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Fluids and Barriers of the CNS
Published in: Fluids and Barriers of the CNS 1/2010

Open Access 01-12-2010 | Research

Characterization of immortalized choroid plexus epithelial cell lines for studies of transport processes across the blood-cerebrospinal fluid barrier

Authors: Juliane Kläs, Hartwig Wolburg, Tetsuya Terasaki, Gert Fricker, Valeska Reichel

Published in: Fluids and Barriers of the CNS | Issue 1/2010

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Abstract

Background

Two rodent choroid plexus (CP) epithelial cell lines, Z310 and TR-CSFB, were compared with primary rat CP epithelial cells and intact CP tissue with respect to transport protein expression, function and tight junction (TJ) formation.

Methods

For expression profiles of transporters and TJ proteins, qPCR and western blot analysis were used. Uptake assays were performed to study the functional activity of transporters and TJ formation was measured by trans-epithelial electrical resistance (TEER) and visualized by electron microscopy.

Results

The expression of known ATP-binding cassette (Abc) transporter and solute carrier (Slc) genes in CP was confirmed by qPCR. Primary cells and cell lines showed similar, but overall lower expression of Abc transporters and absent Slc expression when compared to intact tissue. Consistent with this Mrp1, Mrp4 and P-gp protein levels were higher in intact CP compared to cell lines. Functionality of P-gp and Mrp1 was confirmed by Calcein-AM and CMFDA uptake assays and studies using [3H]bis-POM-PMEA as a substrate indicated Mrp4 function. Cell lines showed low or absent TJ protein expression. After treatment of cell lines with corticosteroids, RNA expression of claudin1, 2 and 11 and occludin was elevated, as well as claudin1 and occludin protein expression. TJ formation was further investigated by freeze-fracture electron microscopy and only rarely observed. Increases in TJ particles with steroid treatment were not accompanied by an increase in transepithelial electrical resistance (TEER).

Conclusion

Taken together, immortalized cell lines may be a tool to study transport processes mediated by P-gp, Mrp1 or Mrp4, but overall expression of transport proteins and TJ formation do not reflect the situation in intact CP tissue.
Appendix
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Metadata
Title
Characterization of immortalized choroid plexus epithelial cell lines for studies of transport processes across the blood-cerebrospinal fluid barrier
Authors
Juliane Kläs
Hartwig Wolburg
Tetsuya Terasaki
Gert Fricker
Valeska Reichel
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Fluids and Barriers of the CNS / Issue 1/2010
Electronic ISSN: 2045-8118
DOI
https://doi.org/10.1186/1743-8454-7-11

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