Nitric oxide in the rat gastrointestinal tract

  • Kenneth J. Price

Student thesis: Doctoral ThesisDoctor of Philosophy

Abstract

This study concerns the nature of nitric oxide synthase (NOS) and the role of nitric oxide (NO) in the rat gastrointestinal tract. The major objectives were (i) to characterise NOS isoforms in the gastric glandular mucosa, (ii) to localise NOS isoforms in the rat gastric glandular mucosa, (iii) to investigate the role of NO in carbachol-stimulated gastric mucus secretion, (iv) to investigate the nature of NOS and small intestine.
Immunoblotting was performed using polyclonal antisera raised against two peptides found in the rat brain NOS sequence and commercial monoclonal antibodies directed against neuronal and endothelial isoforms of NOS. A160kDa band was detected in brain and gastric mucosal samples with antibodies and antisera directed against neuronal NOS sequences, and a 140kDa band was detected in gastric mucosal samples using an anti-endothelial NOS antibody. An intense 160kDa neuronal NOS band was detected in a high-density fraction of gastric mucosal cells separated on a Percoll gradient. Detection of neuronal NOS by a carboxyl-terminal antiserum in samples of brain, but not of gastric mucosa, could be blocked by the peptide (20g/ml) against which the antibody was raised. After affinity purification, recognition of gastric mucosal NOS was blocked by peptide.
Particulate neuronal NOS was found in the brain by immunoblotting while 94% of gastric mucosal enzyme was soluble. Gastric mucosal endothelial NOS was 95% particulate. 95% of NOS activity in the gastric mucosa was due to neuronal NOS.
Paraformaldehyde- and acetone-fixed gastric mucosal sections were subject to
immunocytochemistry using the above antibodies. Neuronal NOS was localised to
the surface mucosal epithelial cells while endothelial NOS was associated with
microvessels at the base of the mucosa and to larger vessels in the submucosa.
Intragastric administration of carbachol or 16, 16-dimethyl prostaglandin E2
increased the thickness of the rat gastric mucus layer. The NOS inhibitor NG-nitro-L-arginine methyl ester dose-dependently, and selectively, prevented the
stimulatory effect of carbachol.
Ca2+-independent NOS activity in rat ileal, jejunal and colonic muscle was
increased after LPS induction. Ca2+-dependent activity was not affected.
Distribution of inducible NOS protein paralleled Ca2+ -independent activity. LPS
treatment did not affect the content of neuronal NOS in colonic muscle.

Date of AwardJan 1997
Original languageEnglish
SupervisorPeter J Hanson (Supervisor)

Keywords

  • nitric oxide
  • NO
  • nitric oxide synthase
  • gastric mucosa
  • mucus
  • lipopolysaccharide
  • colon
  • small intestine

Cite this

Nitric oxide in the rat gastrointestinal tract
Price, K. J. (Author). Jan 1997

Student thesis: Doctoral ThesisDoctor of Philosophy