Some effects of proctolin on the cardiac ganglion of the Maine Lobster, Homarus americanus (Milne Edwards)

The neuropeptide proctolin has excitatory effects on the isolated lobster cardiac ganglion. Selective application to the anterior cell body region produces a dose-dependent (10(-8)--10(-5) M) prolonged depolarization of large anterior cells as well as marked increases in burst frequency and/or duration. In ganglia which have been silenced with tetrodotoxin, proctolin application to anterior cells elicits long-lasting depolarizing responses which are accompanied by a 10-30% increase of the apparent membrane input resistance. Higher proctolin concentrations produce high-frequency trains of driver potentials. It is proposed that a proctolin like peptide may serve a neurohumoral role in the lobster cardiac ganglion and that the anterior motor neurons exhibit endogenous rhythmicity in its presence.     

Functional Organization of the Cardiac Ganglion of the Lobster, Homarus americanus

External recording and stimulation, techniques were used to determine which neurons and interactions are essential for production of the periodic burst discharge in the lobster cardiac ganglion. Burst activity can be modulated by brief single shocks applied to the four small cells, but not by similar stimulation of the five large cells, suggesting that normally one or more small cells primarily determine burst rate and duration. Repetitive electrical stimulation of large cells initiates spike activity in small cells, probably via excitatory synaptic and/or electrotonic connections which may normally act to prolong bursts and decrease burst rate. Transection of the ganglion can result in burst activity in small cells in the partial or complete absence of large cell spike activity, but large cells isolated from small cell excitatory synaptic input by transection or by application of dinitrophenol do not burst. Generally, transections which decrease excitatory feedback to small cells are accompanied by an increase in burst rate, but mean spike frequency over an entire burst cycle stabilizes at the original level within 10–30 min for various groups of cells whose spike-initiating sites are still intact. These and previous results suggest that the system is two layered: one or more small cells generate the burst pattern and impose it on the large cells which are the system's motorneurons.     

Endogenous burst capability in a neuron of the gastric mill pattern generator of the spiny lobster Panulirus interruptus

The gastric system of the lobster stomatogastric ganglion has previously been thought to include no neurons capable of endogenous bursting. We describe conditions under which one of the motorneurons, the CP cell, can burst endogenously in a free-running manner in the absence of other phasic network activity. Isolated preparations of the foregut nervous system were used, and the CP bursting was either spontaneous or was activated by continuous stimulation of an input nerve. Three criteria were applied to establish the endogenous nature of such burst generation in CP: absence of phasic input, reset of the bursting pattern by pulses of current in a characteristic phase-dependent manner, and modulation of burst rate by sustained injected current. (1) The firing of other cells which are known to be related synaptically to CP was monitored in nerve records. These other cells were either silent or fired only tonically. Cross-correlograms showed that CP bursting was not ascribable to phasic activity in these other network cells. (2) A depolarizing current pulse of sufficient strength injected intracellularly between bursts triggered a burst prematurely and reset the subsequent rhythm. A hyperpolarizing pulse during a burst terminated it and reset the subsequent rhythm. Reset behavior was similar to that described for other endogenous bursters. (3) Application of a positive-going ramp current initially caused an increase in burst rate, as described for other endogenous bursters. However, further depolarization caused a slower burst rate due to lengthening of the individual bursts, although mean firing frequency continued to increase throughout the range tested. Such free-running endogenous repetitive bursting appeared to result from the CP's ability to produce slow regenerative depolarizations (“plateau potentials”). When bursting was present, so was the plateau property, as determined by I–V analysis and by the ability of brief current pulses to trigger and terminate bursts. The previous inability to observe endogenous bursting in preparations with central input removed may be due to the usual absence of the plateau property in such preparations.     

A Relaxation Oscillator Description of the Burst-Generating Mechanism in the Cardiac Ganglion of the Lobster,Homarus americanus

Properties of the neural mechanism responsible for generating the periodic burst of spike potentials in the nine ganglion neurons were investigated by applying brief, single shocks to the four small cells with extracellular electrodes placed near the trigger zones of the small cells. The shock elicited a burst if presented during the latter portion of the silent period, terminated a burst during the latter portion of the burst period, and was followed by a newly initiated burst during the early portion of the burst period. The resultant changes in burst and silent period durations were quantitatively described by a second-order non-linear differential equation similar to the van der Pol equation for a relaxation oscillator. The equation also qualitatively described changes in firing threshold of the small cells during the burst cycle. The first derivative of the solution to the equation is similar to slow transmembrane potentials in neurons that are involved in generation of burst activity in other crustacean cardiac ganglia.     

Cholinergic activation of the lobster cardiac ganglion

The frequency of rhythmic burst activity of the isolated lobster cardiac ganglion is increased by exogenously applied acetylcholine and muscarinic agonists. Responses of individual motor neurons isolated from the ganglion by transection consist of a slow depolarization and repetitive bursting. The pharmacological profile of the receptors mediating this response is similar to that of vertebrate neuronal muscarinic receptors. Isolated ganglia incubated in the presence of [3H]-choline (18-19 h) exhibited radiolabelled acetylcholine accumulation. It is suggested that ganglionic excitation may be accomplished by extrinsic or intrinsic activation of muscarinic receptors on the motor neurons.     

Synchronized firing among retinal ganglion cells signals motion reversal

We show that when a moving object suddenly reverses direction, there is a brief, synchronous burst of firing within a population of retinal ganglion cells. This burst can be driven by either the leading or trailing edge of the object. The latency is constant for movement at different speeds, objects of different size, and bright versus dark contrasts. The same ganglion cells that signal a motion reversal also respond to smooth motion. We show that the brain can build a pure reversal detector using only a linear filter that reads out synchrony from a group of ganglion cells. These results indicate that not only can the retina anticipate the location of a smoothly moving object, but that it can also signal violations in its own prediction. We show that the reversal response cannot be explained by models of the classical receptive field and suggest that nonlinear receptive field subunits may be responsible.     

Monoamine pharmacology of the lobster cardiac ganglion

Monoamine agonists and antagonists were applied to the lobster cardiac ganglion in an attempt to clarify the different actions of 5-hydroxytryptamine (5HT) and dopamine (DA) on this rhythmic pattern generator. Experiments were designed to determine whether the similar responses to 5HT and DA applied to the anterior region of the ganglion could be separated by pharmacological approaches, and whether the different responses to 5HT applied to the anterior and posterior regions of the ganglion could be attributed to mediation by different receptors. A small number of the 5HT agonists which were tested mimic the effects of 5HT, in that they increase the frequency of bursting and decrease burst duration when applied to the whole ganglion, but decrease burst frequency and increase burst duration when applied only to the posterior half. Other 5HT agonists decrease frequency and prolong bursts when applied to the whole ganglion. Of the DA agonists tested, none acts as DA itself does. Rather, they mimic the effects of 5HT applied to the posterior ganglion, by slowing bursting and prolonging bursts. The actions of agonists do not correspond in any clear way to the receptor specificities as defined in vertebrates. Most antagonists tested do not show similar specificities to their effects in vertebrates. In particular, most of the DA antagonists tested are more effective in blocking exogenous 5HT than DA. One monoamine agonist directly alters the properties of endogenous burst-organizing potentials (driver potentials) in the motorneurons of the ganglion.     

Studies on re-entrant arrhythmias and ectopic beats in excitable tissues by bifurcation analyses.

A phase-plane bifurcation analysis is a useful way to theoretically understand how various types of arrhythmias may arise from excitable tissues. In this paper, we have performed phase-plane bifurcation analysis to characterize arrhythmogenic states in excitable tissues. To achieve this, we have first formulated a model which is simple enough to be mathematically tractable, yet captures the non-linear features of cardiac excitation and conduction. In this model, single cells are connected in a circular fashion by gap conductances. Each cell carries the following two types of currents: a passive outward current and an inward "excitable" current which contains an activation and an inactivation gate. The activation gate is responsible for the upstroke of action potential and inactivation gate is responsible for the termination of the plateau potential. With this model, we have constructed bifurcation diagrams as a function of a bifurcation parameter. The parameter chosen as the bifurcation parameter has the property of raising maximum diastolic potential while shorting the refractory period. Our analysis revealed the existence of three distinct multi-stable phases in certain ranges of the bifurcation parameter: (1) bistability between a rotor and a quiescent state, (2) bistability between rotor and ectopic beats, and (3) three stable states co-existing among quiescent state, rotor, and ectopic beats. In these three regions, external impulses exert very distinct effects: In region 1, a brief current pulse can annihilate a re-entrant arrhythmia to quiescence. To initiate re-entry from a quiescent tissue, however, it takes two pulses (a primary pulse followed by a premature pulse at a site different from the "primary" site). In region 2, a brief pulse can convert a re-entrant arrhythmia to ectopic beats. To convert the ectopic beats back to circus movement, these beats have to be suppressed by a few brief current pulses to initiate one-way propagation. Depending on the frequency and strength of impulses in region 3, the tissue can switch back and forth among quiescence, circus movement, and ectopic beats. For comparison, we have also included a more complete Beeler-Reuter cardiac cell model in our analysis and obtained essentially the same results. From the behavioral similarities of these models, we conclude that re-entrant and ectopic arrhythmias must be intrinsic properties of excitable tissues and external stimuli can convert one mode of arrhythmia to another in the multistability regions.(ABSTRACT TRUNCATED AT 400 WORDS)     

Identification of two cardioacceleratory neurons in the isopod crustacean, Ligia exotica and their effects on cardiac ganglion cells

We identified two pairs of cardioacceleratory (CA1, CA2) neurons in the central nervous system of the isopod Ligiaexotica and examined their effects on the cardiac ganglion (CG). CA1 neurons had cell bodies in the 2nd thoracic ganglion and had arborizations in the subesophageal ganglion and the 1st and 2nd thoracic ganglia. CA2 neurons had cell bodies in the 3rd thoracic ganglion and had arborizations in the 2nd, 3rd and 4th thoracic ganglia. They sent axons to the heart through the ipsilateral 3rd roots of the ganglia where their cell bodies were located. Repetitive stimulation of the CA1 axon rapidly increased the burst frequency of the CG, and that of CA2 rather slowly. The increased burst rate caused by the CA1 stimulation was significantly higher than that caused by CA2. Overall depolarization of a quiescent CG cell produced by the CA1 stimulation was significantly larger in amplitude than that produced by CA2. Facilitation was obviously seen in the excitatory post-synaptic potentials evoked by the CA1 stimulation. These results show that the synaptic properties of CA1 and CA2 neurons are different, suggesting that they have different functional roles in heart regulation. Accepted: 19 July 1997     

Synchronous Bursting Can Arise from Mutual Excitation, Even When Individual Cells are not Endogenous Bursters

Mutual excitation between two neurons is generally thought to raise the excitation level of each neuron or, if they are both bursty, to act to synchronize their bursts. If only one is bursty, it can induce synchronized bursts in the other cell. Here we show that two nonbursty cells can be induced to burst in synchrony by mutual excitatory synaptic connections, provided the presynaptic threshold for graded synaptic transmission ateach synapse is at a different level. This mechanism may operate in a recently discovered network in the lobster Homarus gammarus.By a duality between presynaptic threshold and injected current, we also show that two identical, nonbursty, mutual excitatory cells could be induced to burst in synchrony by injecting differing amounts of current inthe two cells. Finally we show that differential oscillations betweentwo mutual excitatory cells could be stopped by a slow-tailedhyperpolarizing current pulse into one cell or a slow-taileddepolarizing pulse into the other.     

Neurohormonal alteration of integrative properties of the cardiac ganglion of the lobster Homarus americanus.

The spontaneous burst discharges of isolated lobster (Homarus americanus) cardiac ganglia were recorded with a spaced array of electrodes. Small regions (less than 1 mm) of the ganglion were exposed to the cardioexcitor neurohormone in extracts of pericardial organs (XPO) or to 10(-5) M 5-hydroxytryptamine (5HT). All axons were excited (increased mean firing frequency, f) by both substances, but only by applications in the region between the soma (but excluding it) and proximal site of impulse initiation. Units not so exposed changed their f relatively little despite f increases of as much as threefold in exposed units and changes in burst rate and overall length. Regularity and grouping of all impulse activity into bursts was never disturbed. 5HT increases burst rate at any point of application. The increases are larger if small cells are affected than if only large cells are exposed. Burst length decreases except when the pacemaker is affected. In contrast, XPO affects neither burst rate or length unless small cells are affected. Length is increased if non-pacemaker small cells are affected; both rate and length increase if the pacemaker is affected. The pacemaker usually exhibits an f of intermediate value. Rate changes are not simply related to its f. A small cell can "burst" in the absence of impulses from any other cells. XPO may enhance endogenous "driver potentials," while 5HT may excite by depolarizing at limited sites.     

Excitation-contraction coupling in cardiac muscle of lobster (Homarus americanus); the role of the sarcolemma and sarcoplasmic reticulum

The T-tubules and sarcoplasmic reticulum (SR) serving excitation-contraction (EC) coupling in lobster (Homarus americanus) cardiac muscle are similar to those in mammalian myocardium. Tetanic contraction is elicited by a burst of action potentials from the cardiac ganglion. In this study we evaluated the roles of the sarcolemma and SR in EC coupling of the ostial valve muscle (orbicularis ostii m. or OOM) of lobster heart. The OOM was mounted in a bath with saline on a microscope stage; force was measured by strain gauge. [Ca2+]i was measured using iontophoretically micro-injected fura-2 salt. Peak [Ca+]i, peak tetanic force and time to peak [Ca2+]i increased with that of stimulus train duration (TD), to a maximum at a TD of 500 ms. Force increased with [Ca2+]. Cd2+ reduced force by 90%; ryanodine and caffeine reduced tetanic [Ca2+]i transients by 80% and 70%, and force by 90% and 80%, respectively. Ryanodine, caffeine and cyclopiazonic acid slowed the decline of [Ca2+]i and force during relaxation. Relaxation required [Na+]o. The rate of decline of [Ca2+]i appeared to be a sigmoidal function of the [Ca2+]i and increased for any [Ca2+]i with TD. Inactivity slowed relaxation of force; stimulation accelerated relaxation. These data suggest important contributions of Ca2+ transport both across the sarcolemma and across the SR membrane during EC-coupling of lobster cardiac muscle, while average cytosolic [Ca2+]i regulates the rate of [Ca2+]i elimination during relaxation.     

Bursting as an Effective Relay Mode in a Minimal Thalamic Model

In recent years, accumulating evidence indicates that thalamic bursts are present during wakefulness and participate in information transmission as an effective relay mode with distinctive properties from the tonic activity. Thalamic bursts originate from activation of the low threshold calcium cannels via a local feedback inhibition, exerted by the thalamic reticular neurons upon the relay neurons. This article, examines if this simple mechanism is sufficient to explain the distinctive properties of thalamic bursting as an effective relay mode. A minimal model of thalamic circuit composed of a retinal spike train, a relay neuron and a reticular neuron is simulated to generate the tonic and burst firing modes. The integrate-and-fire-or-burst model is used to simulate the neurons. After discriminating the burst events with criteria based on inter-spike-intervals, statistical indices show that the bursts of the minimal model are stereotypic events. The relation between the rate of bursts and the parameters of the input spike train demonstrates marked nonlinearities. Burst response is shown to be selective to spike-silence-spike sequences in the input spike train. Moreover, burst events represent the input more reliably than the tonic spike in a considerable range of the parameters of the model. In conclusion, many of the distinctive properties of thalamic bursts such as stereotypy, nonlinear dependence on the sensory stimulus, feature selectivity and reliability are reproducible in the minimal model. Furthermore, the minimal model predicts that while the bursts are more frequent in the spike train of the off-center X relay neurons (corresponding to off-center X retinal ganglion cells), they are more reliable when generated by the on-center ones (corresponding to on-center X ganglion cells).     

Nonlinear dynamics of paleocortex manifested in the olfactory EEG

The olfactory bulb is the first central component in a highly sensitive yet markedly stable sensory system. It receives a surge of receptor activity with each inspiration and transmits output as a brief burst of oscillatory activity that is most clearly seen in the EEG. These properties together with the known anatomy and physiology of the bulb are used as design criteria to synthesize, evaluate and solve a set of nonlinear differential equations that represent lumped bulbar dynamics. According to the model bulbar processing is in two stages. In the outer layers the interneurons perform the operations of input range compression, integration, clipping, holding, and bias control. In the inner layers the input surge is converted to a burst, which is transmitted by the mitral cells as a pulse density wave. The phase, frequency duration and amplitude of the wave convey information centrally about both the input and the state of the system. The model suffices to replicate the forms of the EEG burst; the pulse probability distributions conditional on the EEG; the waveforms of averaged evoked potentials (AEPs) and post stimulus time (PST) histograms from the bulb and cortex; and the changes in waveform induced by behavioral control of attentiveness and habituation. It is inferred that with selective attention there is a permanent change in the strength of mutually excitatory connections among excitatory neurons, and that with habituation there is a reversible change in the effectiveness of excitatory synapses. The limitations and deficiencies of the model and the need for centrifugal controls of bulbocortical function are discussed.     

Dual effects of dopamine on the adult heart of the isopod crustacean Ligia exotica

In the adult heart of the isopod crustacean Ligia exotica, the cardiac ganglion acts as the primary pacemaker with the myocardium having a latent pacemaker property. We show several lines of evidence that dopamine modulates the heartbeat of adult L. exotica affecting both pacemaker sites in the heart. Dopamine caused positive chronotropic (frequency increase) and inotropic (amplitude increase) effects on the heartbeat in a concentration dependent manner. The time courses of these effects were considerably different and the inotropic effect appeared later and lasted longer than the chronotropic effect. Dopamine rapidly increased the frequency of the bursting activity in the cardiac ganglion neurons and each impulse burst of the cardiac ganglion was always followed by a heartbeat. Moreover, dopamine slowly increased the amplitude and duration of the action potential plateau (plateau potential) of the myocardium. When the myocardial pacemaker activity was induced by application of tetrodotoxin, which suppresses cardiac ganglion activity, dopamine slowly increased the amplitude and duration of the myocardial plateau potential while decreasing its frequency. These results suggest that dopamine modulates the heartbeat in adult L. exotica producing a dual effect on the two pacemaker sites in the heart, the cardiac ganglion and myocardium.     

Effects of haloperidol and phentolamine on the crustacean cardiac ganglion

Haloperidol (a dopamine D₂ blocker in vertebrates) and phentolamine (an α-adrenergic blocker) alter the pattern of bursting by the isolated cardiac ganglion of the lobster when perfused at concentrations of 10^?6–10^?5 mol/l. Both drugs decrease the frequency of bursting and increase burst duration. They are most effective in slowing the ganglion when applied selectively to the anterior ganglionic trunk, the same region of the ganglion where dopamine (DA) and 5-hydroxytryptamine (5HT) are most effective in speeding up bursting. When exogenous monoamine transmitters are applied in the presence of 3×10^?6 mol/l haloperidol, the effect of 5HT, but not of DA, is significantly reduced. At the same concentration, phentolamine does not suppress the actions of DA, 5HT or noradrenaline (NA). Both haloperidol and phentolamine significantly alter the properties of endogenous burst-organizing potentials (driver potentials) generated by motorneurons in the ganglion. It is possible that the effects of these drugs on bursting reflect alteration of endogenous electrical properties of the constituent neurons, rather than receptor antagonism.     

Activation of command fibres to the stomatogastric ganglion by input from a gastric mill proprioceptor in the crab,Cancer pagurus †

In semi‐intact preparations of the crab Cancer pagurus the normal output from the stomatogastric ganglion (StG) was a regular pyloric cycle (Figure 4). Repeated stimulation of the posterior stomach nerve (psn) of the posterior gastric mill proprioceptor (PSR) often induced series of spontaneous gastric cycles. We were therefore able to describe the normal gastric cycle as recorded in the output nerves from StG and to identify most of the relevant motor neurones by reference to the muscles that they innervate (Figure 10). The gastric cycle output was variable (Figures 5, 6), although in many preparations one complex type of output predominated (Figure 7). The basic feature of the gastric cycle was an alternation of activity between the single cardio‐pyloric neurone (CP) and a complex variable burst in the lateral cardiac (LC), the gastro‐pyloric (GP), the gastric (GM), and other associated neurones. During this normally occurring complex gastric burst significant changes occurred in the pyloric cycle, notably an increase in activity of the pacemaker pyloric dilator (PD) group and an inhibition of the lateral pyloric (LP), inferior cardiac (IC) and ventricular dilator (VD) neurones (Figures 6, 7, 8, 9). These changes are probably associated with an opening of the cardio‐pyloric valve and food passage into the pyloric filter. The gastric output was related to the normally observed movements of the dorsal ossicles of the gastric mill and thus to the operation of the teeth of the mill (Figure 11). Increased input from the PSR is associated with the grinding action of the teeth which is caused by the complex gastric burst (Figure 12).

Stimulation of the psn during an ongoing regular pyloric output caused changes in the cycle which mimicked those occurring during the spontaneous gastric cycle (Figure 13; Table 1). Stimulation of the psn during ongoing gastric activity also affected the gastric units (Figure 14). The input pathway from the PSR is shown to be through the stomatogastric nerve (sgn), the connection between the commissural ganglia and the stomatogastric ganglion (Figure 15). The commissural ganglia are known to receive most of the sensory input from the foregut and PSR input is probably processed there. Recordings from the sgn show that psn stimulation activates a small number of centrally originating units, and that the activity of these units coincides with the pyloric output changes (Figures 15, 16). We therefore label the units command interneurones. Their effects could be mediated by direct connections to only the PD pacemaker neurones of the pyloric cycle. Control experiments showed that PSR input is not necessary for the pyloric output changes to occur during gastric output but that similar output changes can be evoked by input resulting from induced gastric movements (Figure 15(E)). We think that the pyloric cycle output changes are normally controlled by a number of mechanisms at different levels (Figure 17). We cannot easily explain the effects of PSR input on the gastric cycle neurones.

These findings are important because they allow us to study a specific input to the StG without disrupting its normal input‐output pathways to the central nervous system. Further experiments on the system designed to test the assumption that the sgn units are in fact responsible for the pyloric output changes, and to investigate the processing of the PSR input are outlined.     

The Sites of Action of Pericardial Organ Extract and 5-Hydroxytryptamine in the Decapod Crustacean Heart

Responses of isolated, perfused hearts of Homarm americanusto brief, internal application of extracts of pericardial organs(PO's) ofCancer borealis, or 5-hydroxytryptamine (5HT) are verysimilar over a thousand-told range of concentration: an increasein rate and amplitude of beating. These reach their maxima afterwashing out has begun, and recover within ten minutes. Externalapplication is ineffective and the substances do not interactwith effects of stretch stimulation. Intracellular recordingfrom heart muscle fibers reveals facilitation of depolarizationto bursts at heart beat frequencies. There may be some effectof 5HT directly on neuromuscular facilitation. Responses recordedfrom isolated cardiac ganglia show increased burst rate, burstduration, or both. Thresholds and the range of concentrationsfor which coordinated responses are recorded correspond to thosefor perfused hearts. It is concluded that the major sites ofaction of PO extract and 5HT are in the cardiac ganglion. 5HTtachyphlaxis and LSD block effects of 5HT, but not of PO extractor accelerator nerve stimulation. Intracellular recordings fromthe large ganglion cells show no effects on resting, synaptic,or spike potentials. Changes in membrane potential to currentpulses revealed no changes in membrane resistance or in theresistance of the electrotonic pathway between cells. Resultsof selective application to large or small cells suggested thatPO extract may contain a rate-increasing substance and one prolongingthe duration of bursts. The former, and 5HT, may influence pacemakerpotentials; the latter may increase the number of spikes a unitcan produce before becoming refractory.     

Circuits constructed from identified Aplysia neurons exhibit multiple patterns of persistent activity.

We have used identified neurons from the abdominal ganglion of the mollusc Aplysia to construct and analyze two circuits in vitro. Each of these circuits was capable of producing two patterns of persistent activity; that is, they had bistable output states. The output could be switched between the stable states by a brief, external input. One circuit consisted of cocultured L10 and left upper quadrant (LUQ) neurons that formed reciprocal, inhibitory connections. In one stable state L10 was active and the LUQ was quiescent, whereas in the other stable state L10 was quiescent and the LUQ was active. A second circuit consisted of co-cultured L7 and L12 neurons that formed reciprocal, excitatory connections. In this circuit, both cells were quiescent in one stable state and both cells fired continuously in the other state. Bistable output in both circuits resulted from the nonlinear firing characteristics of each neuron and the feedback between the two neurons. We explored how the stability of the neuronal output could be controlled by the background currents injected into each neuron. We observed a relatively well-defined range of currents for which bistability occurred, consistent with the values expected from the measured strengths of the connections and a simple model. Outside of the range, the output was stable in only a single state. These results suggest how stable patterns of output are produced by some in vivo circuits and how command neurons from higher neural centers may control the activity of these circuits. The criteria that guided us in forming our circuits in culture were derived from theoretical studies on the properties of certain neuronal network models (e.g., Hopfield, J. J. 1984. Proc. Natl. Acad. Sci. USA. 81:3088-3092). Our results show that circuits consisting of only two co-cultured neurons can exhibit bistable output states of the form hypothesized to occur in populations of neurons.     

Localization of GABA- and glutamate-like immunoreactivity in the cardiac ganglion of the lobster Panulirus argus

Wholemount immunohistochemical methods were used to examine the localization of γ-aminobutyric acid (GABA) and glutamate within the cardiac system of the Caribbean spiny lobster Panulirus argus. All of the GABA-like immunoreactivity (GABAi) in the cardiac ganglion originated from a single bilateral pair of fibers that entered the heart via the two dorsal nerves. Each GABAi axon bifurcated upon entering the ganglion and gave rise to varicose fibers that surrounded the somata and initial segments of the five large motor neurons. The four small posterior cells did not appear to receive somatic contacts. Double-labeling experiments in which individual motor neurons were injected with Neurobiotin showed that their dendritic processes, which project to muscle bundles adjacent to the ganglion and are thought to respond to stretch, were also accompanied by branches of the GABAi fibers. Glutamate-like immunoreactivity (GLUi) was present in each of the motor neuron cell bodies. In some preparations, GLUi was also detected in large caliber fibers in the major ganglionic nerves. These fibers gave rise to more slender branches that innervated the cardiac muscle bundles. GLUi was also found in the small cell bodies and in fibers surrounding motor neuron somata. Taken together, these findings support previous electrophysiological, pharmacological and anatomical studies indicating that GABA mediates extrinsic inhibition and that glutamate acts as a neuromuscular and intraganglionic transmitter in this system. While axosomatic contacts may play a major role in both transmitter systems, the GABAergic inhibition also appears to involve substantial axodendritic synaptic signaling.