Elsevier

Gene Expression Patterns

Volume 6, Issue 6, August 2006, Pages 581-588
Gene Expression Patterns

Claudin profiling in the mouse during postnatal intestinal development and along the gastrointestinal tract reveals complex expression patterns

https://doi.org/10.1016/j.modgep.2005.12.001Get rights and content

Abstract

Members of the claudin protein family are key regulators of tight junction selectivity and are implicated in influencing development and cellular differentiation in the intestine and other tissues. The goal of the present study was to profile claudin gene expression and protein location during postnatal development of the mouse jejunum and in the adult mouse gut from duodenum to distal colon as a first step in understanding both normal claudin function and the pathologic implications of altered expression patterns. The relative expression of claudins 1–19 and other tight and adherens junction genes was determined by quantitative RT-PCR from six regions of normal mouse intestine and colon. Immunofluorescent localization was performed for claudins 1–5, 7, 8, 10, 12, 15, and 18. Transcripts for claudins 1–5, 7–13, 17, and 18 were all detected in adult intestine, although their relative abundance differed up to 1000-fold within individual segments. In contrast to the unchanging expression and localization of ZO-1, occludin, and JAM, most claudins were expressed in decreasing or increasing gradients or in more complex patterns along the longitudinal axis of the intestine and the crypt to villus/surface differentiation axis. During neonatal development at days 1, 14, 28, and 90 several claudins showed striking increases or decreases in transcript expression as well as changes in tissue localization along the crypt-villus axis. Claudin-19 was only detected at days 1 and 14. This database provides a resource for investigating regional and developmental differences in permselectivity, crypt to villus/surface differentiation and neoplastic changes along the gut and during postnatal development.

Section snippets

Claudin expression patterns in the adult mouse intestine

Quantitative real-time PCR (qRT-PCR) reveals that the adult intestine expresses many different claudins in complex quantitative and spatial patterns. Claudins are expressed variably along both the longitudinal axis and the crypt/villus differentiation axis (Fig. 1, Fig. 3, Fig. 4 and Supplementary Fig. 3 and Table 1). In contrast, transcript levels for the TJ cytoplasmic protein ZO-1 do not vary relative to expression of Eef1a1, which was used as a general reference transcript. Therefore, in

Mice and tissue collection

All animal handling and tissue collection was preformed in accordance with the policies of the Institutional Animal Care Committees of the University of North Carolina-Chapel Hill. Full thickness bowel segments, stripped of omentum, were collected from female C57/Black6 mice 1, 14, 28, and 90 days after birth. For the 90 day adult time-point, samples for RNA isolation and immunofluorescent microscopic localization (IF) were taken from the duodenum, mid-jejunum, mid-ileum, cecum (the cecal tip

Acknowledgments

This work is supported by the National Institutes of Health (NIDDK DK45134), the Broad Medical Research Program and the State of North Carolina. We thank Alan Fanning for helpful discussions.

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