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[Labyrinthine and extralabryinethine influences on the background activity of the spinal interneurons of cats during swaying].

Interneuronal activity in the spinal cord of cats was recorded extracellularly at the lumbosacral level during vestibular stimulation by rocking in the vertical plane. Cats with normal and destroyed labyrinths were used. Four types of interneuronal responses were observed during stimulation. Interneuronal responses to stimulations by rocking in animals with destroyed labyrinths were similar to those in intact animals, but far less pronounced. The results show that responses of spinal cord interneurons to acceleration are under great extralabyrinthine influences

Animals↗

Synaptic convergence of motor and somatosensory cortical afferents onto GABAergic interneurons in the rat striatum.

Cortical afferents to the basal ganglia, and in particular the corticostriatal projections, are critical in the expression of basal ganglia function in health and disease. The corticostriatal projections are topographically organized but also partially overlap and interdigitate. To determine whether projections from distinct cortical areas converge at the level of single interneurons in the striatum, double anterograde labeling from the primary motor (M1) and primary somatosensory (S1) cortices in the rat, was combined with immunolabeling for parvalbumin (PV), to identify one population of striatal GABAergic interneurons. Cortical afferents from M1 and S1 gave rise to distinct, but partially overlapping, arbors of varicose axons in the striatum. PV-positive neurons were often apposed by cortical terminals and, in many instances, apposed by terminals from both cortical areas. Frequently, individual cortical axons formed multiple varicosities apposed to the same PV-positive neuron. Electron microscopy confirmed that the cortical terminals formed asymmetric synapses with the dendrites and perikarya of PV-positive neurons as well as unlabelled dendritic spines. Correlated light and electron microscopy revealed that individual PV-positive neurons received synaptic input from axon terminals derived from both motor and somatosensory cortices. These results demonstrate that, within areas of overlap of functionally distinct projections, there is synaptic convergence at the single cell level. Sensorimotor integration in the basal ganglia is thus likely to be mediated, at least in part, by striatal GABAergic interneurons. Furthermore, our findings suggest that the pattern of innervation of GABAergic interneurons by cortical afferents is different from the cortical innervation of spiny projection neurons.

Animals↗

Firing properties of identified interneuron populations in the mammalian hindlimb central pattern generator.

Little is known about the network structure of the central pattern generator (CPG) controlling locomotor movements in mammals. The present experiments aim at providing such knowledge by focusing on commissural interneurons (CINs) involved in left-right coordination. During NMDA and 5-HT-initiated locomotor-like activity, we recorded intracellularly from caudally or descending projecting L2 and L3 CINs (dCINs) located in the ventromedial area of the lumbar spinal cord in newborn rats. This region is crucial for rhythmic motor output and left-right coordination. The overall sample of dCINs represented a heterogenous population with neurons that fired in all phases of the locomotor cycle and exhibited varying degrees of rhythmicity, from strongly rhythmic to nonrhythmic. Among the rhythmic, putative CPG dCINs were populations that fired in-phase with the ipsilateral or with the contralateral L2 locomotor-like activity. There was a high degree of organization in the dorsoventral location of rhythmic dCINs, with neurons in-phase with the ipsilateral L2 activity located more ventrally. Spikes of rhythmically active dCINs were superimposed on membrane oscillations that were generated predominantly by synaptic input, with little direct contribution from the intrinsic pacemaker hyperpolarization-activated inward current. For both ipsilaterally and contralaterally firing dCINs the dominant synaptic drive was in-phase with the ipsilateral L2 motor activity. This study provides the first characterization of putative CPG interneurons in the mammalian spinal cord. Our results suggest an anatomical and physiological separation of CPG commissural interneurons in the ventral horn and demonstrate that it is possible to target specific interneuron subpopulations in the mammalian locomotor network.

Action Potentials↗

Bursting in leech heart interneurons: cell-autonomous and network-based mechanisms.

Rhythmic activity within the heartbeat pattern generator of the medicinal leech is based on the alternating bursting of mutually inhibitory pairs of oscillator heart interneurons (half-center oscillators). Bicuculline methiodide has been shown to block mutual inhibition between these interneurons and to cause them to spike tonically while recorded intracellularly (Schmidt and Calabrese, 1992). Using extracellular recording techniques, we show here that oscillator and premotor heart interneurons continue to burst when pharmacologically isolated with bicuculline, although the bursting is not robust in some preparations. We propose that a nonspecific leak current introduced by the intracellular microelectrode suppresses endogenous bursting activity to account for the discrepancy with results using intracellular recording. A two-parameter bifurcation diagram (E(leak) vs g(leak)) of a mathematical model of a single heart interneuron shows a narrow stripe of parameter values where bursting occurs, separating large zones of tonic spiking and silence. A similar analysis performed for a half-center oscillator model outlined a much larger area of bursting. Bursting in the half-center oscillator model is also less sensitive to variation in the maximal conductances of voltage-gated currents than in the single-neuron model. Thus, in addition to ensuring appropriate bursting characteristics such as period, phase, and duty cycles, the half-center configuration enhances oscillation robustness, making them less susceptible to random or imposed changes in membrane parameters. Endogenous bursting, in turn, ensures appropriate bursting if the strength of mutual inhibition is weakened and limits the minimum period of the half-center oscillator to a period near that of the single neuron.

Action Potentials↗

Modulatory effects of alpha1-,alpha2-, and beta -receptor agonists on feline spinal interneurons with monosynaptic input from group I muscle afferents.

Previous studies have shown that monoamines may modulate operation of spinal neuronal networks by depressing or facilitating responses of the involved neurons. Recently, activation of interneurons mediating reciprocal inhibition from muscle spindle (Ia) afferents and nonreciprocal inhibition from muscle spindle and tendon organ (Ia/Ib) afferents in the cat was found to be facilitated by noradrenaline (NA). However, which subclass membrane receptors are involved in mediating this facilitation was not established; the aim of the present experiments was to investigate this. Individual Ia- and Ia/Ib-inhibitory interneurons were identified in the cat lumbar spinal cord, and NA agonists were applied close to these neurons by ionophoresis. The agonists included the alpha1-receptor agonist phenylephrine, the alpha2-receptor agonists clonidine and tizanidine, and the beta-receptor agonist isoproterenol. Effects were measured by comparing changes in the number of extracellularly recorded spike potentials evoked by electrical stimulation of muscle nerves and changes in the latency of these potentials before, during, and after application of the tested compounds. Results show that the facilitatory effect of phenylephrine is as strong as that of NA, whereas the facilitatory effect of isoproterenol is weaker. Clonidine depressed activity of both Ia- and Ia/Ib-inhibitory interneurons, whereas tizanidine had no effect. These findings lead to the conclusion that beneficial antispastic effects of clonidine and tizanidine in humans are unlikely to be associated with an enhancement of the actions of Ia- and Ia/Ib-inhibitory interneurons, and the findings also support previous proposals that these compounds exert their antispastic actions via effects on other neuronal populations.

Adrenergic alpha-1 Receptor Agonists↗

Kv3 potassium conductance is necessary and kinetically optimized for high-frequency action potential generation in hippocampal interneurons.

Kv3 channels are thought to be essential for the fast-spiking (FS) phenotype in GABAergic interneurons, but how these channels confer the ability to generate action potentials (APs) at high frequency is unknown. To address this question, we developed a fast dynamic-clamp system (approximately 50 kHz) that allowed us to add a Kv3 model conductance to CA1 oriens alveus (OA) interneurons in hippocampal slices. Selective pharmacological block of Kv3 channels by 0.3 mm 4-aminopyridine or 1 mm tetraethylammonium ions led to a marked broadening of APs during trains of short stimuli and a reduction in AP frequency during 1 sec stimuli. The addition of artificial Kv3 conductance restored the original AP pattern. Subtraction of Kv3 conductance by dynamic clamp mimicked the effects of the blockers. Application of artificial Kv3 conductance also led to FS in OA interneurons after complete K+ channel block and even induced FS in hippocampal pyramidal neurons in the absence of blockers. Adding artificial Kv3 conductance with altered deactivation kinetics revealed a nonmonotonic relationship between mean AP frequency and deactivation rate, with a maximum slightly above the original value. Insertion of artificial Kv3 conductance with either lowered activation threshold or inactivation also led to a reduction in the mean AP frequency. However, the mechanisms were distinct. Shifting the activation threshold induced adaptation, whereas adding inactivation caused frequency-dependent AP broadening. In conclusion, Kv3 channels are necessary for the FS phenotype of OA interneurons, and several of their gating properties appear to be optimized for high-frequency repetitive activity.

Action Potentials↗

[Kainate receptor activation enhances both tonic and phasic GABA-ergic inhibition in CA1 hippocampal interneurons].

Kainate receptor agonists are powerful convulsants and excitotoxins. It has been a lot of controversy around functions of these receptors in the brain. It is shown in this article that kainate enhances evoked GABAergic IPSC (phasic currents) in CA1 interneurons in concentration-dependent manner. The phenomenon is likely to be due to kainate-mediated lowering of the threshold for action potential generation in interneuron axons and increased number of terminals responding to the same stimulus strength. Kainate application also induced an enhancement in tonic GABAergic conductance. This phenomenon can be attributed to massive extracellular GABA accumulation caused by interneuron firing in the presence of kainate. Extracellular GABA also shunts synaptic currents by activating tonic conductance as well as desensitizing synaptic GABAA receptors. Thus, the enhancement of the evoked IPSCs by 1 microM kainate was complicated by early and transient decrease. The kainate receptor-mediated enhancement of GABAergic tonic and phasic signalling to interneurons can contribute to the depression of GABAergic transmission to pyramidal neurons. The consequence of this phenomenon may play a major role in the epileptogenic action of this agent.

Animals↗

CXCR4 regulates interneuron migration in the developing neocortex.

The chemotactic factors directing interneuron migration during cerebrocortical development are essentially unknown. Here we identify the CXC chemokine receptor 4 (CXCR4) in interneuron precursors migrating from the basal forebrain to the neocortex and demonstrate that stromal cell-derived factor-1 (SDF-1) is a potent chemoattractant for isolated striatal precursors. In addition, we show that CXCR4 is present in early generated Cajal-Retzius cells of the cortical marginal zone. In mice with a null mutation in CXCR4 or SDF-1, interneurons were severely underrepresented in the superficial layers and ectopically placed in the deep layers of the neocortex. In contrast, the submeningeal positioning of Cajal-Retzius cells was unaffected. Thus, our findings suggest that SDF-1, which is highly expressed in the embryonic leptomeninx, selectively regulates migration and layer-specific integration of CXCR4-expressing interneurons during neocortical development.

Animals↗

Activation of beta1-adrenoceptors excites striatal cholinergic interneurons through a cAMP-dependent, protein kinase-independent pathway.

The role of noradrenergic neurotransmission was analyzed in striatal cholinergic interneurons. Conventional intracellular and whole-cell patch-clamp recordings were made of cholinergic interneurons in rat brain slice preparations. Bath-applied noradrenaline (NA) (1-300 microm) dose-dependently induced both an increase in the spontaneous firing activity and a membrane depolarization of the recorded cells. In voltage-clamped neurons, an inward current was induced by NA. This effect was not prevented by alpha-adrenoceptor antagonists, whereas it was mimicked by the beta-adrenoceptor agonist isoproterenol and blocked by the beta1 antagonists propranolol and betaxolol. Interestingly, forskolin, activator of adenylate cyclase, mimicked and occluded the membrane depolarization obtained at saturating doses of both dopamine and NA. Accordingly, SQ22,536, a selective adenylate cyclase inhibitor, reduced the response to NA. Analysis of the reversal potential of the NA-induced current did not provide homogeneous results, indicating the involvement of multiple membrane conductances. Because cAMP is known to modulate Ih, the effects of ZD7288, a selective inhibitor of Ih current, were examined on the NA-induced membrane depolarization/inward current. ZD7288 mostly reduced the response to NA. However, both KT-5720 and H-89, selective protein kinase A (PKA) blockers, failed to prevent the excitatory action of NA. Likewise, calphostin C, antagonist of PKC, genistein, inhibitor of tyrosine kinase, and 8-Bromo-cGMP, blocker of PKG, did not affect the response to NA. Finally, double-labeling experiments combining beta1-adrenoceptor and choline acetyltransferase immunocytochemistry by means of confocal microscopy revealed a strong beta1-adrenoceptor labeling on cholinergic interneurons. We conclude that NA depolarizes striatal cholinergic interneurons via beta1-adrenoceptor activation, through a cAMP-dependent but PKA-independent mechanism.

Adenylyl Cyclases↗

Brain-derived neurotrophic factor mediates activity-dependent dendritic growth in nonpyramidal neocortical interneurons in developing organotypic cultures.

Brain-derived neurotrophic factor (BDNF) promotes postnatal maturation of GABAergic inhibition in the cerebral and cerebellar cortices, and its expression and release are enhanced by neuronal activity, suggesting that it acts in a feedback manner to maintain a balance between excitation and inhibition during development. BDNF promotes differentiation of cerebellar, hippocampal, and neostriatal inhibitory neurons, but its effects on the dendritic development of neocortical inhibitory interneurons remain unknown. Here, we show that BDNF mediates depolarization-induced dendritic growth and branching in neocortical interneurons. To visualize inhibitory interneurons, we biolistically transfected organotypic cortical slice cultures from neonatal mice with green fluorescent protein (GFP) driven by the glutamic acid decarboxylase (GAD)67 promoter. Nearly all GAD67-GFP-expressing neurons were nonpyramidal, many contained GABA, and some expressed markers of neurochemically defined GABAergic subtypes, indicating that GAD67-GFP-expressing neurons were GABAergic. We traced dendritic trees from confocal images of the same GAD67-GFP-expressing neurons before and after a 5 d growth period, and quantified the change in total dendritic length (TDL) and total dendritic branch points (TDBPs) for each neuron. GAD67-GFP-expressing neurons growing in control medium exhibited a 20% increase in TDL, but in 200 ng/ml BDNF or 10 mm KCl, this increase nearly doubled and was accompanied by a significant increase in TDBPs. Blocking action potentials with TTX did not prevent the BDNF-induced growth, but antibodies against BDNF blocked the growth-promoting effect of KCl. We conclude that BDNF, released by neocortical pyramidal neurons in response to depolarization, enhances dendritic growth and branching in nearby inhibitory interneurons.

Animals↗

Receptor subtypes involved in the presynaptic and postsynaptic actions of dopamine on striatal interneurons.

By stimulating distinct receptor subtypes, dopamine (DA) exerts presynaptic and postsynaptic actions on both large aspiny (LA) cholinergic and fast-spiking (FS) parvalbumin-positive interneurons of the striatum. Lack of receptor- and isoform-specific pharmacological agents, however, has hampered the progress toward a detailed identification of the specific DA receptors involved in these actions. To overcome this issue, in the present study we used four different mutant mice in which the expression of specific DA receptors was ablated. In D1 receptor null mice, D1R-/-, DA dose-dependently depolarized both LA and FS interneurons. Interestingly, SCH 233390 (10 microm), a D1-like (D1 and D5) receptor antagonist, but not l-sulpiride (3-10 microm), a D2-like (D2, D3, D4) receptor blocker, prevented this effect, implying D5 receptors in this action. Accordingly, immunohistochemical analyses in both wild-type and D1R-/- mice confirmed the expression of D5 receptors in both cholinergic and parvalbumin-positive interneurons of the striatum. In mice lacking D2 receptors, D2R-/-, the DA-dependent inhibition of GABA transmission was lost in both interneuron populations. Both isoforms of D2 receptor, D2L and D2S, were very likely involved in this inhibitory action, as revealed by the electrophysiological analysis of the effect of the DA D2-like receptor agonist quinpirole in two distinct mutants lacking D2L receptors and expressing variable contents of D2S receptors. The identification of the receptor subtypes involved in the actions of DA on different populations of striatal cells is essential to understand the circuitry of the basal ganglia and to develop pharmacological strategies able to interfere selectively with specific neuronal functions.

Animals↗

The roles of local interneurons in the processing of olfactory information in the antennal lobes of the moth Manduca sexta.

The antennal lobe (AL) of the sphinx month Manduca sexta is characterized by a typically glomerular neuropil and two principal classes to neurons local interneurons and projection neurons. The somata of these neurons reside in defined neural cell-body groups in the AL, and the neurons exhibit characteristic patterns of innervation of the glomeruli. Evidence gathered to date indicates that individual antennal olfactory receptor-cell axons project to single glomeruli in the ipsilateral AL and make excitatory, apparently cholinergic synapses with neurites of AL neurons (usually local neurons) innervating the target glomeruli. Much has been learned about the physiology of the projection neurons, but only recently have the physiological properties and functions of the local interneurons been examined systematically through the use of intracellular recording and staining methods. Immunocytochemical studies have shown that most of the local interneurons contain GABA as well as one or more putative neuropeptides. Physiological, pharmacological, and biochemical experiments support the view that GABAergic local interneurons are responsible for inhibitory synaptic inputs to projection neurons that predominate in shaping the activity of projection neurons conveying synaptically processed olfactory information to higher-order centers in the protocerebrum.

Animals↗

[Characteristics of structural organization of interneurons in CA 3 region and mossy fibers of hippocampus].

The paper reviews the current data on the structural peculiarities of the interneurons of hippocampal area CA 3. Special attention is paid to horizontal spiny interneurons located in the stratum lucidum and forming direct contacts with the system of mossy fibers. Their axons terminate at the proximal portions of dendrites and cell body of pyramidal neurons, occasionally they make contacts with the main cells of the hilus and other hippocampal interneurons. Some portion of axon collaterals is in close vicinity to direct and indirect entorhinal-hippocampal connections, indicating their accommodation to selectively suppress mossy fiber input. The latter form the synapses of various types on the principal cells and on various types of interneurons of area CA 3, indicating different mechanisms of transmitter uptake and presynaptic control of their release.

Animals↗

Voltage-dependent nonlinearities in the membrane of locust nonspiking local interneurons, and their significance for synaptic integration.

Some aspects of the electronic and active membrane properties of nonspiking local interneurons were studied in isolated locust thoracic ganglia, using the switched current- and voltage-clamp techniques in neuropilar recordings. The average transmembrane potential (Vr) of the interneurons was -58 +/- 6mV (n = 85), and the input resistance (in the linear region of the current-voltage curve) was 16.5 +/- 8 M omega (n = 19, range 8 to 32 M omega). The membrane and equalizing time constants were estimated from charging curves evoked by the injection of low density hyperpolarizing current pulses from about -80 mV, i.e., from voltages in the linear region of the I-V curve. The curves yielded 2 time constants (tau m and tau l) whose average values were 33.2 +/- 9 msec and 3.3 +/- 1 msec (n = 18), respectively. The mean specific membrane resistance is therefore about 33 k omega.cm2, assuming that the membrane capacitance is ca. 1 microF.cm-2. An outward rectification was always observed upon depolarization from potentials more negative than Vr and was accompanied by a decrease in input resistance and membrane time constant. The "resting" membrane, for example, had a time constant of 26.4 +/- 8 msec (n = 31). This outward rectification was due to at least 2 conductances with different inactivation kinetics, similar to the transient "A" and "delayed-rectifier" potassium conductances. No inward rectification was ever observed upon injection of hyperpolarizing current. In about 60% of the recordings, an active and TTX-resistant depolarizing process could be evoked by rapid depolarization around Vr. The voltage-dependent properties of the membrane of the nonspiking local interneurons had dramatic effects on the shape and time course of natural or evoked unitary PSPs. The half-width of EPSPs, for example, decreased by a factor of 7.5 if the membrane potential was shifted from -93 to -50 mV. When the membrane potential of an interneuron was altered with a triangular current waveform, the reduction of tonically occurring IPSPs depended more on the sign and rate of the induced voltage change than on the absolute transmembrane potential. For 2 identical instantaneous values of membrane potential, for example, the reduction of the PSPs was greater during the depolarizing phase than during the hyperpolarizing phase of the current waveform. The possible nature of the active membrane conductances underlying the nonlinear electrical behavior of the membrane is discussed, together with their functional significance for local circuit synaptic integration.

Action Potentials↗

[Temporal organization of interneuronal relations in the cerebral cortex of the cat in relation to the level of alimentary motivation].

By means of records of multicellular activity, interneuronal relations and their modifications in two cortical zones (Visual and motor) were studied in cats at different levels of alimentary motivation. For quantitative evaluation of interneuronal relations the statistic method of cross-correlation analysis of impulse trains was used in determining the probability of the appearance of the discharge of one neurone after the impulse of the other one. For groups of neurones in both investigated cortical areas, three-neurones microsystems were singled out and their activity was analyzed by temporal parameters of interaction between neurones at the interval of 120 ms, both within one microarea (intraanalyzer connections) and between microareas of two cortical zones. The correlation of temporal parameters of interneuronal connections (temporal delays in the activity of neuronal pairs) changed depending on spatial localization of neurones and functional condition of the animals. The existence is suggested of "informational" (1-30 ms) and "motivational" (90-120 ms) values of interneuronal relations for interanalyser connections.

Animals↗

[Bilateral motor cortex effects on interneurons of the ventral horn of the lumbar segments].

The ventral horn interneurons in the spinal cord L6-7 segments were recorded in anesthetized cats during stimulation of the pericruciate cortex of both cerebral hemispheres. The majority of ventral interneurons are activated by volleys from the contralateral motor cortex and nearly half of them--by volleys from the ipsilateral cortex as well. Two kinds of firing patterns occurred: separate spikes and bursts of spikes. The latency of the 1st type firing was significantly greater than that of the 2nd one. There was a close relation of convergence of the afferent and corticospinal volleys on ventral horn interneurons: neurons activated by ipsilateral afferents only received synaptic influence only from the contralateral motor cortex whereas neurons with ipsilateral as well as contralateral inputs were activated by corticospinal volleys from both hemispheres. The interneurons with bilateral segmental and supraspinal connections seem to be involved in integrative action as important elements of the neuronal mechanism of bilateral motor coordination.

Action Potentials↗

Segmental specialization of a leech swim-initiating interneuron, cell 2051.

The physiological and anatomical properties of an unpaired intersegmental interneuron designated cell 205 are described. Cell 205 is unusual among leech neurons in that it combines a variety of functional properties in a single cell. Constant current depolarization of cell 205 initiates and maintains swimming behavior in semi-intact leeches or the swim motor pattern in brainless, isolated nerve cords. During swim episodes elicited by other stimuli, cell 205 is rhythmically active. Current pulses passed into cell 205 during swimming reset the pattern, indicting that it has access to, or may be a member of, the swim central pattern generator (CPG). Cell 205 is the first interneuron in this system to exhibit both swim-initiating and CPG functions. Individual touch, pressure, and nociceptive primary mechanoreceptor neurons polysynaptically excite cell 205, which, in addition, is coupled electrically to the multimodal S interneuron. These inputs may contribute to the initiation and/or modulation of swimming in response to sensory stimuli. Cell 205 shares some common synaptic inputs and outputs with the only other known swim-initiating interneuron, cell 204, but the two cells differ fundamentally in that cell 204 exerts only a tonic effect on the CPG. No synaptic interactions were found between cells 204 and 205, but their excitatory effects on swimming summate. Unlike other swim neurons which are segmentally repeated, cell 205 generally is present only in segment 9, and numerous lines of evidence suggest that it is, in fact, a segmentally differentiated homolog of cell 204.

Animals↗

Cholinergic, somatostatin-immunoreactive interneurons in the guinea pig intestine: morphology, ultrastructure, connections and projections.

The shape, projection, ultrastructure and chemistry of interneurons that were initially identified by their immunoreactivity for somatostatin in the small intestine of the guinea pig were examined. Somatostatin immunoreactive nerve cell bodies and nerve fibres were located in the myenteric plexus. Simultaneous labelling for 2 antigens revealed that the somatostatin immunoreactive interneurons were also immunoreactive for choline acetyltransferase, but not for calbindin or neuropeptide Y. Cell shapes were determined by immunohistochemistry, ultrastructural analysis and intracellular dye filling. The neurons had large cell bodies (38 x 14 microns) which gave rise to long branched filamentous dendrites and a single axon. The projections of the somatostatin immunoreactive interneurons were determined by analysis of patterns of fibre loss and survival following degenerative section of myenteric nerve pathways and by analysis of individual neurons that were injected intracellularly with dye. The axons projected anally, sometimes after a short oral excursion, and were confined to the myenteric plexus. They gave rise to multiple varicose branches within myenteric ganglia: in the majority of cases the first branch was within 100 microns of the cell body. Synaptic inputs to the cells were examined by light and electron microscopy. All somatostatin immunoreactive neurons received numerous somatostatin immunoreactive inputs on the cell body and all filamentous processes. Ultrastructural investigation indicated that these constituted the majority of all inputs. It is concluded that cholinergic, somatostatin immunoreactive, interneurons have a unique soma morphology and form synaptically connected chains that run anally in the myenteric plexus.

Animals↗