Sx. Deng et al., LUCIFERASE - A SENSITIVE AND QUANTITATIVE PROBE FOR BLOOD-BRAIN-BARRIER DISRUPTION, Journal of neuroscience methods, 83(2), 1998, pp. 159-164
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 B
BB disruption with hypertonic mannitol; control animals received an in
tracarotid inoculation of saline prior to infusion of luciferase. Deli
very of luciferase to brain tissue was then assessed by enzyme assay o
f tissue extracts, and by immunohistochemical staining. Luciferase act
ivity in the brain of mannitol-treated animals was found to be signifi
cantly elevated (approx. sevenfold), when compared to activity in cont
rol (saline-treated) mice. This finding was confirmed by quantitative
immunohistochemical staining of tissue sections, using a luciferase-sp
ecific antibody. These studies showed that there was an eight-fold ele
vation in the level of extravascular luciferase particles within the b
rain of mannitol-treated animals, as compared to controls. Taken toget
her these data show that purified recombinant luciferase can be used a
s a sensitive probe, with which to study the integrity of the BBB. (C)
1998 Elsevier Science B.V. All rights reserved.