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.     

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.     

Pattern generation in the lobster (Panulirus) stomatogastric ganglion

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.     

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.     

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.

     

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.     

Mass spectral comparison of the neuropeptide complement of the stomatogastric ganglion and brain in the adult and embryonic lobster, Homarus americanus

Neuropeptides in the stomatogastric ganglion (STG) and the brain of adult and late embryonic Homarus americanus were compared using a multi-faceted mass spectral strategy. Overall, 29 neuropeptides from 10 families were identified in the brain and/or the STG of the lobster. Many of these neuropeptides are reported for the first time in the embryonic lobster. Neuropeptide extraction followed by liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry enabled confident identification of 24 previously characterized peptides in the adult brain and 13 peptides in the embryonic brain. Two novel peptides (QDLDHVFLRFa and GPPSLRLRFa) were de novo sequenced. In addition, a comparison of adult to embryonic brains revealed the presence of an incompletely processed form of Cancer borealis tachykinin-related peptide 1a (CabTRP 1a, APSGFLGMRG) only in the embryonic brain. A comparison of adult to embryonic STGs revealed that QDLDHVFLRFa was present in the embryonic STG but absent in the adult STG, and CabTRP 1a exhibited the opposite trend. Relative quantification of neuropeptides in the STG revealed that three orcokinin family peptides (NFDEIDRSGFGF, NFDEIDRSGFGFV, and NFDEIDRSGFGFN), a B-type allatostatin (STNWSSLRSAWa), and an orcomyotropin-related peptide (FDAFTTGFGHS) exhibited higher signal intensities in the adult relative to the embryonic STG. RFamide (Arg-Phe-amide) family peptide (DTSTPALRLRFa), [Val¹]SIFamide (VYRKPPFNGSIFa), and orcokinin-related peptide (VYGPRDIANLY) were more intense in the embryonic STG spectra than in the adult STG spectra. Collectively, this study expands our current knowledge of the H. americanus neuropeptidome and highlights some intriguing expression differences that occur during development.     

Cold-resistant changes in heartbeat of the Japanese spiny lobster

Heartbeat in Panulirus japonicus acclimated to 20°C is often augmented during cooling to 15^oC. Augmented contractions of the heart coincided with increasing amplitude of electrocardiogram. In cold saline, a pericardial hormone serotonin (10⁻⁷ M) increased both the amplitude and duration of the heartbeat while another hormone octopamine (10⁻⁶ M) slightly relieved the cold depression of heart rate despite a smaller increase in beat amplitude. In contrast, the application of the cold saline containing F1 (a FMRFamide-related peptide of pericardial hormones, 10⁻⁹ M) maintained the rate and amplitude of the heartbeat around the control level during cold exposure. This suggests that in the presence of F1, the lobster heart becomes cold resistant clearly. We previously reported that the pericardial organs of spiny lobsters are activated by a small fall in body temperature. The ligamental nerves, extensions of the pericardial organs, terminate in the heart beside the ostia and their ends remain in the isolated hearts. Therefore, the ligamental nerve ends might release their hormones into the ventricle with the fall in temperature even in the isolated hearts.     

Threshold determination for olfactory receptors of the spiny lobster

The shallow water caridean shrimps Heptacarpus pictus and H. paludicola are polymorphic in colour pattern. Populations of these species collected over colour‐variable substrates showed the greatest degree of coloration in terms of the proportion of individuals displaying a distinct colour pattern. The frequencies of H. Pictus colour morphs varied significantly between most sampling periods. Apostatic selection by fish predators is suggested as one hypothesis which could explain these changes in morph frequency.

Experiments with Heptacarpus pictus on rapid colour change showed that, although some pigment migration did occur in the chromatosomes studied, the macroscopic appearance of the colour patterns was not altered when shrimps were shifted from black to white backgrounds or vice versa. The results of background choice experiments with H. pictus suggest that these shrimp do not seek out colour backgrounds that would seem to be a matching background in a concealing coloration. This behaviour is consistent with the morphology of the colour patterns which appears to be a disruptive coloration rather than a concealing coloration that closely matches a particular substrate.