Quantitative Atlas of Blood–Brain Barrier Transporters, Receptors, and Tight Junction Proteins in Rats and Common Marmoset

doi:10.1002/jps.23575

Journal of Pharmaceutical Sciences

Volume 102, Issue 9, September 2013, Pages 3343-3355

https://doi.org/10.1002/jps.23575 Get rights and content

ABSTRACT:

The purpose of this study was to determine the protein amounts of blood–brain barrier (BBB) permeability-related transporters, receptors, and tight junction proteins in Sprague Dawley and Wistar rats and common marmoset, and also to investigate inter-species and inter-strain differences across rodents and primates. Quantification of target proteins in isolated brain capillaries was conducted by liquid chromatography–tandem mass spectrometry-based quantitative targeted absolute proteomics, with in silico peptide selection. Most target proteins showed inter-rodent, inter-primate species, and inter-rat strain differences of less than 2-fold. Comparison of rat and human BBB showed that P-glycoprotein, multidrug resistance-associated protein 4, monocarboxylate transporter 1, l-type amino acid transporter, and organic anion transporter 3 exhibited differences of more than two-fold in protein abundance, whereas the amounts of breast cancer resistance protein, glucose transporter 1, and insulin receptor were similar in rat and human. In contrast, the differences between marmoset and human BBB were less than 2-fold for almost all measured proteins. Thus, the molecular basis of BBB functions may be similar in marmoset and human, whereas that of rats shows significant differences. The marmoset may be a good model to access in vivo human BBB permeability characteristics, as an alternative to rat and macaque monkey. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:3343–3355, 2013

Section snippets

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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: Additional Supporting Information may be found in the online version of this article. Supporting Information.

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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

ABSTRACT:

Section snippets

INTRODUCTION

Animals and Reagents

Quantitative Analysis of Membrane Proteins in SD and Wistar Rat Brain Capillaries

DISCUSSION

Acknowledgements

J Pharm Sci

Semin Fetal Neonatal Med

Regul Toxicol Pharmacol

NeuroRx

Epilepsy Res

Neuropharmacology

Pharmacol Res

Neuroscience

Neuroscience

Methods Enzymol

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J Proteomics

Am J Hum Genet

J Pharm Sci

J Pharm Sci

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J Pharmacol Exp Ther

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J Pharmacol Exp Ther

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The blood–brain barrier transport and cerebral distribution of guanidinoacetate in rats: Involvement of creatine and taurine transporters

J Neurochem

Characterization of SAGE Mdr1a (P-gp), Bcrp, and Mrp2 knockout rats using loperamide, paclitaxel, sulfasalazine, and carboxydichlorofluorescein pharmacokinetics

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Research Article: Pharmacokinetics, Pharmacodynamics and Drug Transport and Metabolism
Quantitative Atlas of Blood–Brain Barrier Transporters, Receptors, and Tight Junction Proteins in Rats and Common Marmoset