Harris, H.J. G.K. Wilson, S. G. Hubscher and J. A. McKeating. Hepatology 57:854-5.2013.
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Claudin-1 is predominantly expressed at the apical membrane of hepatocytes in normal liver; however, a minor
pool of claudin-1 is also observed at the basolateral membrane. Claudin-1 is readily detected at the apical surface and is co-localized with CD10, a marker specific for apical membranes. However, basolaterally expressed claudin-1 is more easily discerned when the liver section is co-stained with a marker specific for the hepatic basolateral membrane, such as cytokeratin 8. These co-staining experiments allow us to observe heterogeneous patterns of localization across the liver parenchyma.
Spectral imaging of liver sections allowed us to accurately quantify bound fluorescent antibody irrespective of signal intensity. To achieve this, volumetric imaging of claudin-1 at areas of high (apical) and low (basolateral) expression requires multiple threshold values.
Figure 1: Claudin-1 localization in normal human liver
Representative images of claudin-1 costained with markers specific for apical (CD10) or basolateral (cytokeratin 8) membranes. Heterogeneous patterns of claudin-1 staining at apical and basolateral membranes were observed. The scale bar represents 10 µm.
Figure 2: Differential expression of claudin-1 in apical and basolateral areas. Application of low and high fluorescent intensity thresholds to the images in figure 1 (white boxed areas).
Volumetric images were prepared using Imaris software and show that in purely apical areas (left) claudin-1 (green) associates with CD10 (red), in keeping with a pericanalicular tight junction distribution. In other areas (right) application of low and high fluorescent intensity thresholds identifies both apical tight junction-associated (silver) and basolateral (yellow) pools of claudin-1. Nuclei were stained with DAPI (blue).