Distribution of peptide- and catecholamine-containing neurons in the gastro-intestinal tract of rat and guinea-pig: Immunohistochemical studies with antisera to substance P, vasoactive intestinal polypeptide, enkephalins, somatostatin, gastrin/cholecystokinin, neurotensin and dopamine β-hydroxylase

doi:10.1016/0306-4522(80)90166-9

Neuroscience

Volume 5, Issue 4, April 1980, Pages 689-744

https://doi.org/10.1016/0306-4522(80)90166-9 Get rights and content

Abstract

The distribution of peptide-containing neurons in the oesophagus, stomach and small and large intestine of the rat and the guinea-pig has been studied with the indirect immunofluorescence technique ofCoons & Co-workers (1958) using antisera to substance P, vasoactive intestinal polypeptide (VIP), enkephalin, somatostatin, gastrin and neurotensin. (The gastrin antiserum is to the C-terminal portion and consequently reacts also with cholecystokinin (CCK)-like peptides.) For comparison, the noradrenergic innervation was visualized with antiserum to dopamine β-hydroxylase. For improved visualization of peptide-containing cell bodies, a mitotic inhibitor (colchicine or vinblastine) was applied locally on the different parts of the gastro-intestinal tract of several animals.

Substance P-, VIP-, enkephalin- and somatostatin-like immunoreactivity was observed in all parts of the gastro-intestinal tract studied. Gastrin/CCK had a more limited distribution, especially in the guinea-pig and neurotensin was seen only in certain regions and layers of the rat gastro-intestinal tract.

Immunoreactivity to all peptides except neurotensin was observed both in cell bodies and fibres; immunoreactivity to neurotensin has so far only been seen in nerve fibres. Substance P and enkephalin immunoreactive cells were often numerous in the myenteric plexus, whereas VIP and somatostatin immunoreactive cells were preferentially located in the submucous plexus. Some VIP immunoreactive cells were observed in the lamina propria. Large numbers of especially substance P-, VIP- and enkephalin-containing fibres were often seen in the circular muscle layer and in the two ganglionic plexuses. Substance P immunoreactive fibres formed the densest network in the ganglionic plexuses, whereas VIP immunoreactive fibres constituted the most impressive network in the lamina propria and often extended into the most superficial parts of the mucosa. Enkephalin immunoreactive structures were mainly confined to the circular and longitudinal muscle layers and the myenteric plexus. Somatostatin immunoreactive fibres were mainly found in the ganglionic plexuses.

Peptide-containing fibres, particularly these containing substance P and VIP were often seen along blood vessels, but never with such a density as the noradrenergic (dopamine β-hydroxylase immunoreactive) fibres. No somatostatin or neurotensin immunoreactive fibres were observed in relation to clearly identifiable blood vessels.

The possible coexistence of two peptides in one neuron was studied. For this part of the study the proximal colon and five antisera, namely substance P, VIP, enkephalin. somatostatin and gastrin/CCK antisera were selected. Evidence was obtained for the occurrence of a somatostatin-like and a gastrin/ CCK-like peptide in the same neurons. This may indicate a common precursor for the two peptides in these particular neurons. Each of the substance P-, VIP- and enkephalin-like peptides. on the other hand, seem to be present in different neuronal populations, which were themselves distinct from the somatostatin-gastrin/CCK immunoreactive neurons. In addition, somatostatin immunoreactive neurons different from the gastrin/CCK immunoreactive ones seem to exist. The gastrin/CCK immunoreactive fibres around blood vessels may represent a further, separate population of fibres, since no somatostatin immunoreactive fibres were seen at this location.

The findings indicate the existence of numerous subpopulations of enteric neurons, each characterized by its content of a certain peptide (or peptides). The axons of most of these neurons probably terminate in the wall of the gastro-intestinal tract, but some seem to project to other organs. In addition, some peptide-containing fibres in the gastro-intestinal wall may have an extrinsic origin. The relationship between these peptide-containing neurons and the cholinergic enteric neurons and any of the other non-cholinergic. non-adrenergic inhibitory and excitatory neurons present in the enteric nervous system is not known. It is, however, noteworthy that a somatostatin-like peptide seems to be present in noradrenergic neurons of prevertebral ganglia that project to the intestine. The possibility must be kept in mind that one or more of the peptides in the gut could be localized in neurons that contain other potential transmitters, e.g. acetylcholine.

The wide variety of pharmacological actions of these neuronal peptides on smooth muscle and neurons in the gut and on its blood vessels raises the possibility that some of them may be neurotransmitters.

Reference (246)

ÅlundM. et al.
Depolarization-induced decreases in fluorescence intensity of gastrointestinal quinacrinebinding nerves
Brain Res.
(1979)
ArimuraA. et al.
Radioimmunoassay of somatostatin
Metabolism
(1978)
BlochB. et al.
Immunocytochemical evidence that the same neurons in the human infundibular nucleus are stained with anti-endorphins and antisera of other related peptides
Neuroscience Letters
(1978)
BloomS.R. et al.
Vasoactive intestinal peptide and the watery diarrhoea syndrome
Lancet
(1973)
BryantM.G. et al.
Possible dual role for vasoactive intestinal peptide as gastrointestinal hormone and neurotransmitter substance
Lancet
(1976)
BurnstockG.
Do some nerve cells release more than one transmitter?
Neuroscience
(1976)
CarrawayR. et al.
The isolation of a new hypotensive peptide, neurotensin, from bovine hypothalami
J. biol. Chem.
(1973)
CarrawayR. et al.
The amino acid sequence of a hypothalamic peptide, neurotensin
J. biol. Chem.
(1975)
CocksT. et al.
Effects of neuronal polypeptides on intestinal smooth muscle; A comparison with non-adrenergic, non-cholinergic nerve stimulation and ATP
Eur. J. Pharmacol.
(1979)
CohenM.L. et al.
Somatostatin inhibits adrenergic and cholinergic neurotransmission in smooth muscle
Life Sci.
(1978)

BeleslinD. et al.

The effect of substance P on the responses of the isolated guinea-pig ileum to acetylcholine, nicotine, histamine and 5-hydroxytryptamine

Acta int. pharmacodyn.

(1960)

BennettM.R. et al.

Electrophysiology of the innervation of intestinal smooth muscle

BennettM.R. et al.

Transmission from intramural inhibitory nerves to the smooth muscle of the guinea-pig taenia coli

J. Physiol., Lond.

(1966)

BessonJ. et al.

Vasoactive intestinal peptide (VIP): Tissue distribution in the rat as measured by radioimmunoassay and by radioreceptorassay

Acta endocr., Copnh.

(1978)

BloomF. et al.

Neurons containing β-endorphin in the rat brain exist separately from those containing enkephalin: Immunocytochemical studies

BrazeauP. et al.

Hypothalamic polypeptide that inhibits the secretion of immunoreactive pituitary growth hormone

Science, N.Y.

(1973)

BrodinE. et al.

Tissue concentrations of substance P-like immunoreactivity (SPLI) in various organs in dog, rat and mouse

Acta physiol. scand.

(1980)

BrownM. et al.

Neurotensin-like and bombesin-like peptides: CNS distribution and actions

Nonadrenergic, non-cholinergic autonomic neurotransmission mechanisms

Neurosciences Rec. Progr. Bull.

(1979)

BuryR.W. et al.

A pharmacological investigation of synthetic substance P on the isolated guinea pig ileum

Clin. exp. Pharmacol. Physiol.

(1977)

CampbellG. et al.

Comparative physiology of gastrointestinal motility

CarrawayR. et al.

The amino acid sequence, chemical synthesis and radioimmunoassay of neurotensin

ChangM.M. et al.

Amino-acid sequence of substance P

Nature, New Biol.

(1971)

Chan-PalayV.

Innervation of cerebral blood vessels by norepinephrine, indoleamine, substance P and neurotensin fibers and the leptomeningeal indoleamine axons: Their roles in vasomotor activity and local alterations of brain blood composition

Chan-PalayV. et al.

Serotonin and substance P coexist in neurons of the rats' central nervous system

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Neurons containing β-endorphin in the rat brain exist separately from those containing enkephalin: Immunocytochemical studies

Hypothalamic polypeptide that inhibits the secretion of immunoreactive pituitary growth hormone

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Neurotensin-like and bombesin-like peptides: CNS distribution and actions

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