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D K Hartline

Publications and source records attributed to D K Hartline.

At least 19 recordsLinked to original sources

Effects of soma isolation on outward currents measured under voltage clamp in spiny lobster stomatogastric motor neurons.

1. Outward currents in identified cell types from the pyloric system of the stomatogastric ganglion (STG) of the spiny lobster, Panulirus marginatus, were studied under two-microelectrode voltage clamp. A comparison was made between data from intact cells and somata isolated by ligation of the primary neurite of these monopolar neurons. 2. Despite the elimination of current contributions from the extensive arborizations of STG neurons, few significant differences were found in the mean values of parameters for outward currents between populations of isolated somata and intact cells of a given type. Measurements that showed little difference included magnitude and activation threshold of a calcium-dependent outward current (IJ) and magnitude, activation threshold, voltage dependence, and inactivation time course of A current (IA). Although previous work has suggested that IJ might reside predominantly in the soma, IA is known to be distributed in poorly space-clamped neurite processes. The absence of obvious effects of isolation was thus unexpected. 3. To better understand the mechanisms involved, we used compartmental models derived from reconstructed neurons to simulate the effects of isolation. It was concluded that, for the particular conditions present in stomatogastric neurons, with a large, uniformly distributed outward current conductance activated, even though neurites and axon remain attached, most measured current flows through well-clamped soma membrane. 4. Factors contributing to this result included the outward sign of the current, the large specific conductance activated in these neurons (among the larger reported in somata), and the presence of only a single major process leaving the soma. The potential for serious errors in voltage-clamp measurements from intact cells remains if these conditions are not met.

Animals

Voltage clamp analysis of intact stomatogastric neurons.

Two-electrode voltage clamp of intact, identified pyloric neurons of the spiny lobster stomatogastric ganglion reveals two major outward currents. A rapidly inactivating, tetraethylammonium- (TEA) insensitive, 4-aminopyridine- (4AP) sensitive, outward current resembles IA of molluscan neurons; it activates rapidly on depolarizations above rest (e.g. -45 mV), delaying both the axonal-sodium and the neuropil-calcium spikes which escape voltage-clamp control. We infer that A-current is distributed both in a space clamped region (on or near the soma) and in a non-space clamped region with access to the generators for sodium and calcium spikes. A calcium-dependent outward current, IO(Ca), activates rapidly at clamp steps above -25 mV and inactivates at depolarizing holding voltages. Increasing depolarization results in an increase in both IO(Ca) and firing rate but a reduction in the amplitude of the sodium spike current. Blockage of IO(Ca) with Cd2+ causes little change in spike firing pattern. These observations are consistent with IO(Ca) being activated primarily in the soma and nearby regions which are under good control with a soma voltage clamp (and distant from the Na(+)-spike trigger zone). While the lack of space clamp limits resolution of charging transients and tail currents, the identification of the major current subgroups can still be readily accomplished, and inferences about the location and function of currents can be made which would not be possible if the cells were space clamped or truncated.

4-Aminopyridine

Full-wave rectification from a mixed electrical-chemical synapse.

Electrical and chemical synapses usually reinforce one another, but the pyloric late-to-lateral pyloric (PL-to-LP) neuronal connections in lobster stomatogastric ganglia create an inverted U-shaped transfer function between the two neurons: regardless of whether the PL membrane voltage swings positive or negative, the postsynaptic LP voltage will go negative. When the presynaptic cell voltage goes negative, the effect on the LP voltage is due to electrical coupling. During positive presynaptic voltages, the strong contribution of graded chemical inhibition from the PL to the LP neuron overrides the positive electrical coupling to produce net negativity.

Animals

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. CP bursting during normal gastric mill rhythms, while underlain by plateau potentials, is strongly controlled by network interactions. CP appears not to be well placed in the network to be considered a source of normal gastric rhythmicity. Nevertheless, endogenous bursting in CP may explain some of the partial gastric rhythms seen in behavioral studies, and illustrates one way that cellular properties might contribute to rhythmic behaviors.

Animals

Synaptic regulation of cellular properties and burst oscillations of neurons in gastric mill system of spiny lobsters, Panulirus interruptus.

The properties of neurons in the stomatogastric ganglion (STG) participating in the pattern generator for the gastric mill rhythm were studied by intracellular current injection under several conditions: during ongoing gastric rhythms, in the nonrhythmic isolated STG, after stimulation of the nerve carrying central nervous system (CNS) inputs to the STG, or under Ba2+ or Sr2+. Slow regenerative depolarizations during ongoing rhythms were demonstrated in the anterior median, cardiopyloric, lateral cardiac, gastropyloric, and continuous inhibitor (AM, CP, LC, GP, and CI) neurons according to criteria such as voltage dependency, burst triggering, and termination by brief current pulses, etc. Experiments showed that regenerative-like behavior was not due to synaptic network interactions. The slow regenerative responses were abolished by isolating the stomatogastric ganglion but could be reestablished by stimulating the input nerve. This indicates that certain CNS inputs synaptically induce the regenerative property in specific gastric neurons. Slow regenerative depolarizations were not demonstrable in gastric mill (GM) motor neurons. Their burst oscillations and firing rate were instead proportional to injected current. CNS inputs evoked a prolonged depolarization in GM motor neurons, apparently by a nonregenerative mechanism. All the gastric cells showed prolonged regenerative potentials under 0.5-1.5 mM Ba2+. We conclude that the gastric neurons of the STG can be divided into three types according to their properties: those with a regenerative capability, a repetitively firing type, and a nonregenerative "proportional" type. The cells are strongly influenced by several types of CNS inputs, including "gastric command fibers."

Animals

Graded synaptic transmission between identified spiking neurons.

Graded synaptic transmission between spiking motoneurons of the pyloric group was studied in the stomatogastric ganglion of the spiny lobster, Panulirus interruptus. Intracellular microelectrodes were placed in the cell bodies of both pre- and postsynaptic neurons. Graded synaptic transmission was found between all tested cell pairs that were known to display spike-evoked synaptic transmission, including PD to LP, PD to PE, PD to PL, PL to LP, and LP to PD. Graded synaptic transmission was effective below the threshold for spikes. Thus, it was possible to study the influence of graded synaptic transmission in normally active ganglia without blockage of spikes by tetrodotoxin. PD and LP neurons that were known to produce spike-evoked inhibitory postsynaptic potentials (IPSPs) were also capable of producing inhibitory effects on postsynaptic cells below the threshold for spikes. When tetrodotoxin (TTX) was used to eliminate both spikes and endogenous membrane oscillations, depolarization of presynaptic neurons produced hyperpolarization of postsynaptic cells. The presynaptic response to a current step usually showed a small early peak and a maintained, slightly lower plateau. The postsynaptic response had a delay, then a rise to a pronounced peak, and a roughly exponential decline to a maintained plateau. There was a presynaptic voltage threshold for any postsynaptic response; beyond the threshold, both pre- and postsynaptic peak and plateau responses increased with increasing current. PD neurons normally are depolarized beyond their release threshold in tetrodotoxin and, thus, released transmitter tonically for the many-hour duration of these experiments. Chemical, tonic synaptic transmission, here called graded synaptic transmission, was demonstrated by the presence of the following criteria: 1) reversal in sign of the postsynaptic response, 2) synaptic delay, 3) reversal potential, 4) postsynaptic conductance increase, 5) graded and reversible block by reduction of external Ca2+, and 6) specific graded block of the LP-to-PD synapse without effect on the PD-to-LP synapse by less than 10 microM picrotoxin added to the bathing medium.

Action Potentials

Slow active potentials and bursting motor patterns in pyloric network of the lobster, Panulirus interruptus.

1. Neurons in the central pattern generator for the "pyloric" motor rhythm of the lobster stomatogastric ganglion were investigated for the possible involvement of regenerative membrane properties in their membrane-potential oscillations and bursting output patterns. 2. Evidence was found that each class of pyloric-system neurons can possess a capability for generating prolonged regenerative depolarizations by a voltage-dependent membrane mechanism. Such responses have been termed plateau potentials. 3. Several tests were applied to determine whether a given cell possessed a plateau capability. First among these was the ability to trigger all-or-none bursts of nerve impulses by brief depolarizing current pulses and to terminate bursts in an all-or-none fashion with brief hyperpolarizing current pulses. Tests were made under conditions of a high level of activity in the pyloric generator, often in conjunction with the use of hyperpolarizing offsets to the cell under test to suppress ongoing bursting. 4. For each class, the network of synaptic interconnections among the pyloric-system neurons was shown to not be the cause of the regenerative responses observed. 5. Plateau potentials are viewed as a driving force for axon spiking during bursts and as interacting with the synaptic network in the formation of the pyloric motor pattern.

Animals

A multiaction synapse evoking both EPSPs and enhancement of endogenous bursting.

Selective stimulation of two identified input neurons called the 'IV neurons' has a dual influence on the endogenous bursting activity of certain 'PD' motorneurons in the stomatogastric ganglion of the spiny lobster. The effects include: (i) large, conventional and apparently monosynaptic EPSPs; and (ii) enhancement of the endogenous bursting of the pyloric dilator (PD) cells, seen as an increased amplitude of PD oscillations and a higher spiking rate during bursts. The burst enhancement decayed relatively slowly after stimulation ceased, over seconds or tens-of-seconds, depending on stimulus parameters. Modification of the voltage-dependent membrane properties of the PD cells appeared to underlie this effect. The dual-action nature of the IV-to-PD connection was confirmed by selectively blocking the brief EPSP component with 5 x 10(-4) M curare, under which conditions the burst enhancement still persisted. Data from low-Ca2+ experiments were consistent with a conventional mode of synaptic transmitter release underlying the burst enhancement. Enhancement was found to differ significantly from actions of injected current. The IV inputs appear to act on at least two types of synaptic receptors on PD neurons: a curare-sensitive receptor for the brief conventional EPSP, and a curare-resistant receptor for burst enhancement. Analogies may be drawn to the nicotinic and muscarinic cholinergic receptors of vertebrates. These findings may be considered within the contexts of multiaction synapses, modification of cellular properties, and mechanisms for the CNS activation of motor pattern generators.

Animals

Graded synaptic transmission between spiking neurons.

Graded synaptic transmission occurs between spiking neurons of the lobster stomatogastric ganglion. In addition to eliciting spike-evoked inhibitory potentials in postsynaptic cells, these neurons also release functionally significant amounts of transmitter below the threshold for action potentials. The spikeless postsynaptic potentials grade in amplitude with presynaptic voltage and can be maintained for long periods. Graded synaptic transmission can be modulated by synaptic input to the presynaptic neuron.

Action Potentials

Pattern generation in the lobster (Panulirus) stomatogastric ganglion. I. Pyloric neuron kinetics and synaptic interactions.

There are a number of perspectives gained from a quantitative analysis of the pyloric system which may be applicable to other simple pattern generators: 1. The system is organized around a dominant, endogenously-bursting neuron group, and its properties are tailored to that dominance. In particular, synaptic strengths and firing frequencies of that group appear just sufficient to suppress postsynaptic "follower" cells if the latter are not too highly excited. 2. Repetitive firing properties of follower neurons are such as to facilitate their switch-like mode of activity. This includes pacemaker response nonlinearities, rebound properties, and "burstiness" properties. 3. Proper sequencing of follower cells may be controlled by particular synaptic strengths and time-courses, feedback on the oscillator cells, and functional cellular properties of follower neurons (e.g., rebound; see also next paper). All such properties interact and must be tuned to each other for proper patterns to result.

Adaptation, Physiological

Pattern generation in the lobster (Panulirus) stomatogastric ganglion. II. Pyloric network simulation.

1. Results from the companion paper were incorporated into a physiologically realistic computer model of the three principal cell types (PD/AB, LP, PY) of the pyloric network in the stomatogastric ganglion. Parameters for the model were mostly calculated (sometimes estimated) from experimental data rather than fitting the model to observed output patterns. 2. The initial run was successful in predicting several features of the pyloric pattern: the observed gap between PD and LP bursts, the appropriate sequence of the activity periods (PD, LP, PY), and a substantial PY burst not properly simulated by an earlier model. 3. The major discrepancy between model and observed patterns was the too-early occurrence of the PY burst, which resulted in a much shortened LP burst. Motivated by this discrepancy, additional investigations were made of PY properties. A hyperpolarization-enabled depolarization-activated hyperpolarizing conductance change was discovered which may make an important contribution to the late phase of PY activity in the normal burst cycle. Addition of this effect to the model brought its predictions more in line with observed patterns. 4. Other discrepancies between model and observation were instructive and are discussed. The findings force a substantial revision in previously held ideas on pattern production in the pyloric system. More weight must be given to functional properties of individual neurons and less to properties arising purely from network interactions. This shift in emphasis may be necessary in more complicated systems as well. 5. An example has been provided of the value quantitative modeling can be to network physiology. Only through rigorous quantitative testing can qualitative theories of how the nervous system operates be substantiated.

Animals

Bursting neural networks: a reexamination.

Many of the motor neurons in the lobster (Panulirus interruptus) stomatogastric ganglion exhibit plateau potentials; that is, prolonged regenerative depolarizations resulting from active membrane properties, that drive the neurons to fire impulses during bursts. Plateaus are latent in isolated ganglia but are unmasked by central input. These findings emphasize the role of cellular properties as compared to synaptic wiring in the production of cyclic motor patterns by ensembles of neurons.

Action Potentials

Retrograde invasion of lobster stretch receptor somata in control of firing rate and extra spike patterning.

1. Extra spikes may be interleaved in the otherwise rhythmic discharge pattern of the lobster stretch receptor neuron, about 2 ms after an expected spike. A constant input to the neuron is maintained by injecting current intrasomatically. The axon recovers its excitability while the retrograde invasion of the soma and dendrites is still in progress, which provide electrotonic currents to reexcite the axon. 2. While extra spikes in the axon often arise from a prolonged somatic (dendritic?) depolarization, they may also arise from a delayed retrograde invasion of the soma. 3. Failure of retrograde invasion may cause a sudden jump in the rate of rhythmic discharge, demonstrating the role of the soma-dendritic afterhyperpolarization in the regulation of rhythmic firing rate. 4. The history of repetitive firing is often important. Because extra spikes often first appear during a decline in firing rate, turning on and then off, an additional current may sometimes activate the extra spike mode, thus doubling the resting firing rate in a metastable manner. Another mestastable state is associated with failure of retrograde invasion. 5. Extra spikes augment the high end of the frequency-current curve in some receptor neurons; in other cases, the extra spikes are seen only at low rhythmic firing rates, dropping out as current reaches intermediate values to create a paradoxical negative-sensitivity region (decline in total spikes per second with increasing current). 6. The results suggest that both the extent and the speed of active retrograde invasion of the soma and dendrites are likely candidates for pathophysiological mechanisms, since they may control whether extra spikes are generated.

Animals

Simulation of phase-dependent pattern changes to perturbations of regular firing in crayfish stretch receptor.

A comparison was made of responses of a repetitively firing neuron to perturbations imposed in an ongoing steady firing pattern with those predicted by a model termed the 'active pacemaker model' and by a simpler 'integrate-and-fire' model. The active pacemaker model simulates the kinetics of processes such as active response generation and electrogenic sodium pumping which govern the membrane potential trajectory between a reset-point following an impulse and threshold for the next impulse. Responses of the crayfish stretch receptor neuron (MRO) and of both models were studied for both abrupt steps and square pulses of current, both hyperpolarizing and depolarizing, as a function of phase of the start of the stimulus in the normal interspike interval. Physiological results are consistent with a substantially weaker effect of a perturbation delivered earlier in the interspike interval as compared to the same one given later. The active pacemaker model successfully simulates several of the prominent results of the physiology (though by no means all), while the linear integrate-and-fire model is less successful.

Action Potentials

Simulation of network activity in stomatogastric ganglion of the spiny lobster, Panulirus.

We have compared experimental and model studies on the rhythmic activity of the lobster stomatogastric ganglion. Both the pyloric and gastric mill systems were simulated using a physiologically based network model. In the pyloric simulation the known synaptic connectivity of the 3 principal cell types in the pyloric rhythm was found to be sufficient to produce the correct sequence of cyclic bursting activity over a substantial range of parameter values, even though we did not simulate the known endogenous oscillatory driver potential of one of the cell types. It is not yet known whether the synaptic inhibition in the real system is in the right range to cause cyclic bursting in the absence of the driver potential, but the synaptic connectivity does appear to reinforce the cyclic pattern. Simulations were also done with alternative connectivity schemes to determine which synapses appear essential to generate the correct bursting sequence (in the absence of endogenous bursting activity). Other, more complex, systems simulated were: (1) all 5 cell groups of the pyloric system, and (2) the cells responsible for movement of the lateral teeth in the gastric mill. In both cases good qualitative agreement was achieved between model and real systems.

Action Potentials