Rhythmic patterns evoked in locust leg motor neurons by the muscarinic agonist pilocarpine

1993 ◽  
Vol 69 (5) ◽  
pp. 1583-1595 ◽  
Author(s):  
S. Ryckebusch ◽  
G. Laurent

1. When an isolated metathoracic ganglion of the locust was superfused with the muscarinic cholinergic agonist pilocarpine, rhythmic activity was induced in leg motor neurons. The frequency of this induced rhythm increased approximately linearly from 0 to 0.2 Hz with concentrations of pilocarpine from 10(-5) to 10(-4) M. Rhythmic activity evoked by pilocarpine could be completely and reversibly blocked by 3 x 10(-5) M atropine, but was unaffected by 10(-4) M d-tubocurarine. 2. For each hemiganglion, the observed rhythm was characterized by two main phases: a levator phase, during which the anterior coxal rotator, levators of the trochanter, flexors of the tibia, and common inhibitory motor neurons were active; and a depressor phase, during which depressors of the trochanter, extensors of the tibia, and depressors of the tarsus were active. Activity in depressors of the trochanter followed the activity of the levators of the trochanter with a short, constant interburst latency. Activity in the levator of the tarsus spanned both phases. 3. The levator phase was short compared with the period (0.5-2 s, or 10-20% of the period) and did not depend on the period. The interval between the end of a levator burst and the beginning of the following one thus increased with cycle period. The depressor phase was more variable, and was usually shorter than the interval between successive levator bursts. 4. Motor neurons in a same pool often received common discrete synaptic potentials (e.g., levators of trochanter or extensors of tibia), suggesting common drive during the rhythm. Coactive motor neurons on opposite sides (such as left trochanteral depressors and right trochanteral levators), however, did not share obvious common postsynaptic potentials. Depolarization of a pool of motor neurons during its phase of activity was generally accompanied by hyperpolarization of its antagonist(s) on the same side. 5. Rhythmic activity was generally evoked in both hemiganglia of the metathoracic ganglion, but the intrinsic frequencies of the rhythms on the left and right were usually different. The activity of the levators of the trochanter on one side, however, was strongly coupled to that of the depressors of the trochanter on the other side. 6. The locomotory rhythm was weakly coupled to the ventilatory rhythm such that trochanteral levator activity on either side never occurred during the phase of spiracle opener activity corresponding to inspiration. 7. The rhythmic activity observed in vitro bears many similarities to patterns of neural and myographic activity recorded during walking. The similarities and differences are discussed.

1994 ◽  
Vol 72 (6) ◽  
pp. 2771-2785 ◽  
Author(s):  
S. Ryckebusch ◽  
G. Laurent

1. Rhythmic activity of leg motor neurons could be evoked in isolated locust thoracic ganglia as well as in preparations of two or three connected thoracic ganglia superfused with the muscarinic agonist pilocarpine. Rhythms were always more regular and reliably elicited in single isolated ganglia. When the ganglia were connected, rhythmic activity of leg motor neurons was not usually simultaneously evoked in all six hemiganglia. Typically, some of the hemiganglia were rhythmically active, whereas others showed tonic or highly irregular activity. 2. Action potentials from leg motor neuron pools were recorded extracellularly from motor nerves and cross-correlated with the use of standard algorithms. The following correlations were observed between activities of motor neurons in different hemisegments. 1) Within a segment, trochanteral levators were coactive with contralateral trochanteral depressors. This correlation was strong in the metathoracic ganglion, and weaker in the pro- and mesothoracic ganglia. 2) Coupling between levators on opposite sides of the same segment was variable in the pro- and mesothoracic ganglia, because phase relationships between levators were different in each preparation and could also change during the course of an experiment. In the metathoracic ganglion, levators on opposite sides were never coactive. 3) Trochanteral levators were often active within a short latency of levator bursts in an ipsilateral adjacent hemiganglion. In addition, levators in one segment were often inhibited during levators bursts in the ipsilateral adjacent segment. 4) Trochanteral levators were strongly coupled to ipsilateral adjacent trochanteral depressors, for all three thoracic ganglia. 3. The phase relationships between motor neuron activities revealed by cross-correlation are discussed in the context of what is known about the mechanisms involved in the control of intersegmental coupling during legged locomotion.


1998 ◽  
Vol 79 (3) ◽  
pp. 1396-1408 ◽  
Author(s):  
Stefan Clemens ◽  
Denis Combes ◽  
Pierre Meyrand ◽  
John Simmers

Clemens, Stefan, Denis Combes, Pierre Meyrand, and John Simmers. Long-term expression of two interacting motor pattern-generating networks in the stomatogastric system of freely behaving lobster. J. Neurophysiol. 79: 1396–1408, 1998. Rhythmic movements of the gastric mill and pyloric regions of the crustacean foregut are controlled by two stomatogastric neuronal networks that have been intensively studied in vitro. By using electromyographic recordings from the European lobster, Homarus gammarus, we have monitored simultaneously the motor activity of pyloric and gastric mill muscles for ≤3 mo in intact and freely behaving animals. Both pyloric and gastric mill networks are almost continuously active in vivo regardless of the presence of food. In unfed resting animals kept under “natural-like” conditions, the pyloric network expresses the typical triphasic pattern seen in vitro but at considerably slower cycle periods (2.5–3.5 s instead of 1–1.5 s). Gastric mill activity occurs at mean cycle periods of 20–50 s compared with 5–10 s in vitro but may suddenly stop for up to tens of minutes, then restart without any apparent behavioral reason. When conjointly active, the two networks express a strict coupling that involves certain but not all motor neurons of the pyloric network. The posterior pyloric constrictor muscles, innervated by a total of 8 pyloric (PY) motor neurons, are influenced by the onset of each gastric mill medial gastric/lateral gastric(MG/LG) neuron powerstroke burst, and for one cycle, PY neuron bursts may attain >300% of their mean duration. However, the duration of activity in the lateral pyloric constrictor muscle, innervated by the unique lateral pyloric (LP) motor neuron, remains unaffected by this perturbation. During this period after gastric perturbation, LP neuron and PY neurons thus express opposite burst-to-period relationships in that LP neuron burst duration is independent of the ongoing cycle period, whereas PY neuron burst duration changes with period length. In vitro the same type of gastro-pyloric interaction is observed, indicating that it is not dependent on sensory inputs. Moreover, this interaction is intrinsic to the stomatogastric ganglion itself because the relationship between the two networks persists after suppression of descending inputs to the ganglion. Intracellular recordings reveal that thisgastro-pyloric interaction originates from the gastric MG and LG neurons of the gastric network, which inhibit the pyloric pacemaker ensemble. As a consequence, the pyloric PY neurons, which are inhibited by the pyloric dilator (PD) neurons of the pyloric pacemaker group, extend their activity during the time that PD neuron is held silent. Moreover, there is evidence for a pyloro-gastric interaction, apparently rectifying, from the pyloric pacemakers back to the gastric MG/LG neuron group.


1994 ◽  
Vol 72 (4) ◽  
pp. 1622-1633 ◽  
Author(s):  
D. Cattaert ◽  
A. Araque ◽  
W. Buno ◽  
F. Clarac

1. We investigated the effects of acetylcholine (Ach) on identified motoneurons (MNs) using an in vitro preparation of the crayfish thoracic nervous system. Discontinuous current-clamp and single electrode voltage-clamp recordings from 50 MNs were performed along with micropipette pressure ejection of Ach (or agonists) close to the recording electrode. 2. Localized ejections of relatively large volumes (500–2,500 pl) of Ach (10(-2) M) or of the muscarinic agonist oxotremorine (Oxo, 10(-2)M) onto the MN neuropile region, usually (90% of the cases) induced a slow, alternating rhythmic activity in antagonistic MNs. In other cases (4 experiments), with similar deliveries of Ach or Oxo, MNs developed the ability to fire rhythmically but only when depolarized by sustained current injection. Pressure ejections of smaller volumes (50–200 pl) of Ach (10(-2)M) close to the recorded MN could give rise to a fast (1–2 s) large amplitude (< or = 20 mV) membrane depolarization (12%), a long-lasting (10 s to several minutes) and small (2–5 mV) depolarization (14%), and a combination of the two (74%). These responses appeared to involve different regions of the neurite because they changed when the drug-ejection pipette was displaced in the neuropile. Moreover, fast and long-lasting depolarizing components resulted from a direct effect of Ach onto the MNs because they persisted under tetrodotoxin (TTX, 10(-6)M) and cobalt (Co2+, 5 x 10(-3) M) superfusion. 3. Whereas the membrane resistance decreased during the fast Ach-induced depolarization, it increased during the long-lasting depolarization. The increase in membrane resistance was more pronounced at depolarized potentials more than -55 mV and involve a reduction in K+ conductance. 4. Superfusion with nicotinic and muscarinic antagonists revealed that the fast Ach-induced depolarization involved nicotinic receptors, muscarinic receptors, or both, whereas the slow depolarization was exclusively muscarinic. 5. The Ach-evoked inward currents were studied under voltage clamp. The fast nicotinic component (Inic) increased with hyperpolarizing holding potentials and decreased with depolarizing potentials, reversing at between 10 and 30 mV. The fast muscarinic current (Ifmus) displayed similar characteristics and reversed at about -10 mV. Whereas both fast components were voltage independent, the long-lasting muscarinic component (Ismus) was voltage dependent. The response grew with membrane depolarization, but when the holding potential was hyperpolarized below resting level, the response declined to disappear at about -60 mV and beyond.(ABSTRACT TRUNCATED AT 400 WORDS)


1996 ◽  
Vol 76 (5) ◽  
pp. 3178-3195 ◽  
Author(s):  
R. M. Johnston ◽  
R. B. Levine

1. Larval crawling is a bilaterally symmetrical behavior that involves an anterior moving wave of motor activity in the body wall muscles in conjunction with sequential movements of the abdominal prolegs and thoracic legs. The purpose of this study was to determine whether the larval CNS by itself and without phasic sensory feedback was capable of producing patterned activity associated with crawling. To establish the extent of similarity between the output of the isolated nerve cord and crawling, the motor activity produced in isolated larval nerve cords was compared with the motor activity from freely crawling larvae. 2. When exposed to the muscarinic receptor agonist pilocarpine (1.0 mM), isolated larval nerve cords produced long-lasting rhythmic activity in the motor neurons that supply the thoracic leg, abdominal body wall, and abdominal proleg muscles. The rhythmic activity evoked by pilocarpine was abolished reversibly and completely by bath application of the muscarinic-receptor antagonist atropine (0.01 mM) in conjunction with pilocarpine (1.0 mM), suggesting that the response was mediated by muscarinic-like acetylcholine receptors. 3. Similar to crawling in intact animals, the evoked activity in isolated nerve cords involved bilaterally symmetrical motor activity that progressed from the most posterior abdominal segment to the most anterior thoracic segment. The rhythmic activity in thoracic leg, abdominal proleg, and abdominal body wall motor neurons showed intrasegmental and intersegmental cycle-to-cycle coupling. The average cycle period for rhythmic activity in the isolated nerve cord was approximately 2.5 times slower than the cycle period for crawling in intact larvae, but not more variable. 4. Like crawling in intact animals, in isolated nerve cords, bursting activity in the dorsal body wall motor neurons occurred before activity in ventral/lateral body wall motor neurons within an abdominal segment. The evoked bursting activity recorded from the proleg nerve was superimposed on a high level of tonic activity. 5. In isolated nerve cords, bursts of activity in the thoracic leg levator/extensor motor neurons alternated with bursts of activity in the depressor/flexor motor neurons. The burst duration of the levator/extensor activity was brief and remained relatively steady as cycle period increased. The burst duration of the depressor/ flexor activity occupied the majority of an average cycle and increased as cycle period increased. The phase of both levator/extensor motor nerve activity and depressor/flexor motor nerve activity remained relatively stable over the entire range of cycle periods. The timing and patterning of thoracic leg motor neuron activity in isolated nerve cords quantitatively resembled thoracic leg motor activity in freely crawling larvae. 6. The rhythmic motor activity generated by an isolated larval nerve cord resembled a slower version of normal crawling in intact larvae. Because of the many similarities between activity induced in the isolated nerve cord and the muscle activity and movements of thoracic and abdominal segments during crawling, we concluded that central mechanisms can establish the timing and patterning of the crawling motor pattern and that crawling may reflect the output of a central pattern generating network.


1995 ◽  
Vol 198 (2) ◽  
pp. 435-456 ◽  
Author(s):  
A Büschges ◽  
J Schmitz ◽  
U Bässler

Bath application of the muscarinic agonist pilocarpine onto the deafferented stick insect thoracic nerve cord induced long-lasting rhythmic activity in leg motoneurones. Rhythmicity was induced at concentrations as low as 1x10(-4) mol l-1 pilocarpine. The most stable rhythms were reliably elicited at concentrations from 2x10(-3) mol l-1 to 5x10(-3) mol l-1. Rhythmicity could be completely abolished by application of atropine. The rhythm in antagonistic motoneurone pools of the three proximal leg joints, the subcoxal, the coxo-trochanteral (CT) and the femoro-tibial (FT), was strictly alternating. In the subcoxal motoneurones, the rhythm was characterised by the retractor burst duration being correlated with cycle period, whereas the protractor burst duration was almost independent of it. The cycle periods of the rhythms in the subcoxal and CT motoneurone pools were in a similar range for a given preparation. In contrast, the rhythm exhibited by motoneurones supplying the FT joint often had about half the duration. The pilocarpine-induced rhythm was generated independently in each hemiganglion. There was no strict intersegmental coupling, although the protractor motoneurone pools of the three thoracic ganglia tended to be active in phase. There was no stereotyped cycle-to-cycle coupling in the activities of the motoneurone pools of the subcoxal joint, the CT joint and the FT joint in an isolated mesothoracic ganglion. However, three distinct 'spontaneous, recurrent patterns' (SRPs) of motoneuronal activity were reliably generated. Within each pattern, there was strong coupling of the activity of the motoneurone pools. The SRPs resembled the motor output during step-phase transitions in walking: for example, the most often generated SRP (SRP1) was exclusively exhibited coincident with a burst of the fast depressor trochanteris motoneurone. During this burst, there was a switch from subcoxal protractor to retractor activity after a constant latency. The activity of the FT joint extensor motoneurones was strongly decreased during SRP1. SRP1 thus qualitatively resembled the motoneuronal activity during the transition from swing to stance of the middle legs in forward walking. Hence, we refer to SRPs as 'fictive step-phase transitions'. In intact, restrained animals, application of pilocarpine also induced alternating activity in antagonistic motoneurone pools supplying the proximal leg joints. However, there were marked differences from the deafferented preparation. For example, SRP1 was not generated in the latter situation. However, if the ipsilateral main leg nerve was cut, SRP1s reliably occurred. Our results on the rhythmicity in leg motoneurone pools of deafferented preparations demonstrate central coupling in the activity of the leg motoneurones that might be incorporated into the generation of locomotion in vivo.


1998 ◽  
Vol 79 (1) ◽  
pp. 361-370 ◽  
Author(s):  
Sylvie Baudoux ◽  
Carsten Duch ◽  
Oliver T. Morris

Baudoux, Sylvie, Carsten Duch, and Oliver T. Morris. Coupling of efferent neuromodulatory neurons to rhythmical leg motor activity in the locust. J. Neurophysiol. 79: 361–370, 1998. The spike activity of neuromodulatory dorsal unpaired median (DUM) neurons was analyzed during a pilocarpine-induced motor pattern in the locust. Paired intracellular recordings were made from these octopaminergic neurons during rhythmic activity in hindleg motor neurons evoked by applying pilocarpine to an isolated metathoracic ganglion. This motor pattern is characterized by two alternating phases: a levator phase, during which levator, flexor, and common inhibitor motor neurons spike, and a depressor phase, during which depressor and extensor motor neurons spike. Three different subpopulations of efferent DUM neurons could be distinguished during this rhythmical motor pattern according to their characteristic spike output. DUM 1 neurons, which in the intact animal do not innervate muscles involved in leg movements, showed no change apart from a general increase in spike frequency. DUM 3 and DUM 3,4 neurons produced the most variable activity but received frequent and sometimes pronounced hyperpolarizations that were often common to both recorded neurons. DUM 5 and DUM 3,4,5 neurons innervate muscles of the hindleg and showed rhythmical excitation leading to bursts of spikes during rhythmic activity of the motor neurons, which innervate these same muscles. Sometimes the motor output was coordinated across both sides of the ganglion so that there was alternating activity between levators of both sides. In these cases, the spikes of DUM 5 and DUM 3,4,5 neurons and the hyperpolarization of DUM 3 and DUM 3,4 neurons occurred at particular phases in the motor pattern. Our data demonstrate a central coupling of specific types of DUM neurons to a rhythmical motor pattern. Changes in the spike output of these particular efferent DUM neurons parallel changes in the motor output. The spike activity of DUM neurons thus may be controlled by the same circuits that determine the action of the motor neurons. Functional implications for real walking are discussed.


2002 ◽  
Vol 75 (6) ◽  
pp. 613 ◽  
Author(s):  
Stefano Santabarbara ◽  
Ilaria Cazzalini ◽  
Andrea Rivadossi ◽  
Flavio M. Garlaschi ◽  
Giuseppe Zucchelli ◽  
...  

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Emilia Solomon ◽  
Katie Davis-Anderson ◽  
Blake Hovde ◽  
Sofiya Micheva-Viteva ◽  
Jennifer Foster Harris ◽  
...  

Abstract Background Human induced pluripotent stem cells (iPSC) have opened new avenues for regenerative medicine. Consequently, iPSC-derived motor neurons have emerged as potentially viable therapies for spinal cord injuries and neurodegenerative disorders including Amyotrophic Lateral Sclerosis. However, direct clinical application of iPSC bears in itself the risk of tumorigenesis and other unforeseeable genetic or epigenetic abnormalities. Results Employing RNA-seq technology, we identified and characterized gene regulatory networks triggered by in vitro chemical reprogramming of iPSC into cells with the molecular features of motor neurons (MNs) whose function in vivo is to innervate effector organs. We present meta-transcriptome signatures of 5 cell types: iPSCs, neural stem cells, motor neuron progenitors, early motor neurons, and mature motor neurons. In strict response to the chemical stimuli, along the MN differentiation axis we observed temporal downregulation of tumor growth factor-β signaling pathway and consistent activation of sonic hedgehog, Wnt/β-catenin, and Notch signaling. Together with gene networks defining neuronal differentiation (neurogenin 2, microtubule-associated protein 2, Pax6, and neuropilin-1), we observed steady accumulation of motor neuron-specific regulatory genes, including Islet-1 and homeobox protein HB9. Interestingly, transcriptome profiling of the differentiation process showed that Ca2+ signaling through cAMP and LPC was downregulated during the conversion of the iPSC to neural stem cells and key regulatory gene activity of the pathway remained inhibited until later stages of motor neuron formation. Pathways shaping the neuronal development and function were well-represented in the early motor neuron cells including, neuroactive ligand-receptor interactions, axon guidance, and the cholinergic synapse formation. A notable hallmark of our in vitro motor neuron maturation in monoculture was the activation of genes encoding G-coupled muscarinic acetylcholine receptors and downregulation of the ionotropic nicotinic acetylcholine receptors expression. We observed the formation of functional neuronal networks as spontaneous oscillations in the extracellular action potentials recorded on multi-electrode array chip after 20 days of differentiation. Conclusions Detailed transcriptome profile of each developmental step from iPSC to motor neuron driven by chemical induction provides the guidelines to novel therapeutic approaches in the re-construction efforts of muscle innervation.


2020 ◽  
Vol 2020 (11) ◽  
Author(s):  
David Meltzer ◽  
Allic Sivaramakrishnan

Abstract We derive the Cutkosky rules for conformal field theories (CFTs) at weak and strong coupling. These rules give a simple, diagrammatic method to compute the double-commutator that appears in the Lorentzian inversion formula. We first revisit weakly-coupled CFTs in flat space, where the cuts are performed on Feynman diagrams. We then generalize these rules to strongly-coupled holographic CFTs, where the cuts are performed on the Witten diagrams of the dual theory. In both cases, Cutkosky rules factorize loop diagrams into on-shell sub-diagrams and generalize the standard S-matrix cutting rules. These rules are naturally formulated and derived in Lorentzian momentum space, where the double-commutator is manifestly related to the CFT optical theorem. Finally, we study the AdS cutting rules in explicit examples at tree level and one loop. In these examples, we confirm that the rules are consistent with the OPE limit and that we recover the S-matrix optical theorem in the flat space limit. The AdS cutting rules and the CFT dispersion formula together form a holographic unitarity method to reconstruct Witten diagrams from their cuts.


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