Invertebrate presynaptic inhibition and motor control.
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Animals↗
Biomedical subjects
D Cattaert
Publications and source records attributed to D Cattaert.
Cholinergic control of the walking network in the crayfish Procambarus clarkii.
The output of a neuronal network results generally from both the properties of the component neurons and their synaptic relationships. This article aims at synthesizing various results obtained on the neural network generating locomotion in vitro. In the preparation used, consisting of the last three thoracic ganglia (3-5) along with motor nerves from the 5th leg ganglion to the promotor, remotor, levator and depressor muscles, motor nerve recordings generally revealed only tonic activity in several different motoneurons (MNs). However, rhythmic activity can be obtained by the use of cholinergic agents such as the oxotremorine (Oxo) superfused in the bath (5 x 10(-5) M). If Oxo is pressure-ejected locally in the ganglion, it is possible, depending upon the locus where the drug is applied, to elicit a rhythmic activity restricted to a group of antagonistic MNs. To analyze how cholinergic agents are able to induce such rhythmic activity, very small volumes of drug (50-200 pl), were applied close to the recording electrode. Two types of depolarizing response occurred: a fast large amplitude depolarization (5-20 mV) and a long lasting (10s to several minutes) low amplitude depolarization (1-3 mV). These responses persisted in the presence of TTX and Co(2)+. The transient initial depolarization is a mixed nicotinic and muscarinic voltage-independent response during which the input resistance decreases by 20 to 40%. In contrast, the long lasting component is voltage-dependent, exclusively muscarinic and associated to a 5-10% increase of input resistance due to the closing of a K+ conductance that is active at the resting Vm, and totally suppressed at holding potentials below -70 mV. More generally, K+ currents activated at resting potential are responsible for membrane potential stability. The injection of TEA, a blocker of the K+ currents, through the recording electrode is able to unmask plateaus above a threshold depolarization. These plateaus are TTX-sensitive but persist in the presence of Ca(2)+ channel blockers. Moreover, in 10% of TEA-filled MNs a spontaneous pacemaker activity was revealed. The organization of the locomotor network is also based upon connections between MNs and INs. Within a MN pool, connections are only loosely established, appearing to consist mainly of electrical coupling. Inhibitory synaptic connections between MNs of opposite pools are mediated by chloride channels. However, the neurotransmitter involved could be either GABA or glutamate. Therefore, at the level of a given joint, a basic rhythm occurs due to both motoneuronal membrane properties and motoneuronal connectivity. However, the coordination of all MNs of an entire leg during fictive walking activity requires the involvement of INs. Based upon these data, we propose a two-stage model of the locomotor network organization: a joint motoneuronal level and a whole leg interneuronal level.
Acetylcholine↗
Cd2+ regulation of the hyperpolarization-activated current IAB in crayfish muscle.
The effects of Cd2+ on the hyperpolarization-activated K(+)-mediated current called IAB (Araque, A., and W. Buño. 1994. Journal of Neuroscience. 14:399-408.) were studied under two-electrode voltage-clamp in opener muscle fibers of the crayfish Procambarus clarkii. IAB was reversibly reduced by extracellular Cd2+ in a concentration-dependent manner, obeying the Hill equation with IC50 = 0.452 +/- 0.045 mM and a Hill coefficient of 1 (determined from the maximal chord conductance of IAB). Cd2+ decreased the IAB conductance (GAB) and shifted its voltage dependence towards hyperpolarized potentials in a similar degree, without affecting the slope of the voltage dependence. The IAB activation time constant increased, whereas the IAB deactivation time constant was not modified by Cd2+. The IAB equilibrium potential (EAB) was unmodified by Cd2+, indicating that the selective permeability of IAB channels was not altered. IAB was unaffected by intracellular Cd2+. The Cd(2+)-regulation of IAB did not depend on [K+]o, and the effects of [K+]o on IAB were unchanged by Cd2+, indicating that Cd2+ did not compete with K+. Therefore, Cd2+ probably bound to a different site to that involved in the K+ permeability pathway. We conclude that Cd2+ affected the gating of IAB channels, interfering with their opening but not with their closing mechanism. The results can be explained by a kinetic model in which the binding of Cd2+ to the IAB channels would stabilize the gating apparatus at its resting position, increasing the energy barrier for the transition from the closed to the open channel states.
Animals↗
Chloride conductance produces both presynaptic inhibition and antidromic spikes in primary afferents.
Primary afferents from a crayfish leg proprioceptor display both primary afferent depolarizations (PADs) and antidromic spikes. PADs are generated by activation of GABA receptors and produce presynaptic inhibition, while the antidromic spikes do not elicit any synaptic effect in the postsynaptic neurons. The aim of the present study was to investigate the ionic mechanisms that allow PADs to produce antidromic spikes and to test whether GABA can produce similar effects. Intracellular recordings from the sensory axon terminals within the ganglion where PAD are produced were performed. Lowering the extracellular chloride concentration resulted in an increase in PAD amplitude, which was then capable of producing antidromic spikes. Local application of GABA close to the axon terminal also resulted in production of antidromic spikes. We conclude that antidromic spikes may result from the activation of a GABA-mediated increase in chloride conductance that also produces PADs. Therefore PADs and antidromic spikes may represent two aspects of the same GABAergic inhibitory mechanism that gate sensory transmission.
Action Potentials↗
Nicotinic and muscarinic activation of motoneurons in the crayfish locomotor network.
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)
Acetylcholine↗
MOTOR NEURONES OF THE CRAYFISH WALKING SYSTEM POSSESS TEA+-REVEALED REGENERATIVE ELECTRICAL PROPERTIES
In crustaceans, some motor neurones (MNs) have been shown to be part of the central pattern generator in the stomatogastric system (Harris-Warrick et al. 1992; Moulins, 1990), the swimmeret system (Heitler, 1978) or the walking system (Chrachri and Clarac, 1990). These MNs induce changes in the central rhythm when depolarized and are conditional oscillators in the stomatogastric ganglion. Moreover, in the walking system, rhythmic activity can be triggered by muscarinic cholinergic agonists (Chrachri and Clarac, 1987). We have recently analyzed the role of muscarinic receptors in crayfish walking leg MNs (D. Cattaert and A. Araque, in preparation) and demonstrated that oxotremorine, a muscarinic agonist, evoked long-lasting depolarizing responses associated with an increased input resistance. The outward current blocked by oxotremorine is likely to be carried by K+, as is the case for the M current (IM) in vertebrates (Brown and Adams, 1980). In most neurones, K+ conductances play a principal role in maintaining the membrane potential at rest: for example, IM is active at the resting membrane potential, thus contributing to its maintenance, and the 'delayed-rectifier' (IK) assists the fast repolarization after an action potential. Some K+ conductances are Ca2+-dependent (IK,Ca) and are activated by an increase in internal Ca2+ concentration. In such cases, Ca2+ currents may result in hyperpolarization of the neurone through activation of IK,Ca. In opposition to these K+ currents, the direct effect of Na+ and Ca2+ conductances is to depolarize the neurone. For example, the persistant Na+ current (INap) that is responsible for the slow subthreshold depolarization termed slow pre-potentials (Gestrelius et al. 1983; Leung and Yim, 1991) participates in the formation of pacemaker depolarization (Barrio et al. 1991) and generates plateau-type responses in control conditions (Barrio et al. 1991; Llinas and Sugimori, 1980). Similarly Ca2+ or non-specific (Na+/Ca2+) conductances generate such events in Aplysia californica burster neurones (Adams and Benson, 1985), crustacean cardiac ganglion (Tazaki and Cooke, 1990), insect neurones (Hancox and Pitman, 1991) and crustacean stomatogastric ganglion (Kiehn and Harris-Warrick, 1992). Since crustacean MNs can participate in rhythm production, such depolarizing conductances may exist in most of them and may contribute to the long-lasting MN depolarizations and spike bursts present during locomotion.
Journal Article↗
A new configuration for voltage clamp of axons used to demonstrate nerve conduction blockade by a 2,5-disubstituted pyrrolidine.
An original voltage-clamp technique on axons from crayfish Procambarus clarkii is described in this paper. Its advantages are: a fast dissection leading to the availability of several fibers of different diameters (10-500 microns) that may contain different ion channels; and use of a double-electrode voltage clamp on a chosen fiber with good clamping characteristics (short time clamp and good space clamp, small leak conductance). Because of the absence of exogenous lipidic phase in the superfusion chamber, this technique appears particularly suited to studying how liposoluble neurotoxins affect nerve conduction. This method has been successfully applied to test the effect of a synthetic derivative (2-(1non-8enyl)-5(1non-8enyl)pyrrolidine (Pyr 9)) of ant venom alkaloids from Monomorium species on nerve conduction. We present here evidence of a strong blocking effect on inward current involved in spike conduction. The resting potential of the treated axons did not change and it appears that only the inward current was affected.
Animals↗
Electrical coupling of mechanoreceptor afferents in the crayfish: a possible mechanism for enhancement of sensory signal transmission.
1. Central electrical coupling between primary afferent axons was investigated in an in vitro preparation of the crayfish thoracic locomotor system by using intracellular recordings. 2. Intracellular injection of the dye Lucifer yellow in single afferents resulted in staining of one to three additional afferents through dye-coupling. Three-dimensional confocal imaging of dye-coupled axons revealed a large zone of close apposition that may correspond to the gap junction site. 3. A depolarization preceding the spike in one sensory terminal was shown to facilitate the excitatory postsynaptic potential occurring in postsynaptic motoneurons. Further, a spike in one afferent axon can depolarize other, electrically coupled, axons above spike threshold, resulting in an increased number of active afferents. 4. The electrical coupling occurred between sensory afferents of similar function. It may therefore serve to facilitate sensory signal transmission from functionally homologous afferents onto postsynaptic target neurons.
Action Potentials↗
Presynaptic control as a mechanism of sensory-motor integration.
In studies of central nervous system networks, it is synaptic transmission to the postsynaptic soma-dendritic membrane that has received the most attention, in particular in relation to the analysis of sensory-motor integration. Sensory transmission is gated during ongoing movements in both invertebrates and vertebrates, such that it may be depressed in one phase of a cyclic movement and facilitated in another, in order to optimize the execution of the ongoing motor task. This presynaptic modulation is not limited to sensory afferents, but also occurs in synapses of both excitatory and inhibitory premotor interneurons. The modulation can be mediated by the release of different transmitters at axo-axonal synapses, which activate different types of receptors. In addition, presynaptic sensory axons can be coupled via gap junctions, which under certain conditions may mediate a presynaptic facilitation.
Action Potentials↗
Direct evidence for presynaptic inhibitory mechanisms in crayfish sensory afferents.
1. The central control of sensory inputs from a proprioceptor [chordotonal organ (CO)] in the second joint [coxo-basipodite (CB)] of the fifth leg was studied in crayfish in vitro preparations (Fig. 1A). Simultaneous intracellular recordings from CBCO terminals (CBT) and postsynaptic motoneurons (MNs) were performed along with micropipette pressure ejection or bath application of gamma-aminobutyric acid (GABA), to study the presynaptic mechanisms at work in the CBT (Fig. 1B). 2. Two intracellular recordings were used to show that the spikes never overshoot, and that the more central the recording site within the neuropile, the smaller the spikes (Fig. 2). Only electrotonic conduction occurs, therefore, in the sensory afferents within the ganglion. 3. Pressure ejection of GABA close to the recording site of CBTs in the ganglion (Fig. 3A) gave rise to a membrane depolarization, the reversal potential of which was about -25 mV (Fig. 7), as well as to an increase in the membrane conductance (Fig. 3C) and a decrease in the orthodromic spike amplitude; moreover, it did not elicit either hyperpolarization, or any change in the membrane conductance of the postsynaptic MN (Fig. 3B), which indicates that pressure ejection of GABA affected only a restricted area around the CBT and not the postsynaptic MNs. 4. In CBT, spontaneous primary afferent depolarizations (PADs) occurred irregularly when the activity of the preparation was not rhythmic (Fig. 4A), and in bursts when the preparation displayed fictive locomotion (Fig. 4B). In the latter case, antidromic spikes were sometimes superimposed on PADs (Fig. 4D). The amplitude of the PADs was reduced when picrotoxin (PTX), a GABA antagonist, was applied (Fig. 5), which suggests that GABA may be involved in spontaneous PADs. The reversal potential of PADs was about -25 mV (Figs. 6 and 7). 5. During simultaneous recordings from a CBT and a monosynaptically related MN, GABA applied by pressure ejection close to the CBT (Fig. 8A) completely suppressed the excitatory postsynaptic potentials (EPSPs) elicited by CBT spikes in the MN (Fig. 8, B and D). This was due to a presynaptic mechanism because no change in the membrane potential or membrane conductance was observed in the MN (Fig. 8C) and most of the CBTs associated with a given MN were affected (Fig. 9). 6. Simultaneously recording from a CBT and a monosynaptically related MN demonstrated that, during bouts of PADs, the spike amplitude decreased in proportion to the PAD amplitude (Fig. 10A).(ABSTRACT TRUNCATED AT 400 WORDS)
Animals↗
Central control of the sensory afferent terminals from a leg chordotonal organ in crayfish in vitro preparation.
In an in vitro preparation of the crayfish thoracic locomotor system, intracellular recordings have been performed from terminals of a leg joint coxo-basipodite chordotonal organ (CB). In the terminals, some depolarizing events are observed which display all the characteristics of the primary afferent depolarizations (PADs). PADs reduce the amplitude of orthodromic sensory spikes, and thus correspond to a presynaptic inhibition. PADs are tonic in a tonic preparation, and phasic (phase locked) in a rhythmic preparation. A control of the incoming information from the CB could thus be performed by the central nervous system during fictive locomotion.
Action Potentials↗
Dopamine and motor activity in the lobster Homarus gammarus.
Motor activity similar to agonistic behaviour is obtained after dopamine (DA) injection in lobster. Specially vigorous swimmeret beatings are observed and can be compared to the 'in vitro' motor activity elicited by DA superfusion of the isolated abdominal nervous system. DA-immunoreactive neurons stained by monoclonal antibodies in abdominal ganglia may be involved in swimmeret activation during the agonistic behavior.
Animals↗
Influence of walking on swimmeret beating in the lobster Homarus gammarus.
Influence of walking on swimmeret beating in intact lobsters, Homarus gammarus, has been analyzed using a treadmill experimental device. Belt movement activates both leg stepping and swimmeret beating. The simultaneity of the onset of the two motor systems in this situation is demonstrated to be the result of a startle response initiated when the belt begins to move. This reaction consists of a non-specific motor activity involving several antagonist postural and dynamic muscles. Abdominal extension and vigorous swimmeret beating are the main features of this reaction. The main characteristics of the swimmeret beating as defined by Davis (1969) has been observed here in sequences without walking. However during long walking sequences a very different swimmeret beating pattern occurs. It is suggested that this slow swimmeret beating is completely subordinate to the walking rhythm during sequences of absolute coordination. In more rapid swimmeret beating a relative coordination with leg stepping is very common. The functional meaning of this linkage between legs and swimmerets is discussed.
Animals↗
Functional multimodality of axonal tree in invertebrate neurons.
This review, based on invertebrate neuron examples, aims at highlighting the functional consequences of axonal tree organization. The axonal organization of invertebrate neurons is very complex both morphologically and physiologically. The first part shows how the transfer of information along sensory axons is modified by presynaptic inhibition mechanisms. In primary afferents, presynaptic inhibition is involved in: 1) increasing the dynamic range of the sensory response; 2) processing the sensory information such as increasing spatial and/or temporal selectivity; 3) discriminating environmental information from sensory activities generated by the animal's own movement; and 4) modulating the gain of negative feedback (resistance reflex) during active rhythmic movements such as locomotion. In a second part, the whole organization of other types of neurons is considered, and evidence is given that a neuron may not work as a unit, but rather as a mosaic of disconnected 'integrate-and-fire' units. Examples of invertebrate neurons are presented in which several spike initiating zones exist, such as in some stomatogastric neurons. The separation of a neuron into two functionally distinct entities may be almost total with distinct arborizations existing in different ganglia. However, this functional separation is not definitive and depends on the state of the neuron. In conclusion, the classical integrate-and-fire representation of the neuron, with its dendritic arborization, its spike initiating zone, its axon and axonal tree seems to be no more applicable to invertebrate neurons. A better knowledge of the function of vertebrate neurons would probably demonstrate that it is the case for a large number of them, as suggested by the complex architecture of some reticular interneurons in vertebrates.
Animals↗
Presynaptic inhibition and antidromic discharges in crayfish primary afferents.
The mechanisms of presynaptic inhibition have been studied in sensory afferents of a stretch receptor in an in vitro preparation of the crayfish. Axon terminals of these sensory afferents display primary afferent depolarisations (PADs) mediated by the activation of GABA receptors that open chloride channels. Intracellular labeling of sensory axons by Lucifer yellow combined with GABA immunohistochemistry revealed the presence of close appositions between GABA-immunoreactive boutons and sensory axons close to their first branching point within the ganglion. Electrophysiological studies showed that GABA inputs mediating PADs appear to occur around the first axonal branching point, which corresponds to the area of transition between active and passive propagation of spikes. Moreover, this study demonstrated that whilst shunting appeared to be the sole mechanism involved during small amplitude PADs, sodium channel inactivation occurred with larger amplitude PADs. However, when the largest PADs (>25 mV) are produced, the threshold for spike generation is reached and antidromic action potentials are elicited. The mechanisms involved in the initiation of antidromic discharges were analyzed by combining electrophysiological and simulation studies. Three mechanisms act together to ensure that PAD-mediated spikes are not conveyed distally: 1) the lack of active propagation in distal regions of the sensory axons; 2) the inactivation of the sodium channels around the site where PADs are produced; and 3) a massive shunting through the opening of chloride channels associated with the activation of GABA receptors. The centrally generated spikes are, however, conveyed antidromically in the sensory nerve up to the proprioceptive organ, where they inhibit the activity of the sensory neurons for several hundreds of milliseconds.
Animals↗