| APTS-labeled dextran ladder: a novel tool to characterize cell layer tightness. | |
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MedLine Citation:
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PMID: 16242879 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The aim of this work was the development of an easy manageable analytic system for describing tightness of cell layers in a molecular size dependent manner, which is more precise than currently used ones. Dextrans were labeled by reductive amination with fluorescent 1-aminopyrene-3,6,8-trisulfonate (APTS). This mixture, including internal standard diazepam, was used for transport studies, which were accomplished with an established transwell blood-brain barrier model culturing an immortalized porcine brain microvascular endothelial cell line (PBMEC/C1-2). Samples were analyzed by fluorescence measurements, capillary electrophoresis and RP-LC. Following this approach, a permeability pattern could be achieved including each single fraction from APTS, APTS-glucose to APTS-dextran consisting of 31 glucose units. Permeability coefficients were calculated and ranged from 16.38+/-3.79 microm/min for APTS to 6.07+/-1.23 microm/min for the APTS-dextran with 31 glucose units (diazepam: 67.97+/-7.32 microm/min). All in all, the developed APTS-dextran ladder is an useful tool to characterize cell layer tightness--especially to describe paracellular transport ways and leakiness status of the blood-brain barrier over time--applying a wide range from smaller to larger molecules at the same time in order to refine, e.g. TEER, sucrose or Evans blue measurements. |
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Authors:
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Winfried Neuhaus; Joanna Trzeciak; Regina Lauer; Bodo Lachmann; Christian R Noe |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2005-10-20 |
Journal Detail:
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Title: Journal of pharmaceutical and biomedical analysis Volume: 40 ISSN: 0731-7085 ISO Abbreviation: J Pharm Biomed Anal Publication Date: 2006 Mar |
Date Detail:
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Created Date: 2006-02-20 Completed Date: 2007-05-16 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8309336 Medline TA: J Pharm Biomed Anal Country: England |
Other Details:
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Languages: eng Pagination: 1035-9 Citation Subset: IM |
Affiliation:
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Department of Medicinal/Pharmaceutical Chemistry, University of Vienna, Pharmacy Center, Althanstrasse 14, A-1090 Vienna, Austria. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Blood-Brain Barrier / metabolism* Capillary Permeability Cell Line Dextrans* Electric Impedance Electrophoresis, Capillary Endothelial Cells / metabolism* Fluorescent Dyes Pyrenes* Swine Tight Junctions |
| Chemical | |
Reg. No./Substance:
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0/APTS-dextran; 0/Fluorescent Dyes; 0/Pyrenes; 9004-54-0/Dextrans |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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