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Immunogold localization of tight junctional proteins in normal and osmotically-affected rat blood-brain barrier.
J Mol Histol. 2004 Jun; 35(5):529-39.JM

Abstract

The distribution of molecular components of interendothelial tight junctions (TJs) was studied in rat blood-brain barrier (BBB) microvessels, using immunogold cytochemistry applied to electron microscopy. Samples of rat brains, both normal (unaffected) and osmotically-affected (1, 5, and 30 min after intracarotid infusion of 1.8 M L(+)arabinose), were processed for immunocytochemical localization of TJ-specific integral membrane (occludin, JAM-1, claudin-5) and peripheral (ZO-1) protein molecules. In unaffected interendothelial junctions of control rats the immunosignals (represented by gold particles) for occludin and ZO-1 were of highest, whereas for claudin-5 and JAM-1 were of lower density. At 1 min after infusion, no discernible changes in distribution of junction-associated molecules were noted. At 5 min, however, changes were most conspicuous, and they consisted of segmental attenuation of the endothelial lining and dilatation (opening) of some junctional clefts accompanied by the diminution of the density of immunosignals for TJ-specific transmembrane and peripheral proteins. It was paralleled by disorganization of the spatial relation of these molecules to the junctional complexes. After 30 min, many interendothelial junctions appeared to be still open, whereas other junctions were partially or totally closed. In the opened interendothelial junctions the expression of TJ-associated molecules was weaker than in closed junctions. Our observations indicate that the localization and expression of TJ-specific proteins, especially occludin, and in lower degree claudin-5 and JAM-1, together with the peripheral ZO-1 molecules, are affected by osmotic shock. Presumably, some of these proteins (e.g., occludin, claudin-5 and ZO-1) could be considered sensitive indicators of normal and also of disturbed functional state of the BBB.

Authors+Show Affiliations

Laboratory of Cytochemistry, Department of Developmental Neurobiology, New York State Office of Mental Retardation and Developmental Disabilities, Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA. dobrogowska@msn.comNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15571330

Citation

Dobrogowska, Danuta H., and Andrzej W. Vorbrodt. "Immunogold Localization of Tight Junctional Proteins in Normal and Osmotically-affected Rat Blood-brain Barrier." Journal of Molecular Histology, vol. 35, no. 5, 2004, pp. 529-39.
Dobrogowska DH, Vorbrodt AW. Immunogold localization of tight junctional proteins in normal and osmotically-affected rat blood-brain barrier. J Mol Histol. 2004;35(5):529-39.
Dobrogowska, D. H., & Vorbrodt, A. W. (2004). Immunogold localization of tight junctional proteins in normal and osmotically-affected rat blood-brain barrier. Journal of Molecular Histology, 35(5), 529-39.
Dobrogowska DH, Vorbrodt AW. Immunogold Localization of Tight Junctional Proteins in Normal and Osmotically-affected Rat Blood-brain Barrier. J Mol Histol. 2004;35(5):529-39. PubMed PMID: 15571330.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Immunogold localization of tight junctional proteins in normal and osmotically-affected rat blood-brain barrier. AU - Dobrogowska,Danuta H, AU - Vorbrodt,Andrzej W, PY - 2004/12/2/pubmed PY - 2005/3/26/medline PY - 2004/12/2/entrez SP - 529 EP - 39 JF - Journal of molecular histology JO - J. Mol. Histol. VL - 35 IS - 5 N2 - The distribution of molecular components of interendothelial tight junctions (TJs) was studied in rat blood-brain barrier (BBB) microvessels, using immunogold cytochemistry applied to electron microscopy. Samples of rat brains, both normal (unaffected) and osmotically-affected (1, 5, and 30 min after intracarotid infusion of 1.8 M L(+)arabinose), were processed for immunocytochemical localization of TJ-specific integral membrane (occludin, JAM-1, claudin-5) and peripheral (ZO-1) protein molecules. In unaffected interendothelial junctions of control rats the immunosignals (represented by gold particles) for occludin and ZO-1 were of highest, whereas for claudin-5 and JAM-1 were of lower density. At 1 min after infusion, no discernible changes in distribution of junction-associated molecules were noted. At 5 min, however, changes were most conspicuous, and they consisted of segmental attenuation of the endothelial lining and dilatation (opening) of some junctional clefts accompanied by the diminution of the density of immunosignals for TJ-specific transmembrane and peripheral proteins. It was paralleled by disorganization of the spatial relation of these molecules to the junctional complexes. After 30 min, many interendothelial junctions appeared to be still open, whereas other junctions were partially or totally closed. In the opened interendothelial junctions the expression of TJ-associated molecules was weaker than in closed junctions. Our observations indicate that the localization and expression of TJ-specific proteins, especially occludin, and in lower degree claudin-5 and JAM-1, together with the peripheral ZO-1 molecules, are affected by osmotic shock. Presumably, some of these proteins (e.g., occludin, claudin-5 and ZO-1) could be considered sensitive indicators of normal and also of disturbed functional state of the BBB. SN - 1567-2379 UR - https://www.unboundmedicine.com/medline/citation/15571330/Immunogold_localization_of_tight_junctional_proteins_in_normal_and_osmotically_affected_rat_blood_brain_barrier_ L2 - http://ovidsp.ovid.com/ovidweb.cgi?T=JS&PAGE=linkout&SEARCH=15571330.ui DB - PRIME DP - Unbound Medicine ER -