Colocalization of nitric oxide synthase and NADPH-diaphorase in the myenteric plexus of the rat gut

doi:10.1016/0304-3940(92)90233-W

Neuroscience Letters

Volume 143, Issues 1–2, 31 August 1992, Pages 60-64

https://doi.org/10.1016/0304-3940(92)90233-W Get rights and content

Abstract

The pattern of distribution and colocalization of nitric oxide-synthase (NOS) and NADPH-diaphorase in the myenteric plexus of wholemount preparations of the antrum, duodenum, ileum, caecum, proximal colon and distal colon of the rat were investigated using immunohistochemical and histochemical staining techniques. Almost all the myenteric neurons that were NOS-positive in all regions of the gut examined were also stained for NADPH-diaphorase. However, in the stomach, duodenum and iluem, only a few of the NOS-positive nerve fibres in the tertiary and secondary plexuses and circular muscle layer were also stained for NADPH-diaphorase, whereas in the caecum and distal colon almost all the NOS-positive nerve fibres were also stained for NADPH-diaphorase. The results in the present study are consistent with the view that nitric oxide (NO) has a mediating role in gastrointestinal neurotransmission.

References (23)

A. Belai et al.
Selective damage of intrinsic calcitonin gene-related peptide-like immunoreactive enteric nerve fibres in streptozotocin-induced diabetic rats
Gastroenterology
(1987)
A. Belai et al.
Changes in adrenergic and peptidergic nerves in the submucous plexus of streptozotocin-diabetic rat ileum
Gastroenterology
(1990)
U. Förstermann et al.
Isoforms of nitric oxide synthase: characterisation and purification from different cell types
Biochem. Pharmacol.
(1991)
J.P. Niel et al.
Apamin resistant poststimulus hyperpolarization in the circular muscle of the guineapig ileum
J. Auton. Nerv. Syst.
(1983)
C.W.R. Shuttleworth et al.
Evidence that nitric oxide participates in non-adrenergic inhibitory transmission to intestinal muscle in the guinea-pig
Neuroscience Lett.
(1991)
S.H. Snyder et al.
Nitric oxide as neuronal messenger
Trends Pharmacol. Sci.
(1991)
G.E. Boeckxsteans et al.
Non-adrenergic non-cholinergic relaxation mediated by nitric oxide in the canine ileocolonic junction
Eur. J. Pharmacol.
(1990)
D.S. Bredt et al.
Localization of nitric oxide synthase indicating a neural role for nitric oxide
Nature
(1990)
M. Costa et al.
Apamin distinguishes two types of relaxation mediated by enteric nerves in the guinea-pig gastrointestinal tract
Naunyn-Schmiedebergs Arch. Pharmacol.
(1986)
T.M. Dawson et al.
Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues

R.F. Furchgott et al.

The obligatory role of endothelial cells in the relaxation arterial smooth muscle by acetylcholine

Nature

(1980)

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Colocalization of nitric oxide synthase and NADPH-diaphorase in the myenteric plexus of the rat gut

Abstract

Gastroenterology

Gastroenterology

Biochem. Pharmacol.

J. Auton. Nerv. Syst.

Neuroscience Lett.

Trends Pharmacol. Sci.

Non-adrenergic non-cholinergic relaxation mediated by nitric oxide in the canine ileocolonic junction

Eur. J. Pharmacol.

Localization of nitric oxide synthase indicating a neural role for nitric oxide

Nature

Apamin distinguishes two types of relaxation mediated by enteric nerves in the guinea-pig gastrointestinal tract

Naunyn-Schmiedebergs Arch. Pharmacol.

Nitric oxide synthase and neuronal NADPH diaphorase are identical in brain and peripheral tissues

The obligatory role of endothelial cells in the relaxation arterial smooth muscle by acetylcholine

Nature