Origins of cholinergic inputs to the cell bodies of intestinofugal neurons in the guinea pig distal colon

Integration of function between gut regions is mediated by means of hormones and long neuronal reflex pathways. Intestinofugal neurons, which participate in one of these pathways, have cell bodies within the myenteric plexus and project their axons from the gut with the mesenteric nerves. They form excitatory synapses on neurons in prevertebral ganglia that in turn innervate other gut regions. The aim of the present study was to characterise immunohistochemically the synaptic input to intestinofugal neurons. The cell bodies of intestinofugal neurons that project from the distal colon were labelled with Fast Blue that was injected into the inferior mesenteric ganglia. Varicosities surrounding Fast Blue-labelled neurons were analysed for immunoreactivity for the vesicular acetylcholine transporter, vasoactive intestinal peptide, and bombesin. Most intestinofugal neurons were surrounded by nerve terminals immunoreactive for the vesicular acetylcholine transporter; many of these terminals also contained vasoactive intestinal peptide and bombesin immunoreactivity. This combination of markers occurs in axons of descending interneurons. Extrinsic denervation had no effect on the distribution of cholinergic terminals around intestinofugal neurons. A decrease in the number of vesicular acetylcholine transporter and vasoactive intestinal peptide immunoreactive terminals occurred around nerve cells immediately anal, but not oral, to myotomy operations. Consistent with previous physiological studies, it is concluded that intestinofugal neurons receive cholinergic synaptic input from other myenteric neurons, including cholinergic descending interneurons. Thus, intestinofugal neurons are second, or higher, order neurons in reflex pathways, although physiological data indicate that they also respond directly to distension of the gut wall.     

Synaptic responses evoked by mechanical stimulation of the mucosa in morphologically characterized myenteric neurons of the guinea-pig ileum.

Recordings were made from myenteric neurons of the guinea-pig ileum during reflexes evoked by mechanical stimulation of the mucosa. Impaled neurons were injected with dye (Lucifer yellow or biocytin), and their shapes were determined. All neurons were 5-12 mm from the stimulus, a brush stroke that deformed the mucosal villi. Neurons were classified as S-neurons or AH-neurons (Hirst et al., 1974). About 40% of S-neurons oral to a stimulus responded with bursts of fast EPSPs (average frequency, 15-40 Hz); these neurons were in ascending reflex pathways. About 60% of S-neurons anal to a stimulus responded with similar bursts of fast EPSPs or slow depolarizations; these neurons were in descending pathways. Only 2 of 48 AH-neurons responded, both in descending pathways. Most S-neurons in either ascending or descending pathways received inputs from at least 2 or 3 other neurons. Action potentials evoked during a response averaged 3-10 Hz in frequency, with occasional bursts at up to 100 Hz. The speed of conduction along the reflex pathways was about 0.5 m/sec. All S-neurons were uniaxonal, but they differed in size, dendritic morphology, and projections. The axons of S-neurons injected with biocytin were followed up to 7 mm within the myenteric plexus. Three S-neurons projected to the tertiary plexus and were probably longitudinal muscle motor neurons; 2 of these were in descending pathways. Five S-neurons projected along the intestine and had varicose collaterals in some ganglia. These neurons were probably interneurons; 3 were descending and 2 ascending, and all responded in the appropriate reflex pathway. Many S-neurons had short axons that entered the circular muscle and were probably circular muscle motor neurons. Others projected several millimeters along the intestine before entering the circular muscle or fading beyond detection. From this study, we have been able to deduce the circuits mediating ascending and descending mucosa-to-muscle reflexes. It is concluded that AH-neurons are primary sensory neurons and S-neurons are interneurons and muscle motor neurons in the circuits.     

Targets of myenteric interneurons in the guinea-pig small intestine

Abstract  Polarized outputs of myenteric interneurons in guinea-pig small intestine have been well studied. However, the variety of motility patterns exhibited suggests that some interneuron targets remain unknown. We used antisera selected to distinguish interneuron varicosities and known myenteric neuron types to investigate outputs of three interneuron classes in guinea-pig jejunum; two classes of descending interneurons immunoreactive (IR) for somatostatin (SOM) or nitric oxide synthase (NOS)/vasoactive intestinal peptide (VIP), and one class of ascending interneurons [calretinin/enkephalin (ENK)-IR]. Varicosities apposed to immunohistochemically identified cell bodies were quantified by confocal microscopy. Intrinsic sensory neurons (calbindin-IR) were apposed by few varicosities. Cholinergic secretomotor neurons (neuropeptide Y-IR) were apposed by many SOM-IR varicosities. Longitudinal muscle excitatory motor neurons (calretinin-IR) were apposed by some VIP- and ENK-IR varicosities, but few SOM-IR varicosities. Ascending interneurons (calretinin-IR) were apposed by many varicosities of all types. NOS-IR interneurons and inhibitory motor neurons were apposed by numerous VIP-IR and SOM-IR varicosities. NOS-IR short inhibitory motor neurons were apposed by significantly fewer ENK-IR varicosities than other NOS-IR neurons. Based on the specific chemical coding of ascending (ENK) and descending (SOM) interneurons, we conclude that cholinergic secretomotor neurons and short inhibitory neurons are located in descending reflex pathways, while ascending interneurons and NOS-IR descending interneurons are focal points at which ascending and descending pathways converge.     

Skeletal muscle tone and the misunderstood stretch reflex.

This review presents a revision of long-accepted tenets regarding the genesis of muscle tone in humans. Most discussions liken muscle tone in humans to the reflex tone described by Sherrington in decerebrate animals. That tradition presumes that muscle tone is fully determined by the monosynaptic stretch reflex, that tonic fusimotor activity is necessary for its production in normal humans, and that tonic muscle tone in antigravity leg muscles is responsible for the maintenance of posture. Data reviewed here show that nonreflex, mechanical mechanisms are involved in the maintenance of resting muscle tone; that spinal cord reflex responses are not stereotyped responses, but depend upon the ongoing activity in interneurons upon which inputs from a variety of peripheral sensory receptors and descending fiber systems converge; that long-latency transcortical responses are elicited when a muscle is stretched, and these responses effectively deal with large displacements; and that inertial components and viscoelastic muscle forces can counterbalance small amounts of body sway during quiet standing. In sum, the response to muscle stretch is not fixed and inflexible, but can be adjusted according to the demands of the moment.     

Projections of submucosal neurons to the myenteric plexus of the guinea pig intestine: in vitro tracing of microcircuits by retrograde and anterograde transport

The enteric nervous system (ENS) can mediate reflex activity without input from the brain or spinal cord. The ENS thus contains intrinsic primary afferent neurons that link mucosal sensory receptors with motor neurons in the myenteric plexus. The intrinsic primary afferent neurons of the gut have not yet been identified. Although the submucosal plexus is known to innervate the mucosa, where enteric sensory receptors are located, no submucosal to myenteric projections have previously been found. In order to determine whether such projections exist, the submucosal plexus was examined following the microinjection of a retrograde tracer (Fluoro-Gold or 4-acetoamido, 4'-isothiocyanostilbene-2,2'-disulphonic acid [SITS]) into single myenteric ganglia. In addition, the myenteric plexus was studied following the iontophoretic injection of an anterograde tracer (Phaseolus vulgaris leucoagglutinin; [PHA-L]) into single submucosal ganglia. Ganglia were visualized by use of differential interference contrast optics and were injected from the beveled tip of a glass micropipette; 2.5-3.0 hours were allotted for retrograde and 20-24 hours (under culture conditions) for anterograde transport. In the myenteric plexus, a small number of the neurons of each injected ganglion were fluorescent and additional neurons in distant myenteric ganglia (predominantly orad) were also retrogradely labeled. About five to six submucosal neurons deep to but not directly underneath the injected myenteric ganglion were labeled by Fluoro-Gold or SITS and only rarely was there more than one labeled neuron in a submucosal ganglion. When control injections of Fluoro-Gold were placed into the muscle instead of a ganglion, some myenteric neurons near the injection site became labeled indicating an innervation of the circular muscle by myenteric neurons; however, there was no labeling of neurons in the submucosal plexus. Similarly, if connections between the myenteric and submucosal plexuses were severed before injecting Fluoro-Gold, no submucosal neurons were labeled. Following injection of PHA-L into a single submucosal ganglion, small-diameter axons were labeled in approximately 2 myenteric ganglia as well as in several distant submucosal ganglia (mainly anal and circumferential to the injection site). Additional labeled fibers traveled with blood vessels or surrounded mucosal crypts. It is concluded that submucosal neurons project to the myenteric plexus as well as to the mucosa and to one another. These observations are consistent with the hypothesis that at least some intrinsic enteric primary afferent neurons reside in the submucosal plexus.     

Reflex changes in circular muscle activity elicited by stroking the mucosa: an electrophysiological analysis in the isolated guinea-pig ileum

A preparation of isolated small intestine from the guinea-pig was studied in which reflex responses of the circular muscle were recorded intracellularly when sensory receptors in the mucosa were stimulated mechanically. This preparation was used to examine the properties of mucosa to muscle reflexes that involve non-cholinergic motor neurons innervating the circular muscle. Reproducible stimulation of the mucosa was achieved by stroking with a motor-driven brush. Gentle brush-strokes applied to the mucosa typically evoked inhibitory junction potentials anal to the stimulus and excitatory junction potentials at recording sites oral to the stimulus. Both events were rapid in onset and up to 25 mV in amplitude. The reflexes were blocked by tetrodotoxin (0.5 microM). Junction potentials declined in amplitude with distance from the stimulus, the amplitude of the excitation 15 mm oral to the stimulus was half that at 5 mm from the stimulus, whereas the amplitude of the inhibitory potential at 40-45 mm was about 60% of that at 5-10 mm anal to the stimulus. Hexamethonium (100-200 microM) blocked the ascending excitation but only slightly reduced the descending inhibition. Ascending excitation was blocked by antagonists for substance P receptors in the muscle, and inhibition was substantially reduced by apamin (0.2 microM), both before and after hexamethonium. Both responses were abolished by removal of the mucosa from the stimulus site and when lesions were made through the myenteric plexus between the stimulation and recording sites, but persisted when similar lesions were made through the submucous plexus. It is concluded that there are neurons with mechanoreceptive nerve endings in the mucosa. Stimulation of such sensory neurons leads to activation of pathways in the myenteric plexus that excite motor neurons to the muscle both oral and anal to the stimulation site. The demonstration that mucosa to muscle reflexes can be consistently evoked in the small intestine in vitro provides an opportunity for close analysis of the reflex pathways. Such analysis is not so readily achieved when reflexes are initiated by distension that, by moving the intestine, can dislodge the recording electrode.     

Morphological evidence for novel enteric neuronal circuitry in guinea pig distal colon

The gastrointestinal (GI) tract is unique compared to all other internal organs; it is the only organ with its own nervous system and its own population of intrinsic sensory neurons, known as intrinsic primary afferent neurons (IPANs). How these IPANs form neuronal circuits with other functional classes of neurons in the enteric nervous system (ENS) is incompletely understood. We used a combination of light microscopy, immunohistochemistry and confocal microscopy to examine the topographical distribution of specific classes of neurons in the myenteric plexus of guinea‐pig colon, including putative IPANs, with other classes of enteric neurons. These findings were based on immunoreactivity to the neuronal markers, calbindin, calretinin and nitric oxide synthase. We then correlated the varicose outputs formed by putative IPANs with subclasses of excitatory interneurons and motor neurons. We revealed that calbindin‐immunoreactive varicosities form specialized structures resembling ‘baskets’ within the majority of myenteric ganglia, which were arranged in clusters around calretinin‐immunoreactive neurons. These calbindin baskets directly arose from projections of putative IPANs and represent morphological evidence of preferential input from sensory neurons directly to a select group of calretinin neurons. Our findings uncovered that these neurons are likely to be ascending excitatory interneurons and excitatory motor neurons. Our study reveals for the first time in the colon, a novel enteric neural circuit, whereby calbindin‐immunoreactive putative sensory neurons form specialized varicose structures that likely direct synaptic outputs to excitatory interneurons and motor neurons. This circuit likely forms the basis of polarized neuronal pathways underlying motility.     

Correlation of electrophysiology,shape and synaptic properties of myenteric AH neurons of the guinea pig distal colon

Well-defined correlations between morphology, electrophysiological properties and the types of synaptic inputs received are established for myenteric neurons in the guinea pig ileum. However, in the distal colon, the correlations between AH electrophysiological properties, presence of fast excitatory post-synaptic potentials (EPSPs) and neuronal shape have been inadequately resolved and it is unknown whether any colon neurons receive synaptic inputs that generate sustained excitation. In this work, we have used intracellular recording, dye filling via the recording electrode, and immunohistochemistry to classify distal colon neurons. Neurons (24 of 168) had Dogiel type II morphology and 42% of these were dendritic type II neurons, compared to about 10% in the ileum. All Dogiel type II neurons had AH electrophysiological properties, including a prolonged post-spike after-hyperpolarization (AHP). None of these received fast excitatory post-synaptic potentials, 11 of 22 tested exhibited sustained slow post-synaptic excitation (SSPE) in response to 1 Hz pre-synaptic stimulation and 13 of 15 tested were immunoreactive for calbindin. Neurons (127) had Dogiel type I, filamentous or other uniaxonal cell shape and S type electrophysiology. Neurons of this group had fast excitatory post-synaptic responses to stimulation of synaptic inputs, but did not exhibit a prolonged post-spike after-hyperpolarization or sustained slow post-synaptic excitation. Another group of neurons (17) had both AH electrophysiological characteristics and fast excitatory post-synaptic potentials. These neurons had Dogiel type I, filamentous or other uniaxonal shapes, but none had Dogiel type II morphology and none showed sustained slow post-synaptic excitation. It is concluded that Dogiel type II neurons are all AH neurons and are probably intrinsic sensory neurons that could be involved in long-term changes in excitability in the colon. All other neurons are monoaxonal; these are motor neurons and interneurons, and most are S neurons, electrophysiologically. A small number of monoaxonal neurons display AH electrophysiology and also receive fast excitatory synaptic inputs. These include motor and interneurons, but not sensory neurons.     

Identification of galanin receptor 1 on excitatory motor neurons in the guinea pig ileum

Exogenously administered galanin inhibits cholinergic transmission to the longitudinal muscle and reduces peristaltic efficiency in the guinea pig ileum with a mechanism partially mediated by galanin receptor 1 (GAL-R1). We investigated the effect of exogenous galanin 1-16, which has high affinity for GAL-R1, on the ascending excitatory reflex of the circular muscle elicited by radial distension in isolated segments of guinea pig ileum. We used a three-compartment bath that allows dissecting the ascending pathway into the oral (site of excitatory motor neurons), intermediate (site of ascending interneurons) and caudal compartment (site of intrinsic primary afferent neurons). Galanin 1-16 (0.3-3 micromol L(-1)) applied to the oral compartment inhibited in a concentration-dependent manner the ascending excitatory reflex elicited by the wall distension in the caudal compartment. This effect was antagonized by the GAL-R1 antagonist, RWJ-57408 (1 and 10 micromol L(-1)). By contrast, galanin 1-16 was ineffective when added to the intermediate or caudal compartment up to 3 micromol L(-1). GAL-R1 immunoreactive neurons did not contain neuron-specific nuclear protein, a marker for intrinsic primary afferent neurons. These findings indicate that GAL-R1s are present on motor neurons responsible for the ascending excitatory reflex, but not on ascending interneurons and intrinsic primary afferent neurons.     

Correlation of electrophysiological and morphological characteristics of enteric neurons in the mouse colon

We report on the first correlative study of the electrophysiological properties, shapes, and projections of enteric neurons in the mouse. Neurons in the myenteric plexus of the mouse colon were impaled with microelectrodes containing biocytin, their passive and active electrophysiological properties determined, and their responses to activation of synaptic inputs investigated. Biocytin, injected into the neurons from which recordings were made, was converted to an optically dense product and used to determine the shapes of neurons. By electrophysiological properties, almost all neurons belonged to one of two classes, AH neurons or S neurons. AH neurons had a biphasic repolarization of the action potential, and slow afterhyperpolarizing potentials usually followed the action potentials. S neurons had monophasic repolarizations, no slow afterhyperpolarization, and fast excitatory postsynaptic potentials in response to fibre tract stimulation. By shape, neurons were divided into Dogiel type II (28/136 neurons) and uniaxonal neurons. Dogiel type II neurons had large, smooth-surfaced cell bodies and several long processes that supplied branches within myenteric ganglia. All Dogiel type II neurons had AH electrophysiology; conversely, most AH neurons had Dogiel type II morphology. The majority of uniaxonal neurons had lamellar dendrites, i.e., Dogiel type I morphology. They projected to the circular muscle (circular muscle motor neurons), to the longitudinal muscle (longitudinal muscle motor neurons), and to other myenteric ganglia (interneurons) and in some cases could not be traced to target cells. All S neurons were uniaxonal. A small proportion of uniaxonal neurons (3/70) had AH electrophysiology. Fast excitatory synaptic potentials were only recorded from uniaxonal neurons and were in most cases blocked by nicotinic receptor antagonists. A small component of fast excitatory transmission in some neurons was antagonized by the purine receptor antagonist PPADS. Slow excitatory postsynaptic potentials were observed in both AH and S neurons. Slow inhibitory postsynaptic potentials were recorded from S neurons. We conclude that the major classes of neurons are Dogiel type II neurons with AH electrophysiological properties and Dogiel type I neurons with S electrophysiological properties. The S/Dogiel type I neurons include circular muscle motor neurons, longitudinal muscle motor neurons, and interneurons.     

Mucosal distortion by compression elicits polarized reflexes and enhances responses of the circular muscle to distension in the small intestine.

The possibility that distortion of the mucosa by compression might be a sufficient stimulus to evoke reflex responses in intestinal muscle, and that such reflexes might summate with distension-evoked (stretch) reflexes, was tested in isolated segments of guinea pig small intestine. Opened segments of intestine were pinned flat in an organ bath with, or without, distending balloons embedded in its base. Intracellular microelectrode recordings were taken from the circular muscle oral and anal to sites of application of sensory stimuli. Pressure against the mucosa, which distorts the villi without the wall being stretched, evoked polarized reflex responses in the circular muscle, consisting of excitatory junction potentials oral and inhibitory junction potentials anal to the stimulus. Distension stimuli applied by 6-mm diameter balloons that pushed against either the serosal or the mucosal surface also evoked excitatory junction potentials in the muscle oral to the stimulus and inhibitory junction potentials at anal sites. Response amplitudes were 20% greater when the distending balloon pushed against the mucosal surface. Responses to distension from the serosal side were of 20% greater amplitude when combined with mucosal distortion by compression than without such compression. It is concluded that peristaltic movements that are commonly studied in the small intestine can be consequences of reflexes elicited at the same time from mucosal distortion receptors and from stretch receptors.     

Convergence of reflex pathways excited by distension and mechanical stimulation of the mucosa onto the same myenteric neurons of the guinea pig small intestine.

The effects on morphologically and electrophysiologically characterized myenteric neurons of activation of intestinal reflex pathways were examined in vitro. Opened segments of guinea pig small intestine were pinned serosa down in an organ bath that had two balloons set into its base. A 5-10-mm-wide strip of myenteric plexus between the balloons was exposed from the mucosal side, and neurons were impaled with microelectrodes. Reflex pathways were stimulated by inflation of the balloons to distend the intestinal wall, and by deforming the exposed mucosal villi with a brush. Impaled neurons were classified electrophysiologically as AH-neurons or S-neurons (Hirst et al., 1974) and were injected with biocytin to determine their shapes and projections. None of the 58 AH-neurons responded to distension. In contrast, 63 of 131 S-neurons responded to distension with a burst of fast EPSPs; about one-third of the responding S-neurons received input from ascending reflex pathways, one-third received input from descending reflex pathways, and one-third received input from both ascending and descending pathways. Most neurons in this last group supplied extensive varicose branches to the tertiary plexus and were probably longitudinal muscle motor neurons. Neurons receiving input from only one pathway usually projected in the direction of that pathway; many of these were circular muscle motor neurons. Almost all neurons responding to distension were also excited by deforming the villi. Responses evoked by distension or deforming the mucosa declined when stimuli were repeated at intervals less than 10 sec. This was seen in ascending and descending pathways but was more prominent in the former. Deforming the mucosa evoked a normal response even when the response to repeated distensions had disappeared. It is concluded that distension and deforming the mucosa excite separate populations of sensory neurons to activate reflex pathways that converge onto common motor neurons and probably onto common interneurons.     

Identification of spinal afferent nerve endings in the colonic mucosa and submucosa that communicate directly with the spinal cord: The gut–brain axis

The major sensory nerve pathway between the colon and central nervous system (spinal cord and brain) that underlies the gut–brain axis, is via spinal afferent neurons, with cell bodies in dorsal root ganglia (DRG). Our aim was to identify the sensory nerve endings in the colon that arise from single colorectal-projecting DRG neurons. C57BL/6 mice were anesthetized and lumbosacral L6-S1 DRG injected with dextran biotin. Mice recovered for 7 days. The whole colon was then removed and stained to visualize single axons and nerve endings immunoreactive to calcitonin gene-related peptide (CGRP). Single axons arising from DRG were identified in the distal colon and their morphological features and CGRP immunoreactivity characterized. After entering the colon, single axons ramified rostrally or caudally along many rows of myenteric ganglia with little circumferential displacement, giving off varicose endings in multiple ganglia. Nerve endings arising from two classes of colorectal-projecting DRG neuron were identified. One class was peptidergic neurons that had nerve endings in circular muscle, myenteric ganglia, and submucosa. Another class of nonpeptidergic neurons innervated mucosal crypts, myenteric ganglia, and submucosa. Different morphological types of nerve endings which innervate different anatomical layers of colon can arise from the same axon and sensory neuron in DRG. These findings suggest single peptidergic and nonpeptidergic sensory neurons in DRG are potentially capable of detecting sensory stimuli from different anatomical layers of the colon, via different types of nerve endings.     

Identification of spinal neurons involved in the urethrogenital reflex in the female rat

The urethrogenital (UG) reflex is a spinal sexual reflex that consists of autonomic and somatic nerve activity and vaginal, uterine, and anal sphincter contractions. The UG reflex is under tonic descending inhibition by neurons in the region of the nucleus paragigantocellularis (nPGi). The location of spinal neurons activated by the UG reflex was examined in the female rat using the immediate early gene, c-fos. In addition, the descending inputs from the nPGi onto fos-activated neurons was examined using the anterograde tracer biotin dextran amine injected into the nPGi. The UG reflex resulted in a significant increase in fos-positive nuclei in segments T12-S1, compared with experimental controls in which the UG reflex was not activated. Spinal circuits involved in the UG reflex include neurons relaying afferent information from the pudendal sensory nerve, in the dorsal horn and medial cord of L5-S1. Efferent output includes preganglionic neurons located in the lateral gray of L5-S1 and lateral and medial gray of T13-L2. Spinal interneurons involved in the UG reflex were found close to the preganglionic neurons and in the dorsal horn and intermediate and medial gray of T12-S1. NPGi inputs were found primarily on the autonomic efferents and interneurons in the medial and intermediate gray. These studies demonstrate multisegmental spinal circuits activated with the UG reflex and demonstrate that the descending inhibition from the nPGi is by means of preganglionic and somatic efferents and spinal interneurons closely associated with the efferent output.     

Cluster organization and response characteristics of the giant fiber pathway of the blowfly Calliphora erythrocephala

Intersegmental descending neurons (DNs) link the insect brain to the thoracic ganglia. Iontophoresis of cobalt or fluorescent dyes reveals DNs as uniquely identifiable elements, the dendrites of which are situated within a characteristic region of the lateral deutocerebrum. Here we demonstrate that DNs occur as discrete groups of elements termed DN clusters (DNCs). A DNC is a characteristic combination of neurons that arises from a multiglomerular complex in which the main components of each glomerulus are a characteristic ensemble of sensory afferents. Other neurons involved in the complex are local interneurons, heterolateral interneurons that connect DNCs on both sides of the brain, and neurons originating in higher centers of the brain. We describe the structure, relationships, and projections of eight DNs that contribute to a descending neuron cluster located ventrally in the lateral deutocerebrum, an area interposed between the ventral antennal lobes and the laterally disposed optic lobes. We have named this cluster the GDNC because its most prominent member is the giant descending neuron (GDN), which plays a cardinal role in the midleg "jump" response and which is implicated in the initiation of flight. The GDN and its companion neurons receive primary mechanosensory afferents from the antennae, terminals of wide- and small-field retinotopic neurons originating in the lobula, and endings derived from sensory interneurons that originate in leg neuropil of the thoracic ganglia. We demonstrate that DNs of this cluster share morphological and functional properties. They have similar axon trajectories into the thoracic ganglia, where they invade functionally related neuropils. Neurons of the GDNC respond to identical stimulus paradigms and share similar electrophysiological characteristics. Neither the GDN nor other members of its cluster show spontaneous activity. These neurons are reluctant to respond to unimodal stimuli, but respond to specific combinations of visual and mechanosensory stimulation. These results suggest that in flies groups of morphologically similar DNs responding to context-specific environmental cues may cooperate in motor control.     

Post-stimulus depression of reflex changes in circular muscle activity in the guinea pig small intestine.

The extent and time course of depression of successive reflex responses recorded with intracellular microelectrodes from the circular smooth muscle of the guinea pig small intestine were determined. Two stimuli were used, distension and distortion of the mucosa by compression; these were applied either at the same or at different sites. Excitatory responses oral and inhibitory responses anal to the stimuli were recorded. Post-stimulus depression of both ascending excitatory and descending inhibitory reflexes occurred, but the extent of depression was slightly less for the descending inhibition. A conditioning distension lasting 9 s depressed the excitatory response to a test distension applied 2 s later at the same site by 90%. After 30 s the depression was 50% and test responses were normal if inter-stimulus intervals were increased to 2 min. Increasing the duration of the conditioning stimulus increased the depression. Post-stimulus depression was less for compression stimuli than for distension stimuli and prior mucosal compression had almost no effect on responses to subsequent distension. The post-stimulus depression was greater if conditioning and test stimuli were at the same rather than different sites. For different sites, conditioning stimuli at 15 mm from the recording site (near) depressed responses to stimuli at 30 mm (far) to a greater extent than far stimuli depressed responses to near stimuli. If the conditioning stimulus at 15 mm was maintained until after the far test stimulus was applied, depression of the test response did not occur. It is concluded that the major sites of post-stimulus depression are at the synapses between primary sensory neurons and the first interneurons of reflex pathways, and that post-stimulus depression also occurs at other places in the pathway, presumably at synapses between interneurons or between interneurons and motor neurons.     

Morphology, electrophysiology, and calbindin immunoreactivity of myenteric neurons in the guinea pig distal colon

The morphological and physiological characteristics of myenteric neurons in the guinea pig distal colon were determined using Lucifer yellow- or N-(2-aminoethyl) biotinamide-containing microelectrodes and intracellular recording and staining methods. The neurons in this study (n = 204) were classified on the basis of the shapes of their cell bodies and short processes or dendrites and the number of long processes or axons as Dogiel type I (n = 75 neurons; 36.8%), filamentous (n = 31 neurons; 15.2%), Dogiel type II (n = 38 neurons; 18.6%), and unclassified (n = 60 neurons; 29.4%). All Dogiel type II neurons had action potentials followed by an after-spike hyperpolarization (AH), and most of them (84%) had large, smooth somata and filamentous, short processes in addition to multiple, long processes or axons. Most of Dogiel type I, filamentous, and unclassified neurons (98%) had a single, long process, but four Dogiel type I neurons and one unclassified neuron had two long processes terminating as varicosities within other ganglia or on the surface of longitudinal muscle. The projections of monoaxonal neurons were distributed equally between oral and aboral directions, and most of them received fast excitatory postsynaptic potentials (EPSPs). All of the Dogiel type II neurons and seven Dogiel type I neurons were positive for calbindin immunoreactivity, but three filamentous neurons received fEPSPs, had spikes followed by AH, and were negative for calbindin. The presence of calbindin-immunoreactive(-IR) neurons was quite variable among the ganglia. These results confirm that neither the presence of calbindin immunoreactivity nor the absence of fEPSPs can be used as a predictor of cellular morphology or electrophysiological properties of myenteric neurons in the distal colon.     

Ultrastructural examination of the targets of serotoninn immunoreactive descending interneurons in the guinea pig small intestine

Serotonin neurons are descending interneurons in the myenteric plexus of the guinea pig small intestine. Preembedding single- and double-label immunocytochemistries at the ultrastructural level were used to identify the targets of these serotonin interneurons. Serial ultrathin sections were taken through a myenteric ganglion that had been processed for serotonin immunocytochemistry. The ganglion contained the cell bodies of 69 neurons, including 2 serotonin neurons and 6 neurons with the ultrastructural features of Dogiel type II cells. For each cell body in the ganglion, the number of serotonin inputs (synapses and close contacts) was determined. About 50% of the cell bodies did not receive any serotonin inputs. The most abundant serotonin terminals were related to two targets: other serotonin descending interneurons and a population of neurons with Dogiel type I morphology, but whose neurochemistry and function is unknown. The serotonin inputs to the serotonin cell bodies were located predominantly on the lamellar dendrites. Each of the Dogiel type II neurons received 3 or fewer serotonin inputs, and none of the serotonin inputs to Dogiel type II neurons formed a synapse. Overall, about 40% of the serotonin inputs formed synapses. The serotonin inputs to neurons that received many serotonin inputs were more likely to show synaptic specializations than serotonin inputs to neurons that received few serotonin inputs. Inhibitory motor neurons contain nitric oxide synthase (NOS). At the light microscope level, serotonin nerve fibers do not form dense pericellular baskets around NOS cell bodies. To determine whether there are serotonin input to NOS neurons, serial ultrathin sections were taken through a myenteric ganglion that had been processed for preembedding double-label immunocytochemistry, in which the NOS neurons were labeled with peroxidase-diaminobenzidine and the serotonin neurons with silver-intensified 1 nm gold. Only 1 out of 9 NOS cells examined in serial section received more than 5 serotonin inputs. The results suggest that, in the guinea pig small intestine, the serotonin descending interneurons are not an essential element of the descending inhibitory reflex. © 1995 Wiley-Liss, Inc.     

Reliable Sensory Processing in Mouse Visual Cortex through Cooperative Interactions between Somatostatin and Parvalbumin Interneurons

Intrinsic neuronal variability significantly limits information encoding in the primary visual cortex (V1). However, under certain conditions, neurons can respond reliably with highly precise responses to the same visual stimuli from trial to trial. This suggests that there exists intrinsic neural circuit mechanisms that dynamically modulate the intertrial variability of visual cortical neurons. Here, we sought to elucidate the role of different inhibitory interneurons (INs) in reliable coding in mouse V1. To study the interactions between somatostatin-expressing interneurons (SST-INs) and parvalbumin-expressing interneurons (PV-INs), we used a dual-color calcium imaging technique that allowed us to simultaneously monitor these two neural ensembles while awake mice, of both sexes, passively viewed natural movies. SST neurons were more active during epochs of reliable pyramidal neuron firing, whereas PV neurons were more active during epochs of unreliable firing. SST neuron activity lagged that of PV neurons, consistent with a feedback inhibitory SST→PV circuit. To dissect the role of this circuit in pyramidal neuron activity, we used temporally limited optogenetic activation and inactivation of SST and PV interneurons during periods of reliable and unreliable pyramidal cell firing. Transient firing of SST neurons increased pyramidal neuron reliability by actively suppressing PV neurons, a proposal that was supported by a rate-based model of V1 neurons. These results identify a cooperative functional role for the SST→PV circuit in modulating the reliability of pyramidal neuron activity.SIGNIFICANCE STATEMENT Cortical neurons often respond to identical sensory stimuli with large variability. However, under certain conditions, the same neurons can also respond highly reliably. The circuit mechanisms that contribute to this modulation remain unknown. Here, we used novel dual-wavelength calcium imaging and temporally selective optical perturbation to identify an inhibitory neural circuit in visual cortex that can modulate the reliability of pyramidal neurons to naturalistic visual stimuli. Our results, supported by computational models, suggest that somatostatin interneurons increase pyramidal neuron reliability by suppressing parvalbumin interneurons via the inhibitory SST→PV circuit. These findings reveal a novel role of the SST→PV circuit in modulating the fidelity of neural coding critical for visual perception.     

Early functional organization of spinal neurons in developing lower vertebrates

The spinal neurons in the embryos and young larvae of two amphibians (Xenopus and Triturus) and two fish (Oryzias and Brachydanio) are described and compared. They can be placed into a limited number of common neuron classes: Rohon-Beard sensory, dorsolateral and dorsolateral commissural sensory interneurons, inhibitory ascending interneurons, two classes of inhibitory commissural interneuron, excitatory descending interneurons, motoneurons and possible sensory Kolmer-Agdhur neurons. In Triturus and other urodeles, there are also giant dorsolateral commissural sensory interneurons. The functions of the spinal neurons in simple flexion responses and swimming are considered in relation to evidence mainly from the Xenopus tadpole.