involve cell adhesion. One cell adhesion family of proteins, the cadherin...

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Autores principales: Schaarschuch, Anne, Redies, Christoph, Hertel, Nicole
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5159973/
https://www.ncbi.nlm.nih.gov/pubmed/27978824
http://dx.doi.org/10.1186/s12952-016-0065-9
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author Schaarschuch, Anne
Redies, Christoph
Hertel, Nicole
author_facet Schaarschuch, Anne
Redies, Christoph
Hertel, Nicole
author_sort Schaarschuch, Anne
collection PubMed
description BACKGROUND: Alzheimer’s disease (AD) is characterized by the pathological deposition of amyloid-β (Aβ) protein-containing plaques. Microglia and astrocytes are commonly attracted to the plaques by an unknown mechanism that may involve cell adhesion. One cell adhesion family of proteins, the cadherins, are widely expressed in the central nervous system. Therefore, our study was designed to map the expression of cadherins in AD mouse brains. A particular focus was on plaques because diverse mRNA-species were found in plaques and their surrounding area in brains of AD patients. METHODS: In this study, we used in situ hybridization to visualize cadherin expression in brains of two mouse models for AD (APP/PS1 and APP23). RESULTS: A variable number of plaques was detected in transgenic brain sections, depending on the probe used. Our first impression was that the cadherin probes visualized specific mRNA expression in plaques and that endogenous staining was unaffected. However, control experiments revealed unspecific binding with sense probes. Further experiments with variations in probe length, probe sequence, molecular tag and experimental procedure lead us to conclude that cRNA probes bind generally and in an unspecific manner to plaques. CONCLUSIONS: We demonstrate unspecific binding of cRNA probes to plaques in two mouse models for AD. The widespread and general staining of the plaques prevented us from studying endogenous expression of cadherins in transgenic brain by in situ hybridization. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12952-016-0065-9) contains supplementary material, which is available to authorized users.
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spelling pubmed-51599732016-12-23 Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease Schaarschuch, Anne Redies, Christoph Hertel, Nicole J Negat Results Biomed Research BACKGROUND: Alzheimer’s disease (AD) is characterized by the pathological deposition of amyloid-β (Aβ) protein-containing plaques. Microglia and astrocytes are commonly attracted to the plaques by an unknown mechanism that may involve cell adhesion. One cell adhesion family of proteins, the cadherins, are widely expressed in the central nervous system. Therefore, our study was designed to map the expression of cadherins in AD mouse brains. A particular focus was on plaques because diverse mRNA-species were found in plaques and their surrounding area in brains of AD patients. METHODS: In this study, we used in situ hybridization to visualize cadherin expression in brains of two mouse models for AD (APP/PS1 and APP23). RESULTS: A variable number of plaques was detected in transgenic brain sections, depending on the probe used. Our first impression was that the cadherin probes visualized specific mRNA expression in plaques and that endogenous staining was unaffected. However, control experiments revealed unspecific binding with sense probes. Further experiments with variations in probe length, probe sequence, molecular tag and experimental procedure lead us to conclude that cRNA probes bind generally and in an unspecific manner to plaques. CONCLUSIONS: We demonstrate unspecific binding of cRNA probes to plaques in two mouse models for AD. The widespread and general staining of the plaques prevented us from studying endogenous expression of cadherins in transgenic brain by in situ hybridization. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12952-016-0065-9) contains supplementary material, which is available to authorized users. BioMed Central 2016-12-16 /pmc/articles/PMC5159973/ /pubmed/27978824 http://dx.doi.org/10.1186/s12952-016-0065-9 Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Schaarschuch, Anne
Redies, Christoph
Hertel, Nicole
Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease
title Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease
title_full Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease
title_fullStr Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease
title_full_unstemmed Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease
title_short Unspecific binding of cRNA probe to plaques in two mouse models for Alzheimer’s disease
title_sort unspecific binding of crna probe to plaques in two mouse models for alzheimer’s disease
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5159973/
https://www.ncbi.nlm.nih.gov/pubmed/27978824
http://dx.doi.org/10.1186/s12952-016-0065-9
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