Abstract
Tight junction proteins are integral membrane proteins located apically on epithelial and endothelial cells. They form a selective paracellular barrier restricting the passage of solutes and ions across epithelial and endothelial sheets. In brain endothelial cells, the enrichment of tight junction proteins is one of the unique features of the blood–brain barrier, the physiological boundary that separates the blood from the parenchyma. The predominant tight junction family proteins are the claudins, but several others have been described in recent years including the marvel family, occludin, and lipolysis-stimulated lipoprotein receptor. Together, the tight junctions create a highly electrical-resistant, impermeable paracellular channel that strictly restricts the movement of material from the blood to the parenchyma and vice versa. In this chapter, we will discuss immunohistochemical methods to assess tight junction expression and localization and an ImageJ-based method for quantifying tight junction staining in healthy and diseased states.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sweeney MD et al (2019) Blood-brain barrier: from physiology to disease and Back. Physiol Rev 99(1):21–78
Abbott NJ et al (2010) Structure and function of the blood-brain barrier. Neurobiol Dis 37(1):13–25
Cummins PM (2012) Occludin: one protein, many forms. Mol Cell Biol 32(2):242–250
Wolburg H, Lippoldt A (2002) Tight junctions of the blood-brain barrier: development, composition and regulation. Vasc Pharmacol 38(6):323–337
Morita K et al (1999) Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands. Proc Natl Acad Sci U S A 96(2):511–516
Itoh M et al (1999) Direct binding of three tight junction-associated MAGUKs, ZO-1, ZO-2, and ZO-3, with the COOH termini of claudins. J Cell Biol 147(6):1351–1363
Tornavaca O et al (2015) ZO-1 controls endothelial adherens junctions, cell-cell tension, angiogenesis, and barrier formation. J Cell Biol 208(6):821–838
Nitta T et al (2003) Size-selective loosening of the blood-brain barrier in claudin-5-deficient mice. J Cell Biol 161(3):653–660
Sohet F et al (2015) LSR/angulin-1 is a tricellular tight junction protein involved in blood-brain barrier formation. J Cell Biol 208(6):703–711
Lv J et al (2018) Focusing on claudin-5: a promising candidate in the regulation of BBB to treat ischemic stroke. Prog Neurobiol 161:79–96
Knowland D et al (2014) Stepwise recruitment of transcellular and paracellular pathways underlies blood-brain barrier breakdown in stroke. Neuron 82(3):603–617
Chodobski A, Zink BJ, Szmydynger-Chodobska J (2011) Blood-brain barrier pathophysiology in traumatic brain injury. Transl Stroke Res 2(4):492–516
Greene C, Hanley N, Campbell M (2020) Blood-brain barrier associated tight junction disruption is a hallmark feature of major psychiatric disorders. Transl Psychiatry 10(1):373
Greene C et al (2018) Dose-dependent expression of claudin-5 is a modifying factor in schizophrenia. Mol Psychiatry 23(11):2156–2166
Nishiura K et al (2017) PKA activation and endothelial claudin-5 breakdown in the schizophrenic prefrontal cortex. Oncotarget 8(55):93382–93391
Greene C, Hanley N, Campbell M (2019) Claudin-5: gatekeeper of neurological function. Fluids Barriers CNS 16(1):3
Acknowledgements
This work was supported by grants from Science Foundation Ireland and the Irish Research Council (IRC). The Campbell lab at TCD is also supported by an SFI Centres grant supported in part by a research grant from SFI under grant number 16/RC/3948 and co-funded under the European Regional Development fund by FutureNeuro industry partners.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Greene, C., Campbell, M. (2022). Immunohistochemical Analysis of Tight Junction Proteins. In: Stone, N. (eds) The Blood-Brain Barrier. Methods in Molecular Biology, vol 2492. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2289-6_18
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2289-6_18
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2288-9
Online ISBN: 978-1-0716-2289-6
eBook Packages: Springer Protocols