Expression of multiple Na+/H+ exchanger isoforms in cultured epithelial cells from rat efferent duct and cauda epididymidis

G. P H Leung, Chung Ming Tse, S. B. Cheng Chew, P. Y D Wong

Research output: Contribution to journalArticle

Abstract

Although earlier work has pointed to the presence of Na+/H+ exchangers (NHEs) in the rat epididymis, little is known about the regional distribution of various NHE isoforms and their functions. In the present work, expression of different isoforms of NHE in cultured epithelia of the efferent duct and cauda epdidymidis were studied. Reverse transcription-polymerase chain reaction revealed the presence of NHE1, NHE2, and NHE3, but not NHE4, message in both cultured epithelia. Western blot analysis detected the presence of NHE1 and NHE2 proteins in both cultured epithelia, but NHE3 protein was only detected in the cultured epithelial cells from the efferent duct. Immunohistochemical studies demonstrated that NHE2 was localized in the cytoplasm of the ciliated cells, whereas NHE3 was localized at the apical membrane of the principal cells of the efferent duct. The NHE activities in both cultured epithelia were inhibited by 10 μM HOE-694 (3-methylsulphonyl-4-piperidinobenzoyl guanidine methanesulphonate), a NHE1 inhibitor, by approximately 76%. The HOE-694-resistant NHE activities in the cultured epithelia of efferent duct and cauda epididymidis were completely inhibited by 20 μM S3226 (3-[2-(3-guanidino-2-methyl-3-oxo-propenyl)-5-methyl-phenyl]-N-isopropylidene -2-methyl-acrylamide dihydrochloride), a NHE3 inhibitor, and 300 μM HOE-694 (a dose that can completely block NHE2), respectively. These results indicated that NHE1, NHE2, and NHE3 were expressed in the cultured epithelial cells of the efferent duct, whereas only NHE1 and NHE2 were expressed in the cultured epithelial cells of the cauda epididymidis. It is suggested that NHE1 may provide "housekeeping" functions in both epithelia, whereas NHE2 in the cauda epididymidis and NHE3 in the efferent duct may be involved in Na+ reabsorption and regulation of pH of the luminal fluid.

Original languageEnglish (US)
Pages (from-to)482-490
Number of pages9
JournalBiology of Reproduction
Volume64
Issue number2
StatePublished - 2001
Externally publishedYes

Fingerprint

Sodium-Hydrogen Antiporter
Cultured Cells
Protein Isoforms
Epithelium
Epithelial Cells
3-(2-(3-guanidino-2-methyl-3-oxo-propenyl)-5-methylphenyl)-N-isopropylidene-2-methyl-acrylamide dihydrochloride
Housekeeping
Acrylamide
Epididymis
Reverse Transcription
Cytoplasm
Proteins
Western Blotting
Cell Membrane
Polymerase Chain Reaction
3-methylsulfonyl-4-piperidinobenzoyl guanidine

Keywords

  • Epididymis

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Expression of multiple Na+/H+ exchanger isoforms in cultured epithelial cells from rat efferent duct and cauda epididymidis. / Leung, G. P H; Tse, Chung Ming; Cheng Chew, S. B.; Wong, P. Y D.

In: Biology of Reproduction, Vol. 64, No. 2, 2001, p. 482-490.

Research output: Contribution to journalArticle

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