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Erratum published on 21 April 2017, see Int. J. Mol. Sci. 2017, 18(4), 881.

Open AccessArticle
Int. J. Mol. Sci. 2015, 16(6), 13427-13441; doi:10.3390/ijms160613427

Identification of Site-Specific Stroke Biomarker Candidates by Laser Capture Microdissection and Labeled Reference Peptide

School of Bioscience and Bioengineering, South China University of Technology, Higher Education Mega Center, Guangzhou 510006, China
These authors contributed equally to this work.
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Author to whom correspondence should be addressed.
Academic Editor: David Sheehan
Received: 4 April 2015 / Revised: 1 June 2015 / Accepted: 3 June 2015 / Published: 11 June 2015
(This article belongs to the Collection Advances in Proteomic Research)
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Abstract

The search to date for accurate protein biomarkers in acute ischemic stroke has taken into consideration the stage and/or the size of infarction, but has not accounted for the site of stroke. In the present study, multiple reaction monitoring using labeled reference peptide (LRP) following laser capture microdissection (LCM) is used to identify site-specific protein biomarker candidates. In middle cerebral artery occlusion (MCAO) rat models, both intact and infarcted brain tissue was collected by LCM, followed by on-film digestion and semi-quantification using triple-quadrupole mass spectrometry. Thirty-four unique peptides were detected for the verification of 12 proteins in both tissue homogenates and LCM-captured samples. Six insoluble proteins, including neurofilament light polypeptide (NEFL), alpha-internexin (INA), microtubule-associated protein 2 (MAP2), myelin basic protein (MBP), myelin proteolipid protein (PLP) and 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP), were found to be site-specific. Soluble proteins, such as neuron-specific enolase (NSE) and ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1), and some insoluble proteins, including neurofilament heavy polypeptide (NEFH), glial fibrillary acidic protein (GFAP), microtubule-associated protein tau (MAPT) and tubulin β-3 chain (TUBB3), were found to be evenly distributed in the brain. Therefore, we conclude that some insoluble protein biomarkers for stroke are site-specific, and would make excellent candidates for the design and analysis of relevant clinical studies in the future. View Full-Text
Keywords: laser capture microdissection (LCM); labeled reference peptide (LRP); blood–brain barrier (BBB); stroke; biomarker; multiple reaction monitoring (MRM) laser capture microdissection (LCM); labeled reference peptide (LRP); blood–brain barrier (BBB); stroke; biomarker; multiple reaction monitoring (MRM)
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Lian, T.; Qu, D.; Zhao, X.; Yu, L.; Gao, B. Identification of Site-Specific Stroke Biomarker Candidates by Laser Capture Microdissection and Labeled Reference Peptide. Int. J. Mol. Sci. 2015, 16, 13427-13441.

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