1. ¹Department of Pharmacokinetics, Faculty of Pharmacy, INSERM U705, CNRS UMR 7157, Université Paris Descartes, Université Paris Diderot, Paris, France
2. ²CREMO-CHUL, Faculty of Pharmacy, Laval University, Laval, Quebec, Canada
3. ³Pharmacy, CHU Jean Verdier, AP-HP, Bondy, France

Correspondence: Dr S Cisternino, INSERM U705, CNRS UMR 7157, Université Paris Descartes, Université Paris Diderot, Hôpital F. Widal, 200, rue du Fbg St Denis, Paris 75010, France. E-mail: salvatore.cisternino@jvr.aphp.fr

⁴These authors equally contributed to this work.

Received 7 December 2007; Revised 27 February 2008; Accepted 19 March 2008; Published online 30 April 2008.

Abstract

The in situ mouse brain perfusion method for measuring blood–brain barrier permeability was adapted to assess transport of solutes at the blood–brain and blood–eye barriers. The procedure was checked with radiolabeled markers in oxygenated bicarbonate-buffered fluid infused for 30 to 120 secs via a carotid artery. Vascular flow estimated with diazepam was 2.2-fold lower in the eye than in the brain. The vascular volume and the integrity markers sucrose and inulin indicated that a perfusion flow rate of 2.5 mL/min preserved the physical integrity of these organs. However, the brain vasculature integrity was more sensitive to acute perfusion pressure than the eye vasculature. The functional capacities of blood barriers were assessed with D-glucose; its transport followed Michaelis–Menten kinetics with an apparent K_m of 7.6 mmol/L and a V_max of 23 mol/sec per g in the brain, and a K_m of 22.9 mmol/L and a V_max of 40 mol/sec per g in the eye. The transport of cholesterol to the brain and eye was significantly enhanced by adding the Abca1 inhibitor probucol, suggesting an Abca1-mediated efflux at the mouse brain and eye blood barriers. Thus in situ carotid perfusion is suitable for elucidating transport processes at the blood–brain and blood–eye barriers.