Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 159-164 |
| Number of pages | 6 |
| Journal | Journal of Neuroscience Methods |
| Volume | 83 |
| Issue number | 2 |
| DOIs | |
| State | Published - Sep 1 1998 |
Funding
This work was supported by NIH grants to L.G.E. (RO1 NS37738) and to S.D. (KO4 AI01240). Sophie X. Deng is student in the Medical Scientist Training Program funded by NIH Grant No. 2T32GM07356. The University of Rochester Medical Center is fully accredited by AALAC and all animals in this study were housed according to Animal Welfare Act guidelines.
Keywords
- Blood-brain barrier
- Luciferase
- Mannitol
ASJC Scopus subject areas
- General Neuroscience