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Journal of Negative Results in BioMedicine

, 15:22

First Online: 16 December 2016Received: 27 June 2016Accepted: 14 November 2016DOI: 10.1186-s12952-016-0065-9

Cite this article as: Schaarschuch, A., Redies, C., Hertel, N. et al. J Negat Results BioMed 2016 15: 22. doi:10.1186-s12952-016-0065-9

Abstract

BackgroundAlzheimer’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.

MethodsIn this study, we used in situ hybridization to visualize cadherin expression in brains of two mouse models for AD APP-PS1 and APP23.

ResultsA 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.

ConclusionsWe 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.

KeywordsIn situ hybridization Alzheimer’s disease Cell adhesion Cadherin Plaques Unspecific binding APP-PS1 APP23 AbbreviationsADAlzheimer’s disease

AmyAmygdala

APPAmyloid-β precursor protein

asAntisense

AβAmyloid-β

CA1-3Subdivision 1–3 of the cornu ammonis

CdhCadherin

CxCerebral cortex

DGDentate gyrus

DIGDigoxigenin

FluoFluorescein

HipHippocampus

LVLateral ventricle

PcdhProtocadherin

sSense

TgTransgenic

ThThalamus

wtWild-type

Electronic supplementary materialThe 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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Autor: Anne Schaarschuch - Christoph Redies - Nicole Hertel - Molecular Anatomy and Dysfunction of Mouse Development Group

Fuente: https://link.springer.com/







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