Abstract
The blood vessels that vascularize the central nervous system (CNS) exhibit unique properties, termed the blood-brain barrier (BBB). The BBB allows these blood vessels to tightly regulate the movement of ions, molecules, and cells between the blood and the brain. The BBB is held together by tight junctions of the neighboring endothelial cells of the barrier, more specifically by tight junction proteins (TJPs) which can take the form of either integral transmembrane proteins or accessory cytoplasmic membrane proteins. BBB permeability can furthermore be affected by various factors, including but not limited to TJP expression, size, shape, charge, and type of extravascular molecules, as well as the nature of the vascular beds. The BBB is essential for the proper maintenance of CNS function, and its structural integrity has been implicated in several disorders and conditions. For instance, it has been shown that in the cases of traumatic brain injury (TBI), TBI-associated edema, and increased intracranial pressure are primarily caused by cases of hyperpermeability seen because of BBB dysfunction. Intravital microscopy is one of the most reliable methods for measuring BBB hyperpermeability in rodent models of BBB dysfunction in vivo. Here, we describe the surgical and imaging methods to determine the changes in BBB permeability at the level of the pial microvasculature in a mouse model of TBI using intravital microscopy.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Alluri H, Anasooya Shaji C, Davis ML, Tharakan B (2018) A mouse controlled cortical impact model of traumatic brain injury for studying blood-brain barrier dysfunctions. In: Traumatic and ischemic injury: methods and protocols, Methods in molecular biology, vol 1717. Springer, New York, pp 37–52
Alluri H, Wiggins-Dohlvik K, Davis ML, Huang JH, Tharakan B (2015 Jan 28) Blood-brain barrier dysfunction following traumatic brain injury. Metab Brain Dis 30:1093
Deaglio S, Robson SC (2011) Ectonucleotidases as regulators of purinergic signaling in thrombosis, inflammation, and immunity. Adv Pharmacol 61:301–332
Robinson BD, Isbell CL, Melge AR, Lomas AM, Shaji CA, Mohan CG, Huang JH, Tharakan B (2022) Doxycycline prevents blood-brain barrier dysfunction and microvascular hyperpermeability after traumatic brain injury. Sci Rep 12:5415
Gean AD, Fischbein NJ (2010) Head trauma. Neuroimaging Clin N Am 20:527–556
Kasper C, Yvette C (2015) Traumatic brain injury. In: Annual review of nursing research, vol 33. Springer, New York
Khan M, Im YB, Shunmugavel A, Gilg AG, Dhindsa RK, Singh AK, Singh IJ (2009) Administration of S- nitrosoglutathione after traumatic brain injury protects the neurovascular unit and reduces secondary injury in a rat model of controlled cortical impact. Neuroinflammation 6:32
Kumar P, Shen Q, Pivetti CD, Lee ES, Wu MH, Yuan SY (2009) Molecular mechanisms of endothelial hyperpermeability: implications in inflammation. Expert Rev Mol Med 11:e19
Maas AIR, Stocchetti N, Bullock R (2008) Moderate and severe traumatic brain injury in adults. Lancet Neurol 7:728–741
Marques PE, Oliveira AG, Amaral SS, Nunes-Silva A, Almeida AFS (2012) Intravital microscopy: taking a close look inside the living organisms. Afr J Microbiol Res 6:1603–1614
Masedunskas A, Porat-Shliom N, Tora M, Milberg O, Weigert R (2013) Intravital microscopy for imaging subcellular structures in live mice expressing fluorescent proteins. J Vis Exp 79:50558
O’Connor WT, Smyth A, Gilchrist MD (2011) Animal models of traumatic brain injury: a critical evaluation. Pharmacol Ther 130:106–113
Parikh S, Koch M, Narayan RK (2007) Traumatic brain injury. Int Anesthesiol Clin 45:119–135
Radu M, Chernoff J (2013) An in vivo assay to test blood vessel permeability. J Vis Exp 73:e50062
Shen Q, Wu MH, Yuan SY (2009) Endothelial contractile cytoskeleton and microvascular permeability. Cell Health Cytoskelet 1:43–50
Taqueti VR, Jaffer FA (2013) High-resolution molecular imaging via intravital microscopy: illuminating vascular biology in vivo. Integr Biol (Camb) 5:278–290
Unterberg AW, Stover J, Kress B, Kiening KL (2004) Edema and brain trauma. Neuroscience 129:1021–1029
Xiong Y, Mahmood A, Chopp M (2013) Animal models of traumatic brain injury. Nat Rev Neurosci 14:128–142
Yuan SY (2002) Protein kinase signaling in the modulation of microvascular permeability. Vasc Pharmacol 39:213–223
Acknowledgments
The authors acknowledge the support from National Institutes of Health (NIH) grant 5 SC3 NS127765-02 (BT).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Waithe, O.Y., Shaji, C.A., Childs, E.W., Tharakan, B. (2024). Determination of Blood-Brain Barrier Hyperpermeability Using Intravital Microscopy. In: Tharakan, B. (eds) Vascular Hyperpermeability. Methods in Molecular Biology, vol 2711. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3429-5_10
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3429-5_10
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3428-8
Online ISBN: 978-1-0716-3429-5
eBook Packages: Springer Protocols