J Vet Sci.  2016 Dec;17(4):445-451. 10.4142/jvs.2016.17.4.445.

Claudin-1, -2, -4, and -5: comparison of expression levels and distribution in equine tissues

Affiliations
  • 1Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Korea. ebjeung@chungbuk.ac.kr

Abstract

Claudins, which are known as transmembrane proteins play an essential role in tight junctions (TJs) to form physical barriers and regulate paracellular transportation. To understand equine diseases, it is helpful to measure the tissue-specific expression of TJs in horses. Major equine diseases such as colic and West Nile cause damage to TJs. In this study, the expression level and distribution of claudin-1, -2, -4, and -5 in eight tissues were assessed by Western blotting and immunohistochemistry methods. Claudin-1 was primarily identified in the lung, duodenum, and uterus, claudin-2 was evenly observed in equine tissues, claudin-4 was abundantly detected in the liver, kidney and uterus, and claudin-5 was strongly expressed in the lung, duodenum, ovary, and uterus, as determined by Western blotting method. The localization of equine claudins was observed by immunohistochemistry methods. These findings provide knowledge regarding the expression patterns and localization of equine claudins, as well as valuable information to understand tight junction-related diseases according to tissue specificity and function of claudins in horses.

Keyword

Equus ferus caballus; claudins; paracellular transportation; tight junctions

MeSH Terms

Animals
Blotting, Western/veterinary
Claudins/*genetics/metabolism
Female
*Gene Expression Regulation
Horses/*genetics/metabolism
Immunohistochemistry/veterinary
Male
Tissue Distribution
Claudins

Figure

  • Fig. 1 Protein levels of claudin-1, -2, -4 and -5 in equine tissues. Levels were measured by western blot (A) and were normalized against GAPDH protein (B-E; claudin-1, -2, -4 and -5, respectively). The signal intensity of each band was measured using the NIH Image J software (USA). Data are presented as the means ± standard error of the mean of four equine samples. Liv, Liver; Kid, Kidney; Lu, Lung; Duo, Duodenum; Hrt, Heart; Tes, Testis; Ov, Ovary; Ut, Uterus. *p < 0.01 vs. liver; †p < 0.01 vs. kidney; ‡p < 0.01 vs. lung; §p < 0.01 vs. duodenum; ∥p < 0.01 vs. heart; ¶p < 0.01 vs. ovary; **p < 0.05 vs. uterus.

  • Fig. 2 Localization of claudin-1 in equine tissues via immunohistochemistry. Scale bar = 100 µm.

  • Fig. 3 Localization of claudin-2 in equine tissues via immunohistochemistry. Scale bar = 100 µm.

  • Fig. 4 Localization of claudin-4 in equine tissues via immunohistochemistry. Scale bar = 100 µm.

  • Fig. 5 Localization of claudin-5 in equine tissues via immunohistochemistry. Scale bar = 100 µm.


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