Identification of Site-Specific Stroke Biomarker Candidates by Laser Capture Microdissection and Labeled Reference PeptideReport as inadecuate


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


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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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Academic Editor: David Sheehan

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





Author: Tingting Lian †, Daixin Qu †, Xu Zhao, Lixia Yu and Bing Gao *

Source: http://mdpi.com/



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