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PubMed1987

Morphology of neurosecretory cells in basommatophoran snails homologous with egg-laying and growth hormone-producing cells of Lymnaea stagnalis.

In a light and electron microscope study, neurosecretory cells morphologically homologous with the egg-laying hormone-producing caudodorsal cells (CDC) and growth hormone-producing dorsal cells (DC, light green cells) of the freshwater basommatophoran snail Lymnaea stagnalis have been found in five genera (seven species) of Basommatophora, viz. in Lymnaea palustris and Lymnaea ovata, in Planorbis planorbis and Planorbis vortex, in Planorbarius corneus, in Bulinus truncatus, and in Biomphalaria glabrata. It is concluded that the functions of these cells are homologous as well. The homologies of the respective neuron types regard their locations in the cerebral ganglia, their clustering in groups, location of their neurohemal area (CDC: cerebral commissure; DC: median lip nerves), and ultrastructural characteristics (e.g., abundance of rough endoplasmic reticulum, well-developed Golgi apparatus, presence of two types of neuron-specific secretory granules, and release of granule contents by exocytosis into the hemolymph). In addition, CDC show large electron-dense granules and DC reveal infoldings of the plasma membrane at the abaxonal side of the soma as well as synaptic input. On the other hand, each neuron type shows species-specific characteristics, particularly with regard to the number of cells and the structure of the neurohemal area. Furthermore, the CDC show marked differences between genera in the morphology (especially the mean diameter) of type 2 and, particularly, type 1 secretory granules. The morphology of the two types of secretory granules in the DC differs strongly between species. The possible relation between the morphology and the chemical contents of secretory granules has been discussed.

Animals↗
PubMed1988

The bioelectrical activity of the body wall of the pulmonate freshwater snail Lymnaea stagnalis: effects of neurotransmitters and the sodium influx stimulating neuropeptides.

A method is described to dissect segments of the head skin of Lymnaea stagnalis. These skin segments were used to study the effects of neurotransmitters and of the Lymnaea sodium influx stimulating (SIS) peptides on ion transport, using the Ussing-cell technique. The electrical activity of the segments, with Lymnaea ringer solution at both sides, was low: the mean electrical potential difference (PD) across the skin was 0.7 +/- 0.6 mV (inside positive) at a resistance of 160 +/- 57 ohm.cm2 (inward short-circuit current, SCC, 4.2 +/- 3.0 microA/cm2; n = 25). Acetylcholine, adrenalin, noradrenalin, histamine, dopamine, and GABA, at 10(-5) M, did not affect skin resistance and PD. 5-Hydroxytryptamine-HCl (5-HT), however, in a dose-dependent way, increased the PD, SCC, and, to a much lesser extent, the resistance of skin segments. Extracts of the medium lip nerves, which contain the SIS peptides, had similar effects, but of much longer duration. Effects comparable to those of 5-HT and the SIS peptides could also be brought about by cAMP analogs. The inward current stimulation by 5-HT, SIS peptides, and cAMP was abolished by ouabain. The inward current induced by 5-HT and the SIS peptides was partly (10-25%) inhibited by amiloride. The presence of tetrodotoxin (10(-6) M) did not prevent the inward current stimulation by 5-HT and the SIS peptides.

Amiloride↗
PubMedDate not supplied

The brain of Lymnaea contains a family of FMRFamide-like peptides.

Authentic FMRFamide and two FMRFamide-related heptapeptides were purified from the central nervous system of the fresh water snail Lymnaea stagnalis. The sequences of the heptapeptides were determined as: Ser-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (SDPFLRFamide) and Gly-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (GDPFLRFamide) by modified Edman degradation and enzymatic digestion. Relatively high quantities of the deamidated and therefore non-immunoreactive analogs of these two peptides (SDPFLRF and GDPFLRF) were also found. SDPFLRFamide and GDPFLRFamide were synthesized and were found to be chromatographically and biologically indistinguishable from the natural peptides, confirming the sequences. The log dose-response curves for the chronotropic action of either synthetic peptide on the heart of Lymnaea was very similar to that of FMRFamide. These data indicate that Lymnaea contains a family of FMRFamide-like peptides.

Amino Acid Sequence↗
PubMed1996

Glutamate as a putative neurotransmitter in the mollusc, Lymnaea stagnalis.

Bath-applied glutamate (10-1000 microM) produced excitatory and inhibitory responses on numerous identified neurons of the mollusc Lymnaea stagnalis. Using both in situ and in vitro preparations, glutamate or glutamate agonists produced a depolarization in identified neurons right pedal dorsal 1 and right pedal dorsal 2 and 3. However, attempts to block glutamate-evoked responses with glutamate antagonists were unsuccessful. We examined a potential glutamatergic neuron, visceral dorsal 4. Exogenous application of the peptides (GDPFLRFamide and SDPFLRFamide) could mimic the inhibitory, but not the excitatory effects of visceral dorsal 4 on its postsynaptic cells, implying the presence of a second transmitter. We tested the possibility that glutamate is this second neurotransmitter by using excitatory synapses between visceral dorsal 4 and postsynaptic cells right pedal dorsal 2 and 3, right pedal dorsal 1, visceral F group and right parietal B group neurons. Of all the putative neurotransmitters tested, only glutamate had consistent excitatory effects on these postsynaptic cells. Also, the amplitude of the right pedal dorsal 2 and 3 excitatory postsynaptic potentials was reduced in the presence of N-methyl-D-aspartate and other glutamate agonists, suggesting desensitization of the endogenous transmitter receptor. In conclusion, some identified Lymnaea neurons respond to glutamate via a receptor with novel pharmacological properties. Furthermore, a Lymnaea interneuron may employ glutamate as a transmitter at excitatory synapses.

Animals↗
PubMed1993

Effects of met-enkephalin on electrical coupling between identified neurons in the pulmonate snails, Helix and Lymnaea.

1. The effects of met-enkephalin on electrical coupling between molluscan neurons have been investigated using the isolated brains of Helix pomatia and Lymnaea stagnalis. 2. In the presence of both serotonin and met-enkephalin, non-rectifying electrical coupling is strongly facilitated between identified respiratory neurons in Helix, whilst coupling between putative, serotonin-containing, ciliomotoneurons in Lymnaea is facilitated by met-enkephalin alone. 3. Facilitation of coupling by met-enkephalin is weaker in the strongly coupled neurons, VD1/RPaD2 of Lymnaea. 4. These data suggest that met-enkephalin can modulate different groups of electrically coupled cells and may be involved in coordination of motor patterns.

Animals↗
PubMed2003

In vivo metabolism and organ distribution of a branched 14C-nonylphenol isomer in pond snails, Lymnaea stagnalis L.

The branched isomers of p-nonylphenol (NP) are perceived to be more resistant to biodegradation in aquatic environments as well as to have more estrogen-like toxicity than the straight chain isomers. By use of GC-MS, some of them have been identified and found to exist in higher concentrations in the isomeric compound mixture than the straight chain isomers. The investigations of the distribution and metabolism of these branched isomers in aquatic organisms are therefore considered to be important in understanding the mechanisms of toxicity of NP. A single tertiary isomer of NP, 4(3'-,6'-dimethyl-3'-heptyl)-phenol, was synthesized in the laboratory and used in in vivo studies of its organ distribution and metabolism in Lymnaea stagnalis L., following a constant exposure of the organisms to 14C-NP isomer in water over a period of 8 days at an average exposure concentration of 105 ppb (range: 93-116 ppb). The results obtained clearly showed the distribution and bioconcentration of the isomer residues in various internal organs of Lymnaea after uptake in water and food. Analysis of the extracts of the organ tissues and faeces by HPLC and GC-MS after digestion with Pankreatin/beta-glucuronidase and nitric acid, respectively, showed that the isomer was metabolized by conjugation to glucuronic acid and hydroxylation to a catechol. The findings from these studies and their implications in the biotransformation and estrogenicity of NP in Lymnaea stagnalis L. are further discussed in detail in this paper.

Animals↗
PubMed1997

Diversity in cell specific co-expression of four neuropeptide genes involved in control of male copulation behaviour in Lymnaea stagnalis.

We report here the neuron-specific co-expression of four genes coding for neuropeptides involved in the control of male behaviour. These neurons are located in the anterior lobe of the right cerebral ganglion in the central nervous system of Lymnaea stagnalis and project via the penis nerve to the penial complex. In order to accomplish optimal assurance we applied in situ hybridization, immunocytochemistry and matrix-assisted laser desorption ionization mass spectrometry. The anterior lobe neurons express the gene encoding the amidated tetrapeptide APGWamide. Subsets of these cells are now shown to co-express the APGWamide gene exclusively with one of three other neuropeptide genes, encoding Lymnaea neuropeptide Y, conopressin or pedal peptide, respectively. All four genes are also expressed in other neurons in other centres projecting to the penial complex, but in these cells co-expression was not observed. The neuropeptides encoded by the genes could be identified in the anterior lobe cell bodies on the basis of immunocytochemistry and mass spectrometrical analysis. The neuropeptides APGWamide and Lymnaea neuropeptide Y, which are co-localized in the anterior lobe cells as well as in axons innervating the penis retractor muscle, do not induce muscle contraction but have a modulatory action by affecting the relaxation rate and amplitude of the contraction. APGWamide and conopressin had earlier been suggested to modulate peristalsis of the vas deferens. Thus, it seems that the neurons co-expressing the various combinations of neuropeptide genes in the anterior lobe represent functional units, each acting in the fine tuning of different muscles involved in specific aspects of male copulation behaviour.

Animals↗
PubMed2000

Phyto-adaptogens protect against environmental stress-induced death of embryos from the freshwater snail Lymnaea stagnalis.

The main purpose of the studies presented in this paper is twofold: 1) to evaluate whether phyto-adaptogens (Acanthopanax senticosus and Rhodiola rosea) are able to exert a protective action against stress-induced death of embryos of the pond snail Lymnaea stagnalis; and 2) whether a possible protective action by phyto-adaptogens can be explained by the induction of heat shock proteins. Enhancement in resistance by phyto-adaptogens was studied by applying plant extracts for a period of 20 hours to 3-day old larvae of the pond snail Lymnaea stagnalis. Subsequently they were exposed to a high and toxic dose of different environmental stressors. The following stress conditions were selected: a physical stress condition (heat shock: 43 degrees C for 4 minutes), an oxidative stress condition (superoxide radicals induced by menadione (600 microM for 2 hours)) and heavy metal-induced stress (copper (150 microM for 1 hour) or cadmium (20 microM during 1 hour)). Both Acanthopanax and Rhodiola exert a strong protective action against a lethal heat shock. These adaptogens also significantly protect against the negative effect of superoxide radicals as induced by menadione. With respect to the protective action against exposure to heavy metals a small but significant protection was observed against intoxication with copper or cadmium by the phyto-adaptogens. In summary, there appears to be a difference in efficiency in enhancing resistance to the various stress conditions used (heat shock>menadione>copper>cadmium). Based on the results presented in this paper, we can conclude that phyto-adaptogens are able to enhance the resistance against the different stress conditions tested in developing individuals of Lymnaea. Although the degree to which resistance is enhanced appears to depend on the type of stressor applied, our results confirm the definition of phyto-adaptogens as being universal enhancers of non-specific resistance against different kinds of stress conditions. With respect to the mechanism of enhanced resistance, the question was asked whether this protective action is caused by an induction of heat shock proteins (hsps), which are known to be involved in tolerance and adaptation. The phyto-adaptogens did not induce the synthesis of any of the hsps, nor did they modulate the normal heat shock induced synthesis of these stress proteins. We conclude that it is unlikely that hsps play a major role in obtaining an enhanced state of resistance provided by phyto-adaptogens.

Animals↗
PubMed2005

Identification and functional expression of a family of nicotinic acetylcholine receptor subunits in the central nervous system of the mollusc Lymnaea stagnalis.

We described a family of nicotinic acetylcholine receptor (nAChR) subunits underlying cholinergic transmission in the central nervous system (CNS) of the mollusc Lymnaea stagnalis. By using degenerate PCR cloning, we identified 12 subunits that display a high sequence similarity to nAChR subunits, of which 10 are of the alpha-type, 1 is of the beta-type, and 1 was not classified because of insufficient sequence information. Heterologous expression of identified subunits confirms their capacity to form functional receptors responding to acetylcholine. The alpha-type subunits can be divided into groups that appear to underlie cation-conducting (excitatory) and anion-conducting (inhibitory) channels involved in synaptic cholinergic transmission. The expression of the Lymnaea nAChR subunits, assessed by real time quantitative PCR and in situ hybridization, indicates that it is localized to neurons and widespread in the CNS, with the number and localization of expressing neurons differing considerably between subunit types. At least 10% of the CNS neurons showed detectable nAChR subunit expression. In addition, cholinergic neurons, as indicated by the expression of the vesicular ACh transporter, comprise approximately 10% of the neurons in all ganglia. Together, our data suggested a prominent role for fast cholinergic transmission in the Lymnaea CNS by using a number of neuronal nAChR subtypes comparable with vertebrate species but with a functional complexity that may be much higher.

Amino Acid Sequence↗
PubMedDate not supplied

Activities of glutamate dehydrogenase and aspartate and alanine aminotransferases in freshwater snails Helisoma duryi and Lymnaea natalensis exposed to copper.

In this paper we investigate the potential of glutamate dehydrogenase (GDH) and aspartate and alanine aminotransferases (AST and ALT) as biomarkers of water pollution due to copper in the freshwater snails Helisoma duryi and Lymnaea natalensis. Snails were dosed with copper(II) ion concentrations of 0.01, 0.1 and 1 mg kg(-1) breeding water for a period of 96 h, after which those surviving were shelled. The copper content in the breeding water, in whole snail tissue and in the snail shells was determined at the end of the period of exposure. For enzyme determinations, whole snail tissue was first homogenized and fractionated by centrifugation at 500 g to remove the nuclei. The resulting supernatant was then centrifuged at 10,000 g to give a pellet fraction representing the mitochondrial fraction and a supernatant representing the cytosolic fraction. Copper was very toxic to both snail species at concentrations above 0.2 mg l(-1), with only 3% of the Helisoma and 12% of the Lymnaea surviving at concentrations of approximately 1 mg l(-1). The copper content in the shells and tissues of snails rose with increasing copper concentration in the breeding water, and was 2.1- to 4.9-fold in snails exposed to copper ion at a dose of 1 mg kg(-1) water compared with undosed snails. Similarly, the activities of GDH and AST rose by up to 4.7-fold in the homogenate and the mitochondrial and cytosolic fractions with increasing concentrations of copper. These activities, however, fell at copper concentrations of approximately 1 mg l(-1), which coincided with massive death of snails. Mitochondrial ALT disappeared at copper ion concentrations of approximately 0.2 mg l(-1) for Lymnaea and 1 mg l(-1) for Helisoma, possibly indicating mitochondrial degeneration. These results show that GDH, AST and ALT have the potential to be biomarkers of sublethal copper pollution in these two snail species, since their activities were significantly altered by low copper concentrations.

Alanine Transaminase↗
PubMed1993

Nitric oxide activates buccal motor patterns in Lymnaea stagnalis.

The mollusc, Lymnaea stagnalis, has been used as a model to study the mechanisms of nitric oxide (NO)-dependent processes in the CNS. Putative NO-containing neurones in Lymnaea are localized in the buccal ganglia, predominantly in areas where sensory neurones known to regulate feeding are found. The NO-generating substance, S-nitrosocysteine (S-NC, 5 x 10(-5)-10(-3 M) activates feeding movements of the buccal mass and modulates the activity of buccal motoneurones. An inhibitor of NO synthase, NG-methyl-L-arginine (10(-4) M) decreases the frequency of background buccal movements and has opposite effects to S-NC on the buccal motoneurones. We suggest that NO is a messenger in the CNS of Lymnaea and may be involved in coordination of feeding motor patterns.

Animals↗
PubMed1995

Alternative RNA splicing generates diversity of neuropeptide expression in the brain of the snail Lymnaea: in situ analysis of mutually exclusive transcripts of the FMRFamide gene.

In the CNS of the snail Lymnaea stagnalis, Phe-Met-Arg-Phe-amide (FMRFamide)-like and additional novel neuropeptides are encoded by a common, multi-exon gene. This complex locus, comprising at least five exons, is subject to post-transcriptional regulation at the level of alternative RNA splicing. Our aim was first to analyse the pattern by which exons of this neuropeptide locus combine during splicing of the primary RNA transcript, and second to investigate the functional significance of splicing by mapping the expression and neuronal localization in the CNS of the alternative mRNA transcripts, in the context of defined neuronal networks and single identified neurons. The approach was a combination of comparative in situ hybridization and immunocytochemistry, using a battery of exon-specific oligonucleotides and anti-peptide antisera. The analysis illustrated that exons III, IV and V were always coexpressed and colocalized whereas the expression of exon II was always differential and mutually exclusive. Both sets of exons were, however, coexpressed with exon I: the total number of exon I-expressing neurons was equal to the combined number of neurons expressing exon III/IV/V and neurons expressing exon II. In addition, it was revealed that the extreme 5' of exon II, encoding a potential hydrophobic leader signal, was not expressed in the CNS of Lymnaea but was apparently spliced out during RNA processing. Both mRNA transcripts of the FMRFamide locus, type 1 (exons I/II) and type 2 (exons I/III/IV/V), were translated in the CNS and the resulting protein precursors were also expressed in a mutually exclusive fashion, as were their respective transcripts. The expression of alternative transcripts within identified networks or neuronal clusters was heterogeneous, as exemplified by the cardiorespiratory network. On the basis of this work and a previous cDNA analysis, we put forward a revised model of differential splicing and expression of the FMRFamide gene in the CNS of Lymnaea.

Amino Acid Sequence↗
PubMed1995

Identification, distribution and physiological activity of three novel neuropeptides of Lymnaea: EFLRlamide and pQFYRlamide encoded by the FMRFamide gene, and a related peptide.

We are interested in analysing the detailed modulation of defined neuronal systems by multiple neuropeptides encoded in the FMRFamide locus of the snail Lymnaea. Cloning of the FMRFamide gene has predicted the existence of two novel peptides previously unknown from biochemical analysis, the pentapeptides EFLRlamide and QFYRlamide. These peptides may form part of a new family of peptides sharing the sequence motif -FXRlamide. In this paper we adopt a novel approach to first identify and characterize -FXRlamide-like peptides in extracts from the central nervous system of Lymnaea. By a combination of high-performance liquid chromatography (HPLC) and continuous-flow fast atom bombardment mass spectrometry, we identify three novel peptides: EFLRlamide, pQFYRlamide and pQFLRlamide. The first two are those predicted in exon II of the FMRFamide locus whereas the last is, interestingly, a product which cannot be derived from post-translational modification of the predicted peptides but must be encoded by as yet unidentified nucleotide sequences. A specific antibody raised to EFLRlamide, and immunoreactive to all three peptides, revealed EFLRlamide-like expression throughout the central nervous system in the same cells where exon II is transcribed and the peptide SEEPLY (a post-translational product of exon II) was localized. Additional cells, however, were also identified. Immunoreactivity was mapped in a number of identified neurons in the central nervous system, including two heart cardioexcitatory motoneurons, the Ehe cells (E heart excitors of the visceral ganglion) and penial motoneurons in the right cerebral ganglion. The peripheral tissues (heart and penial complex) that these respective classes of neurons innervate also exhibited EFLRlamide immunoreactivity. The central and peripheral localization of EFLRlamide-like immunoreactivity suggested that EFLRlamide/pQFYRlamide may have an important physiological role in both these peripheral systems as well as in the central nervous system. This was confirmed by physiological experiments that showed that EFLRlamide and pQFYRlamide inhibited many central neurons and in particular the Bgp neurons in the right parietal ganglion. EFLRlamide had complex biphasic effects on the frequency of heart-beat: an initial inhibitory response was followed by a long-lasting increase in the rate of beating. Taken together with earlier work, this study now completes the analysis and localization of the full set of post-translational products of the FMRFamide precursor in Lymnaea and supplies further evidence towards the characterization of the physiological systems which such peptides may modulate in concert.

Amino Acid Sequence↗
PubMed1993

A molecularly defined cardiorespiratory interneuron expressing SDPFLRFamide/GDPFLRFamide in the snail Lymnaea: monosynaptic connections and pharmacology.

1. Neuron-specific expression of alternately spliced exons of the gene encoding the Phe-Met-Arg-Phe-NH2 (FMRFamide) family of neuropeptides and the role of encoded peptides in synaptic transmission were examined in an identified cardiorespiratory interneuron, the visceral white interneuron (VWI), in the snail Lymnaea. 2. In situ hybridization using exon-specific probes showed VWI cytoplasmic expression of the exon encoding the Lymnaea heptapeptides Gly-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (GDPFLRF amide) Ser-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (SDPFLRF amide) but not the exon encoding the tetrapeptides FMRFamide and Phe-Leu-Arg-Phe-NH2 (FLRFamide). 3. The absence of the tetrapeptides (FMRFamide and FLRFamide) in the VWI was indirectly confirmed by the lack of immunoreactivity to a specific antibody raised against the sequence Leu-Tyr. This sequence is present in the Lymnaea tetrapeptide precursor, but not the heptapeptide precursor. 4. The VWI has monosynaptic connections with many identifiable neurons in the CNS. These were excitatory on three clusters of identified neurons [B group (Bgp), E group (Egp), and F group (Fgp)], inhibitory on another cluster [A group (Agp)] or biphasic (excitation followed by inhibition) on a single giant neuron [right pedal dorsal 1 (RPeD1)]. 5. The role of GDPFLRFamide/SDPFLRFamide as putative neurotransmitters was examined by comparing neuronally evoked postsynaptic responses with the effects of focal peptide application. 6. The heptapeptides closely mimicked the inhibitory responses (threshold pressure pipette concentration 10(-9) M) on the Agp cells and RPeD1, including an increase in membrane conductance. FMRFamide was 1 order of magnitude less potent. GDPFLRFamide/SDPFLRFamide, applied either alone or in "cocktails" (combinations of GDPFLRFamide, SDPFLRFamide, FMRFamide, and FLRFamide), did not reproduce the excitatory effect of the VWI on the Bgp, Egp, and Fgp cells. These peptides, applied either together or separately, inhibited the cells. 7. FMRFamide or FLRFamide, but not GDPFLRFamide or SDPFLRFamide, could reproduce the initial depolarizing component of the biphasic response on RPeD1. This only occurred at concentrations of > or = 10(-4) M (10(-3) M was necessary to get spikes on RPeD1) and may not be physiologically significant. 8. We conclude that at least one so far unidentified co-transmitter must be present in the VWI to account for its full range of synaptic responses.

Animals↗
PubMed1995

Dopaminergic transmission between identified neurons from the mollusk, Lymnaea stagnalis.

1. Dopaminergic transmission was investigated in the central nervous system (CNS) of the freshwater snail, Lymnaea stagnalis. 2. The giant pedal neuron, designated as right pedal dorsal one (RPeD1), makes chemical, monosynaptic connections with a number of identified follower cells in the CNS. Previous work has shown that RPeD1 is an interneuron and a important component of the Lymnaea respiratory central pattern generator. In this study, the hypothesis that RPeD1 uses dopamine as its neurotransmitter was tested by chromatographic, pharmacological, and electrophysiological methods. Characterization of RPeD1's transmitter pharmacology is essential to clearly understand its role in Lymnaea. 3. Earlier studies demonstrated that the soma of RPeD1 contains dopamine. This was quantitated in the present study by high-performance liquid chromatography (with electrochemical detection) of isolated RPeD1 somata and growth cones, which yielded 0.8 +/- 0.3 and 0.10 +/- 0.08 pmol of dopamine per soma and growth cone, respectively. 4. Bath or pressure application of dopamine to follower cells of RPeD1, in situ, mimicked the effects of RPeD1 stimulation. Dose-response curves were constructed for the excitatory effect of dopamine on follower cells, visceral dorsal two and three (VD2/3) (ED50 = 39 microM; Hill coefficient = 1.03), and the inhibitory effect of dopamine on follower cell, visceral dorsal four (ED50 = 33 microM; Hill coefficient = 0.92). 5. The following dopamine agonists (100 microM) were tested by bath application: 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (ADTN), apopmorphine, 2-bromo-alpha-ergocryptine, deoxyepinephrine (DE), mesulergine, (-) quinpirole, SKF 38393, and tyramine. Only the general dopamine agonists, ADTN and DE, mimicked RPeD1's effects on its follower cells. 6. When VD2/3 was isolated and plated in vitro, it maintained a depolarizing response to dopamine. This response was reduced by intracellular injection of the G-protein blocker, GDP-beta-S (2 mM in electrode). Similarly, incubation of VD2/3, in vitro for approximately 18 h, with pertussis toxin (PTX; 5 micrograms/ml), the G-protein inactivating exotoxin, also reduced the dopamine response. Injecting GDP or incubating in heat-inactivated PTX did not effect the response. 7. Several dopamine antagonists were used in an attempt to block RPeD1's synapses: chlorpromazine, ergonovine, fluphenazine, haloperidol, 6-hydroxydopamine, SCH 23390, (+/-) sulpiride, and tubocurarine. Only the D-2 dopamine receptor antagonist, (+/-) sulpiride, reversibly blocked synaptic transmission from RPeD1 to its follower cells. Both the (+) and the (-) enantiomer of sulpiride also antagonized synaptic transmission. A dose-inhibition curve for (+/-) sulpiride was constructed (IC50 = 47 microM).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

Halothane-induced synaptic depression at both in vivo and in vitro reconstructed synapses between identified Lymnaea neurons.

1. In the present study we tested the ability of the general anesthetic, halothane, to affect synaptic transmission at in vivo and in vitro reconstructed peptidergic synapses between identified neurons of Lymnaea stagnalis. 2. An identified respiratory interneuron, visceral dorsal 4 (VD4), innervates a number of postsynaptic cells in the central ring ganglia of Lymnaea. Because VD4 has previously been shown to exhibit immunoreactivity for FMRFamide-related peptides, it was hypothesized that these peptides may be utilized by VD4 during synaptic transmission. In the intact, isolated CNS of Lymnaea, we have identified novel connections between VD4 and the pedal A (PeA) cells. We demonstrate that VD4 makes inhibitory connections with the PeA neurons, in particular PeA4, and that these synaptic responses are mimicked by exogenous application of FMRFamide. 3. The synaptic transmission between VD4 and the PeA cells in an intact, isolated CNS preparation was completely blocked in 2%, but not 1% halothanc. Interestingly, the postsynaptic responses (PeA) to exogenous FMRFamide were maintained in the presence of both 1 and 2% halothane. 4. To determine the specificity of the observed responses and to determine the precise synaptic site of anesthetic action, we reconstructed the VD4/PeA synapses in vitro. After isolation from their respective ganglia, both cell types extended processes and established neuritic contact. We demonstrated that not only did the presynaptic neuron reestablish the appropriate inhibitory synapses with the PeA neurons, but that the PeA cells also maintained their responsiveness to exogenous FMRFamide. 5. Superfusion of the in vitro synaptically connected VD4 and PeA cells with 2% halothane completely abolished the synaptic transmission between these cells. However, even higher concentrations of 4% halothane failed to block the responsiveness of the PeA neurons to exogenous FMRFamide. Moreover, both 1 and 2% halothane enhanced the duration of the postsynaptic response to exogenously applied FMRFamide. These data suggest that the halothane-induced depression of synaptic transmission most likely occurred at the presynaptic level. 6. This study provides the first direct evidence that peptidergic transmission in the nervous system may also be susceptible to the actions of general anesthetics. In addition, we utilized a novel approach of in vitro reconstructed synapses for studying the effects of general anesthetics on monosynaptic transmission in the absence of other synaptic influences.

Anesthetics, Inhalation↗
PubMed1991

The whole-body withdrawal response of Lymnaea stagnalis. II. Activation of central motoneurones and muscles by sensory input.

The role of centrally located motoneurones in producing the whole-body withdrawal response of Lymnaea stagnalis (L.) was investigated. The motoneurones innervating the muscles used during whole-body withdrawal, the columellar muscle (CM) and the dorsal longitudinal muscle (DLM) were cells with a high resting potential (-60 to -70 mV) and thus a high threshold for spike initiation. In both semi-intact and isolated brain preparations these motoneurones showed very little spontaneous spike activity. When spontaneous firing was seen it could be correlated with the occurrence of two types of spontaneous excitatory postsynaptic potential (EPSP). One was a unitary EPSP that occasionally caused the initiation of single action potentials. The second was a larger-amplitude, long-duration (presumably compound) EPSP that caused the motoneurones to fire a burst of high-frequency action potentials. This second type of EPSP activity was associated with spontaneous longitudinal contractions of the body in semi-intact preparations. Tactile stimulation of the skin of Lymnaea evoked EPSPs in the CM and DLM motoneurones and in some other identified cells. These EPSPs summated and usually caused the motoneurone to fire action potentials, thus activating the withdrawal response muscles and causing longitudinal contraction of the semi-intact animal. Stimulating different areas of the body wall demonstrated that there was considerable sensory convergence on the side of the body ipsilateral to stimulation, but less on the contralateral side. Photic (light off) stimulation of the skin of Lymnaea also initiated EPSPs in CM and DLM motoneurones and in some other identified cells in the central nervous system (CNS). Cutting central nerves demonstrated that the reception of this sensory input was mediated by dermal photoreceptors distributed throughout the epidermis. The activation of the CM and DLM motoneurones by sensory input of the modalities that normally cause the whole-body withdrawal of the intact animal demonstrates that these motoneurones have the appropriate electrophysiological properties for the role of mediating whole-body withdrawal.

Animals↗
PubMed1999

[Ecology of Lymnaea truncatula Müller, intermediate host of Fasciola hepatica Linné in the microclimate of Tozeur (southeast of Tunisia)].

Systematic visits were carried out in the traditional Tozeur' oasis (Tunisia), in 1997/1998 in order to study the ecology of Lymnaea truncatula as well as its density in relation to habitat characteristics in the hydrographical networks of the oases. Lymnaea truncatula was enumerated on 1 m2 surface by habitat, according to the quadrat method. Samples of soil, irrigation and draining water were also analysed. The density of Lymnaea truncatula detected in autumn was constantly higher. It depends on the nature of the habitat: 54.8% in the secondary irrigation canals, 34.8% in those of draining, 7.6% in the seguia and 2.9% in the principal draining canal. Ecological factors--permanent dampness, suitable luminosity, basic pH of soil and water, temperature between 10 and 28 degrees C--contribute to the multiplicity of snails.

Animals↗