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PubMed1991

Neurons with histaminelike immunoreactivity in the segmental and stomatogastric nervous systems of the crayfish Pacifastacus leniusculus and the lobster Homarus americanus.

We used a polyclonal antiserum against histamine to map histaminelike immunoreactivity (HLI) in whole mounts of the segmental ganglia and stomatogastric ganglion of crayfish and lobster. Carbodiimide fixation permitted both HRP-conjugated and FITC-conjugated secondary antibodies to be used effectively to visualize HLI in these whole mounts. Two interneurons that send axons through the inferior ventricular nerve (ivn) and the stomatogastric nerve to the stomatogastric ganglion had strong HLI, both in crayfish and in lobster. These ivn interneurons were known from other evidence to be histaminergic. The neuropil of the stomatogastric ganglion in both crayfish and lobster contained brightly labeled terminals of axons that entered the ganglion from the stomatogastric nerve. No neuronal cell bodies in this ganglion had HLI. Each segmental ganglion contained at least one pair of neurons with HLI. Some neurons in the subesophageal ganglion and in each thoracic ganglion labeled very brightly. Axons of projection interneurons with strong HLI occurred in the dorsal lateral tracts of each segmental ganglion, and sent branches to the lateral neurophils and tract neurophils of each ganglion. All the labeled neurons were interneurons; no HLI was observed in peripheral nerves.

Animals↗
PubMed1981

Dendritic analysis of lobster stretch receptor neurons: electrotonic properties with single and distributed inputs.

Using steady-state cable analysis as derived by Rall, electrotonic properties of the dendritic trees of the tonic stretch receptor neurons of the spiny lobster. Panulirus interruptus, have been examined. By directly measuring the somatic input resistance and by visualizing the dendritic trees of this neuron by backfilling the axon with cobalt, the electrotonic properties of the dendritic trees have been derived. The Calculated membrane resistivity is 800-3600 omega-cm2. Voltage and current transfer functions were calculated for (a) single dendritic tips the size observed in the cobalt preparation and (b) for processes 2 micrometer or smaller, as observed in electron microscopy. Current transfer to the soma was high in both cases (greater than 80%). Voltage transfer was 22% for large and 4% for small dendrites. When a more natural simultaneous conductance change at the tips of all major dendrites was modeled, voltage transfer was 84% and current transfer 56%. But the dynamic range of the cell (rheobase to saturation) is well-predicted by varying the simultaneous inputs, not by scaling up a single input, thus illustrating that convenient indices of electrotonic properties may not prove useful in appreciating the integrative properties of a neuron.

Action Potentials↗
PubMed1995

Modelling nonlinear integration of synaptic signals by neurones.

Neurones with active conductance on dendrites integrate synaptic signals and modulate generation of axon spikes in a nonlinear way. Owing to experimental difficulties, modelling provides invaluable insight for the comprehension of neurone behaviour particularly when dendrites are excitable. We used experimental data obtained for the Anterior Gastric Receptor neurone (AGR neurone), which controls the lobster gastric mill activity, to derive a set of partial differential equations for the membrane voltage. Simulation showed that upon varying the intensity of stimulation on the dendrite, the response pattern between dendrites and axon activity continuously changes. In addition, when only half of the dendritic tree is active, axon firing exhibits regular oscillations and bursting activity. We discuss these results in relation with the experimental work done on the AGR neurone.

Animals↗
PubMed1984

Monoclonal antibodies to the Ca2+ + Mg2+-dependent ATPase of sarcoplasmic reticulum identify polymorphic forms of the enzyme and indicate the presence in the enzyme of a classical high-affinity Ca2+ binding site.

In order to determine whether polymorphic forms of the Ca2+ + Mg2+-dependent ATPase exist, we have examined the cross-reactivity of five monoclonal antibodies prepared against the rabbit skeletal muscle sarcoplasmic reticulum enzyme with proteins from microsomal fractions isolated from a variety of muscle and nonmuscle tissues. All of the monoclonal antibodies cross-reacted in immunoblots against rat skeletal muscle Ca2+ + Mg2+-dependent ATPase but they cross-reacted differentially with the enzyme from chicken skeletal muscle. No cross-reactivity was observed with the Ca2+ + Mg2+-dependent ATPase of lobster skeletal muscle. The pattern of antibody cross-reactivity with a 100,000 dalton protein from sarcoplasmic reticulum and microsomes isolated from various muscle and nonmuscle tissues of rabbit demonstrated the presence of common epitopes in multiple polymorphic forms of the Ca2+ + Mg2+-dependent ATPase. One of the monoclonal antibodies prepared against the purified Ca2+ + Mg2+-dependent ATPase of rabbit skeletal muscle sarcoplasmic reticulum was found to cross-react with calsequestrin and with a series of other Ca2+-binding proteins and their proteolytic fragments. Its cross-reactivity was enhanced in the presence of EGTA and diminished in the presence of Ca2+. Its lack of cross-reactivity with proteins that do not bind Ca2+ suggests that it has specificity for antigenic determinants that make up the Ca2+-binding sites in several Ca2+-binding proteins including the Ca2+ + Mg2+-dependent ATPase.

Animals↗
PubMed1984

Intramembrane organization of synapses in the lobster stretch receptor organ.

The intramembrane organization of axodendritic and neuromuscular synapses in the lobster stretch receptor organ was investigated by freeze-fracturing. Based on ultrastructural criteria which are known to be correlated with physiological properties, we identified three types of synapse: the inhibitory axodendritic, the inhibitory neuromuscular, and the excitatory neuromuscular synapse. Although these synapses have some features in common, each has a characteristic arrangement of intramembrane particles in both the presynaptic and postsynaptic membranes. All three have, in their presynaptic membranes, aggregates of P-face particles and associated depressions representing sites of synaptic vesicle exocytosis, features which together define active zones. However, in the inhibitory axodendritic synapse the P-face contains short ridges in this region. These ridges may occur singly or in pairs oriented in V-shaped configurations. The ridges are decorated with particles along their entire length. In the inhibitory neuromuscular synapse, no ridges are present. Clusters of particles are present, but they are scattered randomly over a large expanse of presynaptic membrane. In the excitatory neuromuscular synapse, isolated clusters of particles are associated with the P-face and are occasionally located on circular elevations of the membrane. The postsynaptic membrane also shows structural diversity in the three types of synapse. In the inhibitory axodendritic synapse, there is no apparent specialization. However, in the inhibitory neuromuscular synapse, P-face particles are arranged in double rows which are separated by particle-free strips of membrane. In the excitatory neuromuscular synapse, particles are confined to a narrow band that borders the synaptic cleft. This band is demarcated by a single intermittent strand of particles arranged in the direction of the long axis of the muscle fibre. Therefore, intramembrane specializations of both the presynaptic and postsynaptic membranes are sufficiently distinctive that three different types of synapse can be recognized.

Animals↗
PubMed1976

Organization of crustacean neuropil. I. Patterns of synaptic connections in lobster stomatogastric ganglion.

The stomatogastric ganglion of the lobster consists of about thiry neurons, mainly large monopolar cells, which have been well characterized physiologically. This paper presents an anatomical description of this ganglion, emphasizing synaptic connections in the neuropil. The neuron cell bodies are located on the dorsal surface of the ganglion. They send processes into the underlying neuropil mass. The neuropil is differentiated into two regions: a core of coarse neuropil consists of large heavily ensheathed processes; a surrounding region of fine-textured synaptic neuropil consists of smaller unsheather processes. Synapses are found only in synaptic neuropil, not in the core of coarse neuropil. Synaptic contacts, about one million in the entire neuropil, are easily recognized by a set of criteria including presynaptic vesicles and pre- and postsynaptic membrane specializations. Most synaptic contacts invole at least three neural processes, usually one pre- and two postsynaptic elements. Synapses are clustered onto irregular swellings or varicosities on neural processes. These varicosities make both pre- and postsynaptic contacts. Three differenty types of presynaptic profile are recognized. Pyloric dilator, ventricular dilator and lateral posterior gastric neurons belong to type A with clear irregular synaptic vesicles. Lateral pyloric, pyloric, anterior median and dorsal gastric neurons belong to type B with larger clear round vesicles. Many unidentified fibres, presumably stomatogastric nerve afferents, blong to type C with both small clear irregular vesicles and also large dense-core vesicles. The synaptic vesicle types are tentatively correlated with neurotransmitter: type A with acetylcholine, type B with an unknown transmitter, possibly glutamate, and type C with dopamine. The distribution of synaptic contacts on the processes of identified neurons reconstructed from serial section is presented in the following paper.

Animals↗
PubMed1976

Organization of crustacean neuropil. II. Distribution of synaptic contacts on identified motor neurons in lobster stomatogastric ganglion.

Identified neurons in the stomatogastric ganglion of the lobster were examined and reconstructed by serial section electron microscopy. Each motor neuron consists of a soma, a primary process leading directly from the soma to the motor axon which leaves the ganglion, and a group of secondary processes which branch from the primary process and ramify within the neuropil. Synapses are found only on small processes in the synaptic neuropil, never on the primary processes or on larger secondary processes in the coarse neuropil. Nearly every secondary process of every neuron examined makes both pre- and postsynaptic contacts. Hence these neurons are not polarized into distinct pre- and postsynaptic regions but have both input and output distributed over each of the secondary processes in the neuropil. The conncetion between a specific pair of neurons is also distributed over several branches of both the pre- and the postsynaptic neurons. The restriction of synapses to the more distal portions of the secondary processes suggests that no single contact or localized group of contacts can exert an overrriding influence on the neuron by virtue of an especially advantageous position. The close proximity of input and output on most secondary processes suggests that synaptic input may be capable of directly influencing output without the intervention of action potentials. The distribution of specific synapses over several branches of both pre- and postsynaptic neurons suggests that each neuron functions as a whole without differentiation into specialized branches.

Animals↗
PubMed1980

Lobster neuromuscular junctions treated with black widow spider venom: correlation between ultrastructure and physiology.

Black widow spider venom (BWSV) causes marked physiological and morphological alterations at the lobster neuromuscular junction. BWSV is also active at vertebrate neuromuscular junctions but the component which acts on the lobster preparation is different from the one which affects vertebrates. Following exposure to BWSV, lobster neuromuscular junctions showed elevated frequencies of spontaneous miniature synaptic potentials for 15-30 min. Nerve-evoked synaptic potentials became blocked during this period. Subsequently, spontaneous miniature potentials disappeared and less frequent 'giant' spontaneous potentials appeared. Ultrastructural examination of excitatory and inhibitory nerve terminals showed that both types were affected by venom treatment. In untreated terminals, synaptic vesicles were grouped near the dense specialized membranes of the synapses. Soon after venom treatment, the synaptic vesicles were dispersed throughout the terminals and many larger and elongated vesicular structures were apparent. At the time of appearance of 'giant' spontaneous potentials, few synaptic vesicles were seen in the terminals, but large irregular vacuoles were present. Many mitochondria within the nerve terminals were swollen or disrupted, while nearby muscle mitochondria remained normal in size and appearance. Very few presynaptic dense bodies ('active zones') were seen at synapses of affected terminals. The observations are consistent with the hypothesis that BWSV allows an abnormal amount of Ca2+ to enter the nerve terminals, causing the various physiological and morphological changes.

Animals↗
PubMed1979

Location of interganglionic neurons in the stomatogastric system of the spiny lobster.

The stomatogastric ganglion produces distinct and complex patterned output driving the mastication and filtration of food. It does so with a small number of neurons whose properties and interconnections have been extensively examined. The motor patterns are subject to modulation and integration by neurons of other ganglia. This paper reports a search for interneurons of the four interconnecting ganglia of this system, using cobalt chloride backfilling techniques. It was determined that only a small number of neurons may interconnect these ganglia: (1) In the stomatogastric ganglion there are two to three small neurons and six large neurons with neurites projecting anteriorly towards the other ganglia. (2) In the two commissural ganglia there are one to three small neurons whose neurites project to the primary input nerve of the stomatogastric ganglion. (3) In the oesophageal ganglion there are three small neurons whose location would allow them to play a co-ordinating role in the output of all four ganglia.

Animals↗
PubMed1991

Periaxonal ensheathment of lobster giant nerve fibres as revealed by freeze-fracture and lanthanum penetration.

Sheath structure and permeability have been studied in the nerve fibres of lobster (Panulirus argus) walking limbs, in particular the individually ensheathed larger giant fibres, 100-150 microns in diameter, of which there are five or six in a peripheral bundle. They are easily distinguished and can be separated from neighbouring fibre bundles in which smaller giant axons (65-80 microns diameter) and many axons of much smaller diameter (5-15 microns) are ensheathed together. Each of the larger giant axons is enveloped by a Schwann cell layer outside of which is a multilayered sheath consisting of one-cell thick belts of flattened cells and interleaved zones of collagen fibrils and extracellular matrix. The cells in each belt lack basal lamina and, after freeze-fracture, as well as in thin sections, exhibit intercellular gap junctions and incomplete, fascia type, tight junctions; their most striking aspect is an exceedingly large number of exo-endocytic profiles. Permeability to lanthanum chloride in the bathing medium studied before or during fixation both in intact nerves and in nerves with surgically breached (slit) epineurium showed penetration of lanthanum tracer between the cells around the giant fibres, but the electron-dense tracer was excluded from the Schwann cell layer and the periaxonal space unless the epineurium had been slit. The extent of lanthanum diffusion was evaluated by transmission electron microscopy of thin sections and confirmed by X-ray microanalysis (EDAX) of comparable selected areas in such sections. The results indicate structural similarities but distinct permeability differences between the multilayered sheath surrounding the lobster giant axons and the vertebrate nerve perineurium. Other ultrastructural details provided by the freeze-fracture replicas concern the distribution of intramembrane particles in the axolemma and the Schwann and sheath cell membranes.

Animals↗
PubMed1990

Identification of all GABA-immunoreactive neurons projecting to the lobster stomatogastric ganglion.

The stomatogastric ganglion of lobsters (Homarus or Jasus) contains a large number of gamma-aminobutyric acid-immunoreactive processes originating from ten fibres in the single input nerve, the stomatogastric nerve. The cell bodies and axonal pathways of these ten fibres have been identified using gamma-aminobutyric acid immunohistochemistry in combination with Lucifer Yellow staining (double labelling) and nickel chloride backfilling (selective gamma-aminobutyric acid immunoinhibition). It is shown that eight gamma-aminobutyric acid-immunoreactive neurons project to the stomatogastric ganglion: gamma-aminobutyric acid neurons 1 and 2, found posterior to the oesophageal ganglion, entering the stomatogastric nerve via the oesophageal nerve as well as sending an axonal branch into each superior oesophageal nerve; gamma-aminobutyric acid neurons 3 and 4, found anterior to the oesophageal ganglion, each sending an axonal branch into each inferior oesophageal nerve to reach the stomatogastric nerve via the commissural ganglion and the superior oesophageal nerve; and gamma-aminobutyric acid neurons 5 and 6, found in each commissural ganglion, projecting into the stomatogastric nerve via the inferior oesophageal nerve, the oesophageal ganglion and the oesophageal nerve. These gamma-aminobutyric acid-immunoreactive neurons were also characterized by electrophysiological methods coupled with Lucifer Yellow labelling, and their picrotoxin-sensitive effects on several stomatogastric ganglion neurons were demonstrated. The present results provide a firm basis for further studies concerning the physiological significance of one class of neurochemically-defined input neurons to stomatogastric ganglion networks.

Animals↗
PubMed1982

Differentiation of identifiable lobster neuromuscular synapses during development.

The ultrastructure of physiologically identified low and high release synapses arising from a single axon on fibres of the distal accessory flexor muscle (DAFM) in a mature lobster was examined by serial section electron microscopy. Low release neuromuscular terminals located only on the proximal fibre were characterized by large synapses (mean area 2.084 micron2), small presynaptic dense bars (mean are 0.021 micron2) and hence a low (2.3%) ratio of dense bar area to synaptic area. In contrast high output terminals located only on the distal fibre had smaller synapses (mean area 0.625 micron2), large dense bars (mean area 0.066 micron2) and a high (23.9%) ratio of bar area to synaptic area. A similar ratio was consistently found for each synaptic type in several other examples of mature lobsters. Hence it was used as a criterion for determining the point at which differentiation occurs during development. In the first larval stage (24 h old) the innervation was localized and undifferentiated. In the fourth (2 week old) and twelfth (1 y old) stage lobsters, the innervation had proliferated to small bundles of proximal and distal fibres. During development synapses increase in their mean surface area in the proximal fibre while remaining constant in the distal fibre. The mean surface area of the dense bars is similar in all stages except for the proximal fibres of the twelfth stage where it is smaller by 50%. Similarly the ratio fo dense bar area to synaptic area is not significantly different for all stages except for the twelfth stage proximal fibres where it is half the value. Consequently differentiation of low and high release neuromuscular terminals occurs by the twelfth stage with an increase in the mean surface area of synapses and a decrease in the mean surface area of dense bars. This morphological differentiation is enhanced in the mature lobster.

Animals↗
PubMed1982

Arrays of particles in the sarcolemma of the distal accessory flexor muscle of the lobster.

Freeze-fracture of the distal accessory flexor muscle reveals the presence of randomly distributed arrays of P- and E-face particles. The particles are arranged in regularly spaced rows which parallel one another and, in general, the long axis of the muscle fiber. P- and E-face arrays differ in their structure and are apparently complementary. A model explaining the apparent interrelationship of the P- and E-face arrays is presented. The functional significance of the arrays is not known.

Animals↗
PubMed1978

Axonal inclusions in the crab Hemigrapsus nudus.

Light microscopic examination of living giant axons from the walking legs of Hemigrapsus nudus revealed intra-axonal inclusions which were usually several tens of micrometers long and about 5 micron wide. The inclusions were filled with small light-scattering particles. The inclusions were shown, by thin section electron microscopy, to be composed largely 68% by volume) of mitochondria. Each inclusion was surrounded by membrane bounded spaces which are presumed to represent a part of the smooth endoplasmic reticulum. Similar inclusions were not found in the leg axons of a variety of other decapod crustaceans.

Animals↗
PubMed1986

Tactile stimulation of the swimmeret alters motor programs for abdominal posture in the lobster Homarus americanus.

The influence of mechanosensory stimulation of a second segment swimmeret upon the abdominal postural program was examined in an isolated abdominal nerve cord-swimmeret preparation. The swimmeret was stimulated in several different ways to assess the extent of influence exerted on abdominal positioning. Localized tactile stimulation of the swimmeret surface with a mechanical probe usually generated flexion inhibition where the flexor inhibitor (f5) was activated while the small and medium flexor excitors were inhibited. Flexion inhibition was much stronger in females than males. In 50% of the animals a weak flexion excitation was seen. After 3-6 hours the response of one-third of these preparations changed to flexion inhibition. Strong manual stimulation of the swimmeret surface inhibited all of the flexor excitors (f1, f2, f3, f4, and f6) while exciting the inhibitor f5 and increasing extensor activity. Similar extension responses were observed in both sexes. Repeated tactile stimulation of the swimmeret surface elicited a response similar to that evoked during manual stimulation. The strongest extension response was produced at 2 Hz which falls within the normal range of swimmeret beating in intact lobsters. Similar extension responses were also obtained during spontaneous swimmeret beating and rhythmic manual movement of the swimmeret.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Postural interneurons in the abdominal nervous system of lobster. I. Organization, morphologies and motor programs for flexion, extension and inhibition.

Using intracellular recording and dye-filling techniques, a survey of postural interneurons was undertaken by impaling their somata in the 2nd abdominal ganglion of lobster. During the course of study approximately fourty different intersegmental interneurons in this ganglion were sampled. Of these, 8 evoked unique, patterned responses in the postural (superficial) motoneurons; each could be identified morphologically. Five of the 8 interneurons had caudally directed axons; 4 of these projected beyond the 4th abdominal ganglion. The remainder projected rostrally, beyond the 1st abdominal ganglion. The postural interneurons were classified according to the motor program they elicited. Five were flexion producing interneurons (FPIs), one was extension producing (EPI), and two generated only inhibitory motor outputs. All motor responses were bilateral and occurred in several segments, including A2. Two neurons, FPIs 201 and 301, produced the full motor reciprocity that typically is observed when flexion command fibers are stimulated. However, three of the FPIs and the single EPI did not express complete reciprocity in synergistic and antagonistic motoneurons. The results indicate that some interneurons displaying all of the properties of command neurons are located entirely within the abdominal nervous system. The overall organization of posture-evoking interneurons appears to be similar to that found in crayfish, suggesting an even more fundamental homology in the neuronal connectivities of these two species than has been established previously.

Abdomen↗
PubMed1986

Postural interneurons in the abdominal nervous system of lobster. II. Evidence for neurons having both command and driver roles.

The nature of the synaptic relationship between 7 identified postural interneurons and 5 pairs of superficial motoneurons was examined by obtaining dual intracellular recordings from interneuron-motoneuron pairs in the lobster 2nd abdominal ganglion. For six different interneuron-motoneuron pairs EPSPs recorded from motoneurons occurred with a short (1 to 3 ms) fixed latency following each presynaptic spike recorded from the interneuron. This suggests that there is a monosynaptic relationship between these interneurons and motoneurons. Monosynaptic pathways accounted for 27% of all excitatory connections. Preliminary evidence indicates that the monosynaptic potentials are mediated by an excitatory chemical synapse since: all IPSPs occurred with latencies greater than 5 ms, there was no evidence for electrical coupling, and one of the interneurons produced facilitating PSPs. A majority of all monosynaptic connections were made by two of the flexion producing interneurons (FPIs), 201 and 301. The synaptic outputs of these FPIs were similar in that both made monosynaptic connections with a different bilaterally homologous pair of motoneurons. Both also produced larger EPSPs and more vigorous spiking in contralateral members of the bilateral motoneuron pairs. A previous study demonstrated that interneurons 201 and 301 are the only postural interneurons yet identified that express motor programs indistinguishable from command neurons. Taken together, these results suggest that certain intersegmental interneurons share properties with command neurons and driver neurons, and that there may not be a sharp morphological or functional distinction between these two cell types.

Abdomen↗