Pond Snail Lymnaea Stagnalis Research Articles

Contrary Effects of Octopamine Receptor Ligands on Behavioral and Neuronal Changes in Locomotion of Lymnaea

The pond snail Lymnaea stagnalis moves along the sides and bottom of an aquarium, but it can also glide upside down on its back below the water's surface. We have termed these two forms of locomotion "standard locomotion" and "upside-down gliding," respectively. Previous studies showed that standard locomotion is produced by both cilia activity on the foot and peristaltic contraction of the foot muscles, whereas upside-down gliding is mainly caused by cilia activity. The pedal A neurons are thought to receive excitatory octopaminergic input, which ultimately results in increased cilia beating. However, the relationship between locomotory speed and the responses of these neurons to octopamine is not known. We thus examined the effects of both an agonist and an antagonist of octopamine receptors on locomotory speed and the firing rate of the pedal A neurons. We also examined, at the electron and light-microscopic levels, whether structural changes occur in cilia following the application of either an agonist or an antagonist of octopamine receptors to the central nervous system (CNS). We found that the application of an octopamine antagonist to the CNS increased the speed of both forms of locomotion, whereas application of octopamine increased only the firing rate of the pedal A neurons. Microscopic examination of the cilia proved that there were no changes in their morphology after application of octopamine ligands. These data suggest that there is an unidentified octopaminergic neuronal network in the CNS whose activation reduces cilia movement and thus locomotory speed.

Cercaria of the Trematode Plagiorchis Mutationis (Trematoda, Plagiorchiidae) from the Pond Snails, Lymnaea Stagnalis, in Ukraine

Cercaria of the Trematode Plagiorchis Mutationis (Trematoda, Plagiorchiidae) from the Pond Snails, Lymnaea Stagnalis, in Ukraine Morphological characteristics and measurements of Plagiorchis mutationis Panova, 1927 cercaria found for the first time in the intermediate host, pond snail Lymnaea stagnalis in Ukraine are given.

Effect of the Heavy Metals on Biological Characteristics of the Pond Snail (Lymnaea stagnalis L.) from the Water Bodies with Different Rate of Radionuclide Contamination

Effect of the Heavy Metals on Biological Characteristics of the Pond Snail (Lymnaea stagnalis L.) from the Water Bodies with Different Rate of Radionuclide Contamination

Costs of receipt and donation of ejaculates in a simultaneous hermaphrodite

BackgroundSexual conflicts between mating partners can strongly impact the evolutionary trajectories of species. This impact is determined by the balance between the costs and benefits of mating. However, due to sex-specific costs it is unclear how costs compare between males and females. Simultaneous hermaphrodites offer a unique opportunity to determine such costs, since both genders are expressed concurrently. By limiting copulation of focal individuals in pairs of pond snails (Lymnaea stagnalis) to either the male role or the female role, we were able to compare the fecundity of single sex individuals with paired hermaphrodites and non-copulants. Additionally, we examined the investment in sperm and seminal fluid of donors towards feminized snails and hermaphrodites.ResultsCompared to non-mating focal snails, reciprocating individuals as well as male and female copulants experienced a significant fecundity reduction (~40%) after, on average, 3.07 ± 0.12 copulations in their allowed roles (for donors 2.98 ± 0.16 copulations and for recipients 3.14 ± 0.12 copulations). In a single copulation, significantly more sperm was donated to partners that were restricted to mating in the female role than to hermaphrodites, while seminal fluid transfer was unaffected by recipient type.ConclusionsOur data indicate that the costs of mating in both sex functions are high in L. stagnalis. This conclusion is based on fecundity data collected separately for male and female copulants. Male mating costs result from investment in expensive ejaculates, composed of sperm and seminal fluid. For female copulants, fecundity reduction correlated with transferred sperm numbers in the first copulation, while differences in transferred quantities of seminal fluid were not detected. These findings may point toward a "sperm effect" as a novel feature of pond snail reproductive ecology. In conclusion, sex allocation and sexual conflict both contribute to decreased female fecundity in pond snails.

Balanced plasticity and stability of the electrical properties of a molluscan modulatory interneuron after classical conditioning: a computational study

The Cerebral Giant Cells (CGCs) are a pair of identified modulatory interneurons in the Central Nervous System of the pond snail Lymnaea stagnalis with an important role in the expression of both unconditioned and conditioned feeding behavior. Following single-trial food-reward classical conditioning, the membrane potential of the CGCs becomes persistently depolarized. This depolarization contributes to the conditioned response by facilitating sensory cell to command neuron synapses, which results in the activation of the feeding network by the conditioned stimulus. Despite the depolarization of the membrane potential, which enables the CGGs to play a key role in learning-induced network plasticity, there is no persistent change in the tonic firing rate or shape of the action potentials, allowing these neurons to retain their normal network function in feeding. In order to understand the ionic mechanisms of this novel combination of plasticity and stability of intrinsic electrical properties, we first constructed and validated a Hodgkin-Huxley-type model of the CGCs. We then used this model to elucidate how learning-induced changes in a somal persistent sodium and a delayed rectifier potassium current lead to a persistent depolarization of the CGCs whilst maintaining their firing rate. Including in the model an additional increase in the conductance of a high-voltage-activated calcium current allowed the spike amplitude and spike duration also to be maintained after conditioning. We conclude therefore that a balanced increase in three identified conductances is sufficient to explain the electrophysiological changes found in the CGCs after classical conditioning.

Learning-dependent gene expression of CREB1 isoforms in the molluscan brain

Cyclic AMP-responsive element binding protein1 (CREB1) has multiple functions in gene regulation. Various studies have reported that CREB1-dependent gene induction is necessary for memory formation and long-lasting behavioral changes in both vertebrates and invertebrates. In the present study, we characterized Lymnaea CREB1 (LymCREB1) mRNA isoforms of spliced variants in the central nervous system (CNS) of the pond snail Lymnaea stagnalis. Among these spliced variants, the three isoforms that code a whole LymCREB1 protein are considered to be the activators for gene regulation. The other four isoforms, which code truncated LymCREB1 proteins with no kinase inducible domain, are the repressors. For a better understanding of the possible roles of different LymCREB1 isoforms, the expression level of these isoform mRNAs was investigated by a real-time quantitative RT-PCR method. Further, we examined the changes in gene expression for all the isoforms in the CNS after conditioned taste aversion (CTA) learning or backward conditioning as a control. The results showed that CTA learning increased LymCREB1 gene expression, but it did not change the activator/repressor ratio. Our findings showed that the repressor isoforms, as well as the activator ones, are expressed in large amounts in the CNS, and the gene expression of CREB1 isoforms appeared to be specific for the given stimulus. This was the first quantitative analysis of the expression patterns of CREB1 isoforms at the mRNA level and their association with learning behavior.

The distinction between retractor and protractor muscles of the freshwater snail's male organ has no physiological basis

Many animals are equipped with organs that can be everted, a notable example being male copulatory organs. The ability to protrude or evert an organ generally requires protractor and retractor muscles. Male copulatory behaviour of the pond snail Lymnaea stagnalis (L.) involves eversion (protraction) and retraction of the relatively large penis-carrying organ. For this preputium, protractor and retractor muscle bands have been defined, which implies eversion and retraction through the activity of these muscle bands. However, no physiological data are available that confirm that the terms protractor and retractor are appropriate. To test whether eversion and retraction are possible without protractor and/or retractor muscle bands, lesion experiments were performed. The results show that with either one or several muscle bands lesioned, snails were still capable of everting their preputium and using it for copulation. However, the majority of animals that had six or more muscle bands lesioned were unable to retract its preputium. Hence, retractor muscle bands serve their designated function whereas protractor muscle bands do not. We therefore suggest that a different terminology is used in which all muscle bands are retractors and, based on their location, are either called distal or proximal retractors. The findings furthermore indicate that the preputium muscle bands are normally contracted, possibly in a catch state, retaining the organ inside without high-energy expenditure.

Glutamate transporters in the central nervous system of a pond snail

Previous studies on glutamate (GLU) and its receptors in the pond snail Lymnaea stagnalis have suggested that GLU functions as a neurotransmitter in various behaviors, particularly for generation of feeding rhythm. The uptake mechanism of GLU is not yet known in Lymnaea. In the present study, we characterized the GLU transporters and examined their functions in the feeding circuits of the central nervous system (CNS) in Lymnaea. First, measurement of the accumulation of (3)H-labeled GLU revealed the presence of GLU transport systems in the Lymnaea CNS. The highest accumulation rate was observed in the buccal ganglia, supporting the involvement of GLU transport systems in feeding behavior. Second, we cloned two types of GLU transporters from the Lymnaea CNS, the excitatory amino acid transporter (LymEAAT) and the vesicular GLU transporter (LymVGLUT). When we compared their amino acid sequences with those of mammalian EAATs and VGLUTs, we found that the functional domains of both types are well conserved. Third, in situ hybridization revealed that the mRNAs of LymEAAT and LymVGLUT are localized in large populations of nerve cells, including the major feeding motoneurons in the buccal ganglia. Finally, we inhibited LymEAAT and found that changes in the firing patterns of the feeding motoneurons that have GLUergic input were similar to those obtained following stimulation with GLU. Our results confirmed the presence of GLU uptake systems in the Lymnaea CNS and showed that LymEAAT is required for proper rhythm generation, particularly for generation of the feeding rhythm.

Individual growth of Lymnaea stagnalis (Lymnaeidae, Gastropoda): II. Late postlarval ontogeny

Our investigation on the growth of 14 individuals of the great pond snail Lymnaea stagnalis was performed in an aquatic culture at 18 degrees C beginning from the 10th week after hatching until death. It has been demonstrated that the increase in the mollusk mass follows an S-like curve during the whole studied period. Linear growth (conch height) follows a parabolic (convex) curve until the age of 39 weeks. Both weight and linear growth during studied period significantly approximate to the Bertalanffy equation, while the interrelation between mass and conch height corresponds to the allometric equation. The meanings of the coefficients of these equations do not differ significantly in different individuals. At the age of 38 to 39 weeks, all mollusks demonstrate breakage in the curve of linear growth, then followed with abrupt slowing of growth until stopping or even decreasing in size in some cases. Neither the Bertalanffy equation nor the allometric relation describe the linear growth of individuals with ages exceeding 39 weeks.

Atypical guanylyl cyclase from the pond snail Lymnaea stagnalis: cloning, sequence analysis and characterization of expression

Soluble guanylyl cyclases (sGCs) are traditionally recognized as the main molecular receptor for nitric oxide (NO), a gaseous transmitter involved in many functions of the nervous system. Some sGCs are however insensitive to NO and therefore are known as atypical. Although atypical sGCs have been shown to exist in both vertebrate and invertebrate nervous systems, our understanding of their functional role is incomplete. Here we report on the cloning, sequencing and localization of an atypical sGC named Lym-sGCβ3 from the snail Lymnaea stagnalis. We found that Lym-sGCβ3 shares a number of structural characteristics with some previously characterized atypical sGCs including the presence of Tyr140 in the regulatory domain. This residue is thought to be of a critical importance in determining sensitivity of atypical sGCs to oxygen. These findings raise the possibility that Lym-sGCβ3 is an oxygen receptor. The results of our in situ hybridization and RT-PCR experiments support this idea further by showing that Lym-sGCβ3 is expressed in the osphradium, a peripheral sense organ in which oxygen-sensing neurons are located. Also of interest are our observations that many neurons in Lymnaea CNS co-express conventional and atypical sGC subunits. These data are consistent with a possible dominant negative regulatory role of atypical sGC subunits through the formation of heterodimers exhibiting low enzymatic activity.

Embryonic rotational behaviour in the pond snail Lymnaea stagnalis: influences of environmental oxygen and development stage

Responses of freshwater organisms to environmental oxygen tensions (PO 2) have focused on adult (i.e. late developmental) stages, yet responses of embryonic stages to changes in environmental PO 2 must also have implications for organismal biology. Here we assess how the rotational behaviour of the freshwater snail Lymnaea stagnalis changes during development in response to conditions of hypoxia and hyperoxia. As rotation rate is linked to gas mixing in the fluid surrounding the embryo, we predicted that it would increase under hypoxic conditions but decrease under hyperoxia. Contrary to predictions, early, veliger stage embryos showed no change in their rotation rate under hyperoxia, and later, hippo stage embryos showed only a marginally significant increase in rotation under these conditions. Predictions for hypoxia were broadly supported, however, with both veliger and hippo stages showing a marked hypoxia-related increase in their rotation rates. There were also subtle differences between developmental stages, with hippos responding at PO 2s (50% air saturation) greater than those required to elicit a similar response in veligers (20% air saturation). Differences between developmental stages also occurred on return to normoxic conditions following hypoxia: rotation in veligers returned to pre-exposure levels, whereas there was a virtual cessation in embryos at the hippo stage, likely the result of overstimulation of oxygen sensors driving ciliary movement in later, more developed embryos. Together, these findings suggest that the spinning activity of L. stagnalis embryos varies depending on environmental PO 2s and developmental stage, increasing during hypoxia to mix capsular contents and maintain a diffusive gradient for oxygen entry into the capsule from the external environment (“stir-bar” theory of embryonic rotational behaviour).

GABAA- and AMPA-like receptors modulate the activity of an identified neuron within the central pattern generator of the pond snail Lymnaea stagnalis

To examine the neurochemistry underlying the firing of the RPeD1 neuron in the respiratory central pattern generator of the pond snail, Lymnaea stagnalis, we examined electrophysiologically and pharmacologically either "active" or "silent" preparations by intracellular recording and pharmacology. GABA inhibited electrical firing by hyperpolarizing RPeD1, while picrotoxin, an antagonist of GABA(A) receptors, excited silent cells and reversed GABA-induced inhibition. Action potential activity was terminated by 1 mM glutamate (Glu) while silent cells were depolarized by the GluR agonists, AMPA, and NMDA. Kainate exerted a complex triphasic effect on membrane potential. However, only bath application of AMPA desensitized the firing. These data indicate that GABA inhibits RPeD1 via activation of GABA(A) receptors, while Glu stimulates the neuron by activating AMPA-sensitive GluRs.

Environmental rotenone treatment destroys primary receptor neurons in the pond Snaillymnaea Stagnalis

Event Abstract Back to Event Environmental rotenone treatment destroys primary receptor neurons in the pond Snaillymnaea Stagnalis Agnes Vehovszky1*, H. Szabo1 and L. Hernadi1 1 Balaton Limnological Research Institute, HAS, Hungary Environmental rotenon treatment of Lymnaea stagnalis attenuates the DAergic functions, resulting locomotory impairment and decreased feeding behavior in a concentration dependent manner. However, we do not know if the attenuated central DAergic system is responsible for the decreased behavioral responses or rotenone primarily destroys the sensory neurons in the skin or the buccal areas which may lead to the cessation of behaviors. Morphological strudies were carried out to test the effects of environmental rotenon treatment (0.5 uM) on primary sensory neurons located in the oral-buccal areas to determine whether impairment of the sensory system leads to the decreased feeding behavior of treated animals. Retrograde neurobiotin tracing via cerebral and buccal nerves which innervate head areas and the oral-buccal cavity revealed a huge number of primary sensory neurons labeled in the lips, and the oral cavity. The tracing via buccal nerves revealed a huge number of labeled sensory neurons in the bucal cavity and the ventral part of the oral cavity which is also innervated by a cerebral nerve. Tracing via cerebro-buccal connectives revealed that a low number of sensory neurons are labeled in the lips and the oral cavity. In rotenon treated animals the neurobiotin tracing via cerebral and buccal nerves failed to label primary sensory neurons either in the lips and mouth or the oral-buccal cavity suggesting that rotenon treatment destroyed primary sensory neurons in body surfaces exposed to rotenone. Since the activity of these sensory cells is required to the initiation of feeding CPG therefore their degeneration may be responsible for the cessation of Lymnaea food intake. Conference: 12th Meeting of the Hungarian Neuroscience Society, Budapest, Hungary, 22 Jan - 24 Jan, 2009. Presentation Type: Poster Presentation Topic: Pathophysiology and neurology - degenerative disorders Citation: Vehovszky A, Szabo H and Hernadi L (2009). Environmental rotenone treatment destroys primary receptor neurons in the pond Snaillymnaea Stagnalis. Front. Syst. Neurosci. Conference Abstract: 12th Meeting of the Hungarian Neuroscience Society. doi: 10.3389/conf.neuro.01.2009.04.179 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 05 Mar 2009; Published Online: 05 Mar 2009. * Correspondence: Agnes Vehovszky, Balaton Limnological Research Institute, HAS, Tihany, Hungary, agnes@tres.blki.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Agnes Vehovszky H. Szabo L. Hernadi Google Agnes Vehovszky H. Szabo L. Hernadi Google Scholar Agnes Vehovszky H. Szabo L. Hernadi PubMed Agnes Vehovszky H. Szabo L. Hernadi Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Hypoxia-induced modulation of the respiratory CPG

Despite recent advances in our understanding of the neural control of breathing, the precise cellular, synaptic, and molecular mechanisms underlying the generation and modulation of respiratory rhythm remain largely unknown. This lack of fundamental knowledge in the field of neural control of respiration is likely due to the complexity of the mammalian brain where synaptic connectivity between central respiratory neurons, motor neurons and their peripheral counterparts cannot be mapped reliably. We have therefore developed an invertebrate model system wherein the essential elements of the central pattern generator (CPG), the motor neurons and the peripheral chemosensory cells involved in respiratory control have been worked out both in vivo and in vitro. We discuss our recent identification of peripheral, hypoxia sensitive chemoreceptor elements in a sensory organ of the pulmonate freshwater pond snail Lymnaea stagnalis, which provide an excitatory drive to the respiratory CPG neuron RPeD1 via direct chemical synaptic connections. Further studies using this unique invertebrate model system may reveal highly conserved principles of CPG neuromodulation that will remain relevant to more complex mammalian systems.

Upside-Down Gliding ofLymnaea

The pond snail Lymnaea stagnalis can often be observed moving upside down on its back just below the surface of the water. We have termed this form of movement "upside-down gliding." To elucidate the mechanism of this locomotion, we performed a series of experiments involving behavioral analyses and microscopic observations. These experiments were designed (1) to measure the speed of this locomotion; (2) to determine whether the mucus secreted from the foot of Lymnaea repels water, thereby allowing the snail to exploit the surface tension of the water for upside-down gliding; and (3) to observe the beating of foot cilia in this behavior. The beating of these cilia is thought to be the primary driving force for upside-down gliding. Our results demonstrate that upside-down gliding is an efficient active process involving the secretion of mucus that floats up to the water surface to serve as a substrate upon which cilia beat to cause locomotion at the underside of the water surface.

Variations in Functional Activity of the Hormone-Sensitive Adenylate Cyclase System in Tissues of Gastropod Mollusks with Streptozotocin-Induced Diabetes

Treatment of gastropod mollusks of pond snail Lymnaea stagnalis and orb snail Coretus corneus with streptozotocin was followed by an increase in hexose content in the hemolymph and development of the diabetic state (day 1 after treatment). Functional activity of the hormone-sensitive adenylate cyclase system significantly decreased in the muscles and hepatopancreas of mollusks with diabetes. We revealed a decrease in the regulatory effects of biogenic amines and peptide hormones that were realized via stimulatory (octopamine, dopamine, serotonin, tryptamine, and relaxin) and inhibitory G proteins (somatostatin). Disturbances in the hepatopancreas were more pronounced than in the muscle. The severity of disorders in the adenylate cyclase system reached maximum 1 day after streptozotocin treatment. The sensitivity of this system to hormonal and nonhormonal agents was partially restored on days 3 and 5. Hexose content in the hemolymph was elevated after streptozotocin treatment, but returned to normal on day 3. Our results indicate that hyperglycemia is one of the key factors for dysfunction of the adenylate cyclase system in mollusks with the diabetic state.

Different phases of long-term memory require distinct temporal patterns of PKA activity after single-trial classical conditioning

The cAMP-dependent protein kinase (PKA) is known to play a critical role in both transcription-independent short-term or intermediate-term memory and transcription-dependent long-term memory (LTM). Although distinct phases of LTM already have been demonstrated in some systems, it is not known whether these phases require distinct temporal patterns of learning-induced PKA activation. This question was addressed in a robust form of associative LTM that emerges within a matter of hours after single-trial food-reward classical conditioning in the pond snail Lymnaea stagnalis. After establishing the molecular and functional identity of the PKA catalytic subunit in the Lymnaea nervous system, we used a combination of PKA activity measurement and inhibition techniques to investigate its role in LTM in intact animals. PKA activity in ganglia involved in single-trial learning showed a short latency but prolonged increase after classical conditioning. However, while increased PKA activity immediately after training (0-10 min) was essential for an early phase of LTM (6 h), the late phase of LTM (24 h) required a prolonged increase in PKA activity. These observations indicate mechanistically different roles for PKA in recent and more remote phases of LTM, which may underpin different cellular and molecular mechanisms required for these phases.

Isolated neurons as biosensors responding to the release of neuroactive substances

Current neurophysiology has no unambiguous view of the roles of neurotransmitter substances found in the intercellular environment of nervous tissue. The classical doctrine, which restricts the sphere of activity of neurotransmitters to the synaptic cleft, is very obviously inadequate. However, it is difficult to develop suitable alternative concepts because of the inadequacy of the methods used to obtain an objective picture of events occurring in the extrasynaptic space. Methods such as voltometry have been described, which allow the dynamics of a number of neurons to be followed but which have low suitability for studies of neurobiological reality, which is an unstable cocktail of both identified and unidentified neuroactive substances. The method suggested here was developed to overcome this difficulty. It is based on the use of a chemosensitive biosensor ‐ a multisensory isolated neuron which follows circumstances in a selected locus of extracellular space, for example close to a pattern-generating neural network. We have previously reported some preliminary results obtained using this detector [2, 3]; this is the first report on the technical aspects of the method and its possible use for longterm monitoring of the milieu. The method was developed using synaptic preparations from the pond snail Lymnaea stagnalis. Mollusks were reared in aquarium cultures and isolated CNS preparations were made as described in [2], with transection of connectives where necessary. The signal Source. The extracellular medium was tested at the surface of one of the CNS ganglia (usually the buccal or pedal ganglia, less commonly the parietal or visceral ganglia). The outer (thick) coating was surgically removed from the ganglion, leaving the thin layer untouched. Preparations were treated with pronase E (4 mg/ml, 5‐7 min; Sigma) followed by washing with physiological solution. The Biosensor. Neurons used for detection of chemi

Effects of the mixture of diquat and a nonylphenol polyethoxylate adjuvant on fecundity and progeny early performances of the pond snail Lymnaea stagnalis in laboratory bioassays and microcosms

Effects of the bipyridylium herbicide diquat and tank-mix adjuvant Agral ®90 were investigated on various life history traits of the freshwater pulmonate snail Lymnaea stagnalis. Trait expression was measured in simple laboratory bioassays on small size groups of snails, and under more complex, indoor microcosm conditions, on larger groups of snails. Microcosms were provided with sediment, plants, and fish, thus allowing a more complex level of intra and inter-specific interactions to develop. Treatments were performed with substances alone or in mixture, at concentrations ranging from 4.4 to 222.2 μg l −1 for diquat, and from 10 to 500 μg l −1 for Agral 90, under a fixed ratio design. Adult growth was negatively affected by diquat and its mixture with Agral 90 both at the highest concentrations (222.2 and 500 μg l −1, respectively). Fecundity expressed differently in bioassays and microcosms, but no effect of the chemicals could be observed on this trait. Progeny development was impaired by 222.2 μg l −1 diquat and its mixture with 500 μg l −1 Agral 90, as reflected by longer development time and reduced hatching rate of clutches laid by the exposed animals, as compared to the controls. Hatching data suggested that diquat bioavailability was lower in microcosms than under bioassay conditions. Consistently, chemical analysis showed that diquat disappeared more rapidly from the water in microcosms than in bioassays. Moreover, the differential expression of several life history traits under bioassays and microcosms conditions was probably also influenced by the level of intraspecific interaction, which differed among the systems. When significant, the effect of diquat was attenuated by the presence of Agral 90, indicating antagonistic interaction between the two substances. Such a deviation from additivity was partly validated statistically.

Building a house when supplies are minimal: Ca2+ and HCO3− acquisition in freshwater by embryos of the common pond snail Lymnaea stagnalis

Building a house when supplies are minimal: Ca2+ and HCO3− acquisition in freshwater by embryos of the common pond snail Lymnaea stagnalis