PMID- 29553506
OWN - NLM
STAT- MEDLINE
DCOM- 20180622
LR  - 20200226
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 132
DP  - 2018 Feb 26
TI  - An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently 
      Labeled Tracers.
LID - 10.3791/57038 [doi]
LID - 57038
AB  - Blood-brain barrier (BBB) is a specialized barrier that protects the brain 
      microenvironment from toxins and pathogens in the circulation and maintains brain 
      homeostasis. The principal sites of the barrier are endothelial cells of the 
      brain capillaries whose barrier function results from tight intercellular 
      junctions and efflux transporters expressed on the plasma membrane. This function 
      is regulated by pericytes and astrocytes that together form the neurovascular 
      unit (NVU). Several neurological diseases such as stroke, Alzheimer's disease 
      (AD), brain tumors are associated with an impaired BBB function. Assessment of 
      the BBB permeability is therefore crucial in evaluating the severity of the 
      neurological disease and the success of the treatment strategies employed. We 
      present here a simple yet robust permeability assay that have been successfully 
      applied to several mouse models both, genetic and experimental. The method is 
      highly quantitative and objective in comparison to the tracer fluorescence 
      analysis by microscopy that is commonly applied. In this method, mice are 
      injected intraperitoneally with a mix of aqueous inert fluorescent tracers 
      followed by anesthetizing the mice. Cardiac perfusion of the animals is performed 
      prior to harvesting brain, kidneys or other organs. Organs are homogenized and 
      centrifuged followed by fluorescence measurement from the supernatant. Blood 
      drawn from the cardiac puncture just before perfusion serves for normalization 
      purpose to the vascular compartment. The tissue fluorescence is normalized to the 
      wet weight and serum fluorescence to obtain a quantitative tracer permeability 
      index. For additional confirmation, the contralateral hemi-brain preserved for 
      immunohistochemistry can be utilized for tracer fluorescence visualization 
      purposes.
FAU - Devraj, Kavi
AU  - Devraj K
AD  - Institute of Neurology (Edinger Institute), Goethe University Hospital; 
      Pharmazentrum Frankfurt, Institute for General Pharmacology and Toxicology, 
      Goethe University Hospital.
FAU - Guerit, Sylvaine
AU  - Guerit S
AD  - Institute of Neurology (Edinger Institute), Goethe University Hospital.
FAU - Macas, Jakranka
AU  - Macas J
AD  - Institute of Neurology (Edinger Institute), Goethe University Hospital.
FAU - Reiss, Yvonne
AU  - Reiss Y
AD  - Institute of Neurology (Edinger Institute), Goethe University Hospital; 
      Yvonne.Reiss@kgu.de.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20180226
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Blood-Brain Barrier/*metabolism
MH  - Green Fluorescent Proteins/*chemistry
MH  - Mice
MH  - Microscopy, Fluorescence/*methods
PMC - PMC5931381
EDAT- 2018/03/20 06:00
MHDA- 2018/06/23 06:00
PMCR- 2020/02/26
CRDT- 2018/03/20 06:00
PHST- 2018/03/20 06:00 [entrez]
PHST- 2018/03/20 06:00 [pubmed]
PHST- 2018/06/23 06:00 [medline]
PHST- 2020/02/26 00:00 [pmc-release]
AID - 57038 [pii]
AID - 10.3791/57038 [doi]
PST - epublish
SO  - J Vis Exp. 2018 Feb 26;(132):57038. doi: 10.3791/57038.