Lumbar Colonic Nerves Research Articles

Characterisation of parasympathetic ascending nerves in human colon.

In the human large bowel, sacral parasympathetic nerves arise from S2 to S4, project to the pelvic plexus ("hypogastric plexus") and have post-ganglionic axons entering the large bowel near the rectosigmoid junction. They then run long distances orally or aborally within the bowel wall forming "ascending nerves" or "shunt fascicles" running in the plane of the myenteric plexus. They form bundles of nerve fibres that can be distinguished from the myenteric plexus by their straight orientation, tendency not to merge with myenteric ganglia and greater width. To identify reliable marker(s) to distinguish these bundles of ascending nerves from other extrinsic and intrinsic nerves in human colon. Human colonic segments were obtained with informed consent, from adult patients undergoing elective surgery (n = 21). Multi-layer immunohistochemical labelling with neurofilament-H (NF200), myelin basic protein (MBP), von Willebrand factor (vWF), and glucose transporter 1 (GLUT1), and rapid anterograde tracing with biotinamide, were used to compare ascending nerves and lumbar colonic nerves. The rectosigmoid and rectal specimens had 6-11 ascending nerves spaced around their circumference. Distal colon specimens typically had 1-3 ascending nerves, with one located near the mesenteric taenia coli. No ascending nerves were observed in ascending colon specimens. GLUT1 antisera labelled both sympathetic lumbar colonic nerves and ascending nerves in the gut wall. Lumbar colonic nerves joined the myenteric plexus and quickly lost GLUT1 labelling, whereas GLUT1 staining labelled parasympathetic ascending nerves over many centimetres. Ascending nerves can be distinguished in the colorectum of humans using GLUT1 labelling combined with NF200.

Identification of the Visceral Pain Pathway Activated by Noxious Colorectal Distension in Mice

In patients with irritable bowel syndrome, visceral pain is evoked more readily following distension of the colorectum. However, the identity of extrinsic afferent nerve pathway that detects and transmits visceral pain from the colorectum to the spinal cord is unclear. In this study, we identified which extrinsic nerve pathway(s) underlies nociception from the colorectum to the spinal cord of rodents. Electromyogram recordings were made from the transverse oblique abdominal muscles in anesthetized wild type (C57BL/6) mice and acute noxious intraluminal distension stimuli (100–120 mmHg) were applied to the terminal 15 mm of colorectum to activate visceromotor responses (VMRs). Lesioning the lumbar colonic nerves in vivo had no detectable effect on the VMRs evoked by colorectal distension. Also, lesions applied to the right or left hypogastric nerves failed to reduce VMRs. However, lesions applied to both left and right branches of the rectal nerves abolished VMRs, regardless of whether the lumbar colonic or hypogastric nerves were severed. Electrical stimulation applied to either the lumbar colonic or hypogastric nerves in vivo, failed to elicit a VMR. In contrast, electrical stimulation (2–5 Hz, 0.4 ms, 60 V) applied to the rectum reliably elicited VMRs, which were abolished by selective lesioning of the rectal nerves. DiI retrograde labeling from the colorectum (injection sites 9–15 mm from the anus, measured in unstretched preparations) labeled sensory neurons primarily in dorsal root ganglia (DRG) of the lumbosacral region of the spinal cord (L6-S1). In contrast, injection of DiI into the mid to proximal colon (injection sites 30–75 mm from the anus, measured in unstretched preparations) labeled sensory neurons in DRG primarily of the lower thoracic level (T6-L2) of the spinal cord. The visceral pain pathway activated by acute noxious distension of the terminal 15 mm of mouse colorectum is transmitted predominantly, if not solely, through rectal/pelvic afferent nerve fibers to the spinal cord. The sensory neurons of this spinal afferent pathway lie primarily in the lumbosacral region of the spinal cord, between L6 and S1.

The plasticity of the defecation reflex pathway in the enteric nervous system of guinea pigs

The enteric nervous system, the "second" brain, is an independent nervous system that structurally resembles the "first" brain. Appropriate rectal distension elicits rectal (R-R) reflex contractions and simultaneous internal anal sphincter (R-IAS) reflex relaxations that together comprise the defecation reflex. The enteric nervous system, pelvic nerves and lumbar colonic nerves control both reflexes. Using the plasticity of enteric nervous pathways, a new therapy for repairing enteric neural dysfunction could be developed. In vivo experiments were performed on guinea pigs anesthetized with ethyl carbamate. We performed either a lower anterior resection as used for rectal cancer, without damaging the extrinsic nerves or a resection of a 2-cm segment of distal colon, 30 mm orally from the anal verge, with subsequent end-to-end one layer anastomosis of the exposed ends. The recovery of the defecation reflex was found to be the same in both the rectal transection and distal colonic resection procedures. Eight weeks after sectioning the intrinsic reflex nerve pathways in the rectum, the defecation reflex recovered to control levels, accompanied by a regeneration of the reflex pathways. The 5-HT(4) receptor agonist, mosapride (0.5 and 1.0 mg/kg), significantly (P<0.01) enhanced the recovered defecation reflex at this stage. Two weeks after local treatment with brain-derived neurotrophic factor (BDNF: 10(-6) g/ml) at the rectal anastomotic site, the R-IAS reflex relaxations recovered and some bundles of fine nerve fibers were able to be seen interconnecting the oral and anal ends of the myenteric plexus. Also surprisingly, new neurons were found to have generated from neural stem cells at the anastomotic ends. These new neurons had constructed mature enteric neural networks including ganglionic-like structures eight weeks after surgery. These results revealed the plasticity of enteric neurons, allowing the proposal of a new therapy for repairing enteric neural dysfunction at least after surgery.

A new possibility for repairing the anal dysfunction by promoting regeneration of the reflex pathways in the enteric nervous system

Moderate rectal distension elicits recto-rectal reflex contractions and simultaneous recto-internal anal sphincter reflex relaxations that together comprise the defecation reflex. Both reflexes are controlled by 1) pelvic nerves, 2) lumbar colonic nerves, and 3) enteric nervous system. The aim of the present study was to explore a novel approach to repairing the defecation reflex dysfunction by using the plasticity of enteric nervous pathways. Experiments were performed in anesthetized guinea pigs with ethyl carbamate. The rectum 30 mm oral from the anal verge was transected without damage to extrinsic nerves, and subsequent end-to-end one-layer anastomosis was performed. Recovery of the defecation reflex and associated reflex pathways were evaluated. Eight weeks after sectioning of intrinsic reflex nerve pathways in the rectum, the defecation reflex recovered to the control level, accompanied with regeneration of reflex pathways. The 5-HT(4)-receptor agonist mosapride (0.5 and 1.0 mg/kg) significantly (P < 0.01) enhanced the recovered defecation reflex 8 wk after surgery. Two weeks after local treatment with brain-derived neurotrophic factor (BDNF: 10(-6) g/ml) at the rectal anastomotic site, the recto-internal anal sphincter reflex relaxations recovered and some bundles of fine nerve fibers were shown to interconnect the oral and anal ends of the myenteric plexus. These results suggested a possibility for repairing the anal dysfunction by promoting regeneration of the reflex pathways in the enteric nervous system with local application of BDNF.

BDNF-induced facilitation on regeneration of enteric nerve pathways for the defecation reflex in guinea pigs

Moderate rectal distension elicits rectal reflex contractions and simultaneous internal anal sphincter reflex relaxations, composed of defecation reflex. Both reflexes are controlled by (1) sacral excitatory reflexes mediated via pelvic nerves, (2) lumbar inhibitory reflexes mediated via lumbar colonic nerves, which are suppressed by pontine defecation reflex center, (3) intrinsic reflexes mediated via enteric nervous system. The plasticity of the intrinsic reflex pathway has been shown in the guinea pig that underwent chronic destruction of lumbar and sacral cords as follows: after removal of extrinsic reflex pathways, unchanged recto-rectal (R-R) reflex contractions and synchronous recto-IAS (R-IAS) reflex relaxations occurred, indicating intrinsic reflexes are able to compensate the defect of extrinsic reflex pathways.

Colonic motility in the cat. IV. Peripheral pathways mediating the effects induced by hypothalamic and mesencephalic stimulation.

Rostad, H. Colonic motility in the cat. IV. Peripheral pathways mediating the effects induced by hypothalamic and mesencephalic stimulation. Acta physiol. scand. 1973. 89. 154–168. The peripheral pathways by which the hypothalamus and mesencephalon influence the motility of the colon have been studied by combining brain stimulation and peripheral nerve sectioning, and by use of autonomic blocking agents. Excitatory colonic effects induced from the hypothalamic sympatho-inhibitory area with concomitant blood pressure falls were found to be mediated through the sympathetic lumbar colonic nerves, whereas excitatory responses with associated blood pressure rises were conveyed through the latter as well as through the parasympathetic pelvic nerves. On the other hand, the augmentatory colonic effects usually associated with blood pressure rise evoked from the mesencephalon were conveyed through the lumbar colonic nerves only. The responses through the lumbar colonic and pelvic nerves were blocked by guanethidine and atropine, respectively. The augmentatory colonic responses from the hypothalamic sympatho-inhibitory area were blocked by the beta-adrenergic blocking agent propranolol, whereas the effects from the other responsive hypothalamic areas, transmitted through the lumbar colonic nerves, were blocked by alpha-blocker (phenoxybenzaminc), but not by propranolol. This finding support the presence of alpha-excitatory receptors in the colonic wall. Inhibition of colonic motility induced by hypothalamic and mesencephalic stimulation, weir found to be mediated through the lumbar colonic as well as through the splanchnic nerves. The effects were blocked by guanethidine, and those from the responsive hypothalamic zones were also blocked by propranolol.

Morphological characteristics and immunohistochemical detection of nicotinic acetylcholine receptors on intestinofugal afferent neurones in guinea-pig colon.

Intestinofugal afferent neurones (IFANs) provide excitatory synaptic input to abdominal prevertebral ganglion neurones. Input is greatly reduced during blockade of nicotinic acetylcholine receptors (nAChRs) in the wall of the colon, suggesting two projection pathways: a direct pathway without synaptic interruption and an indirect pathway interrupted by at least one nicotinic cholinergic synapse. This study aimed to characterize the morphology of IFANs and examine the distribution of nAChRs on them. We identified IFANs in guinea-pig colon by retrograde labelling with fluorescent tracer DiI placed either on the lumbar colonic nerves in vitro or inferior mesenteric ganglion in vivo. Confocal laser scanning microscopy and computerized image-processing software were used for 3D image reconstruction. Approximately 70% of identified IFANs had Dogiel type I-like morphology, the remainder were Dogiel type II-like. In vivo labelled IFANs were injected with Lucifer Yellow and immunostained for nAChRs using monoclonal antibody MAb35. Approximately 3% of total plasma membrane surface of IFANs with Dogiel type I morphology had MAb35-IR. In contrast, <1% of membrane surface of IFANs with Dogiel type II morphology had MAb35-IR. The finding that IFANs displayed immunostaining for nAChRs suggests the presence of putative nicotinic synapses.

Integrative control of rectoanal reflex in guinea pigs through lumbar colonic nerves.

The aim of the present study was to analyze the neuromodulation of rectoanal reflex activity by lumbar sympathetic nerves in guinea pigs. The mechanical activities of the rectum were recorded with a balloon connected to a pressure transducer, and those of the internal anal sphincter (IAS) were recorded with a custom-made strain gauge force transducer. Gradual and sustained rectal distension evoked the rectoanal reflex, causing cholinergic contractions of the rectum and synchronous nitrergic relaxations of the IAS. Section of the lumbar colonic nerves enhanced both rectal contractions and IAS relaxations. Section of the 13th thoracic cord abolished both rectal contractions and IAS relaxations, but section of the lumbar colonic nerves restored them. Lumbar sympathectomy and pithing sacral cords greatly diminished these rectal contractions and IAS relaxations, but the intrinsic reflex component remained. NG-nitro-L-arginine methyl ester enhanced the intrinsic reflex-mediated contraction of the rectum and abolished reflex-mediated relaxation of the IAS and converted into cholinergic contractions. The present results indicate that the extrinsic lumbar inhibitory outflow causes marked inhibition of the rectoanal reflex via the lumbar colonic nerves.

Effects of extrinsic autonomic inputs on expression of c-Fos immunoreactivity in myenteric neurons of the guinea pig distal colon

c-Fos protein is a nuclear protein coded by c-fos proto-oncogene subsequent to synaptic activation of the neurons. We used immunohistochemical methods to visualize the expression of c-Fos protein in myenteric neurons of the guinea pig distal colon and examined the effects of the extrinsic autonomic inputs on the enteric circuits. No c-Fos immunoreactivity was observed in the colonic segments fixed immediately after removal from the animal body. A number of c-Fos-immunoreactive nuclei of myenteric neurons, however, appeared in all preparations that were incubated in Krebs solution in vitro ( n=10). Application of tetrodotoxin (0.2 μM) abolished the expression of c-Fos-immunoreactivity ( n=6), but hexamethonium (100 μM) failed to decrease the number of c-Fos-positive neurons despite a complete suppression of spontaneous peristaltic movements ( n=5). Neither the electrical stimulation ( n=8) nor the severing of the pelvic nerves ( n=5) changed the number of c-Fos-positive neurons. Application of clonidine, an α 2-agonist, (0.1 μM) abolished the expression of c-Fos protein in all preparations ( n=5), while denervation of the sympathetic fibers in the lumbar colonic and hypogastric nerves in vivo increased the number of c-Fos-positive neurons ( n=5). The results indicate that the enteric circuit in the distal part of the gastrointestinal tract is under tonic inhibition by the sympathetic nervous system from the lumbar spinal cord. c-Fos immunoreactivity expressed in the colonic preparations in vivo might be the results of enhanced activation of non-nicotinic receptors after removal of the sympathetic inhibition.

Colonic Mechanoreceptor Input to Neurons in Guinea Pig Caudal Mesenteric Ganglion

The background activity of pacemaker-like neurons (PLN) of the guinea-pig caudal mesenteric ganglion (CMG) and their reflex responses to colonic distention were studied on combined isolated preparations including the CMG and a colon segment connected with the lumbar colonic nerves. The results allow us to suggest that the “spontaneous” and reflex activity of PLN is of a peripheral origin. Synaptic transmission in the peripheral colonofugal nerve pathways is mediated by acetylcholine and substance P (SP). The SP-induced excitation of PLN probably involves activation of the tachykinin NK3 receptors.

Patterning of sympathetic nerve activity in response to vestibular stimulation

Growing evidence suggests a role for the vestibular system in regulation of autonomic outflow during postural adjustments. In the present paper we review evidence for the patterning of sympathetic nerve activity elicited by vestibular stimulation. In response to electrical activation of vestibular afferents, firing of sympathetic nerves located throughout the body is altered. However, activity of the renal nerve is most sensitive to vestibular inputs. In contrast, high-intensity simultaneous activation of cutaneous and muscle inputs elicits equivalent changes in firing of the renal, superior mesenteric and lumbar colonic nerves. Responses of muscle vasoconstrictor (MVC) efferents to vestibular stimulation are either inhibitory (Type I) or are comprised of a combination of excitation and inhibition (Type II). Interestingly, single MVC units located in the hindlimb exhibited predominantly Type I responses while those located in the forelimb and face exhibited Type II responses. Furthermore, brachial and femoral arterial blood flows were dissociated in response to vestibular stimulation, such that brachial vascular resistance increased while femoral resistance decreased. These studies demonstrate that vestibulosympathetic reflexes are patterned according to both the anatomical location and innervation target of a particular sympathetic nerve, and can lead to distinct changes in local blood flow.

Sympathetic inhibition of ascending and descending interneurones during the peristaltic reflex in the isolated guinea-pig distal colon.

1. We investigated the effects of sympathetic nerve stimulation within ascending and descending reflex pathways underlying the peristaltic reflex in the guinea-pig distal colon. 2. A three-chambered partitioned bath was used to divide a segment of distal colon into stimulation, recording and intermediate regions. The effects of lumbar colonic nerves (LCN) could be localized to the intermediate region by surgical lesions of the mesentery and by application of guanethidine (3 microM) to the stimulation and recording chambers. 3. Brush stroking the mucosa in the anal and oral stimulation chambers elicited a synchronous contraction of the longitudinal muscle (LM) and circular muscle (CM) oral to, and transient relaxation of the LM and CM anal to, the stimulus, respectively. 4. After N omega-nitro-L-arginine (L-NA; 100 microM) in the oral and intermediate chambers, mucosal stimulation in the oral chamber elicited a prolonged descending inhibitory and excitatory complex in both the LM and CM in the anal recording chamber. This was blocked by hexamethonium (300 microM), which did not affect the transient relaxation response recorded in control conditions. 5. Stimulation of the LCN (1200 pulses, 20 Hz), delivered to the intermediate region, abolished the oral contraction and the L-NA-induced anal complex in both the LM and CM, but was without effect on the transient hexamethonium-resistant anal relaxation. These effects of LCN stimulation were reversed by phentolamine (3 microM) or yohimbine (100 nM), but not propranolol (10 microM), when added to the intermediate chamber. 6. LCN stimuli (2-20 Hz, 600 micros pulses) directed to the recording chamber elicited synchronous relaxations in the LM and CM that were unaffected by hexamethonium (300 microM), but were reduced by yohimbine and usually blocked by the further addition of propranolol (10 microM). 7. In conclusion, sympathetic nerve stimulation inhibits orally and anally projecting cholinergic interneurones underlying the peristaltic reflex in the distal colon. In addition, the LM and CM relax synchronously following release of sympathetic neurotransmitter, over a range of stimulus frequencies.

Background and reflex activity of guinea pig caudal mesenteric ganglion neurons

The background activity of the guinea pig caudal mesenteric ganglion (CMG) neurons and their reflex reactions to colonic distension were studied on isolated combined preparations including the CMG and a colon segment connected with the lumbar colonic nerves. In the control, 62% of the neurons under study generated background activity, which consisted of irregular or regular “fast” excitatory postsynaptic potentials (fEPSP) and action potentials (AP). In 27% of the CMG neurons called “pacemaker-like neurons” (PLN), the background activity was represented by highly regular AP never observed in the CMG completely isolated from the distal colon. Reflex responses evoked by colonic distension were recorded from 76% of the units studied. The distension evoked fEPSP and AP in “silent” neurons and increased the background activity. Both the background activity and reflex responses were shown to be due to nicotinic cholinergic transmission. In some neurons, reflex responses (regular AP) were generated as superimposed on a slow depolarization; the latter was insensitive to nicotinic antagonists and either sensitive or insensitive to muscarinic antagonists. It has been concluded that CMG neurons receive nicotinic, muscarinic, and, probably, peptidergic afferent inputs from the distal colon. Although there are no true pacemaker neurons in CMG, some neurons generate pacemaker-like activity of a synaptic origin.

Patterning of somatosympathetic reflexes

In a previous study, we reported that vestibular nerve stimulation in the cat elicits a specific pattern of sympathetic nerve activation, such that responses are particularly large in the renal nerve. This patterning of vestibulosympathetic reflexes was the same in anesthetized and decerebrate preparations. In the present study, we report that inputs from skin and muscle also elicit a specific patterning of sympathetic outflow, which is distinct from that produced by vestibular stimulation. Renal, superior mesenteric, and lumbar colonic nerves respond most strongly to forelimb and hindlimb nerve stimulation (approximately 60% of maximal nerve activation), whereas external carotid and hypogastric nerves were least sensitive to these inputs (approximately 20% of maximal nerve activation). In contrast to vestibulosympathetic reflexes, the expression of responses to skin and muscle afferent activation differs in decerebrate and anesthetized animals. In baroreceptor-intact animals, somatosympathetic responses were strongly attenuated (to <20% of control in every nerve) by increasing blood pressure levels to >150 mmHg. These findings demonstrate that different types of somatic inputs elicit specific patterns of sympathetic nerve activation, presumably generated through distinct neural circuits.

Effects of sympathetic nerve stimulation on membrane potential in the circular muscle layer of mouse distal colon.

Little is known about the effects of sympathetic nerve stimulation on the membrane potential of colonic smooth muscle. In the distal colon of the mouse, intracellular microelectrodes were used to record the effects of lumbar colonic (LCN) and intermesenteric nerve (IMN) stimulation on circular muscle membrane potential in vitro. A two-compartment organ bath was used to selectively perfuse the colon and inferior mesenteric ganglion (IMG). In the presence of nifedipine (1-2 microM) (to the colonic compartment only), spontaneous depolarizations (myoelectric complexes, MCs) were recorded about every 4 min. MCs consisted of a prolonged burst of rapid oscillations in membrane potential (approximately 2 Hz) that were superimposed on a slow depolarization (mean amplitude 12 mV). Single electrical stimuli (600 microseconds duration) delivered to the LCN or IMN did not elicit a detectable change in the membrane potential. However, trains of stimuli (e.g., 60 pulses at 10-20 Hz) to the LCN or IMN during the intervals between MCs evoked a depolarization (with superimposed action potentials in the absence of nifedipine). Trains of stimuli delivered during the plateau phase of the MC reduced or abolished the rapid oscillations, without a further membrane depolarization. The MC period was unaffected by stimulation of the IMN or LCN. Responses were abolished by the selective perfusion of guanethidine (1 microM) to the colon, or by severing the LCN. Hexamethonium (500 microM) (to the colon) abolished MCs, induced sustained depolarization and attenuated the amplitude of the sympathetic depolarizations by 74%. In hexamethonium, sympathetic responses remained attenuated when the membrane of the circular muscle was repolarised by sodium nitroprusside (1 microM). Immunohistochemical studies of the colon revealed intense immunoreactivity for tyrosine hydroxylase in the myenteric plexus but not in the circular muscle layer. It is suggested that responses to sympathetic nerve stimulation in the circular muscle layer of the mouse colon are secondary to actions on the enteric nervous system.

Neuronal circuitry between the inferior mesenteric ganglion and lower intestine of the dog.

Neuronal circuitry between the inferior mesenteric ganglion (IMG) and the distal colon as well as the rectum, forming the intestino-intestinal reflex pathway, was investigated in the dog using immunohistochemistry combined with retrograde tract tracing and denervation experiments. Virtually all IMG neurons were tyrosine hydroxylase (TH)-immunoreactive. Of these ganglionic neurons, about 64% were also immunoreactive for calbindin (Calb), some 35% for neuropeptide Y (NPY), and 2% for vasoactive intestinal peptide (VIP). The retrograde tracer experiments revealed that both Calb/TH neurons and NPY/TH neurons projected to the distal colon and the rectum. In these intestinal walls, Calb/TH positive varicose fibers were found in the myenteric and submucous ganglia as well as in the longitudinal muscle layer, while NPY/TH positive fibers were mainly distributed around the vascular walls. Around Calb/TH neurons of the IMG, abundant varicose nerve fibers immunoreactive for VIP, dynorphin (DYN), calcitonin gene-related peptide (CGRP), enkephalin (ENK), substance P (SP) and bombesin (BOM) were distributed. These immunoreactive fibers disappeared after the total denervation of the IMG. After the application of Fast Blue into the IMG or distal stumps of transected lumbar colonic and hypogastric nerves, retrogradely labeled neurons occurred in the myenteric plexus with increasing density along the distal colon and rectum, and were immunoreactive for VIP, DYN, CGRP, ENK, SP or BOM. Double immunostaining of nerve fibers in the distal stumps of the ligated colonic and hypogastric nerves revealed the presence of viscerofugal fibers containing VIP with DYN and/or CGRP and those containing ENK with SP and/or BOM. These results demonstrate for the first time that the efferent limb of the canine intestino-intestinal reflex arch via the IMG consists of Calb-immunoreactive ganglion neurons projecting to the longitudinal muscles in addition to the enteric plexus of the lower intestine and also of NPY-immunoreactive ganglion neurons projecting to the intestinal blood vessels, and that the afferent limb is composed of at least two discrete groups with different peptide contents, i.e., myenteric neurons containing VIP with DYN and/or CGRP and those containing ENK with SP and/or BOM.

Light, electron, and confocal microscopic study of the mouse superior mesenteric ganglion.

The superior mesenteric ganglion (S.m.g.), a sympathetic prevertebral ganglion, is an integrating center for gastrointestinal reflexes. Many details of its structure are still lacking. In the present study, mouse S.m.g. neurons were studied by light, electron, and confocal microscopy. Neurons had an average of 5-6 primary dendrites. Total dendritic length averaged 963 microns. Confocal microscopy and three-dimensional reconstructed images revealed cell body surface features, precise location where axons and dendrites emerged from it, cell body size, and extent of dendritic projection in three axes. Cell body diameter and dendritic projections were less in the dorsoventral than in the rostrocaudal or mediolateral axes. Cell body surface area and volume averaged 4,271 microns 2 and 4,908 microns 3, respectively. Dendritic surface areas and volumes were 5-6 times larger. Two main neuron types (projecting caudally or rostrally) were distinguished. The former were found throughout the S.m.g., whereas the latter were found only in the cephalad region, comprising about 40% of neurons found there. Rostrally projecting neurons had fewer primary dendrites, fewer total dendritic branches, and shorter total dendritic length than caudally projecting neurons. There were regional differences in percentage of neurons responding to electrical stimulation of left or right hypogastric, lumbar colonic, or left splanchnic nerves but not in nerve fibers connecting the S.m.g. and celiac ganglion. A greater percentage of caudally than rostrally projecting cephalad neurons responded to stimulation of any nerve trunk. These results indicate that the mouse S.m.g. contains at least two distinct types of neurons that differ in their morphology and their source of preganglionic synaptic input.

Distribution and morphological characterization of viscerofugal projections from the large intestine to the inferior mesenteric and pelvic ganglia of the male rat.

Viscerofugal neurons are enteric neurons in the myenteric plexus of the stomach and intestine that project to the prevertebral ganglia as the afferent limb of intestino-intestinal reflexes. This study characterizes viscerofugal projections to the inferior mesenteric ganglion and investigates the possibility of similar projections to the major pelvic ganglia in the male rat. The colon and rectum were examined for retrogradely labelled neurons following the injection of retrograde tracer into the inferior mesenteric or major pelvic ganglia, or following the application of tracer to the caudal end of the cut intermesenteric nerves, or either end of the cut hypogastric nerves. All labelled viscerofugal neurons were found in the myenteric plexus and were often grouped near the mesenteric attachment. The number of viscerofugal neurons projecting to the inferior mesenteric ganglion via the lumbar colonic nerves increases along the length of the large intestine with the maximum number of viscerofugal neurons found in the rectum. Some viscerofugal neurons from the distal colon and rectum reach the inferior mesenteric ganglion via the hypogastric nerves. A similar number and distribution of viscerofugal neurons project via the inferior mesenteric ganglion into the intermesenteric nerves as terminate in the inferior mesenteric ganglion. Very few viscerofugal neurons project to the neurons of the major pelvic ganglia via the rectal nerves, and no viscerofugal neurons project caudally in the hypogastric nerves to these ganglia. The majority of labelled neurons resembled Dogiel type I morphology. Thus the inferior mesenteric ganglion receives a substantial innervation from viscerofugal neurons of the large intestine, with the greatest supply from the distal colon and rectum.(ABSTRACT TRUNCATED AT 250 WORDS)

A novel in vitro technique to study extrinsic neural control of rabbit colonic muscle and epithelial function.

A novel in vitro technique capable of simultaneously measuring distal colonic epithelial potential difference and muscle contraction is described. Under basal conditions, oscillations in both muscle tone and potential difference were observed. Pelvic nerve stimulation was shown to evoke strong "duration" contractile responses in both the longitudinal and circular muscle layers. Additionally, tonic changes in potential difference extending beyond the train of stimuli were observed, suggesting for the first time that colonic ion transport may be influenced by the pelvic nerves. However, it was unclear whether these were direct effects or indirect actions resulting from muscle contractions causing mechanical stimulation of nerves of the submucosal plexus. Lumbar colonic nerve stimulation inhibited spontaneous contractile activity and reduced basal tone in both muscle layers. However, there was no consistent effect of sympathetic nerve stimulation on transepithelial potential difference. Each of the muscle and epithelial effects of sympathetic nerve stimulation was mimicked by exogenous norepinephrine. Based on these data, it is concluded that colonic function is strongly influenced by the extrinsic innervation. Furthermore, relatively long-term modulation of epithelial function can be achieved by short bursts of pelvic nerve activity.

Enteric GABA-containing nerves projecting to the guinea-pig inferior mesenteric ganglion modulate acetylcholine release.

1. The effect of GABA and GABA receptor-modulating drugs on release of [3H]acetylcholine was studied in the guinea-pig inferior mesenteric ganglion. 2. GABA caused a dose-dependent increase in [3H]acetylcholine release during stimulation of the lumbar colonic nerves. Muscimol (10 microM) and diazepam (5 microM) also increased [3H]acetylcholine release during stimulation of the lumbar colonic nerves whereas baclofen (10 microM) had no effect. 3. Bicuculline (20-100 microM) and picrotoxin (50 microM) alone reduced [3H]acetylcholine release during electrical stimulation of the lumbar colonic nerves whereas phaclofen (300 microM) had no effect. 4. Bicuculline (100 microM) significantly decreased whereas diazepam (5 microM) significantly increased distension-induced [3H]acetylcholine release. 5. Colonic distension significantly increased [3H]GABA release in the inferior mesenteric ganglion compared to basal periods when the colon was not distended. Distension-induced release of [3H]GABA resulted from active neuronal transmission from the colon to the inferior mesenteric ganglion, since perfusion of the inferior mesenteric ganglion with tetrodotoxin (1 microM) reduced basal release of [3H]GABA and abolished distension-evoked increases in the release of [3H]GABA. 6. In contrast to its excitatory effects on peripheral colonic afferent cholinergic nerves, exogenous GABA caused a dose-dependent decrease in [3H]acetylcholine release during electrical stimulation of the central lumbar splanchnic nerves. Baclofen (10 microM) also inhibited [3H]acetylcholine release whereas muscimol (10 microM) or diazepam (5 microM) had no effect. Phaclofen (300 microM) antagonized the inhibitory effects of exogenous GABA (10 microM) and of baclofen (10 microM). Bicuculline (100 microM), picrotoxin (50 microM) and phaclofen (300 microM) alone had no effect on [3H]acetylcholine release during splanchnic nerve stimulation. 7. Phaclofen (300 microM) increased [3H]acetylcholine release during simultaneous electrical stimulation of the lumbar colonic nerves and splanchnic nerves and when GABAA receptors were blocked by bicuculline (20 microM). 8. The data suggest that GABAA receptors facilitate release of acetylcholine from peripheral cholinergic mechanosensory nerves projecting from the colon to the inferior mesenteric ganglion and that GABAB receptors inhibit release of acetylcholine from central cholinergic nerves. Enteric GABA-containing nerves projecting to the inferior mesenteric ganglion are mechanosensory. Endogenous release of GABA may act on GABAA receptors to facilitate peripheral cholinergic mechanosensory transmission and/or on GABAB receptors to inhibit central cholinergic transmission.