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P R Benjamin

Publications and source records attributed to P R Benjamin.

At least 73 records · Page 4Linked to original sources

Cardioactive neuropeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) and novel related peptides are encoded in multiple copies by a single gene in the snail Lymnaea stagnalis.

The neuropeptide Phe-Met-Arg-Phe-NH2 (FMRFamide) is a potent cardioactive neuropeptide in Lymnaea stagnalis. Isolation and sequencing of 2 cDNAs and a genomic clone shows that a single gene encodes a precursor protein which contains 9 copies of the FMRFamide peptide, 2 copies of the related peptide Phe-Leu-Arg-Phe-NH2 (FLRFamide), and single copies of the putative pentapeptides Gln-Phe-Tyr-Arg-lle-NH2 (posttranslationally modified to pQFYRlamide) and Glu-Phe-Leu-Arg-lle-NH2 (EFLRlamide). The gene is transcribed in the CNS and gives rise to a single RNA of 1.7 kb in size. The organization of the Lymnaea gene is significant with respect to the evolution of FMRFamide and related peptides in other organisms.

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Appetitive learning in snails shows characteristics of conditioning in vertebrates.

The snail Lymnaea stagnalis can be rapidly conditioned to produce feeding response to a previously neutral tactile stimulus, repeatedly and specifically paired with positive reinforcement (food). The conditioned response has many of the same characteristics seen in appetitive learning in vertebrates. The relative simplicity of the molluscan CNS and the detailed knowledge of the neural circuitry underlying feeding in Lymnaea will facilitate the neurophysiological analysis of appetitive learning as well as stimulus generalization, discriminative learning and generalization of extinction all described in the present paper.

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A comparison of four techniques for mapping the distribution of serotonin and serotonin-containing neurons in fixed and living ganglia of the snail, Lymnaea.

The distribution of serotonin and serotonin-containing neurons was studied in the ganglia of the CNS of the snail Lymnaea stagnalis. Results of the application of three different labelling techniques on wholemount preparations were compared with each other and with the serotonin content of the ganglia, measured by high-performance liquid chromatography. Serotonin immunocytochemistry resulted in the highest number of labelled neurons, but the more recently developed in vivo method of 5,6- or 5,7-dihydroxytryptamine-induced pigmentation also proved to be a reliable technique for the visualization of serotonin-containing cell bodies. In comparison with these two techniques, the glyoxylic acid fluorescence method appeared to be less sensitive. The distribution and number of serotonin-containing neurons and biochemically measured serotonin in specific ganglia showed a close correlation. By combining the results of the three labelling techniques, a detailed map of serotonin-containing neurons was constructed, and this was compared with maps of identified neurons prepared from earlier electrophysiological studies. Previously described serotonergic neurons were consistently found, as well as several new serotonin-containing cell types in the cerebral, visceral and parietal ganglia. A network of serotonin-containing inter- and intraganglionic axon tracts, and thin serotonergic fibres in the perineurium were also demonstrated. This in vivo and in vitro identification of serotonin-containing neurons will facilitate further neurophysiological analysis of serotonergic neural mechanisms in Lymnaea.

5,6-Dihydroxytryptamine↗

Goal-tracking behavior in the pond snail, Lymnaea stagnalis.

The occurrence of goal-tracking, an unconditioned stimulus (US)-directed autoshaping behavior, was studied in open-field tests with control and classically conditioned pond snails, Lymnaea stagnalis. In an appetitive classical conditioning paradigm with a tactile stimulus as conditioned stimulus (CS) and a localized food stimulus as US a conditioned feeding response built up in the experimental but not in the control animals. In the post-training open-field tests the experimental group alone showed an enhanced attraction toward the source of water current in the environment which previously signalled the arrival of the US but did not act as CS in the classical conditioning procedure. We suggest that this stimulus-directed goal-tracking behavior in Lymnaea is the result of a classical-operant interaction, described so far only in vertebrate animals, and that neurophysiological analysis of this behavior is possible in this snail.

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Esophageal mechanoreceptors in the feeding system of the pond snail, Lymnaea stagnalis.

1. We identify esophageal mechanoreceptor (OM) neurons of Lymnaea with cell bodies in the buccal ganglia and axons that branch repeatedly to terminate in the esophageal wall. 2. The OM cells respond phasically to gut distension. Experiments with a high magnesium/low calcium solution suggest that the OM neurons are primary mechanoreceptors. 3. In the isolated CNS preparation, the OM cells receive little synaptic input during the feeding cycle. 4. The OM cells excite the motoneurons active in the rasp phase of the feeding cycle. 5. The OM cells inhibit each of the identified pattern-generating and modulatory interneurons in the buccal ganglia. Experiments with a saline rich in magnesium and calcium suggest that the connections are monosynaptic. 6. Stimulation of a single OM cell to fire at 5-15 Hz is sufficient to terminate the feeding rhythm in the isolated CNS preparation. 7. We conclude that these neurons play a role in terminating feeding behavior.

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The electrotonic location of low-resistance intercellular junctions between a pair of giant neurones in the snail Lymnaea.

The passive electrotonic properties of neurones VD1 and RPD2 in the brain of the snail Lymnaea can be represented by a soma-finite cable model with closed-circuit axon termination. There is a considerable individual variation in input resistance, membrane time constant, electrotonic length and axon-soma conductance ratio, but the average values for these parameters are similar in the two neurones. The cells are tightly coupled by an electrotonic synapse giving an average steady-state coupling coefficient of 0.68 and an average resistance measured between recording sites in the cell bodies of 20 M omega. Calculations using a model consisting of a symmetrical pair of cells with standard values for the electrotonic parameters show that in this system, for a soma-soma resistance of 20 M omega, the junction cannot be more than 0.16 length constants from the cell bodies. Reduction in coupling due to membrane current losses in such short proximal axon segments is insignificant. Intra-axonal recordings indicate that most of the coupling resistance is located at the junction between VD1 and RPD2, which must therefore be closer to the cell bodies than the limiting value of 0.16 length constants assuming an electrical equivalent model which includes the standard electrotonic parameters. If all the soma-soma resistance is located at the junction, then it could be physically a single array of gap-junction particles. Despite its low conductance (1/20 M omega = 50 nS) and possibly small physical dimensions, the electrotonic synapse is more than sufficient to ensure spike synchrony in the two cells.

Action Potentials↗

Interactions of pattern-generating interneurons controlling feeding in Lymnaea stagnalis.

Intracellular recordings were made from rhythm-generating interneurons in the Lymnaea feeding system. The feeding pattern is a three-phase rhythm of interneuronal activity (N1, N2, N3) corresponding to protraction, rasp, and swallow. We describe the firing pattern and anatomy of the premotor interneurons, each of which fires a predominant burst in only one phase of the feeding rhythm. The rhythm can be driven by steady depolarization of N1 cells. The phase of the rhythm is reset by brief stimulation of N2 or N3 interneurons. N1 neurons excite the N2 interneurons, and these in turn inhibit the N1 cells. This recurrent inhibitory pathway can account for the switch from the N1 phase of the feeding cycles to the N2 phase. The endogenous properties of the N2 interneurons are apparently responsible for the termination of N2 bursts. N3 interneurons display postinhibitory rebound (PIR), and this probably contributes to their burst after the end of the N2 inhibitory input. N2 and N3 interneurons inhibit the N1 cells. When the N3 burst dies away, activity in N1 cells resumes under the stimulus of depolarizing current. Interactions between interneurons are mainly by discrete, monophasic postsynaptic potentials, that follow 1:1. They have relatively short latency (2-12 ms) and duration (up to 100 ms). The synaptic connections between the three types of premotor interneurons are sufficient to account for the sequence of activity seen during feeding.

Action Potentials↗

Interactions of the slow oscillator interneuron with feeding pattern-generating interneurons in Lymnaea stagnalis.

We have used intracellular recording from groups of interneurons in the feeding system of the pond snail, Lymnaea stagnalis, to examine the connections of a modulatory interneuron, the slow oscillator (SO), with the network of pattern-generating interneurons (N1, N2, and N3). The SO is an interneuron whose axon branches solely within the buccal ganglia. There is only one such cell in each snail. In half the snails the cell body is in the right buccal ganglion and in the other half in the left buccal ganglion. Stimulation of either the SO or one of the N1 pattern-generating interneurons elicits the feeding rhythm, but of all the buccal neurons, only the SO can drive the feeding rhythm at the frequency seen in the intact snail. The SO makes reciprocal excitatory synapses with the N1 interneurons that drive the protraction of the radula. This ensures strong activation of the feeding system. The SO inhibits the N2 interneurons. Postsynaptic potentials evoked by stimulation of the SO facilitate without spike broadening in the SO. The SO is strongly inhibited by N2 and N3 interneurons, which are active during the retraction phase. This gates any excitatory inputs to the SO, probably preventing protraction of the radula while retraction is underway. The results support the idea of a single interneuron capable of driving a hierarchically organized motor system.

Action Potentials↗

Electrotonic coupling and afterdischarges in the Light Green Cells: a comparison with two other cerebral ganglia neurosecretory cell types in the pond snail. Lymnaea stagnalis.

Electrotonic coupling occurs between the growth hormone-producing Light Green Cells on the same but not the opposite side of the brain of Lymnaea. Long duration inhibitory post synaptic potentials occur spontaneously or can be evoked by nerve or connective stimulation. Afterdischarges, lasting for up to 1 min, are evoked by brief stimulation of the median lip nerves. Activity is confined to cells ipsilateral to the nerve being stimulated. The electrical properties of the Light Green Cells are compared with those of the Caudodorsal Cells and Canopy Cells.

Action Potentials↗

Coupled neuronal oscillators in the snail Lymnaea stagnalis: endogenous cellular properties and network interactions.

The role of endogenous cellular properties and network interactions due to electrotonic coupling were investigated in two bilateral populations of 2-7 peripheral neurones ('Peripheral Bursters') in the snail Lymnaea stagnalis. 1. These cells are endogenously capable of bursting. Their burst frequency does not depend on the level of steady membrane polarization. Short hyperpolarizing current pulses injected during the bursting cycle induce phase advance and no phase delay in subsequent cycles, the phase advance being a function of the phase of stimulus application. Phase response and inter-burst interval curves have been constructed for short hyperpolarizing current pulses. Their shape depends on the intensity and sign of tonically injected current. This property of Peripheral Bursters is one reason for the independence of period duration from membrane polarization. 2. Coordination of burst activity of Peripheral Bursters has been studied as a function of coupling strength: whereas homolateral cells are relatively strongly electrotonically coupled and always burst synchronously, thus forming a single oscillatory unit, heterolateral cells are only weakly electrotonically coupled. Accordingly, they show more variable, though coordinated, patterns of activity. Most commonly, periods with 1:1 entrainment alternate with periods with 1:2 entrainment. The duration and frequency of occurrence of either mode of entrainment are highly variable in different preparations.

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Patterns of activity and axonal projections of the cerebral giant cells of the snail, Lymnaea stagnalis.

1. The paired cerebral giant cells (CGCs) of Lymnaea were studied using electrophysiological and anatomical techniques. 2. A strong electrotonic junction coupling the CGCs is located in the buccal ganglia, several millimetres distal to the cell bodies. This leads to 1:1 spike activity in the two cells. 3. The CGCs receive phasic and tonic inputs which appear to be common to the two cells. Two types of phasic input are synchronous with synaptic input to feeding motoneurones in the buccal ganglia. 4. Morphological studies, using three separate dye injection techniques, confirm the axonal projections shown by electrophysiological methods. The CGCs project ipsilaterally into one lip nerve, the cerebrobuccal connective, and all buccal nerves. 5. Symmetry of anatomy and of firing activity cause the CGCs to function as a single neural unit.

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Synaptic relationships of the cerebral giant cells with motoneurones in the feeding system of Lymnaea stagnalis.

1. The cerebral giant cells (CGCs) of Lymnaea have a tonic, modulatory effect on the intensity of output from feeding motoneurones in the buccal ganglia. 2. Short latency, excitatory and probably monosynaptic connexions occur between the CGCs and three identified feeding motoneurones. Unitary excitatory postsynaptic potentials in these motoneurones, following CGC spikes, are of different sizes and durations, and hence have different summation properties. 3. The CGCs make long latency, excitatory polysynaptic connexions with four other feeding motoneurone types. 4. Bursts of spikes in the CGCs, resulting from phasic synaptic input, synchronous with the feeding cycle, amplify their modulatory effect on burst intensity in feeding motoneurones. 5. Thte for reinforcing their cyclic burst activity.

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The relationship of the central motor pattern to the feeding cycle of Lymnaea stagnalis.

Electromyographic recordings from the buccal muscles of Lymnaea during feeding has shown that there are 4 component phases in the feeding cycle. Cinephotography of feeding cycles has confirmed that these correspond to protraction, 2 phases of retraction, and an inactive phase. The 4 phases of muscle activity can also be related to the cycle of neural activity described previously (Benjamin & Rose, 1979). Thus types 6, 4 group, and type 8 cells are motoneurones involved in protraction and the two retraction phases, while the type 5 cell fires in the inactive period. The combination of physiological and anatomical approaches has led to the suggestion that the single and double input cells described by Benjamin & Rose (1979) are involved with the control of buccal and oesophageal activity respectively.

Action Potentials↗

Central generation of bursting in the feeding system of the snail, Lymnaea stagnalis.

The central generation of burst activity was investigated in the buccal ganglia of Lymnaea. Eight different patterns of burst activity were generated by one or two consecutive phases of compound synaptic potentials resulting from activity of neurones. These inputs acted upon the different endogenous properties of buccal neurones such as post-inhibitory rebound and spike adaptation. Effects of synaptic inputs were reinforced by by electrotonic coupling of some buccal neurones of the same type.

Action Potentials↗

Electrophysiology of "yellow cells," neurosecretory neurones in Lymnaea.

The thirty Yellow Cells of Lymnaea show single, double and other extra spike modes of firing. Yellow Cell bursts consist of various combinations of single, doublet and triplet spikes whose number per burst varies spontaneously. Single spike firing modes of activity can be converted into doublets or bursts (and vice versa) by applying steady currents of the appropriate polarity. Spike activity is basically endogenous although it is modulated by low frequency synaptic input originating from within the brain. Interburst interval is affected by the number of spikes occurring in the preceding burst. This varies spontaneously or can be induced by applying appropriately timed current pulses or occurs following synaptic input. Excitatory synaptic input often induces bursts which far exceed the duration of the input and which are followed by long periods of inhibition.

Action Potentials↗

Environmental osmolarity and neurosecretory neurones in Lymnaea stagnalis (L.).

Cell body volume and Alcian blue-Alcian Yellow staining properties of neurosecretory neurones in the brain of Lymnaea stagnalis were compared for snails kept in de-ionised water and standard tapwater. In the same experiment, the ionic content of the blood, blood volume and body weight and environmental ionic composition were measured. Five days of immersion in de-ionised water resulted in significant decreases in body weight, blood volume and blood, Na+ and Cl- concentrations but no change in blood Ca2+, K+,and HCO3- concentrations, compared with controls. No consistently significant differences across the 5 day period were found in cell body volumes for Dark Green Cells, Yellow Cells or Light Green Cells (used as a control) when these volumes were compared for large numbers of cells from snails kept in de-ionised water and standard tapwater. However, the number of Yellow Cells which could be counted in snails kept in de-ionised water was lower than the number from standard tapwater by day 2 of the experiment and lower for Yellow-green Cells by day 5. We interpret this lower number to be the result of depletion of Alcian blue-Alcian Yellow stained neurosecretory material in these cells which made them impossible to distinguish. This was confirmed by examination of visceral Yellow Cells which could be identified on the basis of known location close to the visceral-right parietal connective.

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