Optimizing Western Blots for the Detection of Endogenous α-Synuclein in the Enteric Nervous System.
Identifieur interne : 001881 ( Ncbi/Merge ); précédent : 001880; suivant : 001882Optimizing Western Blots for the Detection of Endogenous α-Synuclein in the Enteric Nervous System.
Auteurs : Cécile Preterre [France] ; Anne-Gaëlle Corbillé [France] ; Gaëlle Balloy [France] ; Franck Letournel [France] ; Michel Neunlist [France] ; Pascal Derkinderen [France]Source :
- Journal of Parkinson's disease [ 1877-718X ] ; 2015.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : alpha-Synuclein.
- chemical , metabolism : Biomarkers.
- diagnosis : Parkinson Disease.
- innervation : Intestine, Small.
- metabolism : Enteric Nervous System, Intestine, Small.
- methods : Blotting, Western.
- standards : Blotting, Western.
- Animals, Humans, Rats.
Abstract
Alpha-synuclein containing inclusions in neurons, the characteristic pathological lesions of Parkinson's disease (PD), are not limited to the central nervous system, but also affect the enteric nervous system (ENS). This suggests that the ENS offer some potential as a surrogate of central nervous system pathology and that it may represent an original source of biomarkers for PD. However, the usefulness of α-synuclein detection in gastrointestinal biopsies as a biomarker for PD is still unclear, as the different immunohistochemical methods employed to date have led to conflicting results.
DOI: 10.3233/JPD-150670
PubMed: 26599299
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 000431
- to stream PubMed, to step Curation: 000423
- to stream PubMed, to step Checkpoint: 000423
Links to Exploration step
pubmed:26599299Le document en format XML
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<term>Biomarkers (metabolism)</term>
<term>Blotting, Western (methods)</term>
<term>Blotting, Western (standards)</term>
<term>Enteric Nervous System (metabolism)</term>
<term>Humans</term>
<term>Intestine, Small (innervation)</term>
<term>Intestine, Small (metabolism)</term>
<term>Parkinson Disease (diagnosis)</term>
<term>Rats</term>
<term>alpha-Synuclein (analysis)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>alpha-Synuclein</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Biomarkers</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Parkinson Disease</term>
</keywords>
<keywords scheme="MESH" qualifier="innervation" xml:lang="en"><term>Intestine, Small</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Enteric Nervous System</term>
<term>Intestine, Small</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Blotting, Western</term>
</keywords>
<keywords scheme="MESH" qualifier="standards" xml:lang="en"><term>Blotting, Western</term>
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<front><div type="abstract" xml:lang="en">Alpha-synuclein containing inclusions in neurons, the characteristic pathological lesions of Parkinson's disease (PD), are not limited to the central nervous system, but also affect the enteric nervous system (ENS). This suggests that the ENS offer some potential as a surrogate of central nervous system pathology and that it may represent an original source of biomarkers for PD. However, the usefulness of α-synuclein detection in gastrointestinal biopsies as a biomarker for PD is still unclear, as the different immunohistochemical methods employed to date have led to conflicting results.</div>
</front>
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<Title>Journal of Parkinson's disease</Title>
<ISOAbbreviation>J Parkinsons Dis</ISOAbbreviation>
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<ArticleTitle>Optimizing Western Blots for the Detection of Endogenous α-Synuclein in the Enteric Nervous System.</ArticleTitle>
<Pagination><MedlinePgn>765-72</MedlinePgn>
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<Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">Alpha-synuclein containing inclusions in neurons, the characteristic pathological lesions of Parkinson's disease (PD), are not limited to the central nervous system, but also affect the enteric nervous system (ENS). This suggests that the ENS offer some potential as a surrogate of central nervous system pathology and that it may represent an original source of biomarkers for PD. However, the usefulness of α-synuclein detection in gastrointestinal biopsies as a biomarker for PD is still unclear, as the different immunohistochemical methods employed to date have led to conflicting results.</AbstractText>
<AbstractText Label="OBJECTIVE" NlmCategory="OBJECTIVE">Our aim is to propose an optimized immunoblotting method for the detection of endogenous α-synuclein in the healthy ENS that may be used to supplement the immunohistochemical analysis.</AbstractText>
<AbstractText Label="METHODS" NlmCategory="METHODS">Primary culture of rat ENS and homogenates of human small intestine were analyzed by Western Blot using seven different α-synuclein and phospho-α-synuclein antibodies along with two methods that increase α-synuclein retention on blot membranes, namely incubation of the membranes with paraformaldehyde (PFA) or treatment of samples with the crosslinker dithiobis[succinimidylpropionate] (DSP).</AbstractText>
<AbstractText Label="RESULTS" NlmCategory="RESULTS">A moderate improvement in the detection of endogenous enteric α-synuclein was observed following membrane fixation with PFA for only two of the seven antibodies we tested. Immunodetection of total and phosphorylated α-synuclein in the ENS was markedly improved when samples were treated with DSP, regardless of the antibody used.</AbstractText>
<AbstractText Label="CONCLUSIONS" NlmCategory="CONCLUSIONS">Our results demonstrate that the detection of α-synuclein in the gut by Western Blot can be optimized by using methods for enhanced membrane retention of the protein along with the appropriate antibody. Such an optimized protocol opens the way to the development of novel biomarkers for PD that will enable a quantification of α-synuclein in gastrointestinal biopsies.</AbstractText>
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</AffiliationInfo>
<AffiliationInfo><Affiliation>CHU Nantes, Department of Neurology, Nantes, F-44093, France.</Affiliation>
</AffiliationInfo>
<AffiliationInfo><Affiliation>CHU Nantes, Institut des Maladies de l'Appareil Digestif, Nantes, France.</Affiliation>
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<AffiliationInfo><Affiliation>CHU Nantes, Department of Neurology, Nantes, F-44093, France.</Affiliation>
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