Luciferase: a sensitive and quantitative probe for blood-brain barrier disruption.
A novel method for quantitative analysis of blood-brain barrier (BBB) disruption is described, using luciferase as a probe in a murine model system. Purified luciferase was delivered to mouse brain by osmotic BBB disruption with hypertonic mannitol; control animals received an intracarotid inoculation of saline prior to infusion of luciferase. Delivery of luciferase to brain tissue was then assessed by enzyme assay of tissue extracts, and by immunohistochemical staining. Luciferase activity in the brain of mannitol-treated animals was found to be significantly elevated (approx. sevenfold), when compared to activity in control (saline-treated) mice. This finding was confirmed by quantitative immunohistochemical staining of tissue sections, using a luciferase-specific antibody. These studies showed that there was an eight-fold elevation in the level of extravascular luciferase particles within the brain of mannitol-treated animals, as compared to controls. Taken together these data show that purified recombinant luciferase can be used as a sensitive probe, with which to study the integrity of the BBB.
Deng
S X
SX
Department of Medicine, University of Rochester Medical Center, NY 14642, USA.
Panahian
N
N
James
H
H
Gelbard
H A
HA
Federoff
H J
HJ
Dewhurst
S
S
Epstein
L G
LG
D000818:Animals; D000906:Antibodies; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology*; D001923:Brain Chemistry / Q000502:physiology; D004234:Diuretics, Osmotic / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D008099:Liver / Q000737:chemistry; D008156:Luciferases / Q000032:analysis / Q000276:immunology / Q000493:pharmacokinetics*; D008297:Male; D008353:Mannitol / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C
D000906:Antibodies; D004234:Diuretics, Osmotic; D008353:Mannitol; D008156:Luciferases