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Molecular cloning, tissue distribution and ontogenetic expression of sodium proton exchanger isoform 2 (NHE-2) mRNA in the small intestine of pigs

Published online by Cambridge University Press:  01 March 2009

D. Feng
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Ninjing, 210095, PR China College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China
A. Zhi
Affiliation:
College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China Henan Provincial Key Laboratory for Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, PR China
S. Zou
Affiliation:
College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China
X. Zhou
Affiliation:
College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China
J. Zuo
Affiliation:
College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China
Z. Huang
Affiliation:
College of Animal Science, South China Agricultural University, Guangzhou, 510642, PR China
T. Wang*
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Ninjing, 210095, PR China
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Abstract

Molecular cloning, tissue distribution and ontogenetic regulation of sodium/proton exchanger isoform 2 (NHE-2) mRNA expression were evaluated in the pig small intestine during postnatal development. The 2872-bp porcine full cDNA sequence of the NHE-2 (EF672046) cloned in this study showed 80% and 70% homology with known human and mouse gene sequence, respectively. Hydrophobic prediction suggests 13 putative membrane-spanning domains within porcine NHE-2. The porcine NHE-2 mRNA was detected in the brain, liver, kidney, heart, lung, small intestine and muscle. The small intestine had the highest NHE-2 mRNA abundance and the brain, lung and liver had the lowest NHE-2 mRNA abundance (P < 0.05). Along the longitudinal axis, the duodenum had the highest NHE-2 mRNA abundance and the ileum and colon had the lowest NHE-2 mRNA abundance (P < 0.05). The NHE-2 mRNA level was increased from day 1 to day 26 in the duodenum (P < 0.05) and dropped dramatically on day 30 (P < 0.05). There is no difference between day 1 and day 7 (P > 0.05). After day 30, the NHE-2 mRNA level remained the same except on day 90 (P > 0.05). The mRNA expression of NHE-2 was not only differentially regulated by age but also differentially distributed along the small intestine of piglets at early stages and growing stages of life, which may contribute to changes in NHE activity.

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Full Paper
Copyright
Copyright © The Animal Consortium 2008

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Footnotes

a

D. Feng and A. Zhi contributed equally to this study.

References

Ambuhl, PM, Yang, X, Peng, Y, Preisig, PA, Moe, OW, Alpern, RJ 1999. Glucocorticoids enhance acid activation of the Na+/H+ exchanger 3 (NHE-3). Journal of Clinical Investigation 103, 429435.CrossRefGoogle Scholar
Azarani, A, Goltzman, D, Orlowski, J 1995. Parathyroid hormone and parathyroid hormone-related peptide inhibit the apical Na+/H+ exchanger NHE-3 isoform in renal cells (OK) via a dual signaling cascade involving protein kinase A and C. Journal of Biology Chemistry 270, 2000420010.CrossRefGoogle Scholar
Bai, L, Collins, JF, Xu, H, Ghishan, FK 2001. Transcriptional regulation of the rat sodium–hydrogen exchanger isoform 2 (NHE-2) gene by Sp1 transcription factor. American Journal of Physiology-Cell Physiology 280, C1168C1175.CrossRefGoogle ScholarPubMed
Baum, M, Moe, OW, Gentry, DL, Alpern, RJ 1994. Effect of glucocorticoids on renal cortical NHE-3 and NHE-1 mRNA. American Journal of Physiology 267, F437F442.Google ScholarPubMed
Buddington, RK, Elnif, JA, Gardiner, AP, Sangild, PT 2001. Intestinal apical amino acid absorption during development of the pig. American Journal of Physiology Regulatory, Integrative and Comparative Physiology 280, R241R247.CrossRefGoogle ScholarPubMed
Collins, JF, Honda, T, Knobel, S, Bulus, NM, Conary, J, DuBois, R, Ghishan, FK 1993. Molecular cloning, sequencing, tissue distribution, and functional expression of a Na+/H+ exchanger (NHE-2). Proceedings of the National Academy of Sciences of the United States of America 90, 39383942.CrossRefGoogle ScholarPubMed
Collins, JF, Xu, H, Kiela, PR, Zeng, J, Ghishan, FK 1997. Functional and molecular characterization of NHE-3 expression during ontogeny in rat jejunal epithelium. American Journal of Physiology 273, C1937C1946.CrossRefGoogle Scholar
Collins, JF, Kiela, PR, Xu, H, Zeng, J, Ghishan, FK 1998. Increased NHE-2 expression in rat intestinal epithelium during ontogeny is transcriptionally mediated. American Journal of Physiology 275, C1143C1150.CrossRefGoogle Scholar
Dudeja, PK, Rao, DD, Syed, I, Joshi, V, Dahdal, RY 1996. Intestinal distribution of human Na+/H+ exchanger isoforms NHE-1, NHE-2, and NHE-3 mRNA. American Journal of Physiology 271, G483G493.Google ScholarPubMed
Feng, D, Zhou, X, Zuo, J, Zhang, C, Yin, Y, Wang, X, Wang, T 2008. Segmental distribution and expression of two heterodimeric amino acid transporter mRNAs in the intestine of pigs during different ages. Journal of the Science of Food and Agriculture 88, 10121018.CrossRefGoogle Scholar
Livak, KJ, Schmittgen, TD 2001. Analysis of relative gene expression data using real-time PCR quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25, 402408.CrossRefGoogle ScholarPubMed
Lu, G, Moriyama, EN 2004. Vector NTI, a balanced all-in-one sequence analysis suite. Briefings in Bioinformatics 5, 378388.CrossRefGoogle Scholar
Malakooti, J, Dahdal, RY, Schmidt, L, Layden, TJ, Dudeja, PK, Ramaswamy, K 1999. Molecular cloning, tissue distribution, and functional expression of the human Na+/H+ exchanger NHE-2. American Journal of Physiology 277, G383G390.Google Scholar
Moeser, AJ, Blikslager, AT 2007. Mechanisms of porcine diarrheal disease. Journal of the American Veterinary Medical Association 231, 5667.CrossRefGoogle ScholarPubMed
Noel, J, Pouyssgur, J 1995. Hormonal regulation, pharmacology, and membrane sorting of vertebrate Na+/H+ exchanger isoforms. American Journal of Physiology 268, C283C296.CrossRefGoogle ScholarPubMed
Orlowski, J, Grinstein, S 2004. Diversity of the mammalian sodium/proton exchanger SLC9 gene family. Pflügers Archiv: European Journal of Physiology 447, 549565.CrossRefGoogle ScholarPubMed
Orlowski, J, Kandasamy, RA, Shull, GE 1992. Molecular cloning of putative members of the Na/H exchanger gene family. cDNA cloning, deduced amino acid sequence, and mRNA tissue expression of the rat Na/H exchanger NHE-1 and two structurally related proteins. Journal of Biological Chemistry 267, 93319339.CrossRefGoogle ScholarPubMed
Repishti, M, Hogan, DL, Pratha, V, Davydova, L, Donowitz, M 2001. Human duodenal mucosal brush border Na+/H+ exchangers NHE-2 and NHE-3 alter net bicarbonate movement. American Journal of Physiology-Gastrointestinal and Liver Physiology 281, G159G163.CrossRefGoogle Scholar
Rocha, F, Musch, MW, Lishanskiy, L, Bookstein, C, Sugi, K, Xie, Y, Chang, EB 2001. IFN-gamma downregulates expression of Na+/H+ exchangers NHE-2 and NHE-3 in rat intestine and human Caco-2/bbe cells. American Journal of Physiology-Cell Physiology 280, C1224C1232.CrossRefGoogle Scholar
Thwaites, DT, Anderson, CMH 2007. H-coupled nutrient, micronutrient and drug transporters in the mammalian small intestine. Experimental Physiology 92, 603619.CrossRefGoogle ScholarPubMed
Tse, CM, Brant, SR, Walker, MS, Pouyssegur, J, Donowitz, M 1992. Cloning and sequencing of a rabbit cDNA encoding an intestinal and kidney-specific Na+/H+ exchanger isoform (NHE-3). Journal of Biological Chemistry 267, 93409346.CrossRefGoogle ScholarPubMed
Wakabayashi, S, Shigekawa, M, Pouyssegur, J 1997. Molecular physiology of vertebrate Na+/H+ exchangers. Physiology Review 77, 5174.CrossRefGoogle ScholarPubMed
Xu, H, Collins, JF, Bai, L, Kiela, PR, Lynch, RM, Ghishan, FK 2001. Epidermal growth factor regulation of rat NHE-2 gene expression. American Journal of Physiology-Cell Physiology 281, C504C513.CrossRefGoogle Scholar
Zachos, NC, Tse, M, Donowitz, M 2005. Molecular physiology of intestinal Na+/H+ exchange. Annual Review of Physiology 67, 411443.CrossRefGoogle ScholarPubMed