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PubMed2004

Dual regulation of Snail by GSK-3beta-mediated phosphorylation in control of epithelial-mesenchymal transition.

The phenotypic changes of increased motility and invasiveness of cancer cells are reminiscent of the epithelial-mesenchymal transition (EMT) that occurs during embryonic development. Snail, a zinc-finger transcription factor, triggers this process by repressing E-cadherin expression; however, the mechanisms that regulate Snail remain elusive. Here we find that Snail is highly unstable, with a short half-life about 25 min. We show that GSK-3beta binds to and phosphorylates Snail at two consensus motifs to dually regulate the function of this protein. Phosphorylation of the first motif regulates its beta-Trcp-mediated ubiquitination, whereas phosphorylation of the second motif controls its subcellular localization. A variant of Snail (Snail-6SA), which abolishes these phosphorylations, is much more stable and resides exclusively in the nucleus to induce EMT. Furthermore, inhibition of GSK-3beta results in the upregulation of Snail and downregulation of E-cadherin in vivo. Thus, Snail and GSK-3beta together function as a molecular switch for many signalling pathways that lead to EMT.

Amino Acid Motifs↗

PubMed2005

Snail accelerates cancer invasion by upregulating MMP expression and is associated with poor prognosis of hepatocellular carcinoma.

We have previously demonstrated in an in vitro study that Snail increased the invasion activity of hepatoma cells by upregulating matrix metalloproteinase (MMP) gene expression. In the present study, we examined whether Snail gene expression correlates with cancer invasion and prognosis of patients with hepatocellular carcinoma (HCC). Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate Snail, E-cadherin, and MMP mRNA expressions in eight nodule-in-nodule tumours and 47 ordinary HCC tissues. In the nodule-in-nodule tumours, Snail expression significantly increased with tumour dedifferentiation (P=0.047). In the ordinary HCC tissues, Snail expression was significantly correlated with portal vein invasion (P=0.035) and intrahepatic metastasis (P=0.050); it also showed a significant correlation with MT1-MMP expression (r=0.572, P<0.001). In recurrence-free survival, the group with high Snail expression showed significantly poorer prognosis (P=0.035). Moreover, high Snail expression was an independent risk factor for early recurrence after curative resection. During the progression of HCC, Snail expression may be induced and accelerate invasion activity by upregulating MMP expression, resulting in portal invasion, intrahepatic metastasis, and poor prognosis.

Aged↗

PubMed2006

Expression of Snail protein in tumor-stroma interface.

The product of Snail gene is a repressor of E-cadherin transcription and an inductor of the epithelial-to-mesenchymal transition in several epithelial tumor cell lines. In order to examine Snail expression in animal and human tissues, we have raised a monoclonal antibody (MAb) that reacts with the regulatory domain of this protein. Analysis of murine embryos shows that Snail is expressed in extraembryonic tissues and embryonic mesoderm, in mesenchymal cells of lungs and dermis as well as in cartilage. Little reactivity was detected in adult tissues as Snail was not constitutively expressed in most mesenchymal cells. However, Snail expression was observed in activated fibroblasts involved in wound healing in mice skin. Moreover, Snail was detected in pathological conditions causing hyperstimulation of fibroblasts, such as fibromatosis. Analysis of Snail expression in tumors revealed that it was highly expressed in sarcomas and fibrosarcomas. In epithelial tumors, it presented a more limited distribution, restricted to stromal cells placed in the vicinity of the tumor and to tumoral cells in the same areas. These results demonstrate that Snail is present in activated mesenchymal cells, indicate its relevance in the communication between tumor and stroma and suggest that it can promote the conversion of carcinoma cells to stromal cells.

3T3 Cells↗

PubMed2006

The transcription factors Slug and Snail act as repressors of Claudin-1 expression in epithelial cells.

Claudin-1 is an integral membrane protein component of tight junctions. The Snail family of transcription factors are repressors that play a central role in the epithelial-mesenchymal transition, a process that occurs during cancer progression. Snail and Slug members are direct repressors of E-cadherin and act by binding to the specific E-boxes of its proximal promoter. In the present study, we demonstrate that overexpression of Slug or Snail causes a decrease in transepithelial electrical resistance. Overexpression of Slug and Snail in MDCK (Madin-Darby canine kidney) cells down-regulated Claudin-1 at protein and mRNA levels. In addition, Snail and Slug are able to effectively repress human Claudin-1-driven reporter gene constructs containing the wild-type promoter sequence, but not those with mutations in two proximal E-box elements. We also demonstrate by band-shift assay that Snail and Slug bind to the E-box motifs present in the human Claudin-1 promoter. Moreover, an inverse correlation in the levels of Claudin-1 and Slug transcripts were observed in breast cancer cell lines. E-box elements in the Claudin-1 promoter were found to play a critical negative regulatory role in breast cancer cell lines that expressed low levels of Claudin-1 transcript. Significantly, in invasive human breast tumours, high levels of Snail and Slug correlated with low levels of Claudin-1 expression. Taken together, these results support the hypothesis that Claudin-1 is a direct downstream target gene of Snail family factors in epithelial cells.

Animals↗

PubMed2005

Snail-regulated genes in malignant melanoma.

The demonstration that zinc-finger transcriptional repressors can control E-cadherin expression in epithelial cells has provided a new avenue of research in the field of epithelial-mesenchymal transition (EMT). One of these zinc-finger molecules is the transcription factor Snail, which controls gastrulation and neural crest EMT in different species. Additionally, Snail is involved in the development of malignant melanoma where a dramatic change in E-cadherin expression is an important early step for melanoma progression. For this study, a human cancer cDNA array was used which includes genes involved in cancer development and progression. Using the array we compared the gene expression pattern of the melanoma cell line Mel Im with a Mel Im cell clone stable transfected with antisense (as) SNAIL cDNA. We validated the significant differences of the expression of genes on mRNA level. Primarily, we observed changes in the expression of genes involved in EMT. Quantitative real-time polymerase chain reaction showed a down-regulation of MMP-2, EMMPRIN, SPARC, TIMP-1, t-PA, RhoA and Notch4 expression and a re-induction of E-cadherin expression in the as Snail cell clones. Furthermore, we measured the expression of integrin beta3, NM23b and RhoB. Additionally, we investigated whether the selected genes are influenced only through Snail or if E-cadherin can influence the expression of these genes. In summary, all examined genes which are influenced through Snail have a regulatory function in EMT processes as does Snail itself. The Snail target gene E-cadherin has no regulatory function with respect to the selected genes.

Cadherins↗

PubMed2004

Aberrant expression of the transcription factors snail and slug alters the response to genotoxic stress.

Snail and Slug are closely related transcriptional repressors involved in embryonic patterning during metazoan development. In human cancer, aberrant expression of Snail and/or Slug has been correlated with invasive growth potential, a property primarily attributed to their ability to directly repress transcription of genes whose products are involved in cell-cell adhesion, such as E-cadherin, occludin, and claudins. To investigate the molecular mechanisms of alterations in epithelial cell fate mediated by aberrant expression of Snail or Slug, we analyzed the consequences of exogenous expression of these factors in human cancer cells. Aberrant expression of either Snail or Slug led to changes in cell morphology, the loss of normal cell-cell contacts, and the acquisition of invasive growth properties. Snail or Slug expression also promoted resistance to programmed cell death elicited by DNA damage. Detailed molecular analysis revealed direct transcriptional repression of multiple factors with well-documented roles in programmed cell death. Depletion of endogenous Snail by RNA interference led to increased sensitivity to DNA damage accompanied by increased expression of the proapoptotic factors identified as targets of Snail. Thus, aberrant expression of Snail or Slug may promote tumorigenesis through increased resistance to programmed cell death.

Animals↗

PubMed1988

Changes in epilithic communities due to individual and combined treatments of zinc and snail grazing in stream mesocosms.

Effects of 0.5 mg/liter zinc (Zn) and snail grazing (400 snails/m2) on density of dominant algal and protozoan taxa, epilithic glucose respiration, and ash-free dry weight (AFDW) were examined using established (12-day colonization) periphyton communities in flow-through stream mesocosms with four treatments (Zn, snails, Zn and snails, control) for 30 days. Grazing and Zn similarly reduced the abundance of 5 of 10 dominant algal taxa and AFDW during the first 10 days of treatment. Abundance of these taxa and AFDW in grazed (ambient Zn) treatments approached control levels after 10 days as the effect due to snails decreased. Decreasing temperatures may have reduced snail activity. Snails, Zn, and the combination of these treatments contributed to higher rates of glucose respiration per unit AFDW. Protozoan species abundance was reduced to less than half by Zn but was unaffected by snails. Although Zn and snails individually altered structural and functional aspects of this microbial community, the effects when both treatments were combined could not always be inferred from the individual effects. Testing individual and combined variables that affect periphyton with a corresponding assessment of population dynamics, biomass, and community functional attributes will enhance understanding of the overall effects of pollutants on periphyton communities.

Animals↗

PubMed1997

Differential regulation of gastrulation and neuroectodermal gene expression by Snail in the Drosophila embryo.

The initiation of mesoderm differentiation in the Drosophila embryo requires the gene products of twist and snail. In either mutant, the ventral cell invagination during gastrulation is blocked and no mesoderm-derived tissue is formed. One of the functions of Snail is to repress neuroectodermal genes and restrict their expressions to the lateral regions. The derepression of the neuroectodermal genes into the ventral region in snail mutant is a possible cause of defects in gastrulation and in mesoderm differentiation. To investigate such possibility, we analysed a series of snail mutant alleles. We found that different neuroectodermal genes respond differently in various snail mutant background. Due to the differential response of target genes, one of the mutant alleles, V2, that has reduced Snail function showed an intermediate phenotype. In V2 embryos, neuroectodermal genes, such as single-minded and rhomboid, are derepressed while ventral invagination proceeds normally. However, the differentiation of these invaginated cells into mesodermal lineage is disrupted. The results suggest that the establishment of mesodermal cell fate requires the proper restriction of neuroectodermal genes, while the ventral cell movement is independent of the expression patterns of these genes. Together with the data showing that the expression of some ventral genes disappear in snail mutants, we propose that Snail may repress or activate another set of target genes that are required specifically for gastrulation.

Amino Acid Sequence↗

PubMed2001

The Snail protein family regulates neuroblast expression of inscuteable and string, genes involved in asymmetry and cell division in Drosophila.

Delaminated neuroblasts in Drosophila function as stem cells during embryonic central nervous system development. They go through repeated asymmetric divisions to generate multiple ganglion mother cells, which divide only once more to produce postmitotic neurons. Snail, a zinc-finger transcriptional repressor, is a pan-neural protein, based on its extensive expression in neuroblasts. Previous results have demonstrated that Snail and related proteins, Worniu and Escargot, have redundant and essential functions in the nervous system. We show that the Snail family of proteins control central nervous system development by regulating genes involved in asymmetry and cell division of neuroblasts. In mutant embryos that have the three genes deleted, the expression of inscuteable is significantly lowered, while the expression of other genes that participate in asymmetric division, including miranda, staufen and prospero, appears normal. The deletion mutants also have much reduced expression of string, suggesting that a key component that drives neuroblast cell division is abnormal. Consistent with the gene expression defects, the mutant embryos lose the asymmetric localization of prospero RNA in neuroblasts and lose the staining of Prospero protein that is normally present in ganglion mother cells. Simultaneous expression of inscuteable and string in the snail family deletion mutant efficiently restores Prospero expression in ganglion mother cells, demonstrating that the two genes are key targets of Snail in neuroblasts. Mutation of the dCtBP co-repressor interaction motifs in the Snail protein leads to reduction of the Snail function in central nervous system. These results suggest that the Snail family of proteins control both asymmetry and cell division of neuroblasts by activating, probably indirectly, the expression of inscuteable and string.

Animals↗

PubMed1979

Comparison of four species of snails as potential decoys to intercept schistosome miracidia.

Preliminary studies have shown that various species of aquatic snails may be used as decoys or "sponges" to intercept schistosome miracidia, thereby preventing the miracidia from reaching the snails which normally serve as their intermediate host. In this study, four species of snails were evaluated as candidate decoys for field trials: Marisa cornuarietis, Pomacea australis, Helisoma caribaeum, and Tarebia granifera. In the laboratory all four species caused considerable reductions in the proportion of Biomphalaria glabrata infected by miracidia of Schistosoma mansoni. The most effective decoys were M. cornuarietis and H. caribaeum, both of which caused experimental infection levels of 90% to decrease to 25% when five decoy snails were present for each target snail. When ten decoy snails were present for each target snail, the proportion infected decreased to 1%. M. cornuarietis was chosen as the candidate for field trials because it was found more frequently in Puerto Rico than was H. caribaeum. Initial field trials in two ponds showed that M. cornuarietis blocked infections at a ratio of 6 decoys to 1 target snail, confirming the laboratory results. Further studies in flowing water are needed before the technique can be generally evaluated in an endemic area.

Animals↗

PubMed1995

Recovery of Pasteurella multocida from experimentally-exposed freshwater snails.

We determined how long Pasteurella multocida could survive in experimentally-exposed freshwater snails. Physa virginea were collected from the Sacramento National Wildlife Refuge, Glenn County, California (USA), an enzootic site for avian cholera. Exposure to water containing up to 10(7) P. multocida per ml did not produce observable changes or mortality in snails. A minimum of 84 P. multocida per snail was necessary for detection among the normal snail bacterial flora. When snails were exposed to P. multocida in vials containing 10(7) bacteria per ml, P. multocida was detected for up to 72 hours in snails. When uninoculated snails were placed in aquaria containing 10(6) P. multocida per ml, P. multocida was not detected within the snails; further, P. multocida was detected in the water for only 24 hours at this level. Based on these results, we propose that P. virginea is not an effective reservoir for P. multocida.

Animals↗

PubMed2002

[Effect of selective protein kinase C inhibitor on synaptic plasticity of command neurons of defensive behaviour during sensitization in snails].

Polymyxin B (proteinkinase C inhibitor) effects on nociceptive sensitization of semiintact preparation were investigated in LP11 and RP11 snail neurons. It was found that application of sensitizing stimuli to control snail head initiated neural membrane depolarization, increase its excitability as well as depression of neural responses evoked by sensory stimulation during short-term stage. Polymyxin B application suppressed neural responses evoked by sensitizing (nociceptive) stimuli. At the same time changes in neural membrane excitability as well as neural responses evoked by tactile stimulation of snail foot or chemical stimulation of snail head were similar with ones in control snails. Polymyxin Bdid does not change the depression of neural responses evoked by tactile stimulation of snail head during short-term stages of sensitization but significantly suppressed facilitation of neural responses evoked by tactile stimulation of snail head during long-term stage of sensitization. It was suggested that proteinkinase C is involved in regulation of nociceptive mechanisms as well as in plasticity selective induction mechanisms in command neuron synaptic inputs activated by tactile stimulation of snail head.

Animals↗

PubMed2003

[Study on the relationship between Terra-MODIS image and the snail distribution in marshland of Jiangning county, Jiangsu province].

OBJECTIVE: To analyze the relationship between the normalized difference vegetation index (NDVI) and the snail distribution in marshland of Jiangning county in Jiangsu province, and to explore the utility of Terra-MODIS image map in the small scale snail habitats surveillance. METHODS: NDVI were extracted from MODIS image by vector chart of the snail distribution using ArcView 8.1 and ERDAS 8.5 software. The relationship between NDVI and the snail distribution were Investigated using Bivariate correlations and stepwise linear regression. RESULTS: The snail density on marshland was positively correlated with the mean NDVI in the first ten-day of May and the maximum NDVI (N(20max)) in the last ten-day of May. Incidence of pixel with the live snail on marshland was positively correlated with the mean NDVI (N(2mean)) in the first ten-day of May. An equation Y(1) = 0.009 47 x N(20max) (R(2) = 0.73), Y(2) = 0.018 6 x N(2mean) (R(2) = 0.906) was established. CONCLUSION: This study showed that the Terra-MODIS satellite images reflecting the status of the vegetation on marshland in Jiangning county could be applied to the study to supervise the snail habitat. The results suggested that MODIS images could be used to survey the small scale snail habitats on marshland.

Animals↗

PubMed2003

[Application of satellite image for surveillance of vegetation landscapes of Oncomelenia-snail habitats in marshland using unsupervised classification].

OBJECTIVE: To explore the vegetation landscapes in marshland snail habitats using satellite image. METHODS: The false-color composition image from band 2, 3 and 4 of LANDSAT ETM + images was classified in the ERDAS IMAGINE 8.5 to analyze the vegetation types in the marshland of Jiangning county, using the unsupervised classification. The efficiency of classification was evaluated by the transformed divergence. The overlaid layers of the classified vegetation image and the vector layer of snail habitats distribution were used to analyze the relationship between the snail distribution and the landscape types. RESULTS: The land-cover of marshland in LANDSAT ETM234 image in Jiangning county could be classified into 10 types, including water, bare soil, sandy and other landscapes while the transformed divergence analysis showed that there were misclassified pixes between some types especially for the non-continuous types. The study indicated that through adding the NDVI image in the process of classification efficiency of classification and eliminate misclassification in the non-continuous type could be improved. Analysis on the overlaid layer of the vector of snail distribution and the classified image proved that the vegetation covers in marshland snail habitats in Jiangning mainly belonged to type 3, 5 and 6, that responded to the beach with sparse vegetation, exuberant weed and bulrush respectively. The density of snails in the bulrush was higher than that in other 2 landscapes. CONCLUSION: The vegetation type in the marshland snail habitats could be distinguished from the satellite image, which was helpful for the surveillance of snail habitat in marshland and for the prevention of schistosomiasis.

Animals↗

PubMed2006

[Experimental study on compatibility of three species of freshwater snails with Angiostrongylus cantonensis].

OBJECTIVE: To compare the compatibility of three species of freshwater snails, Pomacea canaliculata, Cipangopaludina chinensis, Bellamy a aeruginosa, with Angiostrongylus cantonensis. METHODS: The snails were infected by the first-stage larvae of A. cantonensis under the same conditions. Twenty snails of each species were randomly sampled after exposed to the larvae for 1, 3, 6, 12, 24 hours, respectively. Each group was placed into an aquarium. Each species with same number was established as control. All the aquaria were equipped with a filter, the water temperature was kept at (24 +/- 1) degrees C. In the first two weeks, the number of death was recorded. Later, the snails were successively examined to record the weight and worm burden of each snail. RESULTS: Some snails were dead which mainly happened in the first week postinfection. The death rate and infection rate were not associated with exposure time and snail species. Worm burden of P. canaliculata was significantly higher than the other two species, however, the worm density of P. canaliculata and B. aeruginosa was not significantly different but higher than that of C. chinensis. The worm burden and worm density of P. canaliculata and B. aeruginosa, respectively, were significantly different among five exposure-time groups, but that of C. chinensis was not. CONCLUSION: All the three snail species show a high compatibility with A. cantoneasis. In general, the compatibility of P. canaliculata is superior to the other two species.

Angiostrongylus cantonensis↗

PubMed2000

Discriminant analysis of free fatty acids of some Egyptian snails as a step for schistosomiasis control: new trends.

Schistosomiasis is still the most common occupational health problem of rural workers in Egypt. The use of molluscicides, either chemical or biological, and environmental changing are the most successful methods of snail control. The new trend in Schistosoma control programs is to study the ecological factors that attract miracidium Schistosoma specific snails, to emphasizing to find out new environmental safe control methods. Since Schistosoma worms do not make fatty acids de novo, they require host lipids for survival and to complete their life cycle. Discriminate analysis of the estimated free fatty acids was done in this study in Biomphalaria alexandrina, Biomphalaria glabrata, and Bulinus truncatus, viz. Lymnaea truncatula and Physa acuta (Schistosoma intermediate and non-intermediate respectively). With the objective of determination of the biochemical difference that attract the infestation of Schistosoma miracidium to the target snails, as a step of the new control trends. Caprylic acid (C8:0), and Oleic acid (C18:1) are significantly lower in the tissues of Schistosoma intermediate snails compared to the non-intermediate snails. While, Capric acid (C10:0), Margaric acid (C17:0), and Lenoleic acid (C18:2) of the intermediate snails are significantly higher than that of non-intermediate snails. The percent of correct medical classification of snails are more than 80% according to Caprylic acid, Margaric acid, and Lenoleic acid, the other 15 fatty acids are less than 80%. These three free fatty acids could be chemoattractive of Schistosoma miracidium, and could be used as safe environmental control compounds, which needs further research.

Animals↗

PubMed1994

Sex ratio and susceptibility of the golden apple snail, Pomacea canaliculata.

Golden apple snails, Pomacea canaliculata, were collected at two localities having different ecological environments. In both canal and pond, P. canaliculata males were found more than females during the dry season (summer and winter). In the canal, the male snails were highest in number (86.67%) in May. When rain started, they began decreasing and were lowest at 33.33% in August. Of 575 snails collected, 30.6% were infected by one or more of the three groups of amphistome, distome and echinostome metacercariae. There were two high peaks of infection in April and October, as 60.7% and 68.4%, respectively, during which there were more males than females. The average number of parasites per snail which was highest at 54 was found in the medium-sized males (25 out of 35 males) in October. The number of parasites per snail was significantly correlated with the collected males (p < 0.01), but such relationship was not occurred with the females. Of the females, only the large-sized individuals were infected. In the pond, the female snails were present in much greater numbers than the males during the reproductive time (June-September). The females were highest (94.23%) in August. Only 24 (4.0%) of 605 snails were infected; most of the infected snails were large.

Animals↗

PubMed1987

Alterations in the internal defence system of the pond snail Lymnaea stagnalis induced by infection with the schistosome Trichobilharzia ocellata.

In order to investigate whether the schistosome Trichobilharzia ocellata interferes with defence activities in its snail intermediate host Lymnaea stagnalis, aspects of the immune system of infected snails and of non-infected controls were compared. The elimination of injected live Staphylococcus saprophyticus bacteria starts at a lower rate in infected snails 1 and 5 weeks after exposure to the parasite, but then proceeds faster than in control snails. During the first 3 weeks of infection, when only mother sporocysts are present, the haemocytes of the infected snails have an increased capacity to phagocytose rabbit red blood cells in vitro. From 5 weeks onwards, when mother and daughter sporocysts are present but cercariae are not yet mature, the phagocytic activity decreases to below control level. The number of circulating haemocytes is also higher in infected snails than in controls at this time. Moreover, the cells are larger, have more inclusions and an increased surface area with many long, branched, spiked pseudopods. The development of the parasite is retarded in a subpopulation of snails in which the haemolymph plasma agglutinates erythrocytes with high titres, compared to a subpopulation with low haemagglutinating activity. The haemagglutinating activity in infected snails of the first decreases significantly from 6 weeks onwards.

Animals↗