In situ mouse carotid perfusion model: glucose and cholesterol transport in the eye and brain.
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 sec 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 micromol/sec per g in the brain, and a K(m) of 22.9 mmol/L and a V(max) of 40 micromol/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.
Cattelotte
Julie
J
Department of Pharmacokinetics, Faculty of Pharmacy, INSERM U705, CNRS UMR 7157, Université Paris Descartes, Paris, France.
André
Pascal
P
Ouellet
Mélissa
M
Bourasset
Fanchon
F
Scherrmann
Jean-Michel
JM
Cisternino
Salvatore
S
D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors / Q000378:metabolism; D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D001783:Blood Flow Velocity / Q000187:drug effects / Q000502:physiology; D001812:Blood-Brain Barrier / Q000502:physiology*; D001921:Brain / Q000098:blood supply / Q000378:metabolism*; D002339:Carotid Arteries; D002784:Cholesterol / Q000378:metabolism*; D005123:Eye / Q000098:blood supply / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D051379:Mice; D008955:Models, Cardiovascular; D010477:Perfusion; D011341:Probucol / Q000494:pharmacology
D064286:ATP Binding Cassette Transporter 1; D018528:ATP-Binding Cassette Transporters; D000924:Anticholesteremic Agents; D002784:Cholesterol; D005947:Glucose; D011341:Probucol