A functional in vitro model of rat blood-brain barrier for molecular analysis of efflux transporters

Nicolas Perrière, Salah Yousif, Sylvie Cazaubon, Nathalie Chaverot, Fanchon Bourasset, Salvatore Cisternino, Xavier Declèves, Satoko Hori, Tetsuya Terasaki, Maria Deli, Jean Michel Scherrmann, Jamal Temsamani, Françoise Roux, Pierre Olivier Couraud

研究成果: Article査読

119 被引用数 (Scopus)

抄録

Physiological studies of the blood-brain barrier (BBB) are often performed in rats. We describe the functional characterization of a reproducible in vitro model of the rat BBB and its validation for investigating mechanisms involved in BBB regulation. Puromycin-purified primary cultures of brain endothelial cells, co-cultured with astrocytes in the presence of hydrocortisone (HC) and cAMP, presented low sucrose permeability (≤ 0.1 × 10- 3 cm/min) and high transendothelial electrical resistance (≥ 270 Ω cm2). Expression of specific BBB markers and their transcripts was detected by immunostaining and RT-PCR, respectively: tight junction proteins (claudin-3 and -5, ZO-1 and occludin) and transporters (P-gp, Bcrp and Oatp-2). RT-PCR experiments demonstrated a role of treatment by astrocytes, HC and cAMP in regulation of the transcript level of tight junction proteins (claudin-5 and ZO-1) as well as transporters (Mdr1a, Mrp3, Mrp4, Bcrp, Glut-1), while transcript level of Mdr1b was significantly decreased. The functionality of efflux pumps (P-gp, Mrps and Bcrp) was demonstrated in the presence of specific inhibitors (PSC833, MK571 or Ko143, respectively) by (i) assessing the uptake of the common substrates rhodamine 123 and daunorubicin and (ii) evaluating apical to basolateral and basolateral to apical polarized transport of daunorubicin. In addition, a good correlation (R = 0.94) was obtained between the permeability coefficients of a series of compounds of various lipophilicity and their corresponding in vivo rodent blood-brain transfer coefficients. Taken together, our results provide compelling evidence that puromycin-purified rat brain endothelial cells constitute a reliable model of the rat BBB for physiological and pharmacological characterization of BBB transporters.

本文言語English
ページ(範囲)1-13
ページ数13
ジャーナルBrain research
1150
1
DOI
出版ステータスPublished - 2007 8 31

ASJC Scopus subject areas

  • 神経科学(全般)
  • 分子生物学
  • 臨床神経学
  • 発生生物学

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