I am trying to find the best way to measure the blood brain barrier permeability Invivo (Mice),I tried the Evans blue dye with High molecular weight dextran but I'm still looking for a better method to measure the changes in the blood brain barrier permeability with aging.
Your help is much appreciated.
Hi there,
I am currently drafting a review on this very topic. The available techniques that I have identified for measuring BBB integrity and permeability include: Evans blue, fluorescence, TEER, MRI-Gd, IgG, PAMPA, PET, perfusion CT, horseradish peroxidase, NIR, SPECT, and MRI-ASL.
Regards,
Jerome
Dear Sweilem,
I have worked with BBB a bit and I am still very much interested. Your question is very good, but I think there is no single marker suitable for all purposes. Evans Blue than followed by HRP (horse radish peroxidase) are the classic markers used in testing the BBB markers. A combination of different sized, visualizable dextrans and radiolabeled molecules currently seems to be the most appropriate approach for qualitative and quantitative assessment of barrier integrity. You may discover the best combination of dyes to test the changes in the integrity of BBB associated with aging process.
One thing I have I identified in the past is the expression of TASK or leak potassium channels expressed in the cells lining the BBB. TASK-1 channels also display developmental changes and their expression increases with aging in various cell types. One property that distinguishes TASK-1 from other K channels is that it contains a PDZ domain, which may be involved in linking TASK-1 to other PDZ-containing structural or cytoskeletal, tight and adherens junction proteins such as claudins, occludine, zonula-occludens (ZO), actin and vascular-endothelial cadherin, all of which are critical for controlling permeability across the brain barri ers. Hence, the localization of TASK-1 in brain fluid barriers may be critically involved in responses to various pathological conditions.
Best wishes, Refik
Article The two-pore domain K+ channel TASK-1 is closely associated ...
Dear Dr.Kanjhan
Thank you very much for your very helpful answer and as you suggested that the combination of different sized, visualizable dextrans and radiolabeled molecules might be a good approach.
I will try it and I'll let you know.
Thanks again I really appreciate it.
Sweilem
Hi there,
I am currently drafting a review on this very topic. The available techniques that I have identified for measuring BBB integrity and permeability include: Evans blue, fluorescence, TEER, MRI-Gd, IgG, PAMPA, PET, perfusion CT, horseradish peroxidase, NIR, SPECT, and MRI-ASL.
Regards,
Jerome
Hi Dr.Maller,
Thank you very much and Looking forward for your review.
Best wishes
Sweilem
As for a positive control for BBB leakage or breakage, what would you guys suggest? Thank you very much!
Dear Dr.Campanati
Some studies used Mannitol or other hyperosmotic solutions into the carotid artery works well, but just for a very short time
I am trying now the Lipopolysaccharides( LPS) inejection 1mg/kg which will induce systemic inflammation but you will have side effects throughout the body.
Best wishes
Sweilem
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