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Identification of Claudins by Western Blot and Immunofluorescence in Different Cell Lines and Tissues

Claudins are integral proteins of the TJ. Each epithelia in the organism expresses a unique set of claudins that determines the degree of sealing of the paracellular pathway and the ionic selectivity of the tissue. TJs are dynamic structures whose organization and composition change in resp ...

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Expression and Function of Claudins in Hepatocytes

Tight junctions of hepatocytes play crucial roles in the barrier to keep bile in bile canaliculi away from the blood circulation, which we call the blood–biliary barrier. Tight junction proteins of hepatocytes are regulated by various cytokines and growth factors via distinct signal tra ...

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Analysis of Changes in the Expression Pattern of Claudins Using Salivary Acinar Cells in Primary Culture

Primary saliva is produced from blood plasma in the acini of salivary glands and is modified by ion adsorption and secretion as the saliva passes through the ducts. In rodents, acinar cells of salivary glands express claudin-3 but not claudin-4, whereas duct cells express both claudins-3 and -4. The ...

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Using Molecular Tracers to Assess the Integrity of the Intestinal Epithelial Barrier In Vivo

The examination of the epithelial barrier has been a primary site of focused research for years. Despite the importance of this site to numerous intestinal diseases, the determination of the integrity of this barrier has been clouded by controversies as to the validity of certain techniques a ...

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Development of Biological Tools to Study Claudins in the Male Reproductive Tract

It is estimated that between 12 and 15% of couples are infertile. More than half of these are related to problems associated with male reproductive dysfunction. Of those, 40% occur from idiopathic or unexplained causes. While spermatozoa are formed in the testis, testicular spermatozoa are im ...

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Laboratory Methods in the Study of Endometrial Claudin-4

Immunohistochemistry is a suitable method for the detection of proteins from the Claudin family and several antibodies are commercially available for the detection of Claudin congeners. Immunodetection of Caludin-4 in the paraffin-embedded specimens might be a useful tool for st ...

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In Vivo Imaging of Tight Junctions Using ClaudinEGFP Transgenic Medaka

Tight junctions (TJs) function as a physiological barrier between epithelial and endothelial sheets by restricting the diffusion of fluid through the intercellular space. Recent morphological studies associated with TJs have revealed that the TJ is a dynamic rather than a static str ...

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Claudin Family Proteins in Caenorhabditis elegans

In the last decade, the claudin family of integral membrane proteins has been identified as the major protein component of the tight junctions in all vertebrates. The claudin superfamily proteins also function to regulate channel activity, intercellular signaling, and cell morpholo ...

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Claudins in a Primary Cultured Puffer Fish (Tetraodon nigroviridis) Gill Epithelium

A primary cultured gill epithelium from the model organism Tetraodon nigroviridis (spotted green puffer fish) has been developed for the study of claudin tight junction (TJ) proteins and their potential role in the regulation of paracellular permeability across the gills of fishes. The ...

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Manipulating Claudin Expression in Avian Embryos

Since the discovery of Claudin-1 and -2 by Tsukita and colleagues in the late 1990s , claudin family members have been found to have critical roles in maintaining the integrity of epithelial and endothelial tight junctions . The properties of distinct claudin family members in tight junction p ...

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Anti-claudin-4-Conjugated Highly Luminescent Nanoparticles as Biological Labels for Pancreatic Cancer Sensing

Anti-claudin-4, whose corresponding antigen receptors are known to be overexpressed in both primary and metastatic human pancreatic cancer, is utilized for targeted delivery and imaging of pancreatic cancer. In this protocol, we describe the use of quantum dots (QDs) as sensitive opti ...

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Biochemical Analysis of Claudin-Binding Compatibility

Tissue barrier function is directly mediated by tight junction transmembrane proteins known as claudins. Cells that form tight junctions typically express multiple claudin isoforms, which suggests that heterotypic (head-to-head) binding between different claudin isofo ...

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Dynamics of Claudins Expression in Colitis and Colitis-Associated Cancer in Rat

Claudins comprise a multigene family of 24 species and have been shown to constitute the backbone of tight junction strands in simple epithelial cells and to be directly involved in their barrier functions. Apical-most tight junction protein complexes (TJs) are implicated in inflammat ...

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Electrophysiological Characterization of Claudin Ion Permeability Using Stably Transfected Epithelial Cell Lines

Claudins are tight junction membrane proteins that act as paracellular pores and barriers and regulate epithelial permeability to small ions. A key step in understanding the function of any claudin isoform is the in vitro measurement of its ion permeability and selectivity. Herein, we des ...

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Interactions Between Clostridium perfringens Enterotoxin and Claudins

Clostridium perfringens enterotoxin (CPE), a single polypeptide of approximately 35 kDa in size, is �associated with type A food poisoning and such non-foodborne gastrointestinal diseases as antibiotic-associated diarrhea and sporadic diarrhea. CPE action begins with bindi ...

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The Tight Junction, Intercellular Seal as a Cell Signaling Player: Protocols for Examination of Its Status

Tight junctions (TJs) are intercellular structures in epithelial and endothelial cells, primarily playing �critical roles in cell–cell adhesion. Among their molecular components, claudins are the main constituents as integral membrane proteins, encoded by at least 24 members of a ...

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Biophysical Methods to Probe Claudin-Mediated Adhesion at the Cellular and Molecular Level

Claudins are a family of tetraspan membrane proteins that localize at tight junctions in an epithelial monolayer forming a selective barrier to diffusion of solutes via the intercellular spaces. It is widely accepted that the interaction between the extracellular loops of claudin mol ...

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Detection of Tight Junction Barrier Function In Vivo by Biotin

Tight junctions (TJs) are the most apical component of the junctional complexes in mammalian epithelial cells and form selective paracellular barriers restricting the passage of solutes and ions across the epithelial sheets. Claudins, a TJ integral membrane protein family, play a cri ...

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The Coculture Method to Examine Interactions Between Claudin Isoforms in Tight Junction-Free HEK293 Cells and Tight Junction-Bearing MDCK II Cells

The paracellular transport of water, ions, and small solutes is regulated by tight junctions (TJs) in �epithelial, endothelial, and mesothelial cells. Both the prolonged increase and decrease of the paracellular permeability are involved in various diseases. Claudins, a family of at le ...

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Claudin-4: Functional Studies Beyond the Tight Junction

Claudin-4 is an unusual member of the claudin family; in addition to its role in epithelial tight junction barrier function, it is a receptor for the Clostridium perfringens enterotoxin. We have also found that claudin-4 is regulated in mucosal epithelium M cells, both in increased expression of ...

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