Submucous Neurons Research Articles

Choline acetyltransferase immunoreactivity in the human small and large intestine

BACKGROUND & AIMS: Choline acetyltransferase, an enzyme involved in the synthesis of acetylcholine, is a marker of cholinergic neurons. In this study, the distribution of choline acetyltransferase immunoreactivity in human intestine is described. METHODS: Frozen-section and whole- mount preparations of human small and large bowels were made and labeled with antiserum to choline acetyltransferase. Double labeling with antiserum to neuron-specific enolase enabled the proportion of all neurons that were immunoreactive for choline acetyltransferase to be determined. RESULTS: Nerve fibers, immunoreactive for choline acetyltransferase, were frequent in the circular and longitudinal muscle layers and were widespread in the myenteric and submucous plexuses, but none was observed in the mucosa. Myenteric neurons, immunoreactive for choline acetyltransferase, showed various morphologies, the most common being unipolar and having an irregular outline with several short, lamellar processes. Sixty-four percent of all myenteric neurons were immunoreactive for choline acetyltransferase. Cholinergic submucous neurons were homogeneous in appearance with oval, smooth cell bodies and filamentous dendrites and accounted for 53% of all submucous neurons. A number of cells resembling enteroendocrine cells in the epithelium of the small and large bowels had intense choline acetyltransferase immunoreactivity. CONCLUSIONS: The majority of neurons in human small and large intestines are cholinergic. (Gastroenterology 1996 Aug;111(2):401-8)

Ultrastructural localization of NADPH-diaphorase activity in the submucous ganglia of the guinea-pig intestine after vagotomy.

The reduced nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) activity in the axon terminals presynaptic to the submucous neurons of guinea-pig intestine following unilateral cervical vagotomy was studied by electron microscopy. The reaction product of diaphorase was localized only in the axon terminals that contained predominantly small agranular vesicles, and it was usually deposited around the vesicles. The terminals that contained predominantly large granular or flattened vesicles did not display any signs of diaphorase reactivity. Although there were only few diaphorase-positive submucous neurons in the small intestine, a considerable number of diaphorase-positive axon terminals was observed in the submucous ganglia of the small intestine in the control animals. Ten days after vagotomy, the quantitative study showed that when compared with the control animals, the number of diaphorase-positive terminals in the submucous ganglia of duodenum, mid-small intestine and colon in the vagotomized animals was reduced (P < 0.05). When the NADPH-d-positive terminals were examined in closer detail, it was found that only a small proportion of them showed signs of degeneration as evidenced by the swelling and vacuolation of their contents of mitochondria, with disrupted cristae and clumping of synaptic vesicles. It was therefore concluded that at least some of the diaphorase-positive axon terminals in the submucous ganglia of guinea-pig intestine originated from the vagus nerve.

Localization of choline acetyltransferase and NADPH diaphorase activities in the submucous ganglia of the guinea-pig colon

A combined immunohistochemical and histochemical demonstration of choline acetyltransferase (ChAT) and nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) was carried out, respectively, to determine the localization of the neurotransmitters, acetylcholine and nitric oxide (NO) in the submucous neurons of guinea-pig colon. Almost half of the submucous neurons in the guinea-pig colon exhibited ChAT-immunoreactivity. Some of the ChAT-immunoreactive neurons were also stained for NADPH-d, although most of them showed only weak to moderate diaphorase activity. Many of the submucous neurons displayed exclusively either ChAT or NADPH-d activity. A close spatial relationship was observed between the cholinergic and nitrergic submucous neurons. Thus, in light microscopy, some ChAT-immunoreactive fibres were closely associated with the NADPH-d-positive nerve cell bodies. Ultrastructural study extended the fact that many of the ChAT-immunoreactive terminals made synaptic contacts with the soma of the NADPH-d-positive submucous neurons. A remarkable feature was the demonstration of ChAT and NADPH-d in some of the neurons and their presynaptic axon terminals, suggesting the co-localization of acetylcholine and NO as neurotransmitters in the submucous neurons and their presynaptic axon terminals. It is suggested that the submucous neurons with their specific neurochemical codings would subserve different functions.

ATP-induced currents in submucous plexus neurons of the guinea pig small intestine

ATP-induced membrane durrents in the submucous neurons of the guinea pig small intestine were studied using the whole-cell patch-clamp recording technique. Being applied at −50 mV. ATP activated an inward non-selective cationic current in 68.3% of the investigated neurons. An increase in ATP concentration within the 1–1,000 µM range resulted in the s-like increase in the amplitude of ATP-induced current. The EC50 was 150.0±18.5 µM, while the Hill number was 1.6. The current was selectively activated by ATP and was not blocked by P2 purinoreceptor antagonist suramin (50–300 µM).α,β-Methylene-ATP (100–200 µM) andβ,γ-methylene-ATP (100–200µM), which are P2-purinoreceptor agonists, as well as adenosine (100–300 µM), exerted no effects. Reactive blue 2, if applied up to 4 min, enhanced ATP-induced current, while its longer application partially suppressed this current. In most submucous neurons, acetylcholine (ACh) likewise activated an inward cationic current. The amplitude of ACh-induced current was lower if ACh was applied during a long-lasting application of ATP than if ACh only was applied. Hexamethonium (50 µM), d-tubocurarine (20–40 µM), and trimethaphan (30 µM) completely and reversibly blocked ACh-induced currents, regardless of the presence of ATP, and did not affect ATP-induced currents. The results suggest that ATP-induced currents in submucous neurons are due to activation of a unique type of P2 purinoreceptors, which function in connection with nicotinic ACh receptors.

Do vasoactive intestinal peptide (VIP)- and nitric oxide synthase-immunoreactive terminals synapse exclusively with VIP cell bodies in the submucous plexus of the guinea-pig ileum?

In the submucous plexus of the guinea-pig ileum, previous light-microscopic studies have revealed that vasoactive intestinal peptide (VIP)-immunoreactive and nitric oxide synthase (NOS)-immunoreactive terminals are found predominantly in association with VIP-immunoreactive nerve cell bodies. In this study, double-label immunohistochemistry at the light-microscopic level demonstrated co-localization of NOS-immunoreactivity and VIP-immunoreactivity in axon terminals in submucous ganglia. About 90% of nerve fibres with NOS-immunoreactivity or VIP-immunoreactivity were immunoreactive for both antigens; only about 10% of labelled varicosities contained only NOS-immunoreactivity or VIP-immunoreactivity. The VIP/NOS varicosities were more often seen in the central parts of the ganglia, close to the VIP-immunoreactive cell bodies. Ultrastructural immunocytochemistry with antibodies to VIP was used to determine if NOS/VIP terminals synapse exclusively with VIP-immunoreactive nerve cell bodies. We examined the targets of VIP-immunoreactive boutons in two submucous ganglia from different animals. Serial ultrathin sections were taken through the ganglia after they had been processed for VIP immunocytochemistry. For each cell body, the number of VIP inputs (synapses and close contacts) was determined. The number of VIP-immunoreactive synapses received by the cell bodies of submucous neurons varied from 0-4 and the number of VIP-immunoreactive close contacts varied from 3-10. There was no significant difference between VIP-immunoreactive nerve cell bodies and non-VIP nerve cell bodies in the number of VIP-immunoreactive synapses and close contacts they received. Thus, the implication from light microscopy that NOS/VIP terminals end predominantly on VIP nerve cells was not vindicated by electron microscopy.

Localization of NADPH-diaphorase activity in the submucous plexus of the guinea-pig intestine: light and electron microscopic studies

The localization of reduced nicotinamide adenine dinucleotide phosphate diaphorase in the submucous plexus of duodenum, jejunum, ileum, proximal colon, distal colon and rectum in the guinea-pig was examined histochemically by light and electron microscopy. The majority of reactive submucous neurons displayed features common to either Dogiel type I or type II neurons; some were closely adherent to the outer walls of lymphatic vessels. The use of 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT) at the ultrastructural level showed that nicotinamide adenine dinucleotide phosphate diaphorase is a membrane-associated protein widely distributed in the cells, including the rough endoplasmic reticulum, Golgi apparatus and synaptic vesicles in the axon terminals associated with submucous neurons. On the basis of their diaphorase reactivity or the lack of it, the submucous neuronal somata and their associated terminals were observed to form several different kinds of synaptic configurations. The present quantitative analysis showed that the frequency of reactive submucous neurons in the large intestine was significantly higher than in the small intestine. Based on the ultrastructural localization of the diaphorase reaction product in positive cells, it is speculated that nitric oxide might be synthesized within the neurons. The demonstration of different synaptic configurations in the submucous ganglia suggests that the functional interaction between submucous neurons is extremely complex. Finally, the higher frequency of diaphorase reactive submucous neurons in the large intestine than in the small intestine indicates that submucous neurons in these two gut regions may not play equivalent roles.

Characterization of alkaline phosphatase-reactive neurons in the guinea-pig small intestine

Endogenous alkaline phosphatase activity has been localized histochemically on the surface of enteric neurons of the guinea-pig small intestine by both light and electron microscopy. The enzyme activity was associated with some myenteric neurons that had Dogiel type I morphology, and the histochemical reaction products typically formed a honeycomb-like structure on labelled cell bodies. No Dogiel type II neurons in the myenteric plexus or submucous neurons showed alkaline phosphatase reactivity. Nerve fibres reactive for alkaline phosphatase were present in the myenteric plexus and ran in bundles in the circular muscle and deep muscular plexus. In addition, reactive varicose axons supplied the submucous plexus and non-ganglionated plexus of the mucosa. The results of interruption of the enteric neuronal pathways demonstrated that alkaline phosphatase-reactive myenteric neurons project anally to other myenteric ganglia, to the circular muscle and to the submucous plexus. Sequential enzyme histochemistry showed that virtually all alkaline phosphatase-reactive neurons also contained nitric oxide synthase, revealed by NADPH-diaphorase reactivity. It was estimated that 14–18% of all myenteric neurons showed alkaline phosphatase reactivity. About one-third of nitric oxide synthase-containing myenteric neurons, however, did not contain alkaline phosphatase activity. At the ultrastructural level, alkaline phosphatase activity was associated specifically with the plasma membranes of nerve cell bodies, axons and dendrites of some myenteric neurons. Reactive nerve fibres made close appositions with non-reactive submucous neurons and, within myenteric ganglia, predominantly with other alkaline phosphatase-reactive neurons. In addition to its presence in neurons, alkaline phosphatase reactivity was also present in some endothelial cells in blood vessels in the submucosa and in capillary pericytes. It is concluded, on the basis of the projections and neurochemistry, that in the guinea-pig small intestine alkaline phosphatase activity is associated with nitric oxide synthase-containing neurons which include inhibitory motor neurons to the circular muscle, and anally-directed interneurons to other myenteric and submucous neurons.

Neuroimmune communication in the submucous plexus of guinea pig colon after infection with Trichinella spiralis

Background/Aims: Enteric neuroimmune communication in gastrointestinal hypersensitivity responses includes antigen detection by mast cells and release of chemical messages to the enteric nervous system. The aim of this study was to analyze the electrical and synaptic behavior of neurons in the colonic submucous plexus during exposure to Trichinella spiralis antigen in animals infected earlier with the parasite. Methods: Microelectrodes were used to record in submucous neurons of guinea pig distal colon during application of Trichinella antigen. Results: Neurons in sensitized animals were more excitable than in controls. Hyperexcitability was seen as a greater probability of spontaneous action potential discharge and repetitive firing to depolarizing current or exposure to acetylcholine. Application of histaminergic antagonists reversed the augmented excitability, suggesting endogenously released histamine as a responsible factor. Antigenic exposure increased neuronal excitability and suppressed nicotinic transmission at fast cholinergic synapses only in sensitized animals. Effects on excitability, but not presynaptic inhibitory effects, were blocked by cimetidine. Conclusions: Signaling between mucosal mast cells and the enteric nervous system is involved in colonic anaphylactic responses to sensitizing antigens. Histamine is a paracrine signal in the communication pathway.

Y2‐receptor‐mediated selective inhibition of slow, inhibitory postsynaptic potential in submucous neurones of guinea‐pig caecum

1. The subtype of neuropeptide Y receptor mediating the selective inhibition of the slow inhibitory postsynaptic potential (i.p.s.p.) of submucous neurones in guinea-pig caecum was investigated by use of conventional intracellular electrophysiological recording techniques. 2. Neuropeptide Y (NPY) (1-300 nM) was found to depress or abolish reversibly the slow i.p.s.p. evoked by focal stimulation of internodal fibre tracts. At low concentrations (1-30 nM), a reduction in the duration of the slow i.p.s.p. was often apparent before any inhibition of the amplitude of this synaptic potential. 3. These inhibitory effects of NPY were mimicked by peptide YY (PYY; 0.3-100 nM), NPY13-36 (1-300 nM) and NPY22-36 (10-100 nM); [Leu31,Pro34]NPY ([Pro34]NPY) and bovine pancreatic polypeptide (bPP) were without pre- or postsynaptic effects at concentrations of up to 300 nM. The IC50 +/- s.e. mean values for PYY, NPY, and NPY13-36 were 2.7 +/- 0.3, 7.8 +/- 2.1 and 30 +/- 4.8 nM, respectively, and were significantly different from each other. Thus, the apparent rank order of potency was PYY > NPY > NPY13-36 >> [Pro34]NPY and bPP. 4. In concentrations of up to 300 nM, NPY and its analogues had no depressant effects on the active and passive properties of the impaled neurone and did not affect the amplitude or duration of either cholinergic fast synaptic potentials or non-cholinergic, slow excitatory postsynaptic potentials (e.p.s.ps). Furthermore, none of these peptides altered the amplitude or time-course of changes in membrane potential induced by focal application of acetylcholine or noradrenaline. 5. It is, therefore, concluded that the selective inhibition of the slow i.p.s.p. is mediated by Y2-receptors,located presynaptically on noradrenergic nerve terminals.

Distribution, origin and projections of nitric oxide synthase-containing neurons in gut and pancreas

Nitric oxide has been put forward as an important inhibitory neurotransmitter in the gut. Nitric oxide synthase-containing neurons were visualized by immunocytochemistry using antibodies against neuronal nitric oxide synthase or by β-nicotinamide adenine dinucleotide phosphate diaphorase staining in whole mounts and cryostat sections from the gastrointestinal tract and pancreas of several mamals (mouse, rat, hamster, guinea-pig, cat and man). Nitric oxide synthase-containing neuronal cell bodies were numerous in the myenteric but fewer in the submucous ganglia all along the gut of all species. Varicose nerve terminals formed extensive networks in the circular smooth muscle and the myenteric ganglia. Nitric oxide synthase-containing nerve terminals were frequently found around the Brunner glands in the duodenum; scattered nerve terminals were also found in the gastric and colonie mucosa and around blood vessels in the submucosa all along the gut. In the rat small and large intestine nitric oxide synthase-containing submucous neurons terminated within the mucosa/submucosa and nitric oxide synthase-containing myenteric neurons issued short descending projections, approximately 3 mm, to the smooth muscle and other myenteric ganglia. In the pancreas of all species nitric oxide synthase-containing nerve cell bodies were regularly seen in intrapancreatic ganglia. Positive nerve fibers were mainly found within nerve trunks in interlobular spaces and as delicate fibers within the islets. Double staining for nitric oxide synthase and neuropeptides in intestine and pancreas of rat, guinea-pig and man revealed that only occasionally the nitric oxide synthase-containing nerve cell bodies stored in addition vasoactive intestinal peptide and neuropeptide Y, or enkephalin. However, nitric oxide synthase-containing nerve terminals, particularly those in the circular muscle of the gut, frequently contained vasoactive intestinal peptide/neuropeptide Y (rat and man) or vasoactive intestinal peptide/enkephalin (guinea-pig). In intrapancreatic ganglia few nitric oxide synthase-containing nerve cell bodies were also vasoactive intestinal peptide-immunoreactive. Coexistence of nitric oxide synthase and vasoactive intestinal peptide in nerve terminals could here be detected around blood vessels and interlobular ducts. The distribution of nitric oxide synthase indicates a major role of nitric oxide in the regulation of gut motility; a role in the regulation of blood flow and secretion in both gut and pancreas is also likely.

Neurokinin a mimics the slow excitatory postsynaptic current in submucous plexus neurons of the guinea-pig caecum

Single microelectrode voltage-clamp recordings were made from submucous neurons of the guinea-pig caecum. The slow excitatory postsynaptic current was compared with the currents induced by neurokinin A and substance P. The current induced by neurokinin A (100–300 nM) was associated with a decreased membrane conductance and reversed in polarity between −90 and −100 mV. The neurokinin A current was reduced by Co 2+ (1–2 mM), but was not affected by Cs + (1–2 mM), Ba 2+ (10–100 μM) or low Cl − (20–40 mM) solutions. In about 80% of the neurons, the current induced by substance P (100–300 nM) was associated with a decreased membrane conductance and did not reverse with hyperpolarization of the membrane potential up to −130 mV. The current was reduced by Co 2+ (1–2 mM) and augmented by low Cl − (20–40 mM) solutions, but was not affected by Cs + (1–2 mM) or Ba 2+ (10–100 μM)-containing solutions. In about 20% of the neurons, the substance P current reversed in polarity between −100 and −120 mV. The slow excitatory postsynaptic current elicited by repetitive nerve stimulation (10–40 Hz, three to five pulses) was accompanied by a decreased membrane conductance, and reversed in polarity between −90 and −100 mV. The slow excitatory postsynaptic current was abolished by Co 2+ (1–2 mM) or low Na + (12 mM) solutions, but was not affected by Cs + (1–2 mM), Ba 2+ (10–100 μM) or low Cl − (20–40 mM) solutions. In such neurons, the neurokinin A current was reversed at approximately the same potential at which the slow excitatory postsynaptic current was reversed, while the substance P current was not reversed even by much stronger hyperpolarizations. It was concluded that the neurokinin A current was mainly due to depression of potassium conductances, while the substance P current resulted from both increased anion conductance and decreased potassium conductances. The conductance change underlying the slow excitatory postsynaptic current is similar to that caused by neurokinin A.

Presynaptic inhibition by neuropeptide Y of slow inhibitory synaptic transmission in submucous neurones of guinea-pig caecum.

The occurrence and disposition of immunoreactivity for neuropeptide Y (NPY) was investigated in the submucosa of guinea-pig caecum. There were few NPY-immunoreactive neurones but intraganglionic varicose fibres were so numerous and strongly immunoreactive that some somata may have been obscured. In each neurone tested, exogenous application of NPY (1-100 nM) depressed the duration and amplitude of the slow inhibitory postsynaptic potential. In contrast, NPY (1-300 nM) had no effects on the resting membrane potential, input resistance and fast and slow excitatory synaptic transmission.

Selective presynaptic inhibition by neuropeptide Y of slow IPSP in submucous neurones of guinea-pig caecum

Selective presynaptic inhibition by neuropeptide Y of slow IPSP in submucous neurones of guinea-pig caecum

Elevation of cAMP facilitates noradrenergic transmission in submucous neurons of guinea pig ileum

Slow synaptic excitation and inhibition were studied with intracellular microelectrodes in submucous ganglion cells of the guinea pig ileum. Elevation of adenosine 3',5'-cyclic monophosphate (cAMP) after application of forskolin or the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) resulted in slowly activating depolarization of the membrane potential. The depolarization was associated with increased input resistance, enhanced excitability, and suppression of hyperpolarizing afterpotentials. This occurred in AH/type 2 but not S/type 1 neurons. The action of forskolin or IBMX mimicked slow synaptic excitation in the same neurons. Focal electrical stimulation also evoked slow inhibitory postsynaptic potentials (IPSPs). The amplitude and duration of the IPSPs were increased by forskolin or a membrane-permeant analogue of cAMP. Treatment with phentolamine, yohimbine or idazoxan suppressed the IPSPs before and after potentiation by forskolin, suggesting that the IPSPs were mediated by release of norepinephrine acting at alpha 2-adrenoceptors. Application of adenosine or selective adenosinergic A1 agonists suppressed or abolished the IPSPs. The results suggest that elevation of cAMP facilitates the release of norepinephrine at alpha 2-synapses on submucous neurons of guinea pig small bowel.

Histamine receptors on submucous neurons in guinea pig colon.

Intracellular microelectrodes were used to investigate the actions of histamine in the submucous plexus of the distal colon of the guinea pig. Three effects resulted from application of histamine to submucous neurons. The first was membrane depolarization associated with increased input resistance and augmented excitability. The second was presynaptic suppression of acetylcholine release at nicotinic synapses. The third occurred during long-term application and consisted of recurrent trains of action potentials associated with periodic depolarization of membrane potential. Pharmacological analysis, with selective agonists and antagonists, suggested mediation of the first and third response by postsynaptic histamine H2 receptors. The second response was mediated by presynaptic histamine H3 receptors. These actions of histamine represent a mechanism for neuroimmune signaling between mucosal mast cells and submucous neurons in gastrointestinal type 1 hypersensitivity reactions to allergens.

Submucous neurons are necessary for regeneration of rat jejunal muscle layers

Submucous neurons are necessary for regeneration of rat jejunal muscle layers

Effects of neuropeptide Y on submucous neurons in the guinea-pig small intestine

Effects of neuropeptide Y on submucous neurons in the guinea-pig small intestine

Projections and pathways of submucous neurons to the mucosa of the guinea-pig small intestine.

Double-labelling immunohistochemistry and retrograde transport of the carbocyanine dye, DiI, were used to establish the pathways of submucous neurons to the mucosa of the guinea-pig small intestine. Following the application of DiI to a villus, DiI-labelled nerve cell bodies were found in the submucous plexus up to 8.3 mm circumferentially and 3.8 mm longitudinally. The size of each of the four characterised classes of submucous neurons was determined and their distributions and projections mapped. Cells characterised by vasoactive intestinal polypeptide immunoreactivity accounted for 52% of DiI-labelled cells and had the longest projections. Cells characterised by neuropeptide Y (19%) or by calretinin immunoreactivity (13% of all DiI-labelled neurons) had relatively short projections and cells with substance P immunoreactivity (20%) had intermediate lengths of projection. When DiI was applied directly to the submucous plexus, filled neurons of all classes had significantly shorter projections, indicating that they must run for considerable distances in other pathways to the mucosa, probably via the non-ganglionated plexus. On average, each villus is innervated by at least 70 submucous neurons. From quantitative estimates there are 9 submucous neurons per villus. Thus, each submucous neuron is likely to supply about 8 villi. This demonstrates a high degree of convergence and divergence in the innervation of the mucosa.

Pituitary adenylate cyclase activating polypeptides, PACAP-27 and PACAP-38: stimulators of electrogenic ion secretion in the rat small intestine.

1. The effects of pituitary adenylate cyclase activating polypeptide (PACAP)-27 and PACAP-38 were investigated and compared with vasoactive intestinal polypeptide (VIP) responses in voltage clamped preparations of rat jejunum. Under these conditions electrogenic ion secretion was continuously recorded. 2. PACAP-27 is the most potent secretagogue described thus far, exhibiting a concentration-dependent dual secretory action. At low concentrations it stimulated rapid, transient secretory responses (not seen with either PACAP-38 or VIP) and these were inhibited by tetrodotoxin (TTX). At higher nM concentrations of PACAP-27 more prolonged secretory responses predominated which were insensitive to TTX. 3. In the presence of TTX, the concentration-response curve to PACAP-27 gave an EC50 value of 29.4 +/- 5.4 nM (n = 4) compared with 0.8 +/- 0.1 nM (n = 9) for PACAP-27 alone and 30.6 +/- 5.6 nM (n = 5) for PACAP-38. C-terminal fragments of PACAP-38 were not significantly effective. 4. Blockade of muscarinic and nicotinic receptors partially inhibited the low concentration effects of PACAP-27. Substance P desensitization and capsaicin pretreatment were effective at inhibiting the transient secretory PACAP-27 responses. Evidence is presented for selective, high affinity PACAP-27 receptors on submucous neurones innervating the mucosal region of the rat jejunum.

Differential effect of immunosympathectomy on the expression of rat enteric neurotransmitters

The effect of immunosympathectomy on the pattern of distribution of catecholamine- and peptide-containing nerve fibres and neurones in the myenteric and submucous plexuses of rat ileum was investigated. There was an increase in vasoactive intestinal polypeptide (VIP)-, galanin (GAL)- and substance P-like immunoreactivity in the myenteric plexus of ileum from rats treated with nerve growth facter (NGF) antiserum compared with controls. A similar increase in immunoreactivity was observed in VIP-. GAL- and neuropeptide Y (NPY)-containing submucous neurones and nerve fibres. In contrast, the immunosympathectomy had no effect on the pattern of distribution of catecholamine-, calcitonin gene-related peptide (CGRP)- and NPY-containing nerve fibres in the myenteric plexus or on substance P- and CGRP-containing neurones and nerve fibres of the submucous plexus. The findings of the present study suggest that NGF may differentially regulate the expression of enteric neuropeptides at a postnatal stage of development.