Convergence of noncholinergic afferent neurons in the inferior mesenteric ganglion of the guinea pig

The pattern of noncholinergic innervation of principal ganglionic neurons from the lumbar colonic nerve (LCN) and lumbar splanchnic nerve (LSN) in the inferior mesenteric ganglion (IMG) of the guinea pig was studied with intracellular techniques. Simultaneous stimulation of the LCN and LSN at maximum frequency (20 Hz) and submaximal stimulus voltages (2-4 V) led to summation of slow excitatory postsynaptic potential (EPSPs) indicating convergence of neural input. Summation was observed with submaximal (but not maximal) stimulation parameters in cells with either large or small amplitude maximum slow EPSPs suggesting that each neuron has an individual maximum capacity to depolarize in response to non-cholinergic transmitter substances. The study indicates that neurons originating at both peripheral and central sites converge onto principal ganglionic neurons, thus these ganglionic neurons must perform a significant integrative function in the IMG.     

Cholecystokinin octapeptide depolarizes guinea pig inferior mesenteric ganglion cells and facilitates nicotinic transmission

Cholecystokinin octapeptide (CCK-8) applied either by superfusion (0.1-10 microM) or by pressure ejection elicited a slow depolarization in a portion of inferior mesenteric ganglion cells studied in vitro. The depolarization which persisted in a low Ca2+/high Mg2+ solution, or solution containing cholinergic antagonists, was often associated with a small to moderate increase in neuronal input resistance, and the response was reduced by conditioning hyperpolarization. Nicotinic excitatory postsynaptic potentials were consistently augmented during the course of CCK-8-induced depolarization. Our results, together with findings of the presence of CCK-immunoreactive fibers in the prevertebral ganglia, suggest that the peptide may serve to facilitate nicotinic transmission.     

Neurokinin A depolarizes neurons of the guinea pig inferior mesenteric ganglia

Neurokinin A (NKA) applied either by superfusion (0.1-10 microM) or by pressure ejection evoked a slow membrane depolarization in neurons of the inferior mesenteric ganglia in vitro. The NKA-induced depolarization which was not significantly affected by reducing the Ca concentration or by tetrodotoxin was associated in most cases by a small-to-moderate increase in membrane resistance; however, conditioning hyperpolarization increased rather than decreased the response. The depolarization elicited by NKA was eliminated in the presence of substance P (SP) and the depolarization by SP in the presence of NKA. The results suggest that the actions of NKA on prevertebral neurons are similar to that of SP and that these two peptides may act on the same receptors or share similar ionic channels.     

The ultrastructure of specialized contacts between small intensely fluorescent (SIF) cells in the inferior mesenteric ganglion of the guinea pig

The plasma membranes of small intensely fluorescent (SIF) cells in the inferior mesenteric ganglion of guinea pig have been investigated with thin section electron microscopy and freeze-fracture technique. Gap junctional elements and tight junctional strands were found between adjacent cells in the freeze-fractured material.     

An electrophysiological study of inferior mesenteric ganglion of the dog

Intracellular recordings were made in vitro from 430 sympathetic neurons in the inferior mesenteric ganglion (IMG) of the dog. Ganglion cells had resting membrane potentials between -35 and -70 mV; input resistance (Rin) was approximately 22 M omega. Cell rheobase to depolarizing current was 0.3 nA, and the action potential elicited was 80-100 mV in amplitude followed by an afterhyperpolarization of up to 15 mV in size, which decayed to the resting membrane potential over a range of 50-500 ms. Neurons were classified as either phasic (188 of 280) or tonic (92 of 280) firing cells, depending on their discharge pattern in response to depolarizing current. Two hundred eight of 430 neurons showed continuous electrical activity in the form of spontaneous excitatory postsynaptic potentials, 2-5 mV in amplitude. Continuous electrical activity was unaffected by tetrodotoxin (3 X 10(-6) M) but abolished by hexamethonium (10(-4) M). A small number of cells (21 of 430) adopted a repetitive firing pattern not associated with injury discharge. These cells may have been pacemaking neurons. Stimulation of peripheral and central nerves resulted in multiple synaptic input to ganglion cells. There was marked convergence of excitatory fibers to any one cell. Evoked synaptic potentials were abolished by hexamethonium (10(-4) M). Synaptic input from peripheral and central nerves could not be correlated with location of postganglionic neurons in the IMG. The possibility of neuronal intercommunication and dissemination of central and peripheral commands is discussed.     

Nitric oxide synthase, choline acetyltransferase, catecholamine enzymes and neuropeptides and their colocalization in the anterior pelvic ganglion, the inferior mesenteric ganglion and the hypogastric nerve of the male guinea pig

By the indirect immunofluorescence method, the distribution of nitric oxide synthase (NOS)-like immunoreactivity (LI) and its possible colocalization with neuropeptide immunoreactivities, with two enzymes for the catecholamine synthesis pathway, tyrosine hydroxylase (TH) and dopamine β-hydroxylase (DBH), as well as the enzyme for the acetylcholine synthesis pathway, choline acetyltransferase (ChAT) were studied in the anterior pelvic ganglion (APG), the inferior mesenteric ganglion (IMG) and the hypogastric nerve in the male guinea pig. The analyses were performed on tissues from intact animals, as well as after compression/ligation or cut of the hypogastric nerve. In some cases the colonic nerves were also cut. Analysis of the APG showed two main neuronal cell populations, one group containing NOS localized in the caudal part of the APG and one TH-positive group lacking NOS in its cranial part. The majority of the NOS-positive neurons contained ChAT-LI. Some NOS-positive cells did not contain detectable ChAT, but all ChAT-positive cells contained NOS. NOS neurons often contained peptides, including vasoactive intestinal peptide (VIP), neuropeptide tyrosine (NPY), somatostatin (SOM) and/or calcitonin gene-related peptide (CGRP). Some NOS cells expressed DBH, but never TH. The second cell group, characterized by absence of NOS, contained TH, mostly DBH and NPY and occasionally SOM and CGRP. Some TH-positive neurons lacked DBH. In the IMG, the NOS-LI was principally in nerve fibers, which were of two types, one consisting of strongly immunoreactive, coarse, varicose fibers with a patchy distribution, the other one forming fine, varicose, weakly immunoreactive fibers with a more general distribution. In the coarse networks, NOS-LI coexisted with VIP- and DYN-LI and the fibers surrounded mainly the SOM-containing noradrenergic principal ganglion cells. A network of ChAT-positive, often NOS-containing nerve fibers, surrounded the principal neurons. Occasional neuronal cell bodies in the IMG contained both NOS- and ChAT-LI. Accumulation of NOS was observed, both caudal and cranial, to a crush of the hypogastric nerve. VIP accumulated mainly on the caudal side and often coexisted with NOS. NPY accumulated on both sides of the crush, but mainly on the cranial side, and ENK was exclusively on the cranial side. Neither peptide coexisted with NOS. Both substance P (SP) and CGRP showed the strongest accumulation on the cranial side, possibly partly colocalized with NOS. It is concluded that the APG in the male guinea-pig consists of two major complementary neuron populations, the cholinergic neurons always containing NOS and the noradrenergic neurons containing TH and DBH. Some NOS neurons lacked ChAT and could represent truly non-adrenergic, non-cholinergic neurons. In addition, there may be a small dopaminergic neuron population, that is containing TH but lacking DBH. The cholinergic NOS neurons contain varying combinations of peptides. The noradrenergic population often contained NPY and occasionally SOM and CGRP. It is suggested that NO may interact with a number of other messenger molecules to play a role both within the APG and IMG and also in the projection areas of the APG.     

An intracellular analysis of some intrinsic factors controlling neural output from inferior mesenteric ganglion of guinea pigs

1. In vitro studies were conducted on neurons within the inferior mesenteric ganglion (IMG) of guinea pigs to investigate how intrinsic features of the spike-generating process interact with preganglionic inputs to produce the output firing patterns of these neurons. Intracellular-electrode techniques were used to monitor and control electrical activity of IMG neurons. Preganglionic inputs were activated either synchronously by stimulating an attached nerve trunk or asynchronously by leaving the ganglion attached to a segment of terminal colon and activating the colonic-IMG mechanosensory system. 2. Ninety-seven percent of the neurons studied demonstrated an afterspike hyperpolarization (ASH). The ASH process was activated only by the occurrence of a spike and did not have a synaptically induced component. Further activation of this process was produced by two or more spikes having interspike intervals less than the duration of an ASH following a single spike. An aftertrain hyperpolarization (ATH) resulted from this progressive activation. The amplitude of both the ASH and the ATH decreased when the resting membrane potential was hyperpolarized by current injection or by increasing the external potassium ion concentration. 3. Neuronal excitability was reduced during the ASH. From this observation it was concluded that when IMG neurons operate in the occasional-firing mode, the ASH process prevents output frequency from greatly exceeding the reciprocal of the ASH duration produced by a single spike. 4. Two types of synaptically induced slow depolarizations were observed: a slow, long-latency depolarization and a short-latency depolarization (SLD). These depolarizations differed in their latency, onset, and duration. Both were capable of converting synchronous, preganglionic input from subthreshold (non-spike-activating) to threshold (spike-activating) activity. 5. Neurons having resting potentials more positive than -60 mV were capable of firing in the rhythmic-firing mode; 40% of these neurons demonstrated tonic- and 60% phasic-firing behavior. Frequency-current relations for tonic-discharging neurons were linear from the rhythmic-firing threshold to current levels approximately 2.5 times the threshold value. Minimal frequency for tonic firing and the slope of the linear portion of the frequency-current relation were indirectly related to the duration of the ASH. 6. This study suggests that sympathetic, noradrenergic neurons of the IMG can operate in either the occasional- or rhythmic-firing mode. In the physiologic state in vivo, most IMG neurons probably do not produce action potentials in excess of 10-15 Hz because of their intrinsic properties which regulate firing in both modes of operation.     

Measurements of the DNA amount in mono- and binucleate cells in the celiac superior mesenteric ganglion of the guinea pig

Summary The relative proportion, ultrastructure and DNA-content of the binucleate cells in the celiac superior mesenteric ganglion of the guinea pig was studied using light and electron microscopy as well as computerized image analysis of Feulgen stained cells. The number of mono — versus binucleate cells was found to vary with stage of development with about 40% of the cells being binucleate in adult animals and 50% in late prenatal stage. No difference in ultrastructure was observed between the nuclei of the two cell types. The binucleate cells contain twice the amount of DNA found in the mononucleate cells.     

Sympathetic activity in the recto-rectal reflex of the guinea pig

In the guinea pig, defecation is controlled by the myenteric plexus, whose activity is modulated by the sacral spinal and supraspinal centers. The purpose of this study is to clarify the control of defecation reflex by sympathetic nerves. The propulsive contractions of the rectum produced by rectal distension (recto-rectal excitatory reflex response) were abolished after transection of the Th 13 and/or the L 4 segment. This response was reproduced again after removal of the lumbar segments (L1-4), division of the lumbar dorsal roots (L1-4), the lumbar splanchnic nerves or lumbar colonic nerves (LCN). The frequency of efferent discharges of LCN was increased slightly by rectal distension and remarkably increased after Th 13 and/or L 4 transection. Thus, there occurs during the recto-rectal reflex not only mucosal intrinsic reflex and sacral excitatory reflex via the pelvic nerves but also a lumbar inhibitory reflex via the colonic nerves, whose center may be located in the upper lumbar segments.But, the activity of the inhibitory center was depressed by the supraspinal center, so that an excitatory reflex is produced more dominantly than an inhibitory one in normal animals. All these extrinsic reflexes coordinate the activity of the myenteric plexus in defecation reflex.     

Enkephalin-like immunoreactive principal ganglion cells and nerve fibres in the inferior mesenteric ganglion of the cat

The occurrence and distribution of methionine-enkephalin (ME), leucine-enkephalin (LE) and methionine-enkephalin-Arg6-Gly7-Leu8 (MERGL)-like (LI) immunoreactive material in the inferior mesenteric ganglion (IMG) of the cat were studied by immunohistochemical techniques using the peroxidase-antiperoxidase method. Numerous ME-Li, LE-Li and MERGL-Li immunoreactive fibres with the same distribution pattern were observed. They were varicose and often surrounded closely neighbouring unlabelled ganglion cell bodies. Sometimes they ran in strands between ganglion cells. ME-Li immunoreactive material was detected in a number of cell bodies, the diameter of which was similar to that of unlabelled principal ganglion cell bodies, and which were probably Enk-Li-containing principal ganglion cells. These immunoreactive cells were often surrounded by ME-Li immunoreactive fibres. No LE-Li or MERGL-Li immunoreactive ganglion cell bodies were observed. The presence of ME-Li immunoreactive principal ganglion cells raises the possibility that the Enk-Li immunoreactive fibres present in the IMG may have a prevertebral ganglionic source. The possibility that the Enk-Li material present in nerve fibres might be derived from preproenkephalin-A was suggested by the occurrence of MERGL-Li immunoreactivity.     

Adrenergic nerve elements in the hypogatric ganglion of the guinea pig

Adrenergic nerve elements in the hypogastric ganglion of the guinea pig were studied by fluorescence and electron microscopy. The ganglion contains two main elements exhibiting green to green-yellow fluorescence for catecholamines. One consists of basket-like, simple nerve endings around some of the ganglion cell bodies, and the other of small polyhedral cells with short axonic processes. Electron microscopy revealed the ultrastructure of these elements. The adrenergic nerve endings contain small granular vesicles, about 500 Å in diameter, as well as a few large granular and small agranular vesicles. They were more rare than the cholinergic endings, which contain numerous small agranular vesicles mixed with a few large granular vesicles. Both adrenergic and cholinergic endings submerge to a considerable depth into the cytoplasm of ganglion cells. The ganglion cells send spinous processes into these endings. The fluorescent polyhedral cells are probably in an intermediate position between adrenal medullary cells and ganglion cells. They contain numerous large granular vesicles. Their axonic processes extend for a short distance to end in close topographical relation to capillaries. According to the structure of the granulated vesicles, these cells were classified into four types. The function of the adrenergic elements in ganglionic transmission was discussed.     

Localization of damage in the feline inferior mesenteric ganglion caused by thyroid hypofunction

Bulletin of Experimental Biology and Medicine -     

Excitatory input from the distal colon to the inferior mesenteric ganglion in the guinea-pig

1. Intracellular recordings were obtained from ganglion cells in the guinea-pig inferior mesenteric ganglion (IMG) with a segment of the distal colon attached to the lumbar colonic nerves.2. Continuous electrical activity consisting of excitatory synaptic potentials and action potentials was recorded from ganglion cells in all regions of the IMG.3. The ;spontaneous' synaptic potentials were indistinguishable from those elicited by submaximal stimulation of any of the nerve trunks connected to the IMG.4. The excitatory activity was irreversibly abolished when the lumbar colonic nerves were cut and reversibly abolished when tetrodotoxin (5 x 10(-7) g/ml.) was added to the colon side of a two-compartment organ bath.5. Addition of dihydro-beta-erythroidine (5 x 10(-6) g/ml.) to the ganglion side of the bath abolished the synaptic activity of colonic origin and the synaptic responses to stimulation of any of the nerve trunks connected to the IMG.6. Addition of dihydro-beta-erythroidine (1 x 10(-5) g/ml.) to the colon side of the bath markedly depressed the synaptic input of colonic origin but had no effect on synaptic responses produced by preganglionic nerve stimulation.7. Distension of the colonic segment and the application of 5-HT (1 x 10(-5) g/ml.) to the mucosal surface of the colon increased the frequency of synaptic input.8. The synaptic input from the colon was transiently blocked following repetitive stimulation of any of the nerve trunks connected to the IMG. The discharge of miniature synaptic potentials was unaffected.9. Addition of noradrenaline (1 x 10(-7) to 1 x 10(-6) g/ml.) to the colon side of the bath reduced, and in some cases completely abolished, the synaptic input to the IMG. Phentolamine (1 x 10(-6) g/ml.), when added to the colon side of the bath, blocked the effect of noradrenaline and the transient inhibition following repetitive nerve stimulation.10. Addition of noradrenaline (1 x 10(-4) g/ml.) to the ganglion side of the bath reduced but never abolished the amplitude of the synaptic potentials of colonic origin.11. It was concluded that in the guinea-pig, the IMG is involved in a peripheral reflex whose afferent limit of this reflex consists of the axons of cholinergic neurones within the wall of the colon. Many of these neurones are driven either directly or indirectly by cholinergic synapses. The efferent noradrenergic neurones of the IMG function as a group of inhibitory neurones which depress the activity of the excitatory neurones of the colon which are driving them.     

Differential thermosensitivity of sensory neurons in the guinea pig trigeminal ganglion

Intracellular recordings were employed to study the effects of temperature on membrane properties and excitability in sensory neurons of the intact guinea pig trigeminal ganglion (TG) maintained in vitro. Neurons were classified according to the shape and duration of the action potential into F (short-duration, fast spike) and S (long duration, slow spike with a "hump") types. Most type F (33/34) neurons had axons with conduction velocities >1.5 m/s, while only 30% (6/23) of type S neurons reached these conduction speeds suggesting differences in myelination. Cooling reduced axonal conduction velocity and prolonged spike duration in both neuronal types. In F-type neurons with strong inward rectification. cooling also increased the excitability, augmenting the input resistance and reducing the current firing threshold. These effects were not observed in S-type neurons lacking inward rectification. In striking contrast to results obtained in cultured TG neurons, cooling or menthol did not induce firing in recordings from the acutely isolated ganglion. However, after application of submillimolar concentrations (100 microM) of the potassium channel blocker 4-aminopyridine (4-AP), 29% previously unresponsive neurons developed cold sensitivity. An additional 31% developed ongoing activity that was sensitive to temperature. Only neurons with strong inward rectification (mostly F-type) became thermosensitive. Cooling- and 4-AP-evoked firing were insensitive to intracellular application of 4-AP or somatic membrane hyperpolarization, suggesting that their action was most prominent at the level of the axon. The lack of excitatory actions of low temperature in the excised intact ganglion contrasts with the impulse discharges induced by cooling in trigeminal nerve terminals of the same species, suggesting a critical difference between cold-transduction mechanisms at the level of the nerve terminals and the soma.     

Neuropeptide immunoreactivity of pericellular baskets in the guinea pig trigeminal ganglion

Using immunohistochemical techniques, calcitonin gene-related peptide, substance P and cholecystokinin localize to baskets of fine nerve fibers that completely surround occasional neurons in the guinea pig trigeminal ganglion. These observations, viewed with prior neuroanatomical studies indicating an intrinsic origin for these structures, raise again the possibility of intraganglionic communication within the trigeminal ganglion.     

Presynaptic muscarinic receptors in guinea pig superior cervical ganglion

This study characterizes the presynaptic muscarinic cholinergic receptors associated with the modulation of the electrically-evoked acetylcholine output from guinea pig superior cervical ganglion preincubated with [3H]choline. The M1-selective agonist pilocarpine had no effect while carbachol and oxotremorine strongly decreased the evoked outflow of tritium. Atropine increased such evoked release of [3H]acetylcholine whereas the M1-selective antagonist pirenzepine was ineffective. Moreover, atropine but not pirenzepine antagonized the inhibitory effect of carbachol. These results suggest that the guinea-pig superior cervical ganglion is equipped with presynaptic inhibitory muscarinic receptors of the M2 subtype.     

The effects of physostigmine on synaptic transmission in the inferior mesenteric ganglion of guinea-pigs

1. Synaptic potentials were recorded with intracellular electrodes from cells in the inferior mesenteric ganglion of the guinea-pig.2. Half-widths of the synaptic potentials recorded fell into two groups: type L cells had long synaptic potentials (11.6-15.2 msec) and low thresholds (14.6 mV mean), type S cells had short synaptic potentials (6.1-9.3 msec) and high thresholds (29.9 mV mean).3. Physostigmine (1.2 x 10(-6)M) caused a significant increase in the half-width of both types of synaptic potential.4. Physostigmine caused a significant increase in the half-width of spontaneous synaptic potentials and an increase in their amplitude.5. Repetitive preganglionic stimulation, in the presence of physostigmine, led to a marked and prolonged depolarization in all cells. In most cells repetitive spontaneous firing of action potentials was then observed. This effect was blocked by atropine (1.4 x 10(-7)M).6. The effect of atropine on the half-width in a physostigmine-treated cell was inconsistent: although synaptic potentials in some cells were slightly shortened their half-widths were always greater than the control.7. It is concluded that cholinesterase plays a role in limiting the time course of the synaptic potential, by limiting the duration of action of acetylcholine.