Pattern generation in the lobster (Panulirus) stomatogastric ganglion

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 depolorization-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.     

Motor patterns in the stomatogastric ganglion of the lobster Panulirus argus.

1. Acitivity patterns arising from the thirty cells of the stomatogastric ganglion of Panulirus argus are described for both a semi-intact preparation and an isolated one. 2. The thirty or so cells can be divided so far into two functional groupings: the gastric mill group, with at least ten motor elements, and the pyloric group with at least fourteen. There is some, but not extensive, interaction between groups. 3. The main gastric mill activity is arranged in two sets of elements, each of which is composed of reciprocating elements innervating antagonistic muscles. Thus alternation in activity between the single LC and the two LG neurones results in alternate closing and opening of the lateral teeth; alternation between the four GM and single CP units results in alternate protraction and retraction of the medial tooth. 4. The two sets are phased to each other in such a way that they cause gastric mill teeth to operate effectively to masticate food. 5. The main pyloric activity is arranged in a three-part cycle with each of three sets of units active in sequence. Activity in two PD and one AB unit is followed by bursts in IC and LP units followed in turn by activity in up to seven PY units. Activity in a single VD neurone is locked to this cycle in a more complex pattern.     

Morphology and location of dense-core vesicles in the stomatogastric ganglion of the lobster,Panulirus interruptus

The appearance and distribution of dense-core vesicles in the stomatogastric ganglion of the spiny lobster, Panulirus interruptus, were examined using transmission electron microscopy. Following five fixation techniques, three types of dense-core vesicles were identified on the basis of size and morphology. Type-I vesicles are found in a distinct neuronal fiber system that appears to be involved in chemical transmission within the ganglion. Type-II vesicles occur in nerve processes in the ganglion, in major nerve trunks and in the perineural sheath of the nerves and ganglion. Type-III vesicles are present in all neuronal somata of the ganglion. The distinct morphology and location of the three types of vesicles suggest that their functional roles differ. Furthermore, the histochemical, biochemical and physiological data available for the stomatogastric ganglion indicate that Type-I vesicles may store dopamine.     

Organization of the stomatogastric ganglion of the spiny lobster

Summary The Stomatogastric ganglion ofPanulirus interruptus contains about 30 neurons, and controls the movements of the lobster's stomach. When experimentally isolated, the ganglion continues to generate complex rhythmic patterns of activity in its motor neurons which are similar to those seen in intact animals.In this paper, we describe the synaptic organization of a group of six neurons which drive the stomach's lateral teeth (Figs. 2, 6). This group includes four motor neurons and two interneurons, all but one of which were recorded and stimulated with intracellular microelectrodes.One pair of synergistic motor neurons, LGN and MGN, are electrotonically coupled and reciprocally inhibitory (Figs. 9, 12). A second pair of synergistic motor neurons, the LPGNs, are antagonists of LGN and MGN. The LPGNs are electrotonically coupled (Fig. 14), and are both inhibited by LGN and MGN (Figs. 8, 11). The LPGNs inhibit MGN (Fig. 15) but not LGN. One of the two interneurons in the ganglion, Int 1, reciprocally inhibits both LGN and MGN (Figs. 10, 13). The other interneuron, Int 2, excites Int 1 and inhibits the LPGNs (Fig. 16). The synaptic connections observed in the ganglion are reflected in the spontaneous activity recorded from the isolated ganglion and from intact animals.From the known synaptic organization and observations on the physiological properties of each of the neurons, we have formulated some hypotheses about the pattern-generating mechanism. We found no evidence that any of the neurons are endogenous bursters.We thank D. Kennedy, Eve Marder, and D. Russell for criticizing early drafts of these papers, Nina Pollack and Betty Jorgensen for expert technical assistance, Diane Newsome, SanDee Newcomb, and Pattie Macpherson for typing the many drafts. The authors' research is supported by grant number NS-09322 from N.I.H. and by the Alfred P. Sloan Foundation. B. M. is an NINDS-NIH postdoctoral fellow.     

Organization of the stomatogastric ganglion of the spiny lobster

Summary Three direct synaptic connections occur between neurons in the gastric and pyloric systems of the stomatogastric ganglion ofPanulirus interruptus. Two synapses are inhibitory, and one is electrical. This electrical synapse is both excitatory and inhibitory at different times. These synapses, and others within each system, let the two systems interact under some conditions. The synapses also form multisynaptic pathways which modulate the firing of many neurons in both systems. The consequences of these multisynaptic pathways are described and discussed.I thank Allen I. Selverston, Karen Sigvardt, Eve Marder, David Russell and Mary Chamberlin for criticizing a draft of this paper, Forrest Gompf and Doug Tissdale for technical support, and Nina Pollack and Betty Jorgensen for laboratory assistance. The research was supported by USPHS grant NS-12295 to BM and USPHS grant NS-09322 to Alien I. Selverston. BM was a USPHS NIH Postdoctoral Fellow in A.I. Selverston's laboratory during part of this research and is now a Research Fellow of the Alfred P. Sloan Foundation.     

Electron microscopy of stomatogastric ganglion in the lobster Homarus americanus

The stomatogastric ganglion and two of the associated afferent and efferent nerve trunks (stomatogastric and dorsal ventricular nerves) from Homarus americanus have been examined with light and electron microscopy after glutaraldehyde-osmium tetroxide fixation. The dorsally located neuron somata, rich in ribosomes and glycogen, are encased in multi-layered glial and fibrous sheaths. The synaptic neuropil regions occur scattered throughout the central and ventral part of the ganglion, interspersed amonglarger nerve fibres of extrinsic and intrinsic origin from which the neuropil is derived. Neural processes containing masses of small clear vesicles plus larger dense-core vesicles make apparent synaptic contacts at points of increased membrane density with smaller, non-vesicle-containing or sometimes other vesicle-containing nerve fibres.     

A model for basic pattern generating mechanisms in the lobster stomatogastric ganglion

The stomatogastric ganglion of the lobster contains three central pattern generators-the pyloric, lateral gastric and medial gastric systems. These networks are modelled using a simple neural model in which the only variable parameters are the synaptic potentials and thresholds for each cell. In each case a model network with the appropriate synaptic connections reproduces the main features of the observed output patterns. The basic pattern generating mechanisms are quite different for each of these model networks. For the pyloric and lateral gastric systems our results confirm previously suggested mechanisms. For the medial gastric system we have determined a network which explains pattern generation; no satisfactory mechanism was previously known.     

Behavioral thermoregulation in the spiny lobster Panulirus Argus (latreille)

Ten spiny lobsters (Panulirus argus) were allowed to thermoregulate individually for 3-day periods in an electronic thermoregulatory shuttlebox which allowed them to control water temperatures (and thereby their own body temperatures) by their movements. The range of preferred (voluntarily occupied) temperatures was 25–35°C (mean 29.9°C; mode 30.0°C; median 30.0°C; midpoint 30.0°C; S_k (skewness, Pearson's coefficient) –0.04; s.e.m. 0.19°C; S.D. 2.32°C). The final thermal preferendum (by the gravitation method) in this species is 30°C.     

Differential modulation of chemical and electrical components of mixed synapses in the lobster stomatogastric ganglion

1. Two pairs of neurons in the pyloric network of the spiny lobster, Panulirus interruptus, communicate through mixed graded chemical and rectifying electrical synapses. The anterior burster (AB) chemically inhibits and is electrically coupled to the ventricular dilator (VD); the lateral pyloric (LP) and pyloric (PY) neurons show reciprocal chemical inhibition and electrical coupling. We examined the effects of dopamine (DA), serotonin (5HT) and octopamine (Oct) on these mixed synapses to determine the plasticity possible with opposing modes of synaptic interaction.
2. Dopamine increased net inhibition at all three pyloric mixed synapses by both reducing electrical coupling and increasing chemical inhibition. This reversed the sign of the net synaptic interaction when electrotonic coupling dominated some mixed synapses, and activated silent chemical components of other mixed synapses.
3. Serofonin weakly enhanced LP → PY net inhibition, by reducing electrical coupling without altering chemical inhibition. Serotonin reduced AB→ VD electrical coupling, but variability in its effect on the chemical component made the net effect non-significant.
4. Octopamine enhanced LP→ PY and PY→ LP net inhibition by enhancing the chemical inhibitory component without altering electrical coupling.
5. Differential modulation of chemical and electrical components of mixed synapses markedly changes the net synaptic interactions. This contributes to the flexible outputs that modulators evoke from anatomically defined neural networks.

     

A neurochemical description of the dopaminergic innervation of the stomatogastric ganglion of the spiny lobster

The spiny lobster stomatogastric ganglion has been shown to be innervated by catecholaminergic processes which derive from cells of large central ganglia (Kushner and Maynard, 1977). Biochemical evidence had indicated that the stomatogastric system synthesizes dopamine and not norepinephrine from tritiated tyrosine (Barker, Kushner, and Hooper, 1979). Studies reported here document that the stomatogastric ganglion itself contains dopamine, as measured with a sensitive endogenous assay. Moreover, the ganglion can synthesize dopamine from tritiated tyrosine or DOPA. Additionally, when incubated in tritiated dopamine, the ganglion takes up dopamine and protects it from degradation; this process is inhibited by cocaine. When incubated with 3H-tyrosine, small but measurable amounts of tritiated dopamine were detected in the medium surrounding the ganglion.     

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.     

Osmotic and ionic regulation in the western rock lobster Panulirus longipes (Milne-Edwards)

The western rock lobster, Panulirus longipes (Milne-Edwards) is poikilosmotic over its tolerated salinity range, 25–45 ‰. Blood sodium is accumulated while chloride concentration is reduced. Sodium and chloride vary directly with the external salinity, although maintaining their differences in the same proportions as at normal salinity (36.0 ‰). Calcium is accumulated, ranging from over 150% at salinity 20 ‰ to about 117% at salinity 45 ‰. Potassium concentration is equivalent to the external at normal salinity, but is increased with lowered and decreased with raised salinity. Magnesium is reduced to about one-third that of the external concentration over the salinity range 20–40 ‰, but regulation begins to break down at 45 ‰. Individual ions exhibit, therefore, a range of regulation types, from poikilosmotic to homoiosmotic.Equilibrium for sodium, chloride, and calcium is attained in 10 h at salinities of 25, 30, 40 and 45 ‰ respectively. Rate constants for this exchange are linearly related to salinity differential, and rapid osmotic adjustment is by high permeability, equal in both directions, probably mainly via the gills. Muscle appears to act as a salt pool for sodium, chloride, and potassium but not for magnesium and calcium. Salt-loading causes a slight salt diuresis, the salts being excreted, probably via the gills. Except for calcium, there is no excretion of salt into the gut, but there is evidence of an exchange of chloride with another anion. Magnesium excretion is slow, and in the absence of osmotic stress possibly occurs via the antennal glands. All the ions examined appear to be regulated independently.     

Hemocyanin-derived phenoloxidase activity in the spiny lobster Panulirus argus (Latreille, 1804)

Hemocyanin and phenoloxidase belong to the type-3 copper protein family, sharing a similar active center whereas performing different roles. In this study, we demonstrated that purified hemocyanin (450 kDa) from the spiny lobster Panulirus argus shows phenoloxidase activity in vitro after treatment with trypsin, chymotrypsin and SDS (0.1% optimal concentration), but it is not activated by sodium perchlorate or isopropanol. The optimal pHs of the SDS-activated hemocyanin were 5.5 and 7.0. Hemocyanin from spiny lobster behaves as a catecholoxidase. Kinetic characterization using dopamine, L-DOPA and catechol shows that dopamine is the most specific substrate. Catechol and dopamine produced substrate inhibition above 16 and 2 mM respectively. Mechanism-based inhibition was also evidenced for the three substrates, being less significant for L-DOPA. SDS-activated phenoloxidase activity is produced by the hexameric hemocyanin. Zymographic analysis demonstrated that incubation of native hemocyanin with trypsin and chymotrypsin, produced bands of 170 and 190 kDa respectively, with intense phenoloxidase activity. Three polypeptide chains of 77, 80 and 89 kDa of hemocyanin monomers were identified by SDS-PAGE. Monomers did not show phenoloxidase activity induced by SDS or partial proteolysis.     

Sound production by the western rock lobster Panulirus longipes (Milne Edwards)

Sound characteristics and sound emitting structures of larval, juvenile and adult rock lobsters, Panulirus longipes (Milne Edwards) have been investigated. There is a linear correlation between carapace length and dimensions of the sound producing organ (number and size of plectrum ridges) as well as average duration of a single squeak. The number of pulses per 100 ms squeak, however, decreases asymptotically from $\text{\$}\text{?}$30 in larval forms to about 10 in mature adults. Behavioural changes accompany this process. The energy of a single pulse is distributed equally over the audio frequency range. The same is true for the ‘slow rattle’ sound, produced by mandible-grinding during feeding or moments of extreme agitation. Any predator attacking a rock lobster, is likely to receive injuries from the outstretched, spiny antennae. It is concluded that the antennal sound, accompanying the rock lobsters defence, functions as a warning device aimed primarily at predators, which will learn to associate the sound of rock lobsters with the unpleasant experience of the sharp antennal spines.