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Am J Physiol Gastrointest Liver Physiol 289: G36-G41, 2005. First published February 24, 2005; doi:10.1152/ajpgi.00552.2004
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MUCOSAL BIOLOGY

Subcloning, localization, and expression of the rat intestinal sodium-hydrogen exchanger isoform 8

Hua Xu, Rongji Chen, and Fayez K. Ghishan

Departments of Pediatrics, Physiology, and Nutritional Sciences, Steele Memorial Children’s Research Center, University of Arizona Health Sciences Center, Tucson, Arizona

Submitted 15 December 2004 ; accepted in final form 17 February 2005

Apically expressed intestinal and renal sodium-hydrogen exchangers (NHEs) play a major role in Na+ absorption. Our previous studies on NHE ontogeny have shown that NHE-2 and NHE-3 are expressed at very low levels in young animals. Furthermore, single and/or double NHE-2 and NHE-3 knockout mice display no obvious abnormalities before weaning. These observations suggest that other transporter(s) may be involved in intestinal Na+ absorption during early life. The present studies were designed to clone the novel rat intestinal NHE-8 cDNA and to decipher the NHE-8 protein localization and gene expression pattern during different developmental stages. The rat NHE-8 cDNA has 2,160 bp and encodes a 575-amino acid protein. An antibody against NHE-8 protein was developed. Immunohistochemistry staining indicated apical localization of NHE-8 protein in rat intestinal epithelial cells. The apical localization of NHE-8 was also confirmed by its presence in brush-border membrane and its absence in basolateral membrane preparations. Northern blotting utilizing a NHE-8-specific probe demonstrated higher NHE-8 mRNA expression in young animals compared with adult animals. Western blot analysis revealed a similar pattern. Tissue distribution with multiple human tissue RNA blot showed that NHE-8 was expressed in multiple tissues including the gastrointestinal tract. In conclusion, we have cloned the full-length NHE-8 cDNA from rat intestine and further showed its apical localization in intestinal epithelial cells. We have also shown that NHE-8 gene expression and protein expression were regulated during ontogeny. Our data suggests that NHE-8 may play an important role in intestinal Na+ absorption during early life.

SLC9A8; intestine; ontogeny


Address for reprint requests and other correspondence: F. K. Ghishan, Dept. of Pediatrics, Steele Memorial Children’s Research Center, 1501 N. Campbell Ave., Tucson, AZ 85724 (E-mail: fghishan{at}peds.arizona.edu)




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