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PubMed1980

Morphology and ultrastructure of an identified histamine-containing neuron in the central nervous system of the pond snail, Lymnaea stagnalis L.

An identifiable histamine-containing neuron is located on the anterior dorsal surface of the visceral ganglion of Lymnaea stagnalis L. After [3H]histamine was injected into the perikaryon of this neuron, labelled axonal ramifications were seen in the neuropils of all ganglia except the pedals and right cerebral, and labelled axons occurred in seven nerve trunks. Electron microscopical examination of the perikaryon of the histamine neuron revealed the presence of large aggregations of granulated vesicles (80-200 microns diameter), elaborate endoplasmic reticulum, and numerous mitochondria, Golgi complexes and lysosome-like organelles.

Animals↗
PubMed1989

Voltage-independent barium-permeable channel activated in Lymnaea neurons by internal perfusion or patch excision.

Isolated nerve cells from Lymnaea stagnalis were studied using the internal-perfusion and patch-clamp techniques. Patch excision frequently activated a voltage-independent Ba2+-permeable channel with a slope conductance of 27 pS at negative potentials (50 mM Ba2+). This channel is not seen in patches on healthy cells and, unlike the voltage-dependent Ca channel, is not labile in isolated patches. The activity of the channel in inside-out patches is unaffected by intracellular ATP, Ca2+ below 1 mM or the catalytic subunit of cAMP-dependent protein kinase but is reversibly blocked by millimolar intracellular Ca2+ or Ba2+. The channel can be activated in on-cell patches by either internal perfusion with high Ca2+ or the long-term internal perfusion of low Ca2+ solutions not containing ATP. These channels may carry the inward Ca2+ current which causes a regenerative increase in intracellular Ca+ when snail neurons are perfused with high Ca2+ solutions. High internal Ca2+, or long periods of internal perfusion with ATP-free solutions, induces an increase in a resting (-50 mV) whole-cell Ba2+ conductance. This conductance can be turned off by returning the intracellular perfusate to a low Ca2+ solution containing ATP and Mg2+. The activity of this channel appears to have an opposite dependence on intracellular conditions to that of the voltage-dependent Ca channel.

Animals↗
PubMed1992

The Light Green Cells of Lymnaea: a neuroendocrine model system for stimulus-induced expression of multiple peptide genes in a single cell type.

We review recent experiments showing that the cerebral neuroendocrine Light Green Cells (LGCs) of the freshwater snail, Lymnaea stagnalis, express a family of distinct though related molluscan insulin-related peptide (MIP) genes. The LGCs are involved in the regulation of a wide range of interrelated life processes associated with growth, (energy) metabolism and reproduction. We consider the mechanism of generation of diversity among MIPs, and present evidence that conditions with distinct effects on growth, metabolism and reproduction also can induce distinct patterns of expression of the MIP and schistosomin genes. The stimulus-dependent expression of multiple neuropeptide genes enormously increases the adaptive potential of a peptidergic neuron. We suggest that this contributes significantly to the information-handling capacity of the brain.

Amino Acid Sequence↗
PubMed1978

Chaetogaster limnaei K von Baer 1872 on Lymnaea tomentosa: ingestion of Fasciola hepatica cercariae.

The fresh water mollusc, Lymnaea tomentosa, the intermediate host of the liver fluke is heavily infested under natural conditions with ectocommensal annelid, Chaetogaster limnaei. These annelids which destroy the larval stages of Fasciola hepatica have been observed in the laboratory. The association of C. limnaei with the snail intermediate host is of value in exploiting a control measure against economically important liver fluke disease.

Animals↗
PubMed1978

Ultrastructure, histology, and innervation of the mantle edge of the freshwater pulmonate snails Lymnaea stagnalis and Biomphalaria pfeifferi.

The mantle edge of the freshwater pulmonate snails Lymnaea stagnalis and Biomphalaria pfeifferi was investigated with histochemical and ultrastructural methods. The mantle edge gland, which is involved in shell formation, consists of the periostracal groove and the belt. This belt appears to be composed of various regions. In the area of the periostracal groove a number of subepithelial gland cell types occur; these release their products into the groove. Between the groove cells ciliated free nerve endings terminate; the corresponding perikarya occur in the subepidermal connective tissue. Also in the posterior belt region free nerve endings were observed between the epithelial cells; in addition, a particular type of subepithelial gland cell was found in this area. The epithelial cells of this part of the belt have the ultrastructural characteristics of ion and water transporting cells; they are probably involved in calcium deposition and resorption. The possible role of the free nerve endings and of the subepithelial gland cells is discussed.

Animals↗
PubMed1994

Effects of chronic exposure to cadmium- or lead-enriched environments on ionic currents of identified neurons in Lymnaea stagnalis L.

1. Voltage-activated ionic currents of three identified neurons of Lymnaea stagnalis L. were compared in control snails and in animals having been exposed to a cadmium- or lead-enriched environment for 2 weeks. We determined the presence, amplitude, and changes, if any, in the current-voltage characteristics of calcium and potassium currents in each of the three neurons from each of the three groups of animals. Finally, we have compared the effects of acute administration of Cd2+ or Pb2+ on neurons from control and chronically exposed animals. 2. Chronic exposure to cadmium resulted in a near doubling of the high voltage-activated calcium current. 3. No differences were found in the effects of acute application of Cd2+ or Pb2+ on neurons of pretreated and control animals. Cadmium was a potent blocker of the Ca current in either case, while lead caused only a 20% inhibition of the Ca current in neurons of both control and lead-exposed animals. 4. Potassium currents were affected in both Cd(2+)- and Pb(2+)-exposed animals. While the sustained outward current was not influenced noticeably, the fast K current was affected in different ways in different neurons. Some did not show this current in the controls but expressed it in neurons from the exposed animals. Other neurons showed the current in the controls and its depression in exposed animals. Acute application of cadmium did not modulate the K current, but lead enhanced the peak amplitude of the transient K current in neurons of both exposed and control snails.

Animals↗
PubMed1992

A review of the electrophysiological, pharmacological and single channel properties of heart ventricle muscle cells in the snail Lymnaea stagnalis.

Although a considerable body of information has accumulated describing the pharmacological properties of a wide range of molluscan muscle types, the physiological bases underlying these properties have not been thoroughly investigated. At present, little is known about the types of ion channels and their regulation in molluscan muscle cell membranes. Voltage-clamp, and more recently, patch-clamp techniques have revealed molluscan muscles possess a complex array of channel types with various pharmacological and electrophysiological properties. The gating properties of these channels and their modulation by chemical agents, however, are still poorly understood. This review summarizes some aspects of molluscan muscle function with particular reference to the heart ventricle muscle of the pond snail, Lymnaea stagnalis.

Animals↗
PubMed1996

Transient and sustained expression of FMRFamide-like immunoreactivity in the developing nervous system of Lymnaea stagnalis (Mollusca, Pulmonata).

1. In the present study we have investigated the ontogeny of FMRFamide expression in the snail, Lymnaea stagnalis, from its first appearance to its distribution in young adults. 2. The first FMRFamide-like immunoreactive (FaLI) cells within CNS appear by E45 embryonic stage (premetamorphic veliger). The number of FaLI neurons increases throughout both pre- and post-hatching development. 3. Both transient and sustained expression of FMRFamide-like immunoreactivity by specific sets of neurons occurs. Two cells which transiently express immunoreactivity appear outside the future CNS by the stage E45. Other population of transient FaLI neurons includes bilaterally symmetric groups of cells in the cerebral and pedal ganglia during posthatching stages P1 (hatchlings) to P5 (juveniles). All other immunostained cells which appear during development maintain their transmitter phenotype into adulthood. 4. The possible role of FMRFamide-related peptides in the processes of morpho- and neurogenesis is discussed.

Animals↗
PubMed1997

Characterization of an identified cerebrobuccal neuron containing the neuropeptide APGWamide (Ala-Pro-Gly-Trp-NH2) in the snail Lymnaea stagnalis.

A bilaterally symmetrical pair of cerebrobuccal neurons in Lymnaea stagnalis shows immunoreactivity for the molluscan neuropeptide APGWamide. The neuron somata are whitish in colour and located on the ventral surface of each cerebral ganglion between the roots of the labial nerves. A single axon travels via the ipsilateral cerebrobuccal connective into the buccal ganglia, where it gives rise to fine neuritic branching. Based upon these characteristics, the neuron has been named the cerebrobuccal white cell (CBWC). In isolated CNS preparations, in the absence of feeding motor output, the CBWC is silent and receives few, low amplitude, synaptic inputs. During generation of fictive feeding, the CBWC bursts in phase with cycles of feeding motor output. Tonic or phasic stimulation of CBWC leads to initiation of rhythmic feeding motor output. However, evoked bursts of activity in CBWC, which mimic its normal burst pattern, cannot entrain the buccal rhythm, suggesting that CBWC is not itself a major component of the feeding central pattern generator (CPG). Strong stimulation of CBWC during ongoing feeding motor output leads to a reduction in frequency and/or intensity of the buccal rhythm. Bath application of synthetic APGWamide (10(-7)M-10(-4)M) to the isolated CNS can activate feeding motor output in quiescent preparations after a delay, but disrupts ongoing buccal rhythms. This study represents the first description of a peptidergic cerebrobuccal neuron in the well described gastropod feeding system and also provides new information about the role of a novel molluscan neuropeptide.

Action Potentials↗
PubMed1995

The hybrid modulatory/pattern generating N1L interneuron in the buccal feeding system of Lymnaea is cholinergic.

This study examines neurotransmission between identified buccal interneurons in the feeding system of the snail Lymnaea stagnalis. We compare the pharmacology of the individual synaptic connections from a hybrid modulatory/pattern generating interneuron (N1L) to a pattern generating interneuron (N1M) with that from a modulatory interneuron (SO) to the same follower cell (N1M). The pharmacological properties of the N1L to N1M and the SO to N1M connections closely resemble each other. Both interneurons produce fast cholinergic EPSPs as judged by the blocking effects of cholinergic antagonists hexamethonium, d-tubocurarine and the cholinergic neurotoxin AF-64A. A slower, more complex but non-cholinergic component of the synaptic response is also present after stimulating either the presynaptic N1L or SO interneurons. This second component of the postsynaptic response is not dopaminergic, on the basis of its persistence in the presence of dopaminergic antagonists ergometrine and fluphenazine and the dopaminergic neurotoxin MPP+. We conclude that, although there has been an evolutionary divergence in function, the modulatory SO and the hybrid modulatory/pattern generating N1L are pharmacologically similar. Neither of them contributes directly to dopaminergic modulation of the feeding activity. These neurons also resemble the N1M protraction phase pattern generating neurons which are cholinergic (Elliott and Kemenes, 1992).

Animals↗
PubMed1981

A hormone dependent calcium-binding protein in the mantle edge of the freshwater snail Lymnaea stagnalis.

By means of gel filtration and ion exchange chromatography on calcium-saturated Chelex-100, a calcium-binding fraction was isolated from the mantle edge of the freshwater snail lymnaea stagnalis. This fraction was not present in other tissues. Treatment with trypsin caused a disappearance of the calcium-binding capacity, proving that the active substance in this fraction is a protein (calcium-binding protein; CaBP). Removal of the growth hormone-producing neuroendocrine light green cells resulted in a strong decrease of the amount of CaBP. It is concluded that L. stagnalis possesses a hormone-dependent CaBP, probably responsible for the maintenance of a high calcium concentration in that part of the mantle that produces the outer crystalline layer of the shell.

Animals↗
PubMedDate not supplied

Transmitter-dependent switching of respiratory interneurons to the locomotor rhythm in the pulmonate mollusk Lymnaea.

Cell RPeD1 of the pond snail Lymnaea stagnalis is a dopaminergic neuron known to be involved in generating the respiratory rhythm. This report describes conditions in which RPeD1 follows the rhythm of another central generator, identified as the generator of muscular locomotion. This rhythmic activity of RPeD1 developed in isolated CNS preparations treated with 0.05 mM serotonin (5-hydroxytryptamine, 5-HT) or 0.1 mM 5-hydroxytryptophan (5-HTP). It was coordinated by cyclic synaptic currents with the activities of other pedal neurons and with trains of spikes in the motor (pedal and columellar) nerves. Only the pedal ganglia (both of them) retained the ability to generate 5-HT-dependent cyclic activity after complete isolation, indicating a pedal localization and paired nature for the central generator. In intact animals, injection of 100 microg/g of 5-HTP induced coordinated cyclic movements of the foot and shell which had similar periods and resembled movements during terrestrial (non-ciliary) locomotion. These results, along with the known role of dopamine in generating the respiratory pattern, show that switching of RPeD1 from one central rhythm to another is a transmitter-dependent phenomenon.

5-Hydroxytryptophan↗
PubMed2000

Electrical synapses by guided growth of cultured neurons from the snail Lymnaea stagnalis.

The ability to assemble neuronal networks with designed topology would allow uniquely defined experiments on neurocomputing. We describe a fundamental step, the controlled formation of synapses by guided outgrowth, in vitro for the first time combining simple neuritic geometry with predefined connectivity. We used neurons from the A-clusters in the pedal ganglia of the snail Lymnaea stagnalis. They were cultured on a substrate with linear patterns made by adsorption of brain-derived conditioning factors and photolithography. We induced and observed the frontal collision of two growth cones on narrow lanes. Following such encounters. individual electrical synapses formed that were sometimes strong enough for prolonged presynaptic stimulation to reach the threshold of postsynaptic firing.

Action Potentials↗
PubMed2004

Multilevel inhibition of feeding by a peptidergic pleural interneuron in the mollusc Lymnaea stagnalis.

The pleural interneuron PlB is a white neuron in the pleural ganglion of the snail Lymnaea. We test the hypothesis that it inhibits neurons at all levels of the feeding system, using a combination of anatomy, physiology and pharmacology. There is just one PlB in each pleural ganglion. Its axon traverses the pedal and cerebral ganglia, running into the buccal ganglia. It has neuropilar branches in the regions of the cerebral and buccal ganglia where neurons that are active during feeding also branch. Activation of the PlB blocks fictive feeding, whether the feeding rhythm occurs spontaneously or is driven by a modulatory interneuron. The PlB inhibits all the neurons in the feeding network, including protraction and retraction motoneurons, central pattern generator interneurons, buccal modulatory interneurons (SO, OC), and cerebral modulatory interneurons (CV1, CGC). Only the CV1 interneuron shows discrete 1:1 IPSPs; all other effects are slow, smooth hyperpolarizations. All connections persist in Ca(2+)/Mg(2+)-rich saline, which reduces polysynaptic effects. The inhibitory effects are mimicked by 0.5 to 100 micromol l(-1) FMRFamide, which the PlB soma contains. We conclude that the PlB inhibits neurons in the feeding system at all levels, probably acting though the peptide transmitter FMRFamide.

Action Potentials↗
PubMed2004

Octopamine-containing (OC) interneurons enhance central pattern generator activity in sucrose-induced feeding in the snail Lymnaea.

In the pond snail Lymnaea stagnalis octopamine-containing (OC) interneurons trigger and reconfigure the feeding pattern in isolated CNS by excitation of the central pattern generator. In semi-intact (lip-mouth-CNS) preparations, this central pattern generator is activated by chemosensory inputs. We now test if sucrose application to the lips activates the OC neurons independently of the rest of the feeding central pattern generator, or if the OC interneuron is activated by inputs from the feeding network. In 66% of experiments, sucrose stimulated feeding rhythms and OC interneurons received regular synaptic inputs. Only rarely (14%) did the OC interneuron fire action potentials, proving that firing of OC interneurons is not necessary for the sucrose-induced feeding. Prestimulation of OC neurons increased the intensity and duration of the feeding rhythm evoked by subsequent sucrose presentations. One micromolar octopamine in the CNS bath mimicked the effect of OC interneuron stimulation, enhancing the feeding response when sucrose is applied to the lips. We conclude that the modulatory OC neurons are not independently excited by chemosensory inputs to the lips, but rather from the buccal central pattern generator network. However, when OC neurons fire, they release modulatory octopamine, which provides a positive feedback to the network to enhance the sucrose-activated central pattern generator rhythm.

Animals↗
PubMed2003

Construction of a backcross progeny collection of dextral and sinistral individuals of a freshwater gastropod, Lymnaea stagnalis.

The handedness of gastropods is genetically determined, but the molecular nature of the gene responsible remains unknown. In order to identify the gene by a forward genetic approach, we carried out backcross breeding between dextral and sinistral inbred strains of a freshwater gastropod, Lymnaea stagnalis, cultivated in the laboratory. We used the dextral animals as donor "fathers" and the sinistral animals as recurrent "mothers". Each of the backcross progeny obtained was typed for the chirality of the next generation offspring oviposited by it, because the genotype for the handedness locus emerges as filial chirality. We constructed a collection of DNA and RNA specimens that included about 200 of the backcross F(2) progeny. Success in breeding was confirmed by the expected inheritance behavior of strain-specific DNA markers in the progeny.

Animals↗
PubMed2004

Isolation and evaluation of dextral-specific and dextral-enriched cDNA clones as candidates for the handedness-determining gene in a freshwater gastropod, Lymnaea stagnalis.

The handedness of gastropods is genetically determined. The freshwater gastropod Lymnaea stagnalis is a normally dextral species, but contains minor sinistral populations. The gene responsible for handedness determination in this species is predicted to function maternally and specifically in the dextral-ovipositing snail. In this study, we used differential screening and cDNA subtraction to isolate eight "dextral genes" that are specific to, or enriched in, the dextral-ovipositing strains of L. stagnalis. These genes were promising candidates for the handedness-determining gene. In order to determine whether the true handedness-determining gene was among them, we tested for genetic correlations between the level of expression of each dextral gene and the handedness phenotype, i.e., the chirality of the next generation offspring, by using a collection of backcross F2 progeny of F1 offspring from crosses between dextral and sinistral strains. Although the present study could not identify the handedness-determining molecules, this approach appears to be promising for the isolation of such developmentally important genes.

Amino Acid Sequence↗
PubMed2002

Highland populations of Lymnaea truncatula infected with Fasciola hepatica survive longer under experimental conditions than lowland ones.

A retrospective study was carried out on the experimental infections of Lymnaea truncatula with Fasciola hepatica performed over the last 20 years to determine if the populations of snails living in highland or lowland countries had the same ability to sustain trematode larval development. The six highland populations originated from the Peruvian Altiplano (altitude 2,800 m), the French Alps (2,300 m), and the Massif Central (900-1,400 m), whereas the 13 lowland populations came from different sites located in central France (90-250 m). Bimiracidial infections of 4-mm-high snails were performed to study cercarial shedding and to quantify their redial burden. Compared to lowland populations, snail survival at day 30 post-exposure was significantly higher in the highland L. truncatula (57-75% compared to 31-45%) and their lifespan was greater (a mean of 87-96 days for cercaria-shedding snails compared to 64-77 days). The prevalences of F. hepatica infections, the numbers of free rediae within snail bodies, and the numbers of cercariae did not show any significant differences between highland and lowland snails although the numbers of cercariae were clearly higher in the Peruvian and three French highland populations of L. truncatula. The long survival times of highland snails under laboratory conditions might be an adaptation of these L. truncatula to the more extreme highland climate. The better ability of highland snails to sustain parasite larval development suggests that they would be better intermediate hosts in the life cycle of F. hepatica than lowland populations.

Animals↗