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Deficient neurogenic innervation of the myenteric plexus with normal submucous plexus involving the entire small and large bowel.

A newborn presented with a picture of intestinal obstruction. Multiple biopsies of the small and large bowel showed an unusual neurogenic innervation. The myenteric plexus of Auerbach was severely depleted of ganglion cells and nerve fibers, while the submucous plexus of Meissner was normally innervated. An ileostomy failed to function and extended trial with experimental smooth muscle stimulant (Cisapride) was equally ineffective. The patient was finally treated by a myectomy from the duodenum to the descending colon with a sigmoid colostomy. This procedure, coupled with a Nissen fundoplication, stopped the vomiting and allowed normal defecation through the colostomy. The patient is presently taking increasing increments of oral fluids with a concomitant decrease in the volume of parenteral nutrition. The myectomy initiated marked hypertrophy of the muscularis mucosa. Could this muscular hypertrophy account for the improvement in bowel function? Possible etiology will be discussed. We caution that rectal submucosal suction biopsy alone may be misleading if normal ganglion cells and nerve fibers are found, yet the patient's clinical symptoms fail to improve. A full thickness bowel wall biopsy is then recommended.

Ganglia, Parasympathetic

[The external submucous plexus (Schabadasch) in the small intestine of the swine. I. Form, structure and connections of ganglia and nerve cells].

1. There are described principles of organization of the plexus submucous externus (Schabadasch) in the small intestine of the pig. They can be summarized under the term of polarization of nerve cells. 2. The principle of polarization is realized for instance by the formation of type I and type II cell aggregates and by the specific course of the processes of both cell types. 3. These criteria of organization which are much clearer in the pig than in other animals, recommend the plexus Schabadasch of the pig as a suitable model for further investigations.

Animals

Contractions of the guinea-pig ileum evoked by stimulation of the submucous plexus.

Neural pathways from the submucous plexus to the longitudinal muscle of an adjacent segment of isolated guinea-pig ileum were studied. It was found that electrical field stimulation of a strip of submucosa-submucous plexus produced frequency-dependent longitudinal contractions of an intact segment of intestine lying oral to the point of stimulation. The responses were reduced to less than 10% of control by tetrodotoxin, atropine, morphine and chymotrypsin and by desensitization to substance P (SP). The responses were only inhibited by one-third by hexamethonium and were not affected by desensitization to 5-hydroxytryptamine. The effect of desensitization to SP was reversible, but the effect of chymotrypsin was irreversible. SP-induced desensitization and chymotrypsin did not inhibit the twitch response produced by field stimulation of the whole ileal segment. The same results were observed with preparations made from ileal segments that had been extrinsically denervated. The results suggest that intrinsic neurons with processes in the submucous plexus can excite cholinergic and SP-containing neurons in the myenteric plexus, thereby causing the longitudinal muscle to contract.

Animals

Ultrastructural study of nerve terminals in the submucous plexus and mucous membrane after extirpation of the myenteric plexus.

An attempt is made to separate in the submucous plexus and mucous membrane of the chronically isolated intestine of the cat the fibers originating from the myenteric plexus and those originating from the submucous plexus by secondary degeneration after the extirpation of the myenteric plexus. A considerable part of the nerve processes in the submucous plexus originate from the cells of the myenteric ganglia and establish direct synaptic relationships between the ganglion cells, or are very close to the blood vessels. The numerous intact synapses between the different nerve elements in the submucous plexus point to local reflex connexions within the plexus itself. The degeneration of sub-epithelial nerve elements, containing always clear and dense-core (1,000-1,200 A in diameter) vesicles besides the numerous intact nerve elements, might indicate the presence of real sensory nerve processes in this layer. The terminal fibers remaining intact after removal of the myenteric plexus contain numerous medium-sized dense-core vesicles in potassium permanganate-fixed material. This suggests that there are cells containing 5-hydroxytryptamine in the submucous plexus.

Animals

Neuronal populations in the submucous plexus of the human colon.

The submucous plexus of the human distal colon was studied in order to determine whether or not it contains two or more ganglionated plexuses which can be separately identified. Nerve cells were visualised in sections through the wall of the distal colon, and in wholemount preparations of laminae from the submucous plexus by staining for NADH-diaphorase activity. The submucous plexus appeared to contain three identifiable plexuses: Henle's plexus was located adjacent to the circular muscle layer, Meissner's plexus was located adjacent to the muscularis mucosae and a third intermediate plexus was found which lay closer to the muscularis mucosae than to the circular muscle. In Henle's plexus, there were fewer smaller neurons than in the other plexuses: 15.1% had an area less than 180 microns 2, while in the intermediate plexus and in Meissner's plexus the equivalent figures were 43.1% and 43.2%, respectively. In Meissner's plexus, approximately half the ganglia were associated with single fibre tracts and half with two or more fibre tracts, but in the intermediate plexus and Henle's plexus, approximately three quarters of the ganglia were associated with single fibre tracts and the remaining quarter with multiple tracts.

Adult

Fine structure of the myenteric and submucous plexuses in the stomach of a coral fish, Chelmon rostratus Cuvier.

The fine structure of the teleostean myenteric and submucous plexuses has been studied in the stomach of the coral fish, Chelmon rostratus Cuvier. The myenteric plexus is a prominent loose mesh containing nerve cells, myelinated and unmyelinated axon profiles, vesiculated axon profiles, Schwann cells, collagen and capillaries. Unmyelinated axons greatly outnumber the myelinated ones. Vesiculated axon profiles were uncommon, and synapses were rarely observed. Compared with the myenteric plexus, the submucous plexus was more modest in size; it was devoid of nerve cells and myelin sheaths were only occasionally seen.

Animals

Inhibitory synaptic potentials resulting from alpha 2-adrenoceptor activation in guinea-pig submucous plexus neurones.

Intracellular recordings were obtained from neurones of the guinea-pig submucous plexus. Inhibitory synaptic potentials (i.p.s.p.s) were compared with hyperpolarizations evoked by brief, local applications of noradrenaline and by superfusion with adrenoceptor agonists. Hyperpolarizing potentials elicited by brief applications of noradrenaline were similar to the i.p.s.p. in latency of onset, amplitude, time course, conductance increase, reversal potential and ionic dependence. Both responses were blocked by low concentrations of Ba2+ and quinine. 6-hydroxydopamine selectively and irreversibly abolished the i.p.s.p. and resulted in a complete loss of catecholamine fluorescent nerve fibres in the submucous plexus. The alpha 2-adrenoceptor antagonists, phentolamine, yohimbine and RX781094, reversibly blocked the i.p.s.p. and the noradrenaline hyperpolarization. Prazosin, propranolol, atropine and naloxone had no effect on these responses. Superfusion with noradrenaline and clonidine produced dose-dependent membrane hyperpolarizations. Noradrenaline and clonidine dose-hyperpolarization curves were shifted to the right in a parallel fashion by alpha 2-adrenoceptor antagonists. Determination of the dissociation equilibrium constants for phentolamine, yohimbine and RX781094 showed that the hyperpolarization produced by noradrenaline perfusion is due to alpha 2-adrenoceptor activation. It is concluded that the release of noradrenaline from sympathetic nerves activates post-synaptic alpha 2-adrenoceptors, resulting in the K+ conductance increase which underlies the i.p.s.p. in submucous plexus neurones.

Action Potentials

A non-adrenergic, non-cholinergic slow inhibitory post-synaptic potential in neurones of the guinea-pig submucous plexus.

1. Intracellular recordings were made from neurones in the submucous plexus of guinea-pig ileum and caecum. The responses to electrical stimulation of fibre strands entering the nodes of the plexus were studied. 2. Stimuli comprising trains of pulses (20 Hz, 1-5 s) produced nicotinic excitatory post-synaptic potentials (fast e.p.s.p.s), an adrenergic inhibitory post-synaptic potential (i.p.s.p.), a slow excitatory post-synaptic potential (slow e.p.s.p.) and a fourth, hitherto unnoticed, slow hyperpolarization which followed the slow e.p.s.p. All these responses were abolished by tetrodotoxin or solutions containing a low calcium concentration. 3. The slow hyperpolarization (slow i.p.s.p.) was examined in the presence of blockers of the nicotinic and adrenergic responses, and in conditions in which the slow e.p.s.p. was prevented by desensitizing concentrations of substance P or vasoactive intestinal polypeptide. The slow i.p.s.p. was unaffected by prazosin (0.1-1 microM), propranolol (0.1-1 microM), atropine (1 microM) or naloxone (1 microM). 4. The amplitude and duration of the slow i.p.s.p. increased with increasing numbers of stimulus pulses; it had an amplitude of 17 mV and a duration of 70 s when evoked by a stimulus of 20 Hz for 3 s. 5. The slow i.p.s.p. was associated with a decrease in the input resistance of the cell. It reversed polarity at -90 mV in 4.7 mM-potassium and the extrapolated reversal potential in 0.47 mM-potassium was -145 mV; these findings indicate that the slow i.p.s.p. results from an increase in membrane potassium conductance. 6. The slow i.p.s.p. could still be recorded from submucous plexus neurones in segments of ileum which had been extrinsically denervated 6-11 days previously.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials

Somatostatin increases an inwardly rectifying potassium conductance in guinea-pig submucous plexus neurones.

1. Intracellular recordings were made from neurones in the submucous plexus of the guinea-pig caecum and ileum. 2. Somatostatin hyperpolarized more than 90% of the neurones. The lowest effective concentration was 300 pM and the maximum hyperpolarization (about 30-35 mV) was caused by 30 nM. Under voltage clamp at -60 mV, somatostatin caused outward currents which reached a maximum of 350-700 pA. 3. The hyperpolarization or outward current reversed polarity at a membrane potential (about -90 mV in control solutions) which changed according to the logarithm of the external potassium concentration. 4. The somatostatin current showed inward rectification; when the inward rectification of the resting membrane was prevented by extracellular caesium or rubidium, the inward rectification of the somatostatin current also disappeared. 5. A potassium conductance with the same properties was increased by alpha 2-adrenoceptor agonists and by delta-opioid receptor agonists; however, the effects of somatostatin were unaffected by antagonists at alpha 2- or delta-receptors. The somatostatin analogue, cyclo-aminoheptanoyl-Phe-D-Trp-Lys-(benzyl)Thr, also did not antagonize the actions of somatostatin. 6. The hyperpolarization (or outward current) was unaffected by forskolin, cholera toxin, sodium fluoride, phorbol esters or intracellular application of adenosine 5'-O-(3-thiotriphosphate) (ATP-gamma-S). However, when the recording electrode contained guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) the hyperpolarizations reversed only partially when somatostatin application was discontinued, and repeated applications caused the membrane potential to approach and remain close to the potassium equilibrium potential. 7. It is concluded that somatostatin increases the conductance of a set of inwardly rectifying potassium channels in submucous plexus neurones. The coupling between somatostatin receptor and ion channel involves a guanosine 5'-triphosphate-binding protein, but is not likely to result from changes in intracellular levels of cyclic adenosine 3',5'-monophosphate.

Action Potentials

Peptide-containing neurons in different regions of the submucous plexus of human sigmoid colon.

Specimens of the sigmoid colon were obtained from male and female patients (n = 11) with carcinoma of the colon or rectum and studied immunohistochemically for vasoactive intestinal polypeptide-, somatostatin-, substance P-, neuropeptide Y-, calcitonin gene-related peptide-, met- and leu-enkephalin-, 5-hydroxytryptamine-, and dopamine beta-hydroxylase-containing nerves. In the subdivisions of the submucous plexus (namely, Schabadasch's, Meissner's, and the intermediate plexuses), substance P- and vasoactive intestinal polypeptide-immunoreactive nerve fibers were the most numerous, and equal densities of these nerves were found in all three layers. In contrast, few neuropeptide Y-, met-enkephalin-, leu-enkephalin-, calcitonin gene-related peptide-, somatostatin-, 5-hydroxytryptamine-, and dopamine beta-hydroxylase-immunoreactive nerves were found in these regions. The nerve cell bodies of the submucous plexus contained vasoactive intestinal polypeptide, substance P, leu-enkephalin, somatostatin, and 5-hydroxytryptamine but not neuropeptide Y, met-enkephalin, calcitonin gene-related peptide, and dopamine beta-hydroxylase. Vasoactive intestinal polypeptide-containing nerve cell bodies were found in all three subdivisions. Substance P-, leu-enkephalin-, and somatostatin-immunoreactive nerve cell bodies were found in Schabadasch's plexus and the intermediate region of the submucous plexus, but they were absent from Meissner's plexus; 5-hydroxytryptamine-containing nerve cell bodies were only observed in Schabadasch's plexus. The possible function of the neuropeptide-, dopamine beta-hydroxylase-, and 5-hydroxytryptamine-containing neurons in the different layers of the submucous plexus is discussed.

Adult

Electrophysiological characterization of functionally distinct 5-hydroxytryptamine receptors on guinea-pig submucous plexus.

Intracellular recordings were made from neurons of the guinea-pig submucous plexus and the actions of 5-hydroxytryptamine on the postsynaptic membrane and on evoked synaptic potentials were examined. 5-Hydroxytryptamine produced two types of direct postsynaptic responses: (1) A depolarization associated with a fall in input resistance was observed in all cells. Voltage-clamp and ion substitutions showed that this depolarization resulted primarily from an inward sodium current. This response could be as brief as 30 ms; it showed desensitization and was selectively abolished by 0.2-2 microM ICS 205-930. (2) A depolarization (or inward current) associated with a decreased conductance was observed in about 50% of neurons, usually after the first response was blocked by ICS 205-930. This response was due to a decreased potassium conductance; the minimum time course of this response was 8-10 s. It did not show desensitization and was not sensitive to blockade by currently available antagonists of 5-hydroxytryptamine, nicotinic and/or muscarinic receptors. Higher concentrations of 5-hydroxytryptamine were required to produce the sodium conductance increase than the potassium conductance decrease; 2-methyl-5-hydroxytryptamine was equally effective in producing these responses. 5-Hydroxytryptamine also caused a barrage of "spontaneous" nicotinic excitatory post-synaptic potentials which were sensitive to tetrodotoxin. This response desensitized, was blocked by ICS 205-930 and is presumed to reflect excitation of other cholinergic cell bodies in the plexus by the sodium conductance increase mechanism described. The evoked nicotinic excitatory postsynaptic potential and the adrenergic inhibitory postsynaptic potential were decreased by 5-hydroxytryptamine; a portion of this inhibition showed desensitization and was blocked by ICS 205-930 as well as by the muscarinic receptor antagonists, atropine and pirenzepine. The ICS 205-930-insensitive portion of this inhibition could not be attributed to activation of 5-hydroxytryptamine-1 or 5-hydroxytryptamine-2 receptors. Thus, the following conclusions are drawn: 5-hydroxytryptamine excites submucous plexus neurons by activating two distinct 5-hydroxytryptamine receptors. Activation of the 5-hydroxytryptamine-3 receptor (sensitive to ICS 205-930) produces a depolarization mediated by an increased sodium conductance. The same effect occurring in other cholinergic cell bodies initiates action potentials which are responsible for the 5-hydroxytryptamine-induced release of acetylcholine.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Muscarinic excitation and inhibition of neurons in the submucous plexus of the guinea-pig caecum.

Intracellular recordings were made from neurons in the submucous plexus of the guinea-pig caecum. Muscarinic agonists (acetylcholine, bethanechol and muscarine) depolarized about 70%, and hyperpolarized about 30% of the submucous plexus neurons. Low concentrations of pirenzepine reversibly antagonized both responses. The measured dissociation constants (KD) of 10-30 nM for the depolarizations and 1-3 nM for the hyperpolarizations suggest that each response was mediated by muscarinic M1 cholinoceptors. The muscarinic depolarization and hyperpolarization were associated with a decreased and an increased conductance, respectively, and the reversal potential for the muscarinic responses varied as the potassium concentration varied, always being around the potassium equilibrium potential. In cells depolarized by muscarinic agonists these agents appeared to decrease a potassium conductance that could also be inactivated by substance P. In approximately 30% of the submucous neurons, the slow inhibitory postsynaptic potential, elicited in response to single or repetitive focal stimuli (1-10 pulses at 20-40 Hz), appeared to consist of a large component which was sensitive to the blocking action of idazoxan (100-300 nM) and a small component which was idazoxan-insensitive. The latter (muscarinic slow inhibitory postsynaptic potential) was completely abolished by pirenzepine. The concentrations of pirenzepine which caused a 50% depression ranged from 5 to 20 nM. The muscarinic slow inhibitory postsynaptic potential was increased in amplitude and duration by physostigmine (100-300 nM). The muscarinic slow inhibitory postsynaptic potential was accompanied by a decrease in membrane input resistance, and was reversed in polarity near the potassium equilibrium potential. When muscarine induced a hyperpolarization and/or focal stimulation elicited a muscarinic slow inhibitory postsynaptic potential in the presence of idazoxan (100-300 nM), the intracellular injection of guanosine 5'-O-(3-thiotriphosphate) produced a progressive membrane hyperpolarization during which the muscarinic hyperpolarizing responses were attenuated. It is concluded that the muscarine-induced reduction in potassium conductance is mediated through a muscarinic M1 receptor which has a relatively low affinity for pirenzepine. The muscarine-induced increase in potassium conductance is probably produced by the association of a guanine nucleotide-binding regulatory protein with another muscarinic M1 receptor that has a relatively high affinity for pirenzepine.

Acetylcholine

Motoneurones of the submucous plexus regulate electrical activity of the circular muscle of canine proximal colon.

The hypothesis that the circular muscle of the canine proximal colon receives motor input from neurones in the submucous plexus was tested. Circular muscle cells were impaled with micro-electrodes and submucous plexus neurones were stimulated by electrical field stimulation and microejection of acetylcholine (ACh). In the presence of atropine to block the direct muscarinic effects, microejection of ACh onto the submucosa where intact submucous ganglia were suspended evoked: (i) an inhibitory junction potential (i.j.p.) that reduced the amplitude, duration and rate of rise of the subsequent slow wave; (ii) a slow wave of increased duration following the initial inhibitory response. These responses were enhanced by increasing the volume of ACh administered. Responses to ACh were blocked by hexamethonium, 10(-4) M; d-tubocurarine, 10(-4) M; or tetrodotoxin (TTX), 10(-6) M, suggesting they were neural in origin. Both inhibitory and excitatory responses were the result of non-cholinergic and non-adrenergic nerves. The transmitters mediating these effects are unknown. Removal of the longitudinal muscle, myenteric plexus, and the serosal portion of the circular muscle had no apparent effect on the responses to application of ACh to submucosal ganglia. In these preparations the responses to field stimulation were identical to those produced by ACh. The submucous plexus also provides cholinergic input to the circular muscle. When ACh was discretely applied to the submucosa cholinergic responses were elicited at the muscle cell which were significantly reduced by hexamethonium or TTX. These findings suggest that the cholinergic responses were the result of ACh release by neurones at the effector and not by overflow of the exogenous ACh. Cholinergic responses were also elicited in preparations in which the myenteric plexus had been removed. Slow waves in circular muscle of the proximal colon yield excitation-contraction coupling in the absence of Ca2+ action potentials. Therefore the influence of submucous neurones on electrical slow waves has direct consequences on motor activity. Reduction in the amplitude and duration of slow wave by i.j.p.s. results in reduction in the amplitude and duration of phasic contractions. Excitatory inputs enhance contractions. The data support a new concept: motoneurones emanating from submucous ganglia innervate the circular muscle and provide inhibitory and excitatory inputs to regulate slow wave activity.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine

Two types of neurones lacking synaptic input in the submucous plexus of guinea-pig small intestine.

Intracellular recordings were made from neurones in the submucous plexus of guinea-pig small intestine. Approximately 20% of all neurones studied appeared to lack synaptic input; two types of neurones without synaptic input were identified. The action potential recorded from one of these types of ganglion cells was followed by a prolonged after-hyperpolarization. The after-hyperpolarization was abolished by Co2+ or Mn2+. A Ca2+ component of the evoked action potential was demonstrated after the Na+ component had been abolished by tetrodotoxin (TTX). The other type of neurone was characterized by a high input resistance (220-320 M omega) and spontaneous action potentials which were unaffected by hexamethonium, curare, atropine or TTX. The ionic basis of the action potentials evoked from spontaneously active cells differed from other enteric neurones. These action potentials exhibited a TTX-insensitive Na+ component as well as a Ca2+ component. It is suggested the spontaneously active neurones may be responsible for the ongoing synaptic activity recorded from many submucous plexus neurones in the absence of stimulation.

Action Potentials

Nerve cell density in submucous plexus throughout the gut of cat and opossum.

Quantitative differences in submucous plexus density were sought in cat and opossum gut by examining full-thickness whole mounts of the submucosa stained with silver, and counting ganglia per square centimeter and nerve cell bodies per ganglion in order to compute density of innervation (nerve cell bodies per square centimeter). In the cat, the nerve cell bodies per square centimeter in the 12 named regions were as follows: proximal esophagus, 0; mid-esophagus, 0; distal esophagus, 0; fundus, 84; gastric antrum, 18; duodenum, 5831; jejunum, 4632; ileum, 3191; proximal colon, 1275; mid-colon, 689; distal colon, 359; rectum, 144. In the opossum, values were as follows: proximal esophagus, 37; mid-esophagus, 52; distal esophagus, 84; duodenum, 1812; jejunum, 2234; ileum, 1488; proximal colon, 206; mid-colon, 197; distal colon, 121; rectum, 61. Adequate specimens could not be obtained from opossum stomach. Differences were due more to variations in distribution density of ganglia than in ganglionic size. The relatively dense submucous plexus of the intestine probably is related to the capacity of the intestinal mucosa for peptide secretion as well as to its absorptive function.

Animals

Arterial submucous plexus of duodenum (1st 2.5 cm): vascular etiology in duodenal ulcers.

The arterial submucous plexus in 5 human duodena (1st 2.5 cm of duodenum) were studied by injecting India-ink. The submucous plexus was richly vascular, which excluded the possibility of the end arteries to be the causative factor in production of ulcers in this region. Perhaps, the ulcers in this region may be due to obstruction of blood flow caused by traction of an over filled stomach.

Arteries

Observations on the submucous plexus and mucosal arteries of the dog's stomach and first part of the duodenum.

Arteriolar patterns of the submucous plexus were studied in all areas of the dog's stomach and in the first inch of the duodenum. There appeared to be no poverty of plexus, although in some cases the vessels were somewhat smaller in the pyloric part of the lesser curvature than elsewhere. Mucosal arteries arose from the plexus, and none appeared to have an extramural origin. In man, on the other hand, there is a poverty of the submucous plexus in the 'ulcer region', i.e. in the incisural region of the lesser curvature and in the first inch of the duodenum, associated in some cases with mucosal end arteries of extramural origin. The absence of these features in the dog, which does not suffer from spontaneous chronic ulceration, lends further support to the view that they play a role in the aetiology of the disease in man.

Angiography

Changes in adrenergic and peptidergic nerves in the submucous plexus of streptozocin-diabetic rat ileum.

The effect of streptozocin diabetes on the distribution of adrenergic and peptidergic nerves in the submucous plexus of rat ileum was investigated and compared with the changes in the myenteric plexus of the same region of ileum. There was an increase in the intensity of immunoreactivity in vasoactive intestinal polypeptide- and neuropeptide Y-like immunoreactive nerve fibers and neurons and a decrease in calcitonin gene-related peptide-like immunoreactivity but no change in substance P- and dopamine beta-hydroxylase-like immunoreactivity in the nerve fibers and neurons of the submucous plexus of both 8- and 16-wk streptozocin-diabetic rat ileum. However, in the myenteric plexus of the diabetic rat ileum, there was enlargement of varicosities and an increase followed by a slight decrease in the intensity of immunoreactivity of vasoactive intestinal polypeptide- and dopamine beta-hydroxylase-like immunoreactive nerve fibers and neurons, increased substance P-like immunoreactivity in diabetes at 16 wk, and an initial decrease (at 8 wk) followed by a recovery of calcitonin gene-related peptide-like immunoreactivity at 16 wk, but no change in neuropeptide Y-like immunoreactivity. The markedly different changes in peptidergic and adrenergic nerves between the two enteric plexuses show that diabetic neuropathy induced by streptozocin is not selective and involves factors other than neurotransmitter types.

Adrenergic Fibers