{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T09:31:58Z","timestamp":1791192718863,"version":"4.1.0"},"reference-count":49,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2017,7,11]],"date-time":"2017-07-11T00:00:00Z","timestamp":1499731200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Servier Laboratories (Orl\u00e9ans, France)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Neurochemistry"],"published-print":{"date-parts":[[2017,8]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:label\/><jats:p>Liquid chromatography coupled to tandem mass spectrometry\u2010based targeted absolute protein quantification (in fmol of the analyte protein per \u03bcg of total protein) is employed for the molecular characterization of the blood\u2013brain barrier using isolated brain microvessels. Nevertheless, the heterogeneity of the sample regarding the levels of different cells co\u2010isolated within the microvessels and bovine serum albumin (<jats:styled-content style=\"fixed-case\">BSA<\/jats:styled-content>) contamination (from buffers) are not always evaluated. We developed an unlabeled targeted liquid chromatography coupled to tandem mass spectrometry method to survey the levels of endothelial cells (<jats:styled-content style=\"fixed-case\">EC<\/jats:styled-content>s), astrocytes, and pericytes, as well as <jats:styled-content style=\"fixed-case\">BSA<\/jats:styled-content> contaminant in rat cortical microvessels. Peptide peak identities were evaluated using a spectral library and chromatographic parameters. Sprague\u2013Dawley rat microvessels obtained on three different days were analyzed with this method complemented by an absolute quantification multiple reaction monitoring method for transporter proteins P\u2010gp, Bcrp, and Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup> <jats:styled-content style=\"fixed-case\">ATP<\/jats:styled-content>ase pump using stable isotope labeled peptides as internal standard. Inter\u2010day differences in the cell markers and <jats:styled-content style=\"fixed-case\">BSA<\/jats:styled-content> contamination were observed. Levels of cell markers correlated positively between each other. Then, the correlation between cell marker proteins and transporter proteins was evaluated to choose the best <jats:styled-content style=\"fixed-case\">EC<\/jats:styled-content> marker protein for protein quantification normalization. The membrane protein Pecam\u20101 showed a very high correlation with the <jats:styled-content style=\"fixed-case\">EC<\/jats:styled-content>\u2010specific transporter P\u2010gp (Pearson product\u2010moment correlation coefficient (<jats:italic>r<\/jats:italic>)\u00a0&gt;\u00a00.89) and moderate to high with Bcrp (<jats:italic>r<\/jats:italic>\u00a0\u2265\u00a00.77), that can be found also in pericytes and astrocytes. Therefore, Pecam\u20101 was selected as a marker for the normalization of the quantification of the proteins of endothelial cells.<\/jats:p><\/jats:sec><jats:sec><jats:label\/><jats:p>\n<jats:boxed-text content-type=\"graphic\" position=\"anchor\"><jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/jnc14095-fig-0004-m.png\"><jats:alt-text>image<\/jats:alt-text><\/jats:graphic><\/jats:boxed-text>\n<\/jats:p><\/jats:sec>","DOI":"10.1111\/jnc.14095","type":"journal-article","created":{"date-parts":[[2017,6,5]],"date-time":"2017-06-05T13:42:44Z","timestamp":1496670164000},"page":"597-609","source":"Crossref","is-referenced-by-count":17,"title":["Targeted unlabeled multiple reaction monitoring analysis of cell markers for the study of sample heterogeneity in isolated rat brain cortical microvessels"],"prefix":"10.1111","volume":"142","author":[{"given":"David","family":"Gomez\u2010Zepeda","sequence":"first","affiliation":[{"name":"Inserm UMR\u2010S 1144 Variabilit\u00e9 de la r\u00e9ponse aux psychotropes Paris France"},{"name":"Universit\u00e9 Paris Descartes Paris France"},{"name":"Universit\u00e9 Paris Diderot Paris France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catarina","family":"Chaves","sequence":"additional","affiliation":[{"name":"Inserm UMR\u2010S 1144 Variabilit\u00e9 de la r\u00e9ponse aux psychotropes Paris France"},{"name":"Universit\u00e9 Paris Descartes Paris France"},{"name":"Universit\u00e9 Paris Diderot Paris France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M\u00e9ryam","family":"Taghi","sequence":"additional","affiliation":[{"name":"Inserm UMR\u2010S 1144 Variabilit\u00e9 de la r\u00e9ponse aux psychotropes Paris France"},{"name":"Universit\u00e9 Paris Descartes Paris 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