Neuronal nitric oxide synthase: Expression in rat parietal cells

Shyamal Premaratne, Chun Xue, John M. McCarty, Muhammad Zaki, Robert W. McCuen, Roger A Johns, Wolfgang Schepp, Bruno Neu, Robert Lippman, P. D. Melone, Mitchell L. Schubert

Research output: Contribution to journalArticle

Abstract

Nitric oxide synthases (NOS) are enzymes that catalyze the generation of nitric oxide (NO) from L-arginine and require nicotinamide adenine dinucleotide phosphate (NADPH) as a cofactor. At least three isoforms of NOS have been identified: neuronal NOS (nNOS or NOS I), inducible NOS (iNOS or NOS II), and endothelial NOS (eNOS or NOS II). Recent studies implicate NO in the regulation of gastric acid secretion. The aim of the present study was to localize the cellular distribution and characterize the isoform of NOS present in oxyntic mucosa. Oxyntic mucosal segments from rat stomach were stained by the NADPH-diaphorase reaction and with isoform-specific NOS antibodies. The expression of NOS in isolated, highly enriched (>98%) rat parietal cells was examined by immunohistochemistry, Western blot analysis, and RT-PCR. In oxyntic mucosa, histochemical staining revealed NADPH-diaphorase and nNOS immunoreactivity in cells in the mid-portion of the glands, which were identified as parietal cells in hematoxylin and eosin-stained step sections. In isolated parietal cells, decisive evidence for nNOS expression was obtained by specific immunohistochemistry, Western blotting, and RT-PCR. Cloning and sequence analysis of the PCR product confirmed it to be nNOS (100% identity). Expression of nNOS in parietal cells suggests that endogenous NO, acting as an intracellular signaling molecule, may participate in the regulation of gastric acid secretion.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume280
Issue number2 43-2
StatePublished - Feb 2001

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Nitric Oxide Synthase Type I
Nitric Oxide Synthase
Nitric Oxide Synthase Type II
NADP
Nitric Oxide
Protein Isoforms
Gastric Acid
Polymerase Chain Reaction
Mucous Membrane
Western Blotting
Immunohistochemistry
Glycogen Synthase
Nitric Oxide Synthase Type III
Hematoxylin
Eosine Yellowish-(YS)
Sequence Analysis
Arginine
Organism Cloning
Stomach
Staining and Labeling

Keywords

  • Acid
  • Brain nitric oxide synthase
  • Immunohistochemistry
  • Nicotinamide adenine dinucleotide phosphate-diaphorase
  • Nitric oxide
  • Stomach

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology
  • Physiology (medical)

Cite this

Premaratne, S., Xue, C., McCarty, J. M., Zaki, M., McCuen, R. W., Johns, R. A., ... Schubert, M. L. (2001). Neuronal nitric oxide synthase: Expression in rat parietal cells. American Journal of Physiology - Gastrointestinal and Liver Physiology, 280(2 43-2).

Neuronal nitric oxide synthase : Expression in rat parietal cells. / Premaratne, Shyamal; Xue, Chun; McCarty, John M.; Zaki, Muhammad; McCuen, Robert W.; Johns, Roger A; Schepp, Wolfgang; Neu, Bruno; Lippman, Robert; Melone, P. D.; Schubert, Mitchell L.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 280, No. 2 43-2, 02.2001.

Research output: Contribution to journalArticle

Premaratne, S, Xue, C, McCarty, JM, Zaki, M, McCuen, RW, Johns, RA, Schepp, W, Neu, B, Lippman, R, Melone, PD & Schubert, ML 2001, 'Neuronal nitric oxide synthase: Expression in rat parietal cells', American Journal of Physiology - Gastrointestinal and Liver Physiology, vol. 280, no. 2 43-2.
Premaratne, Shyamal ; Xue, Chun ; McCarty, John M. ; Zaki, Muhammad ; McCuen, Robert W. ; Johns, Roger A ; Schepp, Wolfgang ; Neu, Bruno ; Lippman, Robert ; Melone, P. D. ; Schubert, Mitchell L. / Neuronal nitric oxide synthase : Expression in rat parietal cells. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2001 ; Vol. 280, No. 2 43-2.
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