Research Article: Pharmacokinetics, Pharmacodynamics and Drug Transport and MetabolismQuantitative Atlas of Blood–Brain Barrier Transporters, Receptors, and Tight Junction Proteins in Rats and Common Marmoset
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INTRODUCTION
Liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based quantitative-targeted absolute proteomics (QTAP) has provided a method to understand drug pharmacokinetics in terms of the protein expression amounts of functional proteins, such as transporters, receptors, and enzymes.1 We have found that the mRNA expression levels of CYP enzymes, except for CYP3A4, are not related to the metabolizing activity, and there is no correlation between mRNA and protein expression levels of transporters
Animals and Reagents
Adult SD rats (male, 8 weeks of age) and adult Wistar rats (male, 8 weeks of age) were purchased from CLEA Japan (Tokyo, Japan) (Table 1). Rats were maintained on a 12-h light/dark cycle in a temperature-controlled environment with free access to food and water; they were denied access to only food for 16 h before experiments. All experiments were approved by the Institutional Animal Care and Use Committee in Tohoku University, and were performed in accordance with the guidelines in Tohoku
Quantitative Analysis of Membrane Proteins in SD and Wistar Rat Brain Capillaries
The expression levels of 18 proteins, including 6 ATP-binding cassette (ABC) transporters, 6 solute carrier (SLC) transporters, 6 receptors, tight junction protein, and marker proteins, were analyzed in isolated brain capillaries of the two rat strains (Tables 2 and 4). Both strains showed similar protein expression amounts of 13 molecules in isolated capillaries within less than 2-fold difference (Table 2). The other five molecules were under the limit of quantification. All of the proteins
DISCUSSION
This is the first study to determine the protein expression amounts of transporters, receptors, and tight junction proteins at the BBB in brain capillaries of two strains of rat (SD and Wistar) and marmoset. Comparison with our previous results in mouse, cynomolgus monkey, and human indicates that there are pronounced differences in the protein amounts of those functional proteins between rodents and primates, although there are high degrees of similarity between the rat strains, between mouse
Acknowledgements
We thank Ms. A. Niitomi and N. Handa for secretarial assistance. This study was supported in part by Grant-in-Aids for Scientific Research (A) [KAKENHI: 24249011] from Japan Society for the Promotion of Science (JSPS) and grants for Development of Creative Technology Seeds Supporting Program for Creating University Ventures from Japan Science and Technology Agency (JST). Tetsuya Terasaki and Sumio Ohtsuki are full professors of Tohoku University and Kumamoto University, respectively, and are
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