Acid incubation increases NHE-3 mRNA abundance in OKP cells

M. Amemiya, Y. Yamaji, A. Cano, O. W. Moe, R. J. Alpern

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

With the use of degenerate primers based on conserved amino acid sequences in human, rat, and rabbit Na/H exchanger-3 (NHE-3), a polymerase chain reaction product was obtained from reverse-transcribed OKP (a clonal opossum kidney cell line) mRNA and used to screen an OKP cDNA library. The clone obtained predicted an amino acid sequence that was 86% identical to rat NHE- 3, 33% to NHE-1, 35% to NHE-2, and 30% to NHE-4. Expression of the corresponding cRNA in Xenopus oocytes induced 22Na uptake with ethylisopropylamiloride (EIPA) resistance similar to that of the OKP Na/H antiporter. On RNA blot, the cDNA labeled a 9.5-kb transcript whose abundance was increased 2.2-fold by 24-h incubation of OKP cells at pH 7.0 and 2.5- fold by 24-h incubation at pH 6.8. The acid-induced increase in NHE-3 mRNA was detectable at 12 h and increased further at 24 h. Incubation in acid media caused an increase in EIPA-resistant Na/H antiporter activity that preceded the increase in NHE-3 mRNA. In summary, OKP cells express an NHE-3 transcript that encodes an EIPA resistant Na/H antiporter and is chronically regulated by acid.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume269
Issue number1 38-1
StatePublished - 1995

Fingerprint

Sodium-Hydrogen Antiporter
Messenger RNA
Acids
acids
cells
Rats
Amino Acid Sequence
amino acid sequences
Opossums
Amino Acids
Complementary RNA
Polymerase chain reaction
rats
Xenopus
opossums
kidney cells
sodium-hydrogen antiporter
Reaction products
Gene Library
cDNA libraries

Keywords

  • acidosis
  • ethylisopropylamiloride
  • oocyte expression
  • sodium/proton antiporter

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Agricultural and Biological Sciences(all)

Cite this

Acid incubation increases NHE-3 mRNA abundance in OKP cells. / Amemiya, M.; Yamaji, Y.; Cano, A.; Moe, O. W.; Alpern, R. J.

In: American Journal of Physiology - Cell Physiology, Vol. 269, No. 1 38-1, 1995.

Research output: Contribution to journalArticle

Amemiya, M. ; Yamaji, Y. ; Cano, A. ; Moe, O. W. ; Alpern, R. J. / Acid incubation increases NHE-3 mRNA abundance in OKP cells. In: American Journal of Physiology - Cell Physiology. 1995 ; Vol. 269, No. 1 38-1.
@article{98450eb2d7a34af98051c4ac9bb8d697,
title = "Acid incubation increases NHE-3 mRNA abundance in OKP cells",
abstract = "With the use of degenerate primers based on conserved amino acid sequences in human, rat, and rabbit Na/H exchanger-3 (NHE-3), a polymerase chain reaction product was obtained from reverse-transcribed OKP (a clonal opossum kidney cell line) mRNA and used to screen an OKP cDNA library. The clone obtained predicted an amino acid sequence that was 86{\%} identical to rat NHE- 3, 33{\%} to NHE-1, 35{\%} to NHE-2, and 30{\%} to NHE-4. Expression of the corresponding cRNA in Xenopus oocytes induced 22Na uptake with ethylisopropylamiloride (EIPA) resistance similar to that of the OKP Na/H antiporter. On RNA blot, the cDNA labeled a 9.5-kb transcript whose abundance was increased 2.2-fold by 24-h incubation of OKP cells at pH 7.0 and 2.5- fold by 24-h incubation at pH 6.8. The acid-induced increase in NHE-3 mRNA was detectable at 12 h and increased further at 24 h. Incubation in acid media caused an increase in EIPA-resistant Na/H antiporter activity that preceded the increase in NHE-3 mRNA. In summary, OKP cells express an NHE-3 transcript that encodes an EIPA resistant Na/H antiporter and is chronically regulated by acid.",
keywords = "acidosis, ethylisopropylamiloride, oocyte expression, sodium/proton antiporter",
author = "M. Amemiya and Y. Yamaji and A. Cano and Moe, {O. W.} and Alpern, {R. J.}",
year = "1995",
language = "English (US)",
volume = "269",
journal = "American Journal of Physiology - Heart and Circulatory Physiology",
issn = "0363-6135",
publisher = "American Physiological Society",
number = "1 38-1",

}

TY - JOUR

T1 - Acid incubation increases NHE-3 mRNA abundance in OKP cells

AU - Amemiya, M.

AU - Yamaji, Y.

AU - Cano, A.

AU - Moe, O. W.

AU - Alpern, R. J.

PY - 1995

Y1 - 1995

N2 - With the use of degenerate primers based on conserved amino acid sequences in human, rat, and rabbit Na/H exchanger-3 (NHE-3), a polymerase chain reaction product was obtained from reverse-transcribed OKP (a clonal opossum kidney cell line) mRNA and used to screen an OKP cDNA library. The clone obtained predicted an amino acid sequence that was 86% identical to rat NHE- 3, 33% to NHE-1, 35% to NHE-2, and 30% to NHE-4. Expression of the corresponding cRNA in Xenopus oocytes induced 22Na uptake with ethylisopropylamiloride (EIPA) resistance similar to that of the OKP Na/H antiporter. On RNA blot, the cDNA labeled a 9.5-kb transcript whose abundance was increased 2.2-fold by 24-h incubation of OKP cells at pH 7.0 and 2.5- fold by 24-h incubation at pH 6.8. The acid-induced increase in NHE-3 mRNA was detectable at 12 h and increased further at 24 h. Incubation in acid media caused an increase in EIPA-resistant Na/H antiporter activity that preceded the increase in NHE-3 mRNA. In summary, OKP cells express an NHE-3 transcript that encodes an EIPA resistant Na/H antiporter and is chronically regulated by acid.

AB - With the use of degenerate primers based on conserved amino acid sequences in human, rat, and rabbit Na/H exchanger-3 (NHE-3), a polymerase chain reaction product was obtained from reverse-transcribed OKP (a clonal opossum kidney cell line) mRNA and used to screen an OKP cDNA library. The clone obtained predicted an amino acid sequence that was 86% identical to rat NHE- 3, 33% to NHE-1, 35% to NHE-2, and 30% to NHE-4. Expression of the corresponding cRNA in Xenopus oocytes induced 22Na uptake with ethylisopropylamiloride (EIPA) resistance similar to that of the OKP Na/H antiporter. On RNA blot, the cDNA labeled a 9.5-kb transcript whose abundance was increased 2.2-fold by 24-h incubation of OKP cells at pH 7.0 and 2.5- fold by 24-h incubation at pH 6.8. The acid-induced increase in NHE-3 mRNA was detectable at 12 h and increased further at 24 h. Incubation in acid media caused an increase in EIPA-resistant Na/H antiporter activity that preceded the increase in NHE-3 mRNA. In summary, OKP cells express an NHE-3 transcript that encodes an EIPA resistant Na/H antiporter and is chronically regulated by acid.

KW - acidosis

KW - ethylisopropylamiloride

KW - oocyte expression

KW - sodium/proton antiporter

UR - http://www.scopus.com/inward/record.url?scp=0028855287&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028855287&partnerID=8YFLogxK

M3 - Article

C2 - 7631739

AN - SCOPUS:0028855287

VL - 269

JO - American Journal of Physiology - Heart and Circulatory Physiology

JF - American Journal of Physiology - Heart and Circulatory Physiology

SN - 0363-6135

IS - 1 38-1

ER -