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The suitability of Trochoidea seetzenii of different ages as snail intermediate hosts of Muellerius cf. capillaris (Nematoda:Protostrongylidae).

Larval development of Muellerius cf. capillaris in small (1st-year juveniles, shell diameter 0.96 +/- 0.05 cm) and medium-size (2nd-year juveniles, 1.27 +/- 0.06 cm) Trochoidea seetzenii land snails was significantly more rapid than in large adult snails (1.96 +/- 0.09 cm). Only 9% of the inoculated 1st-stage larvae (400 +/- 20 per snail) were recovered from the large snails as compared to 25% from the medium and small snails. Of the exposed small snails, however, only 35% survived to the end of the experiment (33 days post-infection) as compared to a 97.5% and 98% survival rates for the same period in the medium and large adult snails, respectively.

Animals↗

The prevalence of Fasciola hepatica in its snail intermediate host determined by DNA probe assay.

Accurate snail intermediate host infection prevalence data have the potential to be extremely useful in determining seasonal transmission dynamics of Fasciola hepatica. Because the microscopic techniques currently used lack the sensitivity and specificity necessary to obtain meaningful infection prevalence data, we developed a highly accurate and efficient DNA probe assay. The assay has a sensitivity of 100%, a specificity of > 99%, easily detects a single miracidia and does not cross-hybridize with DNA of Fascioloides magna, Paramphistomum liorchis or Heterobilharzia americana, trematodes that share the same intermediate host and enzootic range as Fasciola hepatica. Using this assay, we determined the prevalence of F. hepatica in its snail intermediate host, Fossaria cubensis, during the second year of a 2-year study on the epizootiology of Fasciola hepatica in Florida. The overall infection prevalence of snails assayed in this study (n = 5246) was 1.5% and ranged from 0.1% to 3.1% for individual cattle ranches. Additionally, infection prevalence differed significantly for successive size groupings of snails, varying from 0% for 1-mm snails to 18.5% for 9- and 10-mm snails. The accuracy and efficiency of the DNA probe assay reported here for determining snail infection prevalence offers an inexpensive alternative to tracer animal studies for determining the epizootiology of F. hepatica.

Animals↗

Chemo-orientation of echinostome cercariae towards their snail hosts: amino acids signal a low host-specificity.

The cercariae of Pseudechinoparyphium echinatum and Echinostoma revolutum approach their intermediate host snails by turning back when swimming in decreasing concentration gradients of snail conditioned water. Host stimuli are small molecular amino compounds, probably amino acids. This response was inhibited reversibly when the cercariae were treated with 0.45 microM-silver nitrate. As silver nitrate binds to the ciliated papillae of the cercarial surface, papillar chemoreceptors may be involved. Snail conditioned water from different snail species stimulated different intensities of chemo-orientation in both species of digeneans. However, when the samples of water conditioned with different snail species or even fish, tadpoles and leeches were diluted to the same total amino acid concentrations, the intensities of cercarial responses were similar. Therefore, some specificity of the chemo-orientation seemed to be achieved only by the total concentration of amino acids and not by particular mixtures of amino acids. In fact, amino acid spectra released by freshwater snails varied not only among different snail species, but also within the same species.

Amino Acids↗

Transfer and effects of cadmium in an experimental food chain involving the snail Helix aspersa and the predatory carabid beetle Chrysocarabus splendens.

The transfer and the toxic effects of Cd were studied in an experimental food chain involving the snail Helix aspersa as prey organism and one of its natural predators, the carabid beetle Chrysocarabus splendens. Juvenile snails were fed plant-based food enriched with 0, 10, 50 and 100 microg g(-1) of Cd, then were offered as prey to beetle larvae from egg hatching to pupation stage. Cd concentrations in snail tissues increased with increasing Cd concentration in food and with duration of exposure. Bioaccumulation factors ranged from 1.87 to 3.39, showing that H. aspersa snails, even in their early life stages, belong to macroconcentrator species for Cd. No significant reduction of snail consumption by beetles was found in exposed groups. Cd concentrations in beetle larvae remained very low (lower than 1 microg g(-1) for all groups), demonstrating a very effective regulation capacity in beetle larvae. However, Cd concentrations in highest exposed groups were higher than those found in control groups. Cd contents in adult beetles were lower than in larvae, showing a loss of Cd during metamorphosis. Despite the low Cd concentrations found in beetles, their exposure to Cd contaminated snails led to 31% of mortality, which occurred only during pupation and for the highest exposure level. No clear sublethal effects were found. These results showed that snails inhabiting heavily polluted areas may represent a risk of secondary poisoning for predatory invertebrates and provided quantitative data on the transfer of Cd between two compartments of a terrestrial food chain.

Animals↗

Ecological factors in schistosome transmission, and an environmentally benign method for controlling snails in a recreational lake with a record of schistosome dermatitis.

The avian schistosomes, Trichobilharzia stagnicolae, T. physellae and Gigantobilharzia sp., that cause Schistosome Dermatitis (Swimmers' Itch) in humans were studied in the laboratory and at Cultus Lake, British Columbia, Canada in relation to the biology and behavior of their intermediate snail hosts, Stagnicola catascopium, Physa sp. and Gyraulus parvus, respectively, and their definite bird hosts. Wind-driven, surface currents were measured. Populations of snails, close to host-bird roosting logs had a very high prevalence of schistosome infections. An experiment that mechanically disturbed the epilithic habitat of the snails using a boat-mounted rototiller or a tractor and rake, eliminated almost all of the snails if the disturbance was done in areas of high snail concentration in shallow areas of the lake during the breeding and early development phase of the snail. It is proposed that the incorporation of snail habitat disturbance into management programs is an effective way to control Schistosome Dermatitis.

Animals↗

Snail-trematode life history interactions: past trends and future directions.

Life history traits expressed by organisms vary due to ecological and evolutionary constraints imposed by their current environmental conditions and genetic heritage. Trematodes often alter the life history of their host snails by inducing parasitic castration. Our understanding of the variables that influence the resulting changes in host growth, fecundity and survivorship is insufficient to confidently predict specific outcomes of novel snail-trematode combinations. In a literature review of the last 30 years, we found 41 publications examining various life history characteristics of trematode-infected snails. These publications reported 113 different field and laboratory experiments involving 30 snail species and 39 trematode species and provided a data set for assessing factors that potentially affect life history outcomes. Analysis of the diverse responses across various snail-trematode systems and experimental conditions teased out general patterns for the expression of host growth, fecundity and survival. These were used to address existing hypotheses and develop several new ones relating the response of snail-trematode interactions to environmental and genetic factors. Finally, we propose directions for future experiments that will better assess the ecological and evolutionary factors influencing snail life history responses to trematode parasitism.

Animals↗

Evolutionary relationships between trematodes and snails emphasizing schistosomes and paragonimids.

Snails and digeneans have been associated for at least 200 million years. Their inter-relationships over such a time-span must have been complex and varied. Few studies have attempted to explore these relationships in the light of knowledge of the phylogeny of both host and parasite groups. Here we focus on two important families of digeneans, the Schistosomatidae and the Paragonimidae, for which molecular phylogenies are available. We investigate the types of evolutionary relationships between host and parasite, operating at different phylogenetic depths, that might explain current host specificity and distributions of both associates. Both families of parasites utilise a number of highly diverged gastropod families, indicating that host extensions have featured in their histories. However, schistosomatids and paragonimids show different patterns of association with their snail hosts. As befits the apparently more ancient group, schistosomatids utilise snails from across a wide phylogenetic range within the Gastropoda. The genus Schistosoma itself has experienced one long-range host switch between pulmonates and caenogastropods. By contrast, paragonimids are restricted to two superfamilies of caenogastropods. Despite these differences, modern schistosomatid species appear to be more host specific than are paragonimids and host additions, at the level of host family, are far less common among species of schistosomatids than among paragonimids. Some species of Paragonimus exhibit remarkably low levels of host specificity, with different populations utilising snails of different families. Existing knowledge relating to the phenomenon will be presented in the context of phylogenies of schistosomatids, paragonimids, and their snail hosts. Discussion focuses on the usefulness of current theories of snail-digenean coevolution for interpreting these findings. In the past, much emphasis has been placed on the idea that digeneans engage in a one-to-one arms race with their snail host. We consider that phylogenetic tracking rather than an arms-race relationship might be a common alternative. Not being bound by the restrictions imposed by an arms race, some digeneans might be able to extend to new host species more easily than the literature suggests. Switches into related host taxa are most likely. However, ecologically equivalent but unrelated gastropod hosts may also be exploited. Given the right ecological setting, digeneans are able to switch across considerable phylogenetic distances. Examples from the Paragonimidae and Schistosomatidae are given.

Animals↗

A molecular role for lysyl oxidase-like 2 enzyme in snail regulation and tumor progression.

The transcription factor Snail controls epithelial-mesenchymal transitions (EMT) by repressing E-cadherin expression and other epithelial genes. However, the mechanisms involved in the regulation of Snail function are not fully understood. Here we show that lysyl-oxidase-like 2 and 3 (LOXL2 and LOXL3), two members of the lysyl-oxidase gene family, interact and cooperate with Snail to downregulate E-cadherin expression. Snail's lysine residues 98 and 137 are essential for Snail stability, functional cooperation with LOXL2/3 and induction of EMT. Overexpression of LOXL2 or LOXL3 in epithelial cells induces an EMT process, supporting their implication in tumor progression. The biological importance of LOXL2 is further supported by RNA interference of LOXL2 in Snail-expressing metastatic carcinoma cells, which led to a strong decrease of tumor growth associated to increased apoptosis and reduced expression of mesenchymal and invasive/angiogenic markers. Taken together, these results establish a direct link between LOXL2 and Snail in carcinoma progression.

Amino Acid Oxidoreductases↗

Hepatocyte growth factor induces cell scattering through MAPK/Egr-1-mediated upregulation of Snail.

Hepatocyte growth factor/scatter factor (HGF) exerts several functions in physiological and pathological processes, among them the induction of epithelial cell scattering and motility. Its pivotal role in angiogenesis, tumor progression, and metastasis is evident; however, the underlying molecular mechanisms are still poorly understood. Here, we demonstrate that HGF induces scattering of epithelial cells by upregulating the expression of Snail, a transcriptional repressor involved in epithelial-mesenchymal transition (EMT). Snail is required for HGF-induced cell scattering, since shRNA-mediated ablation of Snail expression prevents this process. HGF-induced upregulation of Snail transcription involves activation of the mitogen-activated protein kinase (MAPK) pathway and requires the activity of early growth response factor-1 (Egr-1). Upon induction by Egr-1, Snail represses the expression of E-cadherin and claudin-3 genes. It also binds to the Egr-1 promoter and represses Egr-1 transcription, thereby establishing a negative regulatory feedback loop. These findings indicate that Snail upregulation by HGF is mediated via the MAPK/Egr-1 signaling pathway and that both Snail and Egr-1 play a critical role in HGF-induced cell scattering, migration, and invasion.

Base Sequence↗

Snail activation disrupts tissue homeostasis and induces fibrosis in the adult kidney.

During embryonic development, the kidney epithelium originates from cells that undergo a mesenchymal to epithelial transition (MET). The reverse process, epithelium to mesenchyme transition (EMT), has been implicated in epithelial tumor progression and in the fibrosis that leads to end-stage kidney failure. Snail transcription factors induce both natural and pathological EMT, but their implication in renal development and disease is still unclear. We show that Snail genes are downregulated during the MET that occurs during renal development and that this is correlated with Cadherin-16 expression. Snail suppresses Cadherin-16 via the direct repression of the kidney differentiation factor HNF-1beta, a novel route by which Snail disrupts epithelial homeostasis. Indeed, Snail activation is sufficient to induce EMT and kidney fibrosis in adult transgenic mice. Significantly, Snail is also activated in patients with renal fibrosis. Thus, Snail expression is suppressed during renal development and it must remain silent in the mature kidney where its aberrant activation leads to fibrosis.

Animals↗

Regulation of Snail transcription during epithelial to mesenchymal transition of tumor cells.

Expression of Snail transcriptional factor is a determinant in the acquisition of a mesenchymal phenotype by epithelial tumor cells. However, the regulation of the transcription of this gene is still unknown. We describe here the characterization of a human SNAIL promoter that contains the initiation of transcription and regulates the expression of this gene in tumor cells. This promoter was activated in cell lines in response to agents that induce Snail transcription and the mesenchymal phenotype, as addition of the phorbol ester PMA or overexpression of integrin-linked kinase (ILK) or oncogenes such as Ha-ras or v-Akt. Although other regions of the promoter were required for a complete stimulation by Akt or ILK, a minimal fragment (-78/+59) was sufficient to maintain the mesenchymal specificity. Activity of this minimal promoter and SNAIL RNA levels were dependent on ERK signaling pathway. NFkappaB/p65 also stimulated SNAIL transcription through a region located immediately upstream the minimal promoter, between -194 and -78. These results indicate that Snail transcription is driven by signaling pathways known to induce epithelial to mesenchymal transition, reinforcing the role of Snail in this process.

Cell Differentiation↗

Snail induction is an early response to Gli1 that determines the efficiency of epithelial transformation.

Gli family members mediate constitutive Hedgehog signaling in the common skin cancer, basal cell carcinoma (BCC). Snail/Snai1 is rapidly induced by Gli1 in vitro, and is coexpressed with Gli1 in human hair follicles and skin tumors. In the current study, we generated a dominant-negative allele of Snail, SnaZFD, composed of the zinc-finger domain and flanking sequence. In promoter-reporter assays, SnaZFD blocked the activity of wild-type Snail on the E-cadherin promoter. Snail loss-of-function mediated by SnaZFD or by one of several short hairpin RNAs inhibited transformation of RK3E epithelial cells by Gli1. Conversely, enforced expression of Snail promoted transformation in vitro by Gli1, but not by other genes that were tested, including Notch1, ErbB2, and N-Ras. As observed for Gli1, wild-type Snail repressed E-cadherin in RK3E cells and induced blebbing of the cytoplasmic membrane. Induction of a conditional Gli1 transgene in the basal keratinocytes of mouse skin led to rapid upregulation of Snail transcripts and to cell proliferation in the interfollicular epidermis. Established Gli1-induced skin lesions exhibited molecular similarities to BCC, including loss of E-cadherin. The results identify Snail as a Gli1-inducible effector of transformation in vitro, and an early Gli1-responsive gene in the skin.

Animals↗

Wnt-dependent regulation of the E-cadherin repressor snail.

Down-regulation of E-cadherin marks the initiation of the epithelial-mesenchymal transition, a process exploited by invasive cancer cells. The zinc finger transcription factor, Snail, functions as a potent repressor of E-cadherin expression that can, acting alone or in concert with the Wnt/beta-catenin/T cell factor axis, induce an epithelial-mesenchymal transition. Although mechanisms that coordinate signaling events initiated by Snail and Wnt remain undefined, we demonstrate that Snail displays beta-catenin-like canonical motifs that support its GSK3beta-dependent phosphorylation, beta-TrCP-directed ubiquitination, and proteasomal degradation. Accordingly, Wnt signaling inhibits Snail phosphorylation and consequently increases Snail protein levels and activity while driving an in vivo epithelial-mesenchymal transition that is suppressed following Snail knockdown. These findings define a potential mechanism whereby Wnt signaling stabilizes Snail and beta-catenin proteins in tandem fashion so as to cooperatively engage transcriptional programs that control an epithelial-mesenchymal transition.

Amino Acid Sequence↗

Feeding and growth of apple snail Pomacea lineata in the Pantanal wetland, Brazil--a stable isotope approach.

Apple snails Pomacea lineata (SPIX 1827) are widespread in the tropical regions of Brazil as well as in the Pantanal wetland of Mato Grosso in the western part of the country. They have a key position in the Pantanal food web and serve as food for many animals e.g. fishes, birds, and caimans. However, little is known about their feeding preferences and growth rates. Stable isotopes have been used successfully on numerous studies as food source indicator. Therefore, the delta15N and delta13C values of snails from 0.45 to 3.03 cm in length, which were collected in the rainy season from March through May, were analyzed. Snails signatures revealed ambiguous evidence for food preferences. Delta15N and delta13C values ranged between -2.8 and 12.4 per thousand and between -24.2 and -16.4, per thousand respectively. This range of values mirrors the highly variable isotope values of possible food sources comprising C3 and C4 macrophytes. To test whether all common food sources were similarly assimilated, feeding experiments with different diets were conducted. Snail eggs were reared in tanks and offered different but single plants. Snails fed different diets and delta13C values of the food were reflected in the animal tissue. Growth varied considerably in experiments with different diets indicating the preference for certain food sources. Also, the fractionation of nitrogen isotopes between food and animal varied from 0.1 to 17.0 per thousand. The results are explained by different feeding habits, and it is supposed that animals fed either on the plant itself or on bacteria mats growing in the tanks. In an additional experiment juvenile snails were offered one single food with a distinctive C4 grass signature. These snails did not grow detectably, but nevertheless isotope signatures approached to values of the diet.

Animals↗

Structure and function of mitochondria in hepatopancreas cells from metabolically depressed snails.

Mitochondria in cells isolated from the hepatopancreas of aestivating land snails (Helix aspersa) consume oxygen at 30% of the active control rate. The aim of this study was to investigate whether the lower respiration rate is caused by a decrease in the density of mitochondria or by intrinsic changes in the mitochondria. Mitochondria occupied 2% of cellular volume, and the mitochondrial inner membrane surface density was 17 microm(-1), in cells from active snails. These values were not different in cells from aestivating snails. The mitochondrial protein and mitochondrial phospholipid contents of cells were also similar. There was little difference in the phospholipid fatty acyl composition of mitochondria isolated from metabolically depressed or active snails, except for arachidonic acid, which was 18% higher in mitochondria from aestivating snails. However, the activities of citrate synthase and cytochrome c oxidase in mitochondria isolated from aestivating snails were 68% and 63% of control, respectively. Thus the lower mitochondrial respiration rate in hepatopancreas cells from aestivating snails was not caused by differences in mitochondrial volume or surface density but was associated with intrinsic changes in the mitochondria.

Adaptation, Physiological↗

The potential role of CO2 in initiation and maintenance of estivation in the land snail Helix lucorum.

Elevated CO(2) levels are hypothesized to play a role in the initiation and maintenance of estivation in snails through disturbances of acid-base status. The aim of our study was to identify the ambient CO(2) threshold that induces disturbances in acid-base status in the air-breathing land snail Helix lucorum. Acid-base parameters were determined in the hemolymph of snails acclimated to 0.5%, 1%, 2%, 4%, and 8% CO(2) in air for 20 d. In addition, we evaluated the effects of long-term acclimation on metabolic rate and on levels of D-lactate dehydrogenase activity (D-LDH) and of D-lactate in snails after 20 d of exposure to increased CO(2) levels. Helix lucorum proved to be unable to compensate for a decrease in extracellular pH (pH(e)) when acclimated to levels higher than 1% CO(2) in air. The rate of oxygen consumption started to decrease when snails were acclimated to 0.5% CO(2) in air. However, there was no correlation between the drops in pH(e) and in metabolic rate. Long-term acclimation to elevated CO(2) levels induced an increase in the activity of D-LDH with a concomitant accumulation of D-lactate in tissues. This indicates that long-term acclimation to elevated ambient CO(2) levels could reduce the aerobic capacity of land snails and trigger expression of anaerobic pathways of ATP turnover. The threshold levels of ambient CO(2) that induce changes in acid-base status and elicit metabolic depression in adult land snails H. lucorum are higher than the future atmospheric levels that are expected to result from human use of fossil energy resources.

Acclimatization↗

The mesoderm determinant snail collaborates with related zinc-finger proteins to control Drosophila neurogenesis.

The Snail protein functions as a transcriptional regulator to establish early mesodermal cell fate. Later, in germ band-extended embryos, Snail is also expressed in most neuroblasts. Here we present evidence that this expression of Snail is required for central nervous system (CNS) development. The neural function of snail is masked by two closely linked genes, escargot and worniu. Both Escargot and Worniu contain zinc-finger domains that are highly homologous to that of Snail. Although not affecting expression of early neuroblast markers, the deletion of the region containing all three genes correlates with loss of expression of CNS determinants including fushi tarazu, pdm-2 and even-skipped. Transgenic expression of each of the three Snail family proteins can rescue efficiently the fushi tarazu defects, and partially the pdm-2 and even-skipped CNS patterns. These results demonstrate that the Snail family proteins have essential functions during embryonic CNS development, around the time of ganglion mother cell formation.

Amino Acid Sequence↗

Zinc finger domain of Snail functions as a nuclear localization signal for importin beta-mediated nuclear import pathway.

Snail, a DNA-binding zinc finger protein, functions as a transcriptional repressor for genes including E-cadherin during development and the acquisition of tumor cell invasiveness. Human Snail is a 264-amino acid nuclear protein with an amino-terminal basic amino acid-rich domain (SNAG domain) and a carboxyl-terminal DNA-binding domain (zinc finger domain). A series of fusion proteins composed of green fluorescent protein (GFP) and portions of the Snail protein were generated, and their subcellular localization was examined. Fusion of the four zinc fingers to GFP led to the targeting of GFP to the nucleus, demonstrating that the zinc finger domain is sufficient for nuclear localization. Using an in vitro transport system, the nuclear import of Snail was reconstituted by importin (karyopherin) beta in the presence of Ran and NTF2. We further demonstrated that Snail binds directly to importin beta in a zinc finger domain-dependent manner. These results indicate that zinc finger domain of Snail functions as a nuclear localization signal and Snail can be transported into the nucleus in an importin beta-mediated manner.

Animals↗