Method for measurement of the blood-brain barrier permeability in the perfused mouse brain: application to amyloid-beta peptide in wild type and Alzheimer's Tg2576 mice.
The role of transport exchanges of neuroactive solutes across the blood-brain barrier (BBB) is increasingly recognized. To take full advantage of genetically altered mouse models of neurodegenerative disorders for BBB transport studies, we adapted a brain perfusion technique to the mouse. During a carotid brain perfusion with a medium containing sheep red blood cells and mock plasma, the physiological parameters in the arterial inflow, regional cerebral blood flow (14C-iodoantipyrine autoradiography), ultrastructural integrity of the tissue, barrier to lanthanum, brain water content, energy metabolites and lactate levels remain unchanged. Amyloid-beta peptides (Abeta) were iodinated by lactoperoxidase method. Non-oxidized mono-iodinated Abeta monomers were separated by HPLC (as confirmed by MALDI-TOF spectrometry) and used in transport measurements. Transport of intact 125I-Abeta40 across the BBB was time- and concentration-dependent in contrast to negligible 14C-inulin uptake. In 5-6 months old Alzheimer's Tg2576 mice, Abeta40 BBB transport was increased by >eight-fold compared to age-matched littermate controls, and was mediated via the receptor for advanced glycation endproducts. We conclude the present arterial brain perfusion method provides strictly controlled environment in cerebral microcirculation suitable for examining transport of rapidly and slowly penetrating molecules across the BBB in normal and transgenic mice.
Department of Neurosurgery, Frank P. Smith Neurosurgical Research Laboratory, Center of Aging & Developmental Biology, University of Rochester Medical Center, 601 Elmwood Avenue, Box 670, Rochester, NY 14642, USA.
Barbra
B
LaRue
Elizabeth
E
Hogg
Abhay
A
Sagare
Suzana
S
Jovanovic
Lawrence
L
Maness
Calvin
C
Maurer
Rashid
R
Deane
Berislav V
BV
Zlokovic
D000255:Adenosine Triphosphate / Q000378:metabolism; D000544:Alzheimer Disease / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000378:metabolism / Q000493:pharmacokinetics*; D000818:Animals; D001345:Autoradiography / Q000379:methods; D001812:Blood-Brain Barrier / Q000378:metabolism* / Q000648:ultrastructure; D001923:Brain Chemistry / Q000187:drug effects / Q000502:physiology; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D002560:Cerebrovascular Circulation / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004195:Disease Models, Animal; D007839:Functional Laterality; D017127:Glycation End Products, Advanced; D006801:Humans; D007456:Iodine Isotopes / Q000493:pharmacokinetics; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic / Q000502:physiology; D008854:Microscopy, Electron / Q000379:methods; D010446:Peptide Fragments / Q000378:metabolism / Q000493:pharmacokinetics*; D010477:Perfusion / Q000379:methods; D010539:Permeability; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; D012039:Regional Blood Flow / Q000502:physiology
D016229:Amyloid beta-Peptides; D017127:Glycation End Products, Advanced; D007456:Iodine Isotopes; D010446:Peptide Fragments; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; C078592:amyloid beta-protein (1-40); C075222:amyloid beta-protein (1-42); D019344:Lactic Acid; D000255:Adenosine Triphosphate