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Nonspecific snail muscle adenylate deaminase: simplified purification, characterization and use for the preparation of deamino derivatives of NAD, NADH and AMP-P(NH)P.

Chromatography on phosphocellulose P-11 under conditions different from those applicable for deaminases specific to 5' AMP resulted in homogeneous preparation of snail foot muscle enzyme. Snail deaminase is a tetramer with molecular weight of 240,000, composed of four apparently identical subunits. Its amino acid composition is remarkably different from deaminases of higher animals, it was not inhibited by EDTA, but zinc became inhibitory to the snail enzyme. Unlike deaminases specific to 5' AMP, nonspecific deaminase is not a zinc-containing enzyme. It was adopted further for the preparation of hypoxanthine derivatives of adenosine-containing nucleotides such as NAD, NADH, AMP-P(NH)P, AMP, ADP and ATP.

AMP Deaminase↗

Sexual selection on land snail shell ornamentation: a hypothesis that may explain shell diversity.

BACKGROUND: Many groups of land snails show great interspecific diversity in shell ornamentation, which may include spines on the shell and flanges on the aperture. Such structures have been explained as camouflage or defence, but the possibility that they might be under sexual selection has not previously been explored. PRESENTATION OF THE HYPOTHESIS: The hypothesis that is presented consists of two parts. First, that shell ornamentation is the result of sexual selection. Second, that such sexual selection has caused the divergence in shell shape in different species. TESTING THE HYPOTHESIS: The first part of the hypothesis may be tested by searching for sexual dimorphism in shell ornamentation in gonochoristic snails, by searching for increased variance in shell ornamentation relative to other shell traits, and by mate choice experiments using individuals with experimentally enhanced ornamentation. The second part of the hypothesis may be tested by comparing sister groups and correlating shell diversity with degree of polygamy. IMPLICATIONS OF THE HYPOTHESIS: If the hypothesis were true, it would provide an explanation for the many cases of allopatric evolutionary radiation in snails, where shell diversity cannot be related to any niche differentiation or environmental differences.

Animals↗

Organization of the peripheral fly eye: the roles of Snail family transcription factors in peripheral retinal apoptosis.

The periphery of the fly eye contains a number of concentrically arranged cellular specializations that are induced by Wingless (Wg) signaling from the surrounding head capsule (HC). One of these is the pigment rim (PR), which is a thick layer of pigment cells that lies directly adjacent to the HC and completely circumscribes the rest of the retina. Many of the cells of the PR are derived from presumptive pigment cells that previously surrounded peripheral ommatidia that subsequently died. Here, we describe the Wg-elicited expression of Snail family transcription factors in the eye periphery that directs the ommatidial death and subsequent PR formation. These transcription factors are expressed only in a subset of the ommatidial cells not including the photoreceptors. Yet, the photoreceptors die and, thus, a non-autonomous death signal is released from the Snail-family-expressing cells that direct the death of the photoreceptors. In addition, Wg also elicits a similar peripheral expression of Notum, an enzyme that limits the extent of Wg signaling. Furthermore, we describe a later requirement for Snail family proteins in the 2 degrees and 3 degrees pigment cells throughout the main body of the eye.

Animals↗

Conserved and divergent roles for members of the Snail family of transcription factors in the chick and mouse embryo.

The members of the Snail family of zinc-finger transcription factors have been implicated in the formation of distinct tissues within the developing vertebrate and invertebrate embryo. Two members of this family have been described in higher vertebrates, Snail (Sna) and Slug (Slu), where they have been implicated in the formation of tissues such as the mesoderm and the neural crest. We have isolated the mouse homologue of the Slu gene enabling us to analyse and compare the amino acid sequences and the patterns of expression of both Sna and Slu in the chick and mouse. We have detected features in the sequences that allow the unequivocal ascription of any family member to the Sna or Slu subfamilies and we have observed that, during early stages of development, many of the sites of Slu and Sna expression in the mouse and chick embryo are swapped. Later in development, the sites of expression of Slu and Sna are conserved between these two species. These data, together with the data available in other species, lead us to propose that Slu and Sna arose as a duplication of an ancestor gene and that an extra duplication in the fish lineage has given rise to two Sna genes. Furthermore, several early sites of Slu and Sna expression have been swapped in the avian lineage. Our analysis of the Snail family may also shed new light on the origin of the neural crest.

Animals↗

The transcription factor Slug represses E-cadherin expression and induces epithelial to mesenchymal transitions: a comparison with Snail and E47 repressors.

Transcriptional repression mechanisms have emerged as one of the crucial processes for the downregulation of E-cadherin expression during development and tumour progression. Recently, several E-cadherin transcriptional repressors have been characterized (Snail, E12/E47, ZEB-1 and SIP-1) and shown to act through an interaction with proximal E-boxes of the E-cadherin promoter. We have analyzed the participation of another member of the Snail family, Slug, and observed that it also behaves as a repressor of E-cadherin expression. Stable expression of Slug in MDCK cells leads to the full repression of E-cadherin at transcriptional level and triggers a complete epithelial to mesenchymal transition. Slug-induced repression of E-cadherin is mediated by its binding to proximal E-boxes, particularly to the E-pal element of the mouse promoter. Detailed analysis of the binding affinity of different repressors to the E-pal element indicates that Slug binds with lower affinity than Snail and E47 proteins. These results, together with the known expression patterns of these factors in embryonic development and carcinoma cell lines, support the idea that the in vivo action of the different factors in E-cadherin repression can be modulated by their relative concentrations as well as by specific cellular or tumour contexts.

Animals↗

The actions of FMRFamide-like peptides on visceral and somatic muscles of the snail Helix aspersa.

Phe-Met-Arg-Phe-NH2 (FMRFamide) and pyroGlu-Asp-Pro-Phe-Leu-Arg-Phe-NH2 (pQDPFLRFamide) occur in the ganglia and tissues of the snail, Helix aspersa. This report describes the effects of these two neuropeptides on five visceral organs or somatic muscles isolated from the snail (Table 1). The epiphallus, as well as the rest of the male reproductive tract, was contracted by both FMRFamide and pQDPFLRFamide, and the threshold was usually below 5 X 10(-9) mol l-1 (Fig. 1). Both peptides also reduced the resting tone of the crop and decreased the force and frequency of its rhythmic activity; FMRFamide is about 10 times more potent (Fig. 4). In contrast, pQDPFLRFamide was about 100 times more potent than FMRFamide as a cardioexcitatory agent (Fig. 5). The actions of the peptides on the pharyngeal and tentacle retractor muscles were markedly different: FMRFamide primarily contracted these muscles; and pQDPFLRFamide usually had no effect alone, but relaxed or diminished contractions induced by FMRFamide and acetylcholine (ACh) (Figs 6, 8, 9). Other analogues of FMRFamide were tested, but none was as effective a relaxing agent as pQDPFLRFamide. The effects of FMRFamide and pQDPFLRFamide on all of the preparations could be distinguished from those produced by ACh and 5-hydroxytryptamine (5-HT); thus the actions of the neuropeptides were not mediated by cholinergic or serotonergic neurones. The stimulation of the musculature in the male reproductive tract and the inhibition of motility of the digestive system by FMRFamide and pQDPFLRFamide implicate these peptides in the control of reproductive behaviour. The effectiveness of pQDPFLRFamide in relaxing the retractor muscles and as a cardioexcitatory agent led to the hypothesis that this heptapeptide and FMRFamide, acting at distinct receptors, cooperate to regulate the excitability and contractility of the snail's musculature between the extremes of aestivation and active locomotion.

Acetylcholine↗

Bisphenol A induces superfeminization in the ramshorn snail Marisa cornuarietis(Gastropoda: Prosobranchia) at environmentally relevant concentrations.

Previous investigations have shown that bisphenol A (BPA) induces a superfeminization syndrome in the freshwater snail Marisa cornuarietis at concentrations as low as 1 microg/L. Superfemales are characterized by the formation of additional female organs, enlarged accessory sex glands, gross malformations of the pallial oviduct, and a stimulation of egg and clutch production, resulting in increased female mortality. However, these studies were challenged on the basis of incomplete experimentation. Therefore, the objective of the current approach was to bridge several gaps in knowledge by conducting additional experiments. In an initial series of experiments, study results from the reproductive phase of the snails were evaluated in the sub-micrograms per liter range. Before and after the spawning season, superfemale responses were observed [NOEC (no observed effect concentration) 7.9 ng/L, EC10 (effective concentration at 10%) 13.9 ng/L], which were absent during the spawning season. A further experiment investigated the temperature dependence of BPA responses by exposing snails at two temperatures in parallel. The adverse effect of BPA was at least partially masked at 27 degrees C (EC10 998 ng/L) when compared with 20 degrees C (EC10 14.8 ng/L). In M. cornuarietis, BPA acts as an estrogen receptor (ER) agonist, because effects were completely antagonized by a co-exposure to tamoxifen and Faslodex. Antiandrogenic effects of BPA, such as a significant decrease in penis length at 20 degrees C, were also observed. Competitive receptor displacement experiments indicate the presence of androgen- and estrogen-specific binding sites. The affinity for BPA of the estrogen binding sites in M. cornuarietis is higher than that of the ER in aquatic vertebrates. The results emphasize that prosobranchs are affected by BPA at lower concentrations than are other wildlife groups, and the findings also highlight the importance of exposure conditions.

Animals↗

Speciation and gene flow between snails of opposite chirality.

Left-right asymmetry in snails is intriguing because individuals of opposite chirality are either unable to mate or can only mate with difficulty, so could be reproductively isolated from each other. We have therefore investigated chiral evolution in the Japanese land snail genus Euhadra to understand whether changes in chirality have promoted speciation. In particular, we aimed to understand the effect of the maternal inheritance of chirality on reproductive isolation and gene flow. We found that the mitochondrial DNA phylogeny of Euhadra is consistent with a single, relatively ancient evolution of sinistral species and suggests either recent "single-gene speciation" or gene flow between chiral morphs that are unable to mate. To clarify the conditions under which new chiral morphs might evolve and whether single-gene speciation can occur, we developed a mathematical model that is relevant to any maternal-effect gene. The model shows that reproductive character displacement can promote the evolution of new chiral morphs, tending to counteract the positive frequency-dependent selection that would otherwise drive the more common chiral morph to fixation. This therefore suggests a general mechanism as to how chiral variation arises in snails. In populations that contain both chiral morphs, two different situations are then possible. In the first, gene flow is substantial between morphs even without interchiral mating, because of the maternal inheritance of chirality. In the second, reproductive isolation is possible but unstable, and will also lead to gene flow if intrachiral matings occasionally produce offspring with the opposite chirality. Together, the results imply that speciation by chiral reversal is only meaningful in the context of a complex biogeographical process, and so must usually involve other factors. In order to understand the roles of reproductive character displacement and gene flow in the chiral evolution of Euhadra, it will be necessary to investigate populations in which both chiral morphs coexist.

Animals↗

Photoperiod and temperature interaction in the determination of reproduction of the edible snail, Helix pomatia.

Snails were kept in self-cleaning housing chambers in an artificially controlled environment. Mating was frequent under long days (18 h light) and rare under short days (8 h light) regardless of whether the snails were kept at 15 degrees C or 20 degrees C. An interaction between photoperiod and temperature was observed for egg laying. The number of eggs laid (45-50/snail) and the frequency of egg laying (90-130%) were greater in long than in short days (16-35/snail and 27-77%) but a temperature of 20 degrees C redressed, to some extent, the inhibitory effect of short days. At both temperatures only long photoperiods brought about cyclic reproduction over a period of 16 weeks, confirming the synchronizing role of photoperiod on the neuroendocrine control of egg laying in this species of snail.

Animals↗

Monoamines in the pedal plexus of the land snail Megalobulimus oblongus (Gastropoda, Pulmonata).

In molluscs, the number of peripheral neurons far exceeds those found in the central nervous system. Although previous studies on the morphology of the peripheral nervous system exist, details of its organization remain unknown. Moreover, the foot of the terrestrial species has been studied less than that of the aquatic species. As this knowledge is essential for our experimental model, the pulmonate gastropod Megalobulimus oblongus, the aim of the present study was to investigate monoamines in the pedal plexus of this snail using two procedures: glyoxylic acid histofluorescence to identify monoaminergic structures, and the unlabeled antibody peroxidase anti-peroxidase method using antiserum to detect the serotonergic component of the plexus. Adult land snails weighing 48-80 g, obtained from the counties of Barra do Ribeiro and Charqueadas (RS, Brazil), were utilized. Monoaminergic fibers were detected throughout the pedal musculature. Blue fluorescence (catecholamines, probably dopamine) was observed in nerve branches, pedal and subepithelial plexuses, and in the pedal muscle cells. Yellow fluorescence (serotonin) was only observed in thick nerves and in muscle cells. However, when immunohistochemical methods were used, serotonergic fibers were detected in the pedal nerve branches, the pedal and subepithelial plexuses, the basal and lateral zones of the ventral integument epithelial cells, in the pedal ganglion neurons and beneath the ventral epithelium. These findings suggest catecholaminergic and serotonergic involvement in locomotion and modulation of both the pedal ganglion interneurons and sensory information. Knowledge of monoaminergic distribution in this snail s foot is important for understanding the pharmacological control of reflexive responses and locomotive behavior.

Animals↗

Nutritional status of four species of giant land snails in Nigeria.

Four species of African giant land snails (Archachatina marginata (ovum) Pfeiffer, Archachatina marginata (saturalis) Philippi, Achatina achatina and Limicolaria spp.) were assessed for their proximate and mineral compositions aimed at establishing their nutritive values on wet weight basis. Analysis of muscle revealed that composition of crude protein varied from 18.66%+/-0.57% in Limicolaria spp. and 20.56%+/-0.05% in Archachatina marginata (ovum) Pfeiffer; moisture content was 76.56%+/-0.04% in Archachatina marginata (ovum) Pfeiffer and 78.68%+/-0.68% in Limicolaria spp. and ash was 1.34%+/-0.02% in Achatina achatina and 1.44%+/-0.01% in Archachatina marginata (ovum) Pfeiffer. These values were statistically different from each other (P<0.05). Carbohydrate and fat content were generally low. Crude fibre was not detected in any of the species. The concentrations of zinc, iron, manganese, magnesium, calcium, phosphorus, sulphur, potassium and sodium in the flesh of the snails were determined. Values of iron, magnesium, calcium, phosphorus, potassium and sodium were consistently high while cobalt, copper and lead were not detected. Snails complement the required trace and minor elements needed for proper growth and development in human being, so it is recommended for regular consumption.

Animals↗

Miracidia of an Egyptian strain of Schistosoma mansoni differentiate between sympatric snail species.

The host-finding behavior of miracidia of 2 strains of Schistosoma mansoni from Egypt and Brazil was studied by recording their responses to snail-conditioned water (SCW) from the Egyptian sympatric snails, Biomphalaria alexandrina, Physa acuta, Lymnaea cailliudi, and Balinus truncatus, as well as from Biomphalaria arabica and Biomphalaria glabrata. Miracidia of the Egyptian strain significantly preferred SCW from their compatible hosts B. alexandrina and B. arabica and showed no or a weak response to SCW from the other sympatric species, whereas miracidia of the Brazilian strain did not differentiate between SCW from different snail species.

Animals↗

Methods for toxicity assessment of contaminated soil by oral or dermal uptake in land snails: metal bioavailability and bioaccumulation.

Using two biological characteristics of the land snail (subspecies Helix aspersa aspersa and Helix aspersa maxima), i.e., soil eating and crawling on wet surfaces, methods were developed to assess the bioavailability of heavy metals (Cd, Cr, Pb, and Zn) from soil and aqueous leachates of soil. Measurement of heavy-metal concentrations in soil, leachates, and soft tissues of snails showed that contamination from ingested soil is the major route for metal uptake. Generally, the concentrations of metals were greater in the viscera than in the foot and rose with the proportion of spiked soil (Sl) in the diet. After ingestion of soil, bioaccumulation factors in the viscera were over two for Cd (3.5-27) and Zn (1.7-4.4) for both subspecies but were below one for Cr (0.04-0.6, except for H. aspersa maxima: 1.21) and Pb (0.02-0.23). Contamination by leachate Sl mainly caused accumulation of Cr in the tissues. Determining the metal burden per snail enables an estimation to be made of the risk of secondary poisoning encountered by predators. The present study provides data on the bioavailability of metals in soils and on the relationships between bioaccumulation and sublethal effects.

Animals↗

Piscivorous behavior of a temperate cone snail, Conus californicus.

Most of the more than 500 species of predatory marine snails in the genus Conus are tropical or semitropical, and nearly all are thought to be highly selective regarding type of prey. Conus californicus Hinds, 1844, is unusual in that it is endemic to the North American Pacific coast and preys on a large variety of benthic organisms, primarily worms and other molluscs, and also scavenges. We studied the feeding behavior of C. californicus in captivity and found that it regularly killed and consumed live prickleback fishes (Cebidichthys violaceus and Xiphister spp.). Predation involved two behavioral methods similar to those employed by strictly piscivorous relatives. One method utilized stings delivered by radular teeth; the other involved engulfing the prey without stinging. Both methods were commonly used in combination, and individual snails sometimes employed multiple stings to subdue a fish. During the course of the study, snails became aroused by the presence of live fish more quickly, as evidenced by more rapid initiation of hunting behavior. Despite this apparent adaptation, details of prey-capture techniques and effectiveness of stings remained similar over the same period.

Animals↗

Radix auricularia coreana: natural snail host of Clinostomum complanatum in Korea.

An epidemiological survey was carried out to determine the first intermediate host of Clinostomum complanatum among freshwater snails in Korea. Two species of snails belonging to the family Lymnaeidae were collected in Kaum-ji (pond), Uisong-gun, Kyongsangbuk-do. Twelve (0.9%) out of 1,273 Radix auricularia coreana examined were found to liberate cercariae of C. complanatum, which were identified by morphological characteristics and experimental infections in freshwater fish. Pseudorasbora parva. The cercariae were brevifurcate and clinostomatoid. They had a transparent dorsal fin, a well developed penetrating organ and a pair of eye spots. The body measured 119-147 x 33-36 microns, tail stem, 275-370 x 19-26 microns, and furcae, 72-104 microns. Rediae were demonstrated in the infected snail after crushing. Redia, 527-1,630 microns long and 121-368 microns wide, contained 10-45 germ balls and cercariae in various developmental stages. The metacercariae recovered from fish experimentally infected with C. complanatum cercariae were morphologically identical to those from naturally infected fish.

Animals↗

The electrical potential difference through the foot epithelium of the snail Achatina achatina, Lameere during mechanical and chemical stimulation.

An important electrophysiological variable--the transepithelial potential difference reflects the electrogenic transepithelial ion currents, which are produced and modified by ion transport processes in polarized cells of epithelium. These processes result from coordinated function of transporters in apical and basolateral cell membranes and have been observed in all epithelial tissues studied so far. The experiments were performed on isolated specimens of snail foot. In the experiments, the baseline transepithelial electrical potential difference--PD, changes of transepithelial difference during mechanical stimulation--dPD and the transepithelial resistance were measured with an Ussing apparatus. A total of 60 samples of foot ventral surface of 28 snails were studied. The transepithelial electrical potential difference of isolated foot ranged from -6.0 to 10.0 mV under different experimental conditions. Mechanical stimulation of foot ventral surface caused changes of electrogenic ion transport, observed as transient hyperpolarization (electrical potential difference became more positive). When the transepithelial electrical potential difference decreased during stimulation, the reaction was described as depolarization. When amiloride and bumetanide were added to the stimulating fluid so that the sodium and chloride ion transport pathways were inhibited, prolonged depolarization occurred. Under the influence of different stimuli: mechanical (gentle rinsing), chemical (changes of ion concentrations) and pharmacological (application of ion inhibitors), transient changes of potential difference (dPD) were evoked, ranging from about -0.7 to almost 2.0 mV. Changes in transepithelial potential difference of the pedal surface of the snail's foot related to these physiological stimuli are probably involved in the locomotion of the animal and are under control of the part of the nervous system in which tachykinin related peptides (TRP) act as transmitters.

Animals↗

Conditioning of tentacle lowering in the snail (Helix aspersa): acquisition, latent inhibition, overshadowing, second-order conditioning, and sensory preconditioning.

In a series of related experiments, we studied associative phenomena in snails (Helix aspersa), using the conditioning procedure of tentacle lowering. Experiments 1A and 1B demonstrated a basic conditioning effect in which the pairing of an odor (apple) as the conditioned stimulus (CS) with the opportunity to feed on carrot as the unconditioned stimulus (US) made snails exhibit increased levels of tentacle lowering in the presence of the CS. Experiments 2 and 3 showed that the magnitude of the conditioning was reduced when snails were exposed to the CS prior to the conditioning trial (a latent inhibition effect). Experiment 4 examined the effects produced by pairing a compound CS (apple-pear) with food presentations and demonstrated the existence of an overshadowing effect between the two odors. Experiment 5 revealed that pairing one CS with another previously conditioned stimulus increased tentacle lowering to the new CS (a second-order conditioning effect). Finally, Experiment 6 showed that pairing two odors prior to conditioning of one of them promoted an increase in tentacle lowering in response to the other (a sensory preconditioning effect). The results are discussed in terms of an associative analysis of conditioning and its implications for the study of cognition in invertebrates.

Animals↗

Cross-talk of WNT and FGF signaling pathways at GSK3beta to regulate beta-catenin and SNAIL signaling cascades.

WNT and FGF signaling pathways cross-talk during a variety of cellular processes, such as human colorectal carcinogenesis, mouse mammary tumor virus (MMTV)-induced carcinogenesis, E2A-Pbx-induced leukemogenesis, early embryogenesis, body-axis formation, limb-bud formation, and neurogenesis. Canonical WNT signals are transduced through Frizzled receptor and LRP5/6 coreceptor to downregulate GSK3beta (GSK3B) activity not depending on Ser 9 phosphorylation. FGF signals are transduced through FGF receptor to the FRS2-GRB2-GAB1-PI3K-AKT signaling cascade to downregulate GSK3beta activity depending on Ser 9 phosphorylation. Because GSK3beta-dependent phosphorylation of beta-catenin and SNAIL leads to FBXW1 (betaTRCP)-mediated ubiquitination and degradation, GSK3beta downregulation results in the stabilization and the nuclear accumulation of beta-catenin and SNAIL. Nuclear beta-catenin is complexed with TCF/LEF, Legless (BCL9 or BCL9L) and PYGO (PYGO1 or PYGO2) to activate transcription of CCND1, MYC, FGF18 and FGF20 genes for the cell-fate determination. Nuclear SNAIL represses transcription of CDH1 gene, encoding E-cadherin, to induce the epithelial-mesenchymal transition (EMT). Mammary carcinogenesis in MMTV-Wnt1 transgenic mice is accelerated by MMTV infection due to MMTV integration around Fgf3-Fgf4 or Fgf8 loci, and mammary carcinogenesis in MMTV-Fgf3 transgenic mice due to MMTV integration around Wnt1-Wnt10b locus. Coactivation of WNT and FGF signaling pathways in tumors leads to more malignant phenotypes. Single nucleotide polymorphism (SNP) and copy number polymorphism (CNP) of WNT and FGF signaling molecules could be utilized as screening method of cancer predisposition. cDNA-PCR, microarray or ELISA reflecting aberrant activation of WNT and FGF signaling pathways could be developed as novel cancer-related biomarkers for diagnosis, prognosis, and therapy. Cocktail therapy using WNT and FGF inhibitors, such as small-molecule compounds and human neutralizing antibodies, should be developed to increase the efficacy of chemotherapy through the inhibition of recurrence by destructing cancer stem cells.

Animals↗