Role of Ih in differentiating the dynamics of the gastric and pyloric neurons in the stomatogastric ganglion of the lobster, Homarus americanus.

The hyperpolarization-activated inward cationic current (Ih) is known to regulate the rhythmicity, excitability, and synaptic transmission in heart cells and many types of neurons across a variety of species, including some pyloric and gastric mill neurons in the stomatogastric ganglion (STG) in Cancer borealis and Panulirus interruptus However, little is known about the role of Ih in regulating the gastric mill dynamics and its contribution to the dynamical bifurcation of the gastric mill and pyloric networks. We investigated the role of Ih in the rhythmic activity and cellular excitability of both the gastric mill neurons (medial gastric, gastric mill) and pyloric neurons (pyloric dilator, lateral pyloric) in Homarus americanus Through testing the burst period between 5 and 50 mM CsCl, and elimination of postinhibitory rebound and voltage sag, we found that 30 mM CsCl can sufficiently block Ih in both the pyloric and gastric mill neurons. Our results show that Ih maintains the excitability of both the pyloric and gastric mill neurons. However, Ih regulates slow oscillations of the pyloric and gastric mill neurons differently. Specifically, blocking Ih diminishes the difference between the pyloric and gastric mill burst periods by increasing the pyloric burst period and decreasing the gastric mill burst period. Moreover, the phase-plane analysis shows that blocking Ih causes the trajectory of slow oscillations of the gastric mill neurons to change toward the pyloric sinusoidal-like trajectories. In addition to regulating the pyloric rhythm, we found that Ih is also essential for the gastric mill rhythms and differentially regulates these two dynamics.

Coordination of distinct but interacting rhythmic motor programs by a modulatory projection neuron using different co-transmitters in different ganglia.

While many neurons are known to contain multiple neurotransmitters, the specific roles played by each co-transmitter within a neuron are often poorly understood. Here, we investigated the roles of the co-transmitters of the pyloric suppressor (PS) neurons, which are located in the stomatogastric nervous system (STNS) of the lobster Homarus americanus. The PS neurons are known to contain histamine; using RT-PCR, we identified a second co-transmitter as the FMRFamide-like peptide crustacean myosuppressin (Crust-MS). The modulatory effects of Crust-MS application on the gastric mill and pyloric patterns, generated in the stomatogastric ganglion (STG), closely resembled those recorded following extracellular PS neuron stimulation. To determine whether histamine plays a role in mediating the effects of the PS neurons in the STG, we bath-applied histamine receptor antagonists to the ganglion. In the presence of the antagonists, the histamine response was blocked, but Crust-MS application and PS stimulation continued to modulate the gastric and pyloric patterns, suggesting that PS effects in the STG are mediated largely by Crust-MS. PS neuron stimulation also excited the oesophageal rhythm, produced in the commissural ganglia (CoGs) of the STNS. Application of histamine, but not Crust-MS, to the CoGs mimicked this effect. Histamine receptor antagonists blocked the ability of both histamine and PS stimulation to excite the oesophageal rhythm, providing strong evidence that the PS neurons use histamine in the CoGs to exert their effects. Overall, our data suggest that the PS neurons differentially utilize their co-transmitters in spatially distinct locations to coordinate the activity of three independent networks.

Short-term synaptic plasticity compensates for variability in number of motor neurons at a neuromuscular junction.

We studied how similar postsynaptic responses are maintained in the face of interindividual variability in the number of presynaptic neurons. In the stomatogastric ganglion of the lobster, Homarus americanus, the pyloric (PY) neurons exist in variable numbers across animals. We show that each individual fiber of the stomach muscles innervated by PY neurons received synaptic input from all neurons present. We performed intracellular recordings of excitatory junction potentials (EJPs) in the muscle fibers to determine the consequences of differences in the number of motor neurons. Despite the variability in neuron number, the compound electrical response of muscle fibers to natural bursting input was similar across individuals. The similarity of total synaptic activation was not due to differences in the spiking activity of individual motor neurons across animals with different numbers of PY neurons. The amplitude of a unitary EJP in response to a single spike in a single motor neuron also did not depend on the number of PY neurons present. Consequently, the compound EJP in response to a single stimulus that activated all motor axons present was larger in individuals with more PY neurons. However, when axons were stimulated with trains of pulses mimicking bursting activity, EJPs facilitated more in individuals with fewer PY neurons. After a few stimuli, this resulted in depolarizations similar to the ones in individuals with more PY neurons. We interpret our findings as evidence that compensatory or homeostatic regulatory mechanisms can act on short-term synaptic dynamics instead of absolute synaptic strength.

Mass spectrometric characterization and physiological actions of novel crustacean C-type allatostatins.

The crustacean stomatogastric ganglion (STG) is modulated by numerous neuropeptides that are released locally in the neuropil or that reach the STG as neurohormones. Using 1,5-diaminonaphthalene (DAN) as a reductive screening matrix for matrix-assisted laser desorption/ionization (MALDI) mass spectrometric profiling of disulfide bond-containing C-type allatostatin peptides followed by electrospray ionization quadrupole time-of-flight (ESI-Q-TOF) tandem mass spectrometric (MS/MS) analysis, we identified and sequenced a novel C-type allatostatin peptide (CbAST-C1), pQIRYHQCYFNPISCF-COOH, present in the pericardial organs of the crab, Cancer borealis. Another C-type allatostatin (CbAST-C2), SYWKQCAFNAVSCFamide, was discovered using the expressed sequence tag (EST) database search strategy in both C. borealis and the lobster, Homarus americanus, and further confirmed with de novo sequencing using ESI-Q-TOF tandem MS. Electrophysiological experiments demonstrated that both CbAST-C1 and CbAST-C2 inhibited the frequency of the pyloric rhythm of the STG, in a state-dependent manner. At 10(-6)M, both peptides were only modestly effective when initial frequencies of the pyloric rhythm were >0.8Hz, but almost completely suppressed the pyloric rhythm when applied to preparations with starting frequencies <0.7Hz. Surprisingly, these state-dependent actions are similar to those of the structurally unrelated allatostatin A and allatostatin B families of peptides.

Spectral analyses reveal the presence of adult-like activity in the embryonic stomatogastric motor patterns of the lobster, Homarus americanus.

The stomatogastric nervous system (STNS) of the embryonic lobster is rhythmically active prior to hatching, before the network is needed for feeding. In the adult lobster, two rhythms are typically observed: the slow gastric mill rhythm and the more rapid pyloric rhythm. In the embryo, rhythmic activity in both embryonic gastric mill and pyloric neurons occurs at a similar frequency, which is slightly slower than the adult pyloric frequency. However, embryonic motor patterns are highly irregular, making traditional burst quantification difficult. Consequently, we used spectral analysis to analyze long stretches of simultaneous recordings from muscles innervated by gastric and pyloric neurons in the embryo. This analysis revealed that embryonic gastric mill neurons intermittently produced pauses and periods of slower activity not seen in the recordings of the output from embryonic pyloric neurons. The slow activity in the embryonic gastric mill neurons increased in response to the exogenous application of Cancer borealis tachykinin-related peptide 1a (CabTRP), a modulatory peptide that appears in the inputs to the stomatogastric ganglion (STG) late in larval development. These results suggest that the STG network can express adult-like rhythmic behavior before fully differentiated adult motor patterns are observed, and that the maturation of the neuromodulatory inputs is likely to play a role in the eventual establishment of the adult motor patterns.

Understanding circuit dynamics using the stomatogastric nervous system of lobsters and crabs.

Studies of the stomatogastric nervous systems of lobsters and crabs have led to numerous insights into the cellular and circuit mechanisms that generate rhythmic motor patterns. The small number of easily identifiable neurons allowed the establishment of connectivity diagrams among the neurons of the stomatogastric ganglion. We now know that (a) neuromodulatory substances reconfigure circuit dynamics by altering synaptic strength and voltage-dependent conductances and (b) individual neurons can switch among different functional circuits. Computational and experimental studies of single-neuron and network homeostatic regulation have provided insight into compensatory mechanisms that can underlie stable network performance. Many of the observations first made using the stomatogastric nervous system can be generalized to other invertebrate and vertebrate circuits.

Central pattern generating neurons simultaneously express fast and slow rhythmic activities in the stomatogastric ganglion.

Neuronal firing patterns can contain different temporal information. It has long been known that the fast pyloric and the slower gastric motor patterns in the stomatogastric ganglion of decapod crustaceans interact. However, the bidirectional influences between the pyloric rhythm and the gastric mill rhythm have not been quantified in detail from preparations that spontaneously express both patterns in vitro. We found regular and stable spontaneous gastric and pyloric activity in 71% of preparations of the isolated stomatogastric nervous system of the lobster, Homarus americanus. The gastric [cycle period: 10.96 +/- 2.67 (SD) s] and pyloric (cycle period: 1.35 +/- 0.18 s) patterns showed bidirectional interactions and coordination. Gastric neuron firing showed preferred phases within the reference frame of the pyloric cycle. The relative timing and burst parameters of the pyloric neurons systematically changed within the reference frame of the gastric cycle. The gastric rhythm showed a tendency to run at cycle periods that were integer multiples of the pyloric periods, but coupling and coordination between the two rhythms were variable. We used power spectra to quantify the gastric and pyloric contributions to the firing pattern of each individual neuron. This provided us with a way to analyze the firing pattern of each gastric and pyloric neuron type individually without reference to either gastric or pyloric phase. Possible functional consequences of these network interactions for motor output are discussed.

Red pigment concentrating hormone strongly enhances the strength of the feedback to the pyloric rhythm oscillator but has little effect on pyloric rhythm period.

The neuropeptide, red pigment concentrating hormone (RPCH), strengthened the inhibitory synapse from the lateral pyloric (LP) neuron to the pyloric dilator (PD) neurons in the pyloric network of the stomatogastric ganglion (STG) of the lobster, Homarus americanus. RPCH produced several-fold increases in the amplitude of both action potential-mediated and non-impulse-mediated transmission that persisted for as long as the peptide remained present. Because the LP to PD synapse is the only feedback to the pacemaker kernel of the pyloric network, which consists of the electrically coupled two PD neurons and the anterior burster (AB) neuron, it might have been expected that strengthening the LP to PD synapse would increase the period of the pyloric rhythm. However, the period of the pyloric rhythm increased only transiently in RPCH, and a transient increase in cycle period was observed even when the LP neuron was hyperpolarized. Phase response curves were measured using the dynamic clamp to create artificial inhibitory inputs of variable strength and duration to the PD neurons. Synaptic conductance values seen in normal saline were ineffective at changing the pyloric period throughout the pyloric cycle. Conductances similar to those seen in 10(-6) M RPCH also did not evoke phase resets at phases when the LP neuron is typically active. Thus the dramatic effects of RPCH on synaptic strength have little role in modulation of the period of the pyloric rhythm under normal operating conditions but may help to stabilize the rhythm when the cycle period is too slow or too fast.

Animal-to-animal variability in motor pattern production in adults and during growth.

Which features of network output are well preserved during growth of the nervous system and across different preparations of the same size? To address this issue, we characterized the pyloric rhythms generated by the stomatogastric nervous systems of 99 adult and 12 juvenile lobsters (Homarus americanus). Anatomical studies of single pyloric network neurons and of the whole stomatogastric ganglion (STG) showed that the STG and its neurons grow considerably from juvenile to adult. Despite these changes in size, intracellularly recorded membrane potential waveforms of pyloric network neurons and the phase relationships in the pyloric rhythm were very similar between juvenile and adult preparations. Across adult preparations, the cycle period and number of spikes per burst were not tightly maintained, but the mean phase relationships were independent of the period of the rhythm and relatively tightly maintained across preparations. We interpret this as evidence for homeostatic regulation of network activity.

Inhibitory synchronization of bursting in biological neurons: dependence on synaptic time constant.

Using the dynamic clamp technique, we investigated the effects of varying the time constant of mutual synaptic inhibition on the synchronization of bursting biological neurons. For this purpose, we constructed artificial half-center circuits by inserting simulated reciprocal inhibitory synapses between identified neurons of the pyloric circuit in the lobster stomatogastric ganglion. With natural synaptic interactions blocked (but modulatory inputs retained), these neurons generated independent, repetitive bursts of spikes with cycle period durations of approximately 1 s. After coupling the neurons with simulated reciprocal inhibition, we selectively varied the time constant governing the rate of synaptic activation and deactivation. At time constants 400 ms), bursts became phase-locked in a fully overlapping pattern with little or no phase lag and a shorter period. During the in-phase bursting, the higher-frequency spiking activity was not synchronized. If the circuit lacked a robust periodic burster, increasing the time constant evoked a sharp transition from out-of-phase oscillations to in-phase oscillations with associated intermittent phase-jumping. When a coupled periodic burster neuron was present (on one side of the half-center circuit), the transition was more gradual. We conclude that the magnitude and stability of phase differences between mutually inhibitory neurons varies with the ratio of burst cycle period duration to synaptic time constant and that cellular bursting (whether periodic or irregular) can adopt in-phase coordination when inhibitory synaptic currents are sufficiently slow.

The localization of two voltage-gated calcium channels in the pyloric network of the lobster stomatogastric ganglion.

Voltage-gated calcium channels are critical to all aspects of nervous system function, with differing roles within the neuronal somata, at synaptic terminals, and at the neuromuscular junction. We have developed antibodies against two voltage-gated Ca(2+) channel genes from the spiny lobster, Panulirus interruptus, which are homologous to the Drosophila Ca1A (a P/Q-type channel) and Ca1D (an L-type channel) genes. Using these antibodies, we have found that each channel shows unique patterns of localization within the stomatogastric nervous system. Both antibodies stain somata of most of the neurons in the pyloric network to varying degrees. The high degree of variability in staining intensity within individual pyloric cell classes supports the hypothesis of Golowasch et al. (1999a,b) that individual cells can vary in their composition of ionic currents and still have similar firing properties. Anti-Ca1A stains structures in the neuropil, some of which are terminals of axons descending from higher ganglia; however, the majority of these are neither neurites nor blood vessels, but may instead be glial cells or other support elements. Anti-Ca1A labeling was also prominent in the peripheral axons of pyloric motoneurons as they enter muscles, indicating that this channel may be involved in regulation of synaptic transmission onto the foregut muscles. Anti-Ca1D does not label neurites in the neuropil of the stomatogastric ganglion. It stains glial cells in the stomatogastric ganglion in the region of their nuclei, presumably from protein being produced in the perinuclear rough endoplasmic reticulum, en route to the glial cell periphery. While anti-Ca1D labeling is seen in a patchy distribution along peripheral pyloric axons, it was never seen near the muscle. We conclude that the localization of these two calcium channels is tightly controlled within the stomatogastric nervous system, but we cannot conclusively demonstrate that Ca1A and/or Ca1D channels play roles in synaptic integration within the stomatogastric ganglion.

Quantification of gastric mill network effects on a movement related parameter of pyloric network output in the lobster.

It has long been known that gastric mill network activity (cycle period 5-10 s) alters pyloric network output (cycle period approximately 1 s), but these effects have not been quantified. Many pyloric muscles extract gastric mill timed variations in pyloric motor neuron firing, and consequently produce gastric mill timed movements even though no gastric mill neurons innervate them. Determining pyloric behavior therefore requires detailed description of gastric mill effects on pyloric neural output. Pyloric muscle activity correlates well with motor neuron overall spike frequency (OSF, burst spike number divided by cycle period). We quantified OSF variation of all pyloric neurons as a function of time into the gastric mill cycle [as measured from the beginning of Gastric Mill (GM) neuron bursts] in the lobster, Panulirus interruptus. No repeating pattern within individual gastric mill cycles of Lateral Pyloric (LP) and Ventricular Dilator (VD) neuron OSF was visually apparent. Averaged data showed that VD and LP neuron OSF decreased (approximately 0.5 and 1.5 Hz, respectively) at the beginning of each gastric mill cycle. Visually apparent patterns of OSF waxing and waning within each gastric mill cycle were present for the Inferior Cardiac (IC), Pyloric Dilator (PD), and Pyloric (PY) neurons. However, when averaged as a function of phase or delay in the gastric mill cycle, the average changes were smaller than those in individual gastric mill cycles because when the OSF variations occurred varied considerably in different gastric mill cycles. We therefore used a "pattern-based" analysis in which an identifying characteristic of each neuron's repeating OSF variation pattern was defined as pattern pyloric cycle zero. The pyloric cycles in each repetition of the OSF variation pattern were numbered relative to the zero cycle, and averaged to create an average OSF variation profile. The zero cycle delays relative to GM neuron burst beginning were then averaged to determine when in the gastric mill cycle the profile occurred. This technique preserved the full extent of pyloric neuron OSF changes. Maximum PY neuron OSF occurred within the GM neuron burst, whereas maximum IC and PD neuron OSF occurred during the GM neuron interburst interval. Despite these changes, pyloric cycling did not phase lock with gastric mill activity, nor were an integer number of pyloric cycles present in each gastric mill cycle. In addition to providing data necessary to predict pyloric movement, this work shows how pattern-based analysis can successfully quantify interactions between nonphase-locked networks.

Alternate splicing of the shal gene and the origin of I(A) diversity among neurons in a dynamic motor network.

The pyloric motor system, in the crustacean stomatogastric ganglion, produces a continuously adaptive behavior. Each cell type in the neural circuit possesses a distinct yet dynamic electrical phenotype that is essential for normal network function. We previously demonstrated that the transient potassium current (I(A)) in the different component neurons is unique and modulatable, despite the fact that the shal gene encodes the alpha-subunits that mediate I(A) in every cell. We now examine the hypothesis that alternate splicing of shal is responsible for pyloric I(A) diversity. We found that alternate splicing generates at least 14 isoforms. Nine of the isoforms were expressed in Xenopus oocytes and each produced a transient potassium current with highly variable properties. While the voltage dependence and inactivation kinetics of I(A) vary significantly between pyloric cell types, there are few significant differences between different shal isoforms expressed in oocytes. Pyloric I(A) diversity cannot be reproduced in oocytes by any combination of shal splice variants. While the function of alternate splicing of shal is not yet understood, our studies show that it does not by itself explain the biophysical diversity of I(A) seen in pyloric neurons.

RCPH modulation of a multi-oscillator network: effects on the pyloric network of the spiny lobster.

The neuropeptide red pigment concentrating hormone (RPCH), which we have previously shown to activate the cardiac sac motor pattern and lead to a conjoint gastric mill-cardiac sac pattern in the spiny lobster Panulirus, also activates and modulates the pyloric pattern. Like the activity of gastric mill neurons in RPCH, the pattern of activity in the pyloric neurons is considerably more complex than that seen in control saline. This reflects the influence of the cardiac sac motor pattern, and particularly the upstream inferior ventricular (IV) neurons, on many of the pyloric neurons. RPCH intensifies this interaction by increasing the strength of the synaptic connections between the IV neurons and their targets in the stomatogastric ganglion. At the same time, RPCH enhances postinhibitory rebound in the lateral pyloric (LP) neuron. Taken together, these factors largely explain the complex pyloric pattern recorded in RPCH in Panulirus.

Nonlinear behavior of sinusoidally forced pyloric pacemaker neurons.

Periodic current forcing was used to investigate the intrinsic dynamics of a small group of electrically coupled neurons in the pyloric central pattern generator (CPG) of the lobster. This group contains three neurons, namely the two pyloric dilator (PD) motoneurons and the anterior burster (AB) interneuron. Intracellular current injection, using sinusoidal waveforms of varying amplitude and frequency, was applied in three configurations of the pacemaker neurons: 1) the complete pacemaker group, 2) the two PDs without the AB, and 3) the AB neuron isolated from the PDs. Depending on the frequency and amplitude of the injected current, the intact pacemaker group exhibited a wide variety of nonlinear behaviors, including synchronization to the forcing, quasiperiodicity, and complex dynamics. In contrast, a single, broad 1:1 entrainment zone characterized the response of the PD neurons when isolated from the main pacemaker neuron AB. The isolated AB responded to periodic forcing in a manner similar to the complete pacemaker group, but with wider zones of synchronization. We have built an analog electronic circuit as an implementation of a modified Hindmarsh-Rose model for simulating the membrane potential activity of pyloric neurons. We subjected this electronic model neuron to the same periodic forcing as used in the biological experiments. This four-dimensional electronic model neuron reproduced the autonomous oscillatory firing patterns of biological pyloric pacemaker neurons, and it expressed the same stationary nonlinear responses to periodic forcing as its biological counterparts. This adds to our confidence in the model. These results strongly support the idea that the intact pyloric pacemaker group acts as a uniform low-dimensional deterministic nonlinear oscillator, and the regular pyloric oscillation is the outcome of cooperative behavior of strongly coupled neurons, having different dynamical and biophysical properties when isolated.

Topology selection by chaotic neurons of a pyloric central pattern generator.

The pyloric Central Pattern Generator (CPG) in the lobster has an architecture in which every neuron receives at least one connection from another member of the CPG. We call this a "non-open" network topology. An "open" topology, where at least one neuron does not receive synapses from any other CPG member, is found neither in the pyloric nor in the gastric mill CPG. Here we investigate a possible reason for this topological structure using the ability to perform a biologically functional task as a measure of the efficacy of the network. When the CPG is composed of model neurons that exhibit regular membrane voltage oscillations, open topologies are as able to maximize this functionality as non-open topologies. When we replace these models by neurons which exhibit chaotic membrane voltage oscillations, the functional criterion selects non-open topologies. As isolated neurons from invertebrate CPGs are known in some cases to undergo chaotic oscillations, this suggests that there is a biological basis for the class of non-open network topologies that we observe.

Mechanisms underlying stabilization of temporally summated muscle contractions in the lobster (Panulirus) pyloric system.

Muscles are the final effectors of behavior. The neural basis of behavior therefore cannot be completely understood without a description of the transfer function between neural output and muscle contraction. To this end, we have been studying muscle contraction in the well-investigated lobster pyloric system. We report here the mechanisms underlying stabilization of temporally summating contractions of the very slow dorsal dilator muscle in response to motor nerve stimulation with trains of rhythmic shock bursts at a physiological intraburst spike frequency (60 Hz), physiological cycle periods (0.5-2 s), and duty cycles from 0.1 to 0.8. For temporal summation to stabilize, the rise and relaxation amplitudes of the phasic contractions each burst induces must equalize as the rhythmic train continues. Stabilization could occur by changes in rise duration, rise slope, plateau duration, and/or relaxation slope. We demonstrate a generally applicable method for quantifying the relative contribution changes in these characteristics make to contraction stabilization. Our data show that all characteristics change as contractions stabilize, but their relative contribution differs depending on stimulation cycle period and duty cycle. The contribution of changes in rise duration did not depend on period or duty cycle for the 1-, 1.5-, and 2-s period regimes, contributing approximately 30% in all cases; but for the 0.5-s period regime, changes in rise duration increased from contributing 25% to contributing 50% as duty cycle increased from 0.1 to 0.8. At all cycle periods decreases in rise slope contributed little to stabilization at small duty cycles but increased to contributing approximately 80% at high duty cycles. The contribution of changes in plateau duration decreased in all cases as duty cycle increased; but this decrease was greater in long cycle period regimes. The contribution of changes in relaxation slope also decreased in all cases as duty cycle increased; but for this characteristic, the decrease was greatest in fast cycle period regimes, and in these regimes at high duty cycles these changes opposed contraction stabilization. Exponential fits to contraction relaxations showed that relaxation time constant increased with total contraction amplitude; this increase presumably underlies the decreased relaxation slope magnitude seen in high duty cycle, fast cycle period regimes. These data show that changes in no single contraction characteristic can account for contraction stabilization in this muscle and suggest that predicting muscle response in other systems in which slow muscles are driven by rapidly varying neuronal inputs may be similarly complex.

Modeling observed chaotic oscillations in bursting neurons: the role of calcium dynamics and IP3.

Chaotic bursting has been recorded in synaptically isolated neurons of the pyloric central pattern generating (CPG) circuit in the lobster stomatogastric ganglion. Conductance-based models of pyloric neurons typically fail to reproduce the observed irregular behavior in either voltage time series or state-space trajectories. Recent suggestions of Chay [Biol Cybern 75: 419-431] indicate that chaotic bursting patterns can be generated by model neurons that couple membrane currents to the nonlinear dynamics of intracellular calcium storage and release. Accordingly, we have built a two-compartment model of a pyloric CPG neuron incorporating previously described membrane conductances together with intracellular Ca2+ dynamics involving the endoplasmic reticulum and the inositol 1,4,5-trisphosphate receptor IP3R. As judged by qualitative inspection and quantitative, nonlinear analysis, the irregular voltage oscillations of the model neuron resemble those seen in the biological neurons. Chaotic bursting arises from the interaction of fast membrane voltage dynamics with slower intracellular Ca2+ dynamics and, hence, depends on the concentration of IP3. Despite the presence of 12 independent dynamical variables, the model neuron bursts chaotically in a subspace characterized by 3-4 active degrees of freedom. The critical aspect of this model is that chaotic oscillations arise when membrane voltage processes are coupled to another slow dynamic. Here we suggest this slow dynamic to be intracellular Ca2+ handling.

Interacting biological and electronic neurons generate realistic oscillatory rhythms.

Small assemblies of neurons such as central pattern generators (CPG) are known to express regular oscillatory firing patterns comprising bursts of action potentials. In contrast, individual CPG neurons isolated from the remainder of the network can generate irregular firing patterns. In our study of cooperative behavior in CPGs we developed an analog electronic neuron (EN) that reproduces firing patterns observed in lobster pyloric CPG neurons. Using a tuneable artificial synapse we connected the EN bidirectionally to neurons of this CPG. We found that the periodic bursting oscillation of this mixed assembly depends on the strength and sign of the electrical coupling. Working with identified, isolated pyloric CPG neurons whose network rhythms were impaired, the EN/biological network restored the characteristic CPG rhythm both when the EN oscillations are regular and when they are irregular.

Monoamine control of the pacemaker kernel and cycle frequency in the lobster pyloric network.

The monoamines dopamine (DA), serotonin (5HT), and octopamine (Oct) can each sculpt a unique motor pattern from the pyloric network in the stomatogastric ganglion (STG) of the spiny lobster Panulirus interruptus. In this paper we investigate the contribution of individual network components in determining the specific amine-induced cycle frequency. We used photoinactivation of identified neurons and pharmacological blockade of synapses to isolate the anterior burster (AB) and pyloric dilator (PD) neurons. Bath application of DA, 5HT, or Oct enhanced cycle frequency in an isolated AB neuron, with DA generating the most rapid oscillations and Oct the slowest. When an AB-PD or AB-2xPD subnetworks were tested, DA often reduced the ongoing cycle frequency, whereas 5HT and Oct both evoked similar accelerations in cycle frequency. However, in the intact pyloric network, both DA and Oct either reduced or did not alter the cycle frequency, whereas 5HT continued to enhance the cycle frequency as before. Our results show that the major target of 5HT in altering the pyloric cycle frequency is the AB neuron, whereas DA's effects on the AB-2xPD subnetwork are critical in understanding its modulation of the cycle frequency. Octopamine's effects on cycle frequency require an understanding of its modulation of the feedback inhibition to the AB-PD group from the lateral pyloric neuron, which constrains the pacemaker group to oscillate more slowly than it would alone. We have thus demonstrated that the relative importance of the different network components in determining the final cycle frequency is not fixed but can vary under different modulatory conditions.