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PubMed1999

The effects of hypercapnia on force and rate of contraction and intracellular pH of perfused ventricles from the land snail Helix lucorum (L.).

The effects of hypercapnia, together with low and high levels of extracellular Ca(2+), on heart activity and intracellular pH were examined in isolated perfused hearts from the land snail Helix lucorum. In addition, the intracellular level of Ca(2+) was determined in slices of ventricles superfused with both normal and hypercapnic salines, containing low and high concentrations of Ca(2+), to investigate whether low extracellular pH affects the entry of Ca(2+) into the heart cells. We also examined the effect of a saline that simulated the composition of the haemolymph of snails after estivating for 3 months on the heart activity and intracellular pH. The results showed that hypercapnia causes decreases in the rate and force of heart contraction, and these are more pronounced in the presence of low levels of extracellular Ca(2+). Moreover, the present results indicate that Ca(2+) maintains the contractility of the heart muscle under acidic conditions and seems to act by competing with protons for the Ca(2+ )binding sites on sarcolemma. The negative effect of hypercapnia on heart activity appears to be due to a reduction in extracellular pH rather than to changes in intracellular pH.

Journal Article↗
PubMed1999

Effects of anoxia on the extra- and intracellular acid-base status in the land snail helix lucorum (L.): lack of evidence for a relationship between pyruvate kinase down-regulation and acid-base status

The aims of the present study were to describe a possible correlation between the regulation of the key glycolytic enzyme pyruvate kinase and the acid-base status in the haemolymph and in several other tissues of land snails during anoxia. To illustrate whether such a relationship exists, we determined (i) the acid-base variables in the haemolymph and tissues of the land snail Helix lucorum, (ii) the kinetic properties of pyruvate kinase from several tissues and (iii) the levels of the anaerobic end-products d-lactate and succinate in the haemolymph and tissues of aerobic and anoxic Helix lucorum. The results showed that the pH of haemolymph (pHe) decreased significantly over the first 20 h of anoxia and then recovered slowly towards control values. A similar pattern was observed for intracellular pH (pHi), which decreased significantly over the first 16 h of anoxia and slowly returned towards control levels. The reduction and recovery of pHi and pHe seem to reflect the rate of anaerobic metabolism. The main anaerobic end-products, d-lactate and succinate, accumulated rapidly during the initial stages of anoxia and more slowly as anoxia progressed. The decrease in the rate of accumulation of anaerobic end-products during prolonged anoxia was due to the conversion of tissue pyruvate kinase to a less active form. The results demonstrate a correlation between pyruvate kinase down-regulation and the recovery of acid-base status in the haemolymph and the tissues of land snails during anoxia.

Journal Article↗
PubMed1998

Cross-reactivity between terrestrial snails (Helix species) and house-dust mite (Dermatophagoides pteronyssinus). I. In vivo study.

Clinical reports have suggested an unusual frequency in the number of patients with food allergy to snails who are also allergic to the house-dust mite (HDM). As allergy to HDM is one of the most frequent sensitizations in atopic patients of Western countries, evaluation of the relevance of the concomitant sensitization to Dermatophagoides pteronyssinus and to snails is an important consideration. To evaluate the responsibility of different snail components and of snail mites for inducing in vivo hypersensitivity in patients allergic to HDM, the in vivo reactivity of patients with clinical symptoms after ingestion of snails was assessed by skin prick tests with extracts and hemolymph from four different Helix species snails, and extracts from the snail parasitic mite, Riccardoella limacum. In addition, to obtain epidemiologic data on cosensitization to HDM and snails in allergic patients, the frequency of snail sensitization and its relationship to HDM sensitization were determined in a population of 169 allergic children. All patients allergic to snails had positive skin prick tests to the snail extracts and none to R. limacum extract. The number of positive skin reactions did not significantly differ whatever the species, snail part, or heating procedure used. The strongest reactions were obtained with Helix pomatia (Burgundy snail). Among the 169 prospectively tested children, 38 had a positive prick test to snail extracts; 79% of the snail-sensitized children had sensitization to HDM; and 31% of the children allergic to HDM were found to be sensitized to snails. These results show that snail components, and not the mite R. limacum, were responsible for the in vivo hypersensitivity. These snail components reacting in vivo are present in different parts of snails, including the hemolymph. One-third of the children allergic to HDM were sensitized to snails without any previous ingestion of snails: this observation suggests that HDM was the sensitizing agent and that the cross-reaction could be clinically relevant in countries where eating snails is common.

Adolescent↗
PubMedDate not supplied

[Effect of 5,6-dihydroxytryptamine and 6-hydroxydopamine on the maze behavior of the snail Helix lucroum].

One-trial Y-maze test in snails did not reveal any predominance of the right or left alley choice. The choice of the right alley (but not of the left one) was accompanied by a slowing down of forward motion. After the administration of 10 mg/kg of 6-hydroxydopamine 24 hr before the test, right alley choice without slowing down became more frequent. Administration of 5,6-dihydroxytrypamine (10 mg/kg) caused a reduction of motion rate along all parts of the maze and an increase in frequency of the orienting head movements towards the alley to be chosen.

5,6-Dihydroxytryptamine↗
PubMed1986

Immunocytochemical and ultrastructural evidence for supra- and subesophageal localization of the dorsal-body cells of the snail Helix aspersa.

Dorsal-body endocrine cells (DBEC) of the snail were studied by means of immunocytochemical and electron microscopic methods at different times of the reproductive cycle. They specifically bind the anti-methionine-enkephalin (vertebrate--opioid-pentapeptide) antibody and are located not only near the cerebral ganglia but also in the connective tissue surrounding the subesophageal ganglia. Ultrastructural characteristics of these subesophageal cells, however, confirm their clear identity with the previously described supraesophageal cells. The quantitative variations of their immunoreactive content allow us to postulate a likely involvement in reproductive physiology (mating and egg laying). These observations prove that the distribution of the classical "dorsal-body cells" is more extensive than has been admitted until now and that they synthesize methionine-enkephalin-like substance(s).

Animals↗
PubMed1994

[Influence of brain grafts on growth restoration of snail (Helix aspersa) deprived of the midbrain].

Microsurgical removal of the mesocerebrum from the brain of juvenile snails stopped their growth whereas intracerebral implantation of desheathed cerebral ganglia (CG) re-established it. When the animals were grafted with CG from very young snails growth was much more stimulated than with CG from donors of the same age or from adults. Furthermore, young CG of juvenile fast growing specimens of the large species (Helix aspersa maxima) induced a higher growth rate than the CG of the ordinary small garden snails (Helix aspersa aspersa). Labelling of the neural grafts with the vital fluorescent dye fast blue enabled us to follow the repopulation of the lesioned area of the brain of the host during the functional integration of the implanted neurons into the circuits that control growth in snails.

Age Factors↗
PubMed1998

Changes in the reproductive system of the snail Helix aspersa caused by mucus from the love dart.

The function of the love dart in certain species of terrestrial snails is unknown. In Helix aspersa, the dart is a sharp calcareous structure that is used to pierce the partner's skin during courtship. When expelled, the dart is covered with a thick mucus. The hypothesis tested here is that the mucus contains a biologically active substance. Extracts of the digitiform glands that produce this mucus were applied to parts of the reproductive system in vitro. The extracts triggered an initial reconfiguration of the copulatory canal that caused the bursa tract diverticulum to become more accessible to the spermatophore. The reconfiguration of the copulatory canal also closed off the tract leading to the bursa copulatrix, a sperm-digesting organ. A few minutes after the initial contraction, the peristaltic contractions in the diverticulum became significantly more frequent. This latter effect continued for at least 1 h, provided that the mucus extract remained in the saline bath. The minimum effective dosage was less than the 2.2 mg of mucus transferred with the dart. Sperm competition is expected in Helix aspersa since multiple matings occur before eggs are laid. By influencing the female organs involved in the processing of foreign sperm, the dart shooter may increase the chance that his sperm will fertilise eggs.

Animals↗
PubMed1990

Dissociation of sexual arousal and sexual proclivity in the garden snail, Helix aspersa.

Sexual arousal (intensity of courtship) and sexual proclivity (tendency to court) in Helix aspersa can be reliably measured using externally observable correlates. Snails with sexual proclivity are significantly more likely to turn toward an anesthetized conspecific after contacting it than are sexually unreceptive snails. Sexual arousal can be inferred from the stage of a snail's genital eversion, which appears only during courtship. The higher the stage of the eversion, the shorter the time required to complete introductory courtship behavior and the higher the rate of successful copulation, the fewer the number of breaks and pauses during courtship, and the longer the time a snail will spend in contact with an anesthetized conspecific. Sexual proclivity has no effect on feeding or locomotory behavior; however, sexual arousal inhibits feeding and increases locomotor activity. Snails that were allowed daily contact with conspecifics required less time to complete introductory courtship behavior relative to snails that were isolated from conspecifics for 1 week. This suggests that daily contact increases sexual arousal. A greater percentage of isolated snails exhibited courtship behavior than did snails which had experienced daily conspecific contact. This suggests that isolation increases sexual proclivity. These differences indicate that sexual arousal is not merely due to an increase in sexual proclivity.

Animals↗
PubMedDate not supplied

[Generalized and signal-specific long-term nociceptive sensitization in the snail Helix lucorum].

Neurophysiological and behavioural correlates of the long-term sensitization were investigated in Helix lucorum small. Application of 10% quinine solution on the snail's head initiated a long-term (for more than 24 hours) facilitation of defensive reactions. The behavioural effects correlated with the changes in evoked and spontaneous activity of L-PP11 neurons, i.e., facilitation of synaptic components in responses to testing stimulation, increase in membrane excitability and depolarization. Efficacy of sensitization depended on the duration of the experimental procedure. After daily training site-specific and modality-specific effects dominated (i.e., more expressed facilitation of responses to testing stimulation of the sensitized body site than that of the other sites or to testing stimulation of the same modality as sensitizing stimulation). After 3 days of training the effects of general sensitization were predominantly observed, i.e., facilitation of neuronal responses to stimulation of different modalities and body sites, depolarization of the membrane and increase in its excitability.

Animals↗
PubMed1986

Mannitol oxidase: partial purification and characterization of the membrane-bound enzyme from the snail Helix aspersa.

Mannitol oxidase, a membrane-bound oxidase has been purified 250-fold from snail digestive gland tissue. The activity is solubilized by a number of ionic, non-ionic, and zwitterionic detergents. Purification of the solubilized enzyme was by polyethylene glycol fractionation and column chromatography using anionic exchange resins, hydroxylapatite, and gel filtration. The enzyme is stabilized by glycerol and remains active for at least one week at -20 degrees. Hydrogen peroxide is the oxygen reduction product and a mannose/hydrogen peroxide stoichiometry of 0.86 was found. D-Arabinitol and D-mannitol were the most active substrates of those tested. Results with these and other substrates suggest that the configuration around carbons-2 and -4 is critical for binding and reactivity. The apparent Km for D-mannitol is 6 mM and for oxygen, 40 microM. The pH optimum for the enzyme is between 8 and 8.5 and the isoelectric point is 5.4-5.6.

Alcohol Oxidoreductases↗
PubMed1991

Extrusion of calcium from a single isolated neuron of the snail Helix pomatia.

Simultaneous optical measurements of extra- and intracellular Ca2+ concentrations were carried out on isolated snail neurons injected iontophoretically with Ca2+. The fluorescent indicator Fura-2 was used to measure intracellular concentration of free Ca, and the absorbant indicator Antipyrylazo III to measure changes in extracellular calcium concentration in the micro-chamber containing the cell. The velocity of Ca2+ extrusion from a single cell has been shown to be in accordance with the level of free Ca in the neuronal cytoplasm. After an increase in intracellular free Ca by iontophoretic injection from a microeletrode to 0.2-0.5 microM, the velocity of Ca2+ extrusion from the neuron was approximately 0.3-4.6 microM/sec per cell volume. During caffeine-induced calcium-dependent calcium release of Ca2+ from intracellular stores a stimulation of calcium extrusion took place, reaching the velocity of 5.0 microM/sec per cell volume.

Animals↗
PubMed1979

Microelectrode investigations of learning phenomena in snail (Helix pomatia) neurones.

We have examined changes of postsynaptic potentials and of pattern activity of the identified silent and oscillatory snail neurones in Helix pomatia during conditioning. Local changes of EPSP or IPSP have been recorded during association following the first stimulus in the silent cells, whereas spike discharges could be observed in response to the 2nd stimulus. In the oscillatory neurones changes of pattern activity have been recorded following the 2nd stimulus, while the first stimulus proved to be ineffective. The formation of temporary connections of snail neurones seemed to be a specific phenomenon, because it was necessary to pair stimuli of different inputs for the development of these modifications. These plastic changes seemed to depend on the interstimulus as well as on the intertrial intervals. Our experimental data underline the probable role of the stimulus parameters and of the electrical properties of neurones during the formation of learned neuronal responses.

Action Potentials↗
PubMedDate not supplied

[The neuronal mechanisms of site-specific nociceptive sensitization in the snail Helix lucorum].

Electrophysiological effects of site-specific sensitization after application of strong quinine solution onto snail head or snail mantle were studied in Helix lucorum LPl1 and RPl1 interneurons. Sensitizing stimulation (strong quinine solution) applied onto the snail head or mantle produced synapse-specific facilitation of neural responses evoked by tactile stimulation: synaptic facilitation was more expressed in neural responses evoked by tactile stimulation of the sensitized body site. Synaptic facilitation of the neural responses evoked by tactile stimulation of the snail head was observed 1 hour after corresponding sensitizing stimulation. Two definite phases were separated in the synaptic facilitation. The first phase persisted for 1 hour and was characterized by a "peak-shaped" increase of the EPSP square, in the second phase the EPSP facilitation was maintained at a relatively constant level for more than 3 hours. The synaptic facilitation of the neural responses to tactile stimulation of the snail mantle began 40-60 min later than that of the responses to the stimulation of the head and was characterized by the absence of the first phase. It was suggested that specific character of synaptic facilitation in a command interneuron is determined by peculiarities of genetical regulation of different neural inputs.

Animals↗
PubMed1995

Effects of low doses of caffeine on [Ca2+]i in voltage-clamped snail (Helix aspersa) neurones.

1. We have measured cytosolic free Ca2+ concentrations ([Ca2+]i) in voltage-clamped snail neurones using fura-2. Transient increases in [Ca2+]i were induced by depolarizing voltage steps of 20-60 mV for 0.1-10 s from a holding potential of -50 or -60 mV. 2. Low doses of caffeine, 0.2-1 mM, increased the size of the [Ca2+]i transients by both increasing the peak and producing an undershoot. 3. Ryanodine, an inhibitor of Ca2+ release from the intracellular Ca2+ stores, and cyclopiazonic acid (CPA), an inhibitor of the Ca(2+)-ATPase of the intracellular Ca2+ stores, both reduced the size of the [Ca2+]i transients and blocked the effects of caffeine on the transients. 4. The effects of caffeine and CPA were greater on transients produced by long, small, rather than short, large depolarizations. This suggests that calcium-induced calcium release (CICR) played a greater role in the [Ca2+]i increase resulting from longer, smaller depolarizations. 5. Increasing the extracellular pH from 7.5 to over 9, which inhibits the plasmalemmal Ca(2+)-H(+)-ATPase, increased the resting [Ca2+]i level. Depolarization-induced [Ca2+]i transients became much larger while the two effects of caffeine remained. CPA was ineffective at high pH. 6. In some experiments the increase in basal [Ca2+]i caused by alkaline pH was reduced by 0.2 or 0.5 mM caffeine. The increase in basal [Ca2+]i caused by maintained depolarization was reduced, after a transient increase, by 0.5 mM caffeine. Both reduction and increase were blocked by CPA. 7. We conclude that low doses of caffeine can increase uptake by intracellular Ca2+ stores. Caffeine could also release Ca2+ from ryanodine-insensitive Ca(2+)-ATPase-dependent stores as well as facilitating normal ryanodine-sensitive CICR.

Animals↗
PubMed1997

[In vivo and in vitro study of the effect of somatostatin and somatostatin-immunoreactive substance on the histology of the mantle edge in adult and juvenile snails (Helix aspersa)].

We have demonstrated structural differences between the mantle edge gland of young, adult and adult shell repairing snails. In vivo and in vitro, somatostatin-14 and somatostatin-immunoreactive substance (SSI) induce a juvenile status of the gland in adults and an activation of juvenile ones. These results suggest that they could regulate the growth and the repair of the shell and that the mantle edge gland might be considered as one of their tissue targets.

Animals↗