Iron is an essential element for metabolic processes intrinsic to life, but the properties that make iron a necessity also make it potentially deleterious. To avoid harm, iron homeostasis is achieved through iron transport, storage and... more
Iron is an essential element for metabolic processes intrinsic to life, but the properties that make iron a necessity also make it potentially deleterious. To avoid harm, iron homeostasis is achieved through iron transport, storage and regulatory proteins. The functions of some of these molecules are well described, for example transferrin and ferritin, whereas the roles of others remain unclear. The past decade has seen the identification of new molecules involved in iron metabolism, such as divalent metal transporter-1, and hepcidin. The present review aims at surveying the studies carried out on some of the most important genes involved in transport and storage of iron in Antarctic Notothenioidei, a dominating fish group endowed of a number of striking adaptive characters, including reduced (or absence of) hematocrit. This unique peculiarity among vertebrates makes this fish group a suitable system to studying the relationship between hemoglobin and iron metabolism and to understanding the adaptive changes occurred in Antarctic fish metabolism during their evolution to avoid the deleterious effects of iron overload in the absence of hemoglobin. The results summarised here indicate that the loss of hemoglobin in the most specialized group of Antarctic notothenioids, belonging to the Channychthyidae family, is accompanied by remodulation of the iron metabolism.
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Research Interests: Genetics, Reproduction, Biology, Medicine, Lizards, and 7 moreFemale, Animals, Metallothionein, Cadmium, Lizard, Molecular sciences, and Oviparity
The possible role of metallothionein (MT) in metal homeostasis has been investigated in growing oocytes and eggs of the lizard Podarcis sicula. Chromatographic analysis does not reveal the presence of MT in both ovary and eggs, the only... more
The possible role of metallothionein (MT) in metal homeostasis has been investigated in growing oocytes and eggs of the lizard Podarcis sicula. Chromatographic analysis does not reveal the presence of MT in both ovary and eggs, the only metal-binding proteins detected being represented by high molecular mass components. De novo synthesis of MT could be observed in the ovary of cadmium-treated lizards. A cDNA encoding MT was obtained from the liver of P. sicula by RT-PCR followed by a RACE strategy, using primers designed on consensus motifs of vertebrate MT. In spite of the lack of MT in the ovary of untreated animals, Northern blot analysis demonstrates that the maternal untranslated MT transcript is expressed constitutively in the ovary in all the periods of the ovarian cycle. MT mRNA content increases during the oocyte growth, reaching the highest level in ovulated eggs, concomitantly with the accumulation of zinc and copper. Our findings suggest that maternal MT mRNA accumulates in the egg and is translated sometime during development to cope with the future needs of the growing embryo. The appearance of MT after cadmium treatment suggests that the block that makes the oocytarian MT mRNA untranslatable is removed by the metal.
Research Interests: Biology, Medicine, Oogenesis, Sequence alignment, Liver, and 15 moreCopper, Lizards, Female, Animals, Metallothionein, Superoxide Dismutase, Molecular cloning, Amino Acid Sequence, Base Sequence, Ovary, Oocytes, Biochemistry and cell biology, Molecular Sequence Data, ovum, and Paediatrics and reproductive medicine
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Research Interests: Evolutionary Biology, Phylogenetics, Molecular Evolution, Biology, Medicine, and 15 moreBiological Sciences, Phylogeny, Sequence alignment, Animals, Metallothionein, Molecular Phylogenetics and Evolution, Gene Duplication, Vertebrate, Phylogenetic Tree, Protein Conformation, Amino Acid Sequence, Base Sequence, Likelihood Functions, Triturus, and Molecular Sequence Data
Research Interests: Biology, Medicine, Biological Sciences, Humans, Animals, and 6 moreVertebrates, Estrogen, Male, Nonylphenol, Estrogens, and Vitellogenin
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Research Interests: Evolutionary Biology, Genetics, Phylogenetics, Molecular Evolution, Biology, and 15 moreMedicine, PCR, Animals, Metallothionein, Phylogenetic analysis, Evolutionary History, Molecular Phylogenetics and Evolution, Gene, Gene Duplication, Bp, Molecular cloning, cDNA, Amino Acid Sequence, Base Sequence, and Molecular Sequence Data
Research Interests: Genetics, Biology, Medicine, Gene expression, Oogenesis, and 15 moreMolecular Mechanics, Biological Sciences, Cell line, Humans, Copper, Lizards, Animals, Growth and development, Membrane transport proteins, Molecular cloning, Biological Process, Amino Acid Sequence, Base Sequence, Oocytes, and Molecular Sequence Data
Research Interests: Fisheries, Bioaccumulation, Biology, Environmental Monitoring, Environmental Chemistry, and 14 moreAquaculture, Medicine, Multidisciplinary, Heavy metals, Animals, Lead, Metallothionein, Fish Farming, Cadmium, Environmental monitoring and assessment, Perciformes, Gill, Environmental Pollution Assessment and Monitoring, and Tissue distribution
Research Interests: Biology and Sea Urchin
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Organisms living in the Southern Ocean are exposed to strong environmental constraints, especially temperature. The Perciform suborder Notothenioidei, today largely endemic to the Antarctic, has developed numerous cold-adapted characters.... more
Organisms living in the Southern Ocean are exposed to strong environmental constraints, especially temperature. The Perciform suborder Notothenioidei, today largely endemic to the Antarctic, has developed numerous cold-adapted characters. The most striking peculiarity of this group of fish is the drastic reduction of hemoglobin content in their blood. This condition gives rise to a low metabolic demand of iron. Recently, we have undertaken a study to add new insights on iron metabolism in hemoglobinless fish. By taking advantage to our previous studies on transferrins from Antarctic Notothenioids, in this article we compared the evolutionary properties of these sequences to those from other fish groups and mammals. Our findings based on analysis of dN/dS ratio and on the results of the relative rate ratio test suggest that the transferrin gene has undergone positive selection in fish but not in mammals. The results of type I functional divergence provide statistical evidence for shifted evolutionary rate after speciation. Finally, when superimposed onto the three-dimensional structure of transferrin, the sites identified as responsible of the divergence in Notothenioids appear to cluster preferentially at non-iron binding locations, occupying surface locations that might affect the rate of iron binding and/or release in a cold environment.
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Research Interests: Biology, Cell Biology, Medicine, Animals, Zinc, and 12 moreMetallothionein, Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis, Gel Permeation Chromatography, Egg Proteins, Sea Urchins, Sea Urchin, Paracentrotus lividus, Species Specificity, Oocytes, Protein Binding, Cytoplasm, and Biochemistry and cell biology
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Pharmaceuticals are a class of emerging environmental contaminants. Gadolinium (Gd) is a metal of the lanthanide series of the elements whose chelates are commonly employed as contrast agents for magnetic resonance imaging, and... more
Pharmaceuticals are a class of emerging environmental contaminants. Gadolinium (Gd) is a metal of the lanthanide series of the elements whose chelates are commonly employed as contrast agents for magnetic resonance imaging, and subsequently released into the aquatic environment. We investigated the consequences of sea urchin embryo exposure to sublethal Gd concentrations, comparing the effects on the development of four phylogenetically and geographically distant species: two Mediterranean species, Paracentrotus lividus and Arbacia lixula, and two species living in the East coast of Australia, Heliocidaris tuberculata and Centrostephanus rodgersii. Measures of the Gd and Ca content inside embryos by ICP-MS showed a time- and dose-dependent Gd increase, in parallel with a reduction of the Ca amount, suggesting that Gd is able to compete with Ca for binding to the Ca channels. In all the four species, we observed a general delay of embryo development at 24h post-fertilization, and a strong inhibition of skeleton growth at 48h, with species-specific threshold levels of sensitivity. Further experiments were carried out on P. lividus embryos. Removal of Gd after 24h caused partial recovery of embryo development 48h post fertilization. We detected an increase of the LC3 autophagic marker at 24 and 48h, while confocal microscopy studies confirmed the increased number of autophagosomes and autophagolysosomes. In contrast, immunofluorescence studies showed no apoptotic induction. RT-PCR gene expression analysis showed the misregulation of several genes implicated both in the skeletogenic and the left-right axis specification networks, including: transcription factors (Alx-1, Nodal), signaling molecules (univin, VEGF, VEGF-R, FGF) and skeletal matrix proteins (p16, p19 and msp130). Taken together, the results pose serious questions on the hazard of Gd in the marine environment and indicate that Gd is able to affect three different levels of the stress response in sea urchin embryos: morphogenesis, survival strategies such as autophagy, and gene expression
Research Interests: Toxicology, Biology, Development, Skeletogenesis, Embryo, and 3 moreToxicity, Gadolinium, and Sea Urchin
Studies of congenital malformations and their etiology in wild animals are a powerful tool when considered in a holistic/overall evaluation of the “one health status”. Reptiles, according to their habitat and biological features, can be... more
Studies of congenital malformations and their etiology in wild animals are a powerful tool when considered in a holistic/overall evaluation of the “one health status”. Reptiles, according to their habitat and biological features, can be promising predictors of environmental contamination. However, research in this area remains scarce and dispersed. The purpose of this review is to compile some documentation that describes or discusses the teratogenic effects of pesticides on reptiles. Further research in this area is urgently needed to implement concrete measures to improve ecosystem health and stop the present decline of reptile populations.
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The teratogenic effects of thermal stress were studied in the oviparous Italian wall lizard Podarcis sicula. To this purpose, the eggs were exposed to continuous or temporary cold (15°C) or warm (30°C) stresses and the effects were... more
The teratogenic effects of thermal stress were studied in the oviparous Italian wall lizard Podarcis sicula. To this purpose, the eggs were exposed to continuous or temporary cold (15°C) or warm (30°C) stresses and the effects were analysed at the cytological and molecular levels. The results demonstrated the lethality of the continuous regimes and of the warm temporary regime, no matter if given at early (5 days) or late (15 days) stages of development. Temporary cold stress also resulted in lethality, but only if given in the early stage; later, in fact, it resulted in an abnormal development, with marked alterations in the encephalic vesicles, in the eyes and the trunk organs. By in situ hybridization, it was demonstrated that these alterations were often correlated with changes in HSP70 expression. In conclusion, our data indicate that Podarcis embryos have a limited potential to tolerate thermal changes, especially warm ones. The average predicted temperature increase of 2-4°C in the next few decades could therefore represent a real threat for lizard populations living in temperate areas.
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Research Interests: Botany, Chemistry, Photosynthesis, Multidisciplinary, RuBisCO, and 3 moreCatalase, Ascorbic Acid, and Fagaceae
Research Interests: Environmental Toxicology, Biology, Apoptosis, Medicine, Oogenesis, and 15 moreBiological Sciences, Environmental Sciences, Lizards, Female, Animals, Metallothionein, Cadmium, Lizard, CHEMICAL SCIENCES, Ovarian Follicle, Environmental Pollutants, Cell Proliferation, Environmental, Cell count, and Cadmium Chloride
The present report is an attempt to investigate the influence of intraperitoneal and dietary cadmium exposure on the distribution of cadmium accumulation and induction of metallothionein gene expression in different tissues of the lizard... more
The present report is an attempt to investigate the influence of intraperitoneal and dietary cadmium exposure on the distribution of cadmium accumulation and induction of metallothionein gene expression in different tissues of the lizard Podarcis sicula. Cadmium accumulation in liver, kidney, ovary, brain and intestine was measured by atomic absorption spectrometry. Metallothionein gene induction was determined by dot blot analyses on the total RNA extracted from the same organs. Our data indicate that cadmium exposure results in significant cadmium uptake, but the patterns of this uptake varies with organ and exposure route. After a single intraperitoneal treatment, concentrations of cadmium and metallothionein transcript are positively correlated in kidney, liver and ovary. Following a dietary cadmium treatment, a positive correlation between the increase of metallothionein mRNA and cadmium accumulation is found in intestine, ovary and kidney, while no correlation is present in liver and brain.
Research Interests: RNA, Biology, Medicine, Gene expression, In Situ Hybridization, and 15 moreBiological Sciences, Environmental Sciences, Kidney, Lizards, Chronic Disease, Female, Animals, Metallothionein, Atomic Absorption Spectrometry, Cadmium, Ovary, Cadmium Poisoning, Acute Disease, Tissue distribution, and Medical and Health Sciences
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Given their strong site fidelity and low mobility, lizards have been identified as potential model species for terrestrial vertebrate ecotoxicology since they can provide useful information about land-based pollution. Lizards can be... more
Given their strong site fidelity and low mobility, lizards have been identified as potential model species for terrestrial vertebrate ecotoxicology since they can provide useful information about land-based pollution. Lizards can be directly exposed to fertilizers, pesticides and other chemical substances used in agriculture and livestock breeding through various routes: ingestion of soil and contaminated food, dermal exposure, inhalation, maternal transfer to eggs, and absorption by eggs of contaminants from environments. The advantage of using Podarcis sicula as a bioindicator of estrogen pollution is that its reproductive cycle is well known, both in males and females. Males, in particular, are excellent sentinels since they are able to synthesize vitellogenin (VTG) when exposed to an estrogenic environment; hence, VTG is commonly used as a biomarker of pollution from estrogenic compounds, mainly in aquatic or semi-aquatic species. In this chapter, we present data collected using P. sicula as an experimental model for monitoring soil health status in both intensive and organic farming, as well as for the study of damage caused at tissutal and cellular levels following environmental contamination by estrogenic compounds.
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Soil contaminants (herbicides, pesticides, and heavy metals) are among the main causes of change in terrestrial ecosystems. These substances lead to a general loss of biodiversity, both of flora and fauna and being able to biomagnify and... more
Soil contaminants (herbicides, pesticides, and heavy metals) are among the main causes of change in terrestrial ecosystems. These substances lead to a general loss of biodiversity, both of flora and fauna and being able to biomagnify and pass through the food chain, they can endanger the survival of terrestrial vertebrates at the top of this chain. This review analyzes the risks associated with exposure to glyphosate, the active principle of many herbicide products, for the reproductive health of the field lizard (Podarcis siculus) potentially exposed to the substance in its natural habitat; therefore, introducing it as a possible model organism. Data demonstrate that glyphosate is toxic for this animal, affecting the health of the reproductive organs, both in males and females, and of the liver, the main detoxifying organ and closely involved in the female reproductive process. Sharing structural and functional characteristics of these organs with many other vertebrates, the inform...
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Pituitary adenylate cyclase-activating peptide (PACAP) is a neuropeptide involved in multiple functions, including vertebrate reproduction. Recently, we reported the presence of PACAP in the testis of Italian wall lizard Podarcis sicula... more
Pituitary adenylate cyclase-activating peptide (PACAP) is a neuropeptide involved in multiple functions, including vertebrate reproduction. Recently, we reported the presence of PACAP in the testis of Italian wall lizard Podarcis sicula during reproductive period (May-June). Herein we investigated the PACAP mRNA expression and the localization of PACAP/PACAP receptor system, in the other periods of the Podarcis reproductive cycle, namely in summer stasis, early autumnal resumption, mid-autumnal resumption, winter stasis, and spring resumption. Using biomolecular and immunohistochemical investigations, we demonstrated that PACAP mRNA was widely expressed in all germ and somatic cells; in summer stasis (July-August) and early autumnal resumption (September) in particular, the mRNA was always found in Sertoli cells while was transiently expressed in germ and in Leydig cells. Differently from the mRNA, the protein was always present in germ and somatic cells independently from the reproductive cycle phase. As PACAP, the PAC1 receptor was always present in the testis, except for the summer stasis (July-August) and the early autumnal resumption (September), when PACAP was lacking in germ and somatic cells (Leydig and Sertoli cells). The present results strongly suggest that PACAP/PAC1 receptor system is widely represented during the reproductive cycle of male lizard. The possible involvement of PACAP/PACAP receptor system in the control of spermatogenesis is discussed.
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Glyphosate (Gly) is a broad-spectrum herbicide widely used thanks to its high efficiency and low toxicity. However, evidence exists of its toxic effects on non-target organisms. Among these, the animals inhabiting agricultural fields are... more
Glyphosate (Gly) is a broad-spectrum herbicide widely used thanks to its high efficiency and low toxicity. However, evidence exists of its toxic effects on non-target organisms. Among these, the animals inhabiting agricultural fields are particularly threatened. Recent studies demonstrated that exposure to Gly markedly affected the morphophysiology of the liver and testis of the Italian field lizard Podarcis siculus. The present study aimed to investigate the effects of the herbicide on the female reproductive system of this lizard in order to have a full picture of Gly-induced reproductive impairment. The animals were exposed to 0.05 and 0.5 μg/kg of pure Gly by gavage for 3 weeks. The results demonstrated that Gly, at both doses tested, profoundly interfered with ovarian function. It induced germ cells’ recruitment and altered follicular anatomy by anticipating apoptotic regression of the pyriform cells. It also induced thecal fibrosis and affected oocyte cytoplasm and zona pelluc...
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doi: 10.3389/fncel.2014.00212 Haptoglobin increases with age in rat hippocampus and modulates Apolipoprotein E mediated cholesterol trafficking in neuroblastoma cell lines
Fish and mammalian metallothioneins (MTs) differ in the amino acid residues placed between their conserved cysteines. We have expressed the MT of an Antarctic fish, Notothenia coriiceps, and characterized it by means of multinuclear NMR... more
Fish and mammalian metallothioneins (MTs) differ in the amino acid residues placed between their conserved cysteines. We have expressed the MT of an Antarctic fish, Notothenia coriiceps, and characterized it by means of multinuclear NMR spectroscopy. Overall, the architecture of the fish MT is very similar to that of mammalian MTs. However, NMR spectroscopy shows that the dynamic behaviour of the two domains is markedly different. With the aid of absorption and CD spectroscopies, we studied the conformational and electronic features of fish and mouse recombinant Cd-MT and the changes produced in these proteins by heating. When the temperature was increased from 20 to 90°C, the Cd-thiolate chromophore absorbance at 254nm of mouse MT was not modified up to 60°C, whereas the absorbance of fish MT decreased significantly starting from 30°C. The CD spectra also changed quite considerably with temperature, with a gradual decrease of the positive band at 260nm that was more pronounced for fish than for mouse MT. The differential effect of temperature on fish and mouse MTs may reflect a different stability of metal-thiolate clusters of the two proteins. Such a conclusion is also corroborated by results showing differences in metal mobility between fish and mouse Zn-MT.
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Abstract: Cadmium is a highly polluting heavy metal known to have undesirable effects on health in both animals and humans, targeting the kidneys, the liver and the vascular system. A wide spectrum of deleterious effects has been reported... more
Abstract: Cadmium is a highly polluting heavy metal known to have undesirable effects on health in both animals and humans, targeting the kidneys, the liver and the vascular system. A wide spectrum of deleterious effects has been reported also on the reproductive organs and the developing embryo. Cadmium in fact is a strong endocrine disruptor that interferes with functioning of endogenous receptors and hormones causing detrimental effects on offspring production and survival. In spite of the wide number of studies carried out in laboratory mammals, data on cadmium effects on gonadic tissues, fertility and reproduction of wild terrestrial vertebrates are still limited. In particular, information on the consequences of environmental cadmium exposure on reptiles survival and biodiversity is particularly scanty. Reptiles are presently considered highly susceptible to a number of environmental pollutants and this has contributed to the global decline of several wild populations of turtl...
Fertility rates in animals have shown a progressive decrease in recent decades, and reproductive toxicity is considered an important regulatory endpoint in health hazard assessment [...]
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The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant. The sea urchin embryo has long been used to study the... more
The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant. The sea urchin embryo has long been used to study the effects induced by metals, including V. In this study we used an integrated approach that correlates the biological effects on embryo development with proteolytic activities of gelatinases that could better reflect any metal-induced imbalances. V-exposure caused morphological/morphometric aberrations, mainly concerning the correct distribution of embryonic cells, the development of the skeleton, and the embryo volume. Moreover, V induced a concentration change in all the gelatinases expressed during embryo development and a reduction in their total proteolytic activity. The presence of three MMP-like gelatinases (MMP-2, -9, and -14) was also demonstrated and their levels depended on V-concentration. In particular, the MMP-14-like protein modified its ex...
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Dottorato di ricerca in biologia evoluzionistica e del differenziamento. 8. ciclo. A.a. 1992-95. Relatore e coordinatore S. FilosaConsiglio Nazionale delle Ricerche - Biblioteca Centrale - P.le Aldo Moro, 7, Rome; Biblioteca Nazionale... more
Dottorato di ricerca in biologia evoluzionistica e del differenziamento. 8. ciclo. A.a. 1992-95. Relatore e coordinatore S. FilosaConsiglio Nazionale delle Ricerche - Biblioteca Centrale - P.le Aldo Moro, 7, Rome; Biblioteca Nazionale Centrale - P.za Cavalleggeri, 1, Florence / CNR - Consiglio Nazionale delle RichercheSIGLEITItal
Research Interests: Zinc and Sea Urchin
In aquatic organisms, cadmium exposure occurs from ovum to death and the route of absorption is particularly wide, being represented by skin, gills and gastrointestinal tract, through which contaminated water and/or preys are ingested. It... more
In aquatic organisms, cadmium exposure occurs from ovum to death and the route of absorption is particularly wide, being represented by skin, gills and gastrointestinal tract, through which contaminated water and/or preys are ingested. It is known that cadmium interferes with the gut; however, less information is available on cadmium effects on an important component of the gut, namely goblet cells, specialized in mucus synthesis. In the present work, we studied the effects of two sublethal cadmium concentrations on the gut mucosa of Danio rerio. Particular attention was paid to changes in the distribution of glycan residues, and in metallothionein expression in intestinal cells. The results show that cadmium interferes with gut mucosa and goblet cells features. The effects are dose- and site-dependent, the anterior gut being more markedly affected than the midgut. Cadmium modifies the presence and/or distribution of glycans in the brush border and cytoplasm of enterocytes and in th...
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Research Interests: Biology, Medicine, Biological Sciences, Humans, Animals, and 6 moreVertebrates, Estrogen, Male, Nonylphenol, Estrogens, and Vitellogenin
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Research Interests: Kinetics, Biology, Magnetic Resonance Spectroscopy, Medicine, Biological Sciences, and 15 moreCircular Dichroism, Escherichia coli, Mice, Animals, Metallothionein, Biochemical, Cadmium, Recombinant DNA, CHEMICAL SCIENCES, Perciformes, Protein Conformation, Absorbance, Amino Acid Sequence, Molecular Sequence Data, and Medical and Health Sciences
The growing presence of lanthanides in the environment has drawn the attention of the scientific community on their safety and toxicity. The sources of lanthanides in the environment include diagnostic medicine, electronic devices,... more
The growing presence of lanthanides in the environment has drawn the attention of the scientific community on their safety and toxicity. The sources of lanthanides in the environment include diagnostic medicine, electronic devices, permanent magnets, etc. Their exponential use and the poor management of waste disposal raise serious concerns about the quality and safety of the ecosystems at a global level. This review focused on the impact of lanthanides in marine organisms on reproductive fitness, fertilization and embryonic development, using the sea urchin as a biological model system. Scientific evidence shows that exposure to lanthanides triggers a wide variety of toxic insults, including reproductive performance, fertilization, redox metabolism, embryogenesis, and regulation of embryonic gene expression. This was thoroughly demonstrated for gadolinium, the most widely used lanthanide in diagnostic medicine, whose uptake in sea urchin embryos occurs in a time- and concentration-...
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The expansion of agriculture produces a steady increase in habitat fragmentation and degradation due to the increased use of pesticides and herbicides. Habitat loss and alteration associated with crop production play an important role in... more
The expansion of agriculture produces a steady increase in habitat fragmentation and degradation due to the increased use of pesticides and herbicides. Habitat loss and alteration associated with crop production play an important role in reptile decline, among which lizards are particularly endangered. In this study, we evaluated testicular structure, steroidogenesis, and estrogen receptor expression/localization after three weeks of oral exposure to glyphosate at 0.05 and 0.5 μg/kg body weight every other day in the field lizard Podarcis siculus. Our results show that glyphosate affected testicular morphology, reduced spermatogenesis, altered gap junctions and changed the localization of estrogen receptors in germ cells, increasing their expression; the effects were mostly dose-dependent. The result also demonstrates that glyphosate, at least at these concentrations, did not influence steroidogenesis. Overall, the data indicate that this herbicide can disturb the morphophysiology o...
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Vanadium toxicology is a topic of considerable importance as this metal is widely used in industrial and biomedical fields. However, it represents a potential emerging environmental pollutant because wastewater treatment plants do not... more
Vanadium toxicology is a topic of considerable importance as this metal is widely used in industrial and biomedical fields. However, it represents a potential emerging environmental pollutant because wastewater treatment plants do not adequately remove metal compounds that are subsequently released into the environment. Vanadium applications are limited due to its toxicity, so it is urgent to define this aspect. This metal is associated with sea urchin embryo toxicity as it perturbs embryogenesis and skeletogenesis, triggering several stress responses. Here we investigated its bioaccumulation and the correlation with cellular and molecular developmental pathways. We used cytotoxic concentrations of 1 mM and 500 μM to perform quantitative analyses, showing that vanadium accumulation interferes with calcium uptake during sea urchin development and provokes a disruption in the biomineralization process. At the end of the whole treatment, the accumulation of vanadium was about 14 and 8 ...
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Cadmium is a persistent contaminant accumulated in the environment from both anthropogenic and natural sources. Every year, large quantities of this metal are released in the different environmental compartments and may pose a significant... more
Cadmium is a persistent contaminant accumulated in the environment from both anthropogenic and natural sources. Every year, large quantities of this metal are released in the different environmental compartments and may pose a significant threat to the biota exposed. Intracellular damage caused by cadmium exposure includes protein denaturation, lipid peroxidation, generation of reactive oxygen species and DNA strand breaks. Many studies have also demonstrated that this ion has a teratogenic or lethal effect on embryos, related to the dose and exposure time. In spite of the wide number of studies carried out in laboratory mammals, data on cadmium effects on fertility, reproduction and embryonic development of wild terrestrial vertebrates are still limited. In particular, information on the consequences of environmental cadmium exposure on reptiles survival and biodiversity are particularly scanty. Reptiles are presently considered highly susceptible to a number of environmental pollu...
The biological mechanisms linking nutrition and antioxidants content of the diet with cardiovascular protection are subject of intense investigation. It has been demonstrated that dietary supplementation with cow, donkey or human milk,... more
The biological mechanisms linking nutrition and antioxidants content of the diet with cardiovascular protection are subject of intense investigation. It has been demonstrated that dietary supplementation with cow, donkey or human milk, characterized by distinct nutritional properties, triggers significant differences in the metabolic and inflammatory status through the modulation of hepatic and skeletal muscle mitochondrial functions. Cardiac mitochondria play a key role for energy-demanding heart functions, and their disfunctions is leading to pathologies. Indeed, an altered heart mitochondrial function and the consequent increased reactive oxygen species (ROS) production and inflammatory state, is linked to several cardiac diseases such as hypertension and heart failure. In this work it was investigated the impact of the milk consumption on heart mitochondrial functions, inflammation and oxidative stress. In addition, it was underlined the crosstalk between mitochondrial metabolic...
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Given their strong site fidelity and low mobility, lizards have been identified as potential model species for terrestrial vertebrate ecotoxicology since they can provide useful information about land-based pollution. Lizards can be... more
Given their strong site fidelity and low mobility, lizards have been identified as potential model species for terrestrial vertebrate ecotoxicology since they can provide useful information about land-based pollution. Lizards can be directly exposed to fertilizers, pesticides and other chemical substances used in agriculture and livestock breeding through various routes: ingestion of soil and contaminated food, dermal exposure, inhalation, maternal transfer to eggs, and absorption by eggs of contaminants from environments. The advantage of using Podarcis sicula as a bioindicator of estrogen pollution is that its reproductive cycle is well known, both in males and females. Males, in particular, are excellent sentinels since they are able to synthesize vitellogenin (VTG) when exposed to an estrogenic environment; hence, VTG is commonly used as a biomarker of pollution from estrogenic compounds, mainly in aquatic or semi-aquatic species. In this chapter, we present data collected using P. sicula as an experimental model for monitoring soil health status in both intensive and organic farming, as well as for the study of damage caused at tissutal and cellular levels following environmental contamination by estrogenic compounds.
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Environmental chemicals can be introduced by consuming contaminated foods. The environmental chemical dichlorodiphenyldichloroethylene (DDE), a persistent metabolite of dichlorodiphenyltrichloroethane (DDT), can affect spermatogenesis.... more
Environmental chemicals can be introduced by consuming contaminated foods. The environmental chemical dichlorodiphenyldichloroethylene (DDE), a persistent metabolite of dichlorodiphenyltrichloroethane (DDT), can affect spermatogenesis. Our study aims to evaluate, by using spectrophotometric analyses, western blot, and immunohistochemistry, the adaptive responses in testis of adult rats treated with a non-toxic dose of DDE, alone or in association with a high-fat diet (HFD). Four experimental groups were performed: N (normal diet); D (HFD); D + DDE (HFD + DDE); N + DDE (normal diet + DDE). D group showed a reduction in antioxidant capacity, and increases in lipid peroxidation, apoptosis, and proliferation associated with morphological impairment. A reduction in androgen receptor (AR) and serum testosterone levels were also found. DDE-treated groups exhibited higher lipid peroxidation levels compared to N and D, associated with pronounced defect in antioxidant capacity, apoptosis, cel...
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Exploiting the annotation of the western clawed frog Silurana tropicalis genome, we identified a new metallothionein (MT) gene, exhibiting all the features to be considered an active gene, but with an atypical coding region, showing only... more
Exploiting the annotation of the western clawed frog Silurana tropicalis genome, we identified a new metallothionein (MT) gene, exhibiting all the features to be considered an active gene, but with an atypical coding region, showing only 17 cysteine residues instead of the canonical 20 cysteines of vertebrate metallothioneins and two anomalous cysteine triplets. However, the presence of a gene in the genome does not ensure its effective expression. By using conventional and Real-Time PCR analyses, we demonstrated that this atypical MT is constitutively expressed throughout the life cycle of the African clawed frog Xenopus laevis; moreover, this gene is highly expressed in the adult liver, the major site of MT expression and synthesis in vertebrates. To our knowledge, the X. laevis MT described in this paper is the first sequence of a vertebrate MT showing only 17 cysteine residues, arranged in two Cys-Cys-Cys motifs. Phylogenetic analyses also demonstrated that the atypical X. laevi...
Research Interests: RNA, Computational Biology, Aging, Biological Sciences, Phylogeny, and 12 moreAnimals, Metallothionein, Biological evolution, Molecular cloning, Xenopus laevis, Cysteine, Protein isoforms, Amino Acid Sequence, Base Sequence, Gene Expression Regulation, Molecular Sequence Data, and real time polymerase chain reaction
The cleidoic eggs of oviparous reptiles are protected from the external environment by membranes and a parchment shell permeable to water and dissolved molecules. As a consequence, not only physical but also chemical insults can reach the... more
The cleidoic eggs of oviparous reptiles are protected from the external environment by membranes and a parchment shell permeable to water and dissolved molecules. As a consequence, not only physical but also chemical insults can reach the developing embryos, interfering with gene expression. This review provides information on the impact of the exposure to cadmium contamination or thermal stress on gene expression during the development of Italian wall lizards of the genus Podarcis. The results obtained by transcriptomic analysis, although not exhaustive, allowed to identify some stress-reactive genes and, consequently, the molecular pathways in which these genes are involved. Cadmium-responsive genes encode proteins involved in cellular protection, metabolism and proliferation, membrane trafficking, protein interactions, neuronal transmission and plasticity, immune response, and transcription regulatory factors. Cold stress changes the expression of genes involved in transcriptiona...
The growing number of studies on metallothioneins (MTs), cysteine-rich metal-binding proteins, have been disclosing new functions of these proteins. Thanks to their inducibility, they were considered to play a pivotal role in regulating... more
The growing number of studies on metallothioneins (MTs), cysteine-rich metal-binding proteins, have been disclosing new functions of these proteins. Thanks to their inducibility, they were considered to play a pivotal role in regulating trace metals homeostasis and in detoxification from heavy metals; nowadays, it is known that they are involved in various physiological and pathological processes, such as regulation of apoptosis, elimination of free radicals, and protection of nucleic acids against toxic insults. MT induction has been demonstrated following stress factors other than heavy metals, such as endocrine-disrupting chemicals, insecticides, and herbicides. However, retrieved data are often controversial: in some cases, xenobiotics elicit MT expression and synthesis; under different conditions, they lead to a decrease in cellular MT content. This review describes the MT response to dichlorodiphenyltrichloroethane (DDT) contamination in mammalian tissues. In particular, atten...
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The potential toxicity of glyphosate, a widely used broad-spectrum herbicide, is currently a great matter of debate. As vertebrate insectivores, lizards protect plants from herbivorous insects increasing plant biomass via the trophic... more
The potential toxicity of glyphosate, a widely used broad-spectrum herbicide, is currently a great matter of debate. As vertebrate insectivores, lizards protect plants from herbivorous insects increasing plant biomass via the trophic cascade and represent an important link between invertebrates and higher predators. A negative effect of glyphosate on lizards’ survival could have major impacts at the ecological levels. In this study, we investigated the effects of the exposure to low doses of glyphosate on the liver of the wall lizard Podarcis siculus, a suitable bioindicator of soil pollution. Two different doses of pure glyphosate (0.05 and 0.5 μg/kg body weight) were orally administered every other day for 3 weeks to sexually mature males and females. The results demonstrated that both doses, despite being very low, are toxic for the liver that showed clear signs of suffering, regardless of sex. The histological analysis provided a scenario of severe hepatic condition, which degen...
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Spermatogenesis is a fascinating biological process aiming to generate haploid spermatozoa from diploid spermatogonia through a specific hormonal network between gonadotropins and steroids. Increasing evidence suggests that the primary... more
Spermatogenesis is a fascinating biological process aiming to generate haploid spermatozoa from diploid spermatogonia through a specific hormonal network between gonadotropins and steroids. Increasing evidence suggests that the primary female sex hormone estrogen plays an active role in this process. This research points out on the role of estrogen during lizard spermatogenesis by using three experimental approaches: (1) exposure to an analogue of nonsteroidal estrogen as Clomiphene citrate that acts both as estrogen agonist and antagonist; (2) exposure to the gonadotropin FSH; and (3) exposures to FSH followed by Clomiphene. Histological and immunohistochemical results demonstrate that in the lizardduring the mating period, Clomiphene as well as FSH determines the breakdown of spermatogenesis and the epididymal regression, presumably through estrogens input as indirectly demonstrated by the appearance of ERand vitellogenin in the liver. The ability of Clomiphene to restore the gona...
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Chelates of Gadolinium (Gd), a lanthanide metal, are employed as contrast agents for magnetic resonance imaging and are released into the aquatic environment where they are an emerging contaminant. We studied the effects of... more
Chelates of Gadolinium (Gd), a lanthanide metal, are employed as contrast agents for magnetic resonance imaging and are released into the aquatic environment where they are an emerging contaminant. We studied the effects of environmentally relevant Gd concentrations on the development of two phylogenetically and geographically distant sea urchin species: the Mediterranean Paracentrotus lividus and the Australian Heliocidaris tuberculata. We found a general delay of embryo development at 24h post-fertilization, and a strong inhibition of skeleton growth at 48h. Total Gd and Ca content in the larvae showed a time- and concentration-dependent increase in Gd, in parallel with a reduction in Ca. To investigate the impact of Gd on the expression of genes involved in the regulation of skeletogenesis, we performed comparative RT-PCR analysis and found a misregulation of several genes involved in the skeletogenic and left-right axis specification gene regulatory networks. Species-specific di...
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The effects of an exposure to cadmium chloride 0.47μM for 150days were studied in kidneys of juveniles Sparus aurata by a multidisciplinary approach so to correlate uptake and detoxification potential to changes in brush border and... more
The effects of an exposure to cadmium chloride 0.47μM for 150days were studied in kidneys of juveniles Sparus aurata by a multidisciplinary approach so to correlate uptake and detoxification potential to changes in brush border and glycocalyx sugar composition. Results demonstrated that cadmium concentration in kidney significantly increased from day 30 reaching a plateau on day 120 while metallothioneins reached a peak on day 90 and by day 120 were already decreasing to control values. Cytological damage was extensive on day 90, clearly detectable at both structural and ultrastructural levels, in tubular cells and brush-border. Staining with a panel of four lectins revealed a significant increase in N-Ac-Gal and a decrease in mannose in the glycocalyx and the tubular basal membranes. From day 120, when cadmium concentration was high and metallothionein concentration decreasing, a clear recovery was observed in tubular cells morphology and sugar composition. Possible significance of...
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The "sex-hormone" estrogen-17β promotes several cognitive functions and is a master regulator of brain bioenergetics via the estrogen receptors α and β (ERα and ERβ). In this work, by using Real-Time PCR analysis, we... more
The "sex-hormone" estrogen-17β promotes several cognitive functions and is a master regulator of brain bioenergetics via the estrogen receptors α and β (ERα and ERβ). In this work, by using Real-Time PCR analysis, we evaluated the effect of aging and high fat diet (HFD) on ERα and ERβ expression in rat hippocampus and cortex. In young rats, ERβ is abundant in cortex and ERα in hippocampus. During the aging, in the cortex, we observe a general decrease in ERα and ERβ expression; in hippocampus ERα increases and ERβ decreases. ER expression patterns in rat brain are also affected by the administration of an HFD. In cortex, after 4weeks of HFD, ERβ transcripts are down-regulated, whereas ERα levels remain unchanged; after 12weeks, both ERα and ERβ expression is up-regulated. In the hippocampus, the level of ERβ transcripts does not change following HDF, whereas ERα expression is affected by HDF, in a time-dependent manner: it increases after the 4-week treatment and decreases after 12weeks. Possible involvements of these receptors in the control of cortex and hippocampus functions during aging and in the modulation of energetic metabolism and feeding behaviour are discussed.
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Metallothioneins (MTs) are an evolutionary conserved multigene family of proteins whose role was initially identified in binding essential metals. The physiological role of MT, however, has been revealed to be more complex than expected,... more
Metallothioneins (MTs) are an evolutionary conserved multigene family of proteins whose role was initially identified in binding essential metals. The physiological role of MT, however, has been revealed to be more complex than expected, since not only are MTs able to bind to toxic heavy metals, but many isoforms have shown specialized and alternative functions. Within this uncertainty, the information available on MTs in non-mammalian vertebrates, particularly in neglected tetrapods such as the reptiles, is even more scant. In this review, we provide a summary of the current understanding on metallothionein presence and function in the oviparous lizard Podarcis sicula, highlighting the results obtained by studying MT gene expression in most representative adult and embryonic tissues. The results demonstrate that in adults, cadmium induces MT transcription in a dose- and tissue-specific manner. Thus, the MT mRNAs appear, at least in some cases, to be an unsuitable tool for detecting...
Research Interests: Genetics, Reproduction, Lizards, Female, Animals, and 4 moreMetallothionein, Cadmium, Molecular sciences, and Oviparity
In this study, our aim was to determine whether caspase 3 plays a role, during previtellogenesis, in the ovarian follicular epithelium of the lizard Podarcis sicula. We investigated the presence and localization of proform and active... more
In this study, our aim was to determine whether caspase 3 plays a role, during previtellogenesis, in the ovarian follicular epithelium of the lizard Podarcis sicula. We investigated the presence and localization of proform and active caspase 3 by enzyme assay, Western blotting and immunocytochemistry. In parallel, a fragment of caspase 3 was cloned for the first time in this species, sequenced and used for in situ hybridization to localize messengers and analysed by a phylogenetic survey to shed light on its homology with reptilian caspases. Results demonstrated that: (1) the follicle cells expressed a caspase of the 3/7 group and the mRNA for caspase 3 was transcribed in the stem phase and was completely translated during cell differentiation; (2) the proform protein was stored during the differentiated (nurse) stage and activated at the end of previtellogenesis provoking the degeneration of cells; (3) the predicted protein sequence, although partial, had a strong similarity with the known reptilian caspases 3. The epithelial cells of the ovarian follicle, therefore, do not employ caspase 3 during the nurse stage but, instead, prepare for apoptosis long before the process actually begins. The relevance of this strategy is discussed.
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In oviparous vertebrates, vitellogenin (VTG) is the major yolk precursor synthesized in the liver of sexually mature females during the reproductive period. In males, the VTG gene is silent, but it may be activated by estradiol-17β (E2)... more
In oviparous vertebrates, vitellogenin (VTG) is the major yolk precursor synthesized in the liver of sexually mature females during the reproductive period. In males, the VTG gene is silent, but it may be activated by estradiol-17β (E2) or estrogen-like substances. Until now, extra-hepatic expression and synthesis of VTG after estrogen exposure has been reported only for aquatic vertebrates. This study demonstrates the ability of testis and epididymis of the terrestrial oviparous lacertid Podarcis sicula to synthesize VTG following E2 exposure. The results of in situ hybridization and immunohistochemistry analysis show the presence of both VTG mRNA and protein in these districts besides the known induction in the liver. The possible contemporaneous uptake of the E2-induced hepatic VTG by means of the specific vitellogenin receptor has been also evaluated. Finally, histological analysis shows that the E2-treatment during the mating season impairs spermatogenesis.
Research Interests: Physiology, Zoology, Lizards, Female, Animals, and 5 moreMale, tESTIS, Estrogens, Veterinary Sciences, and Epididymis
The teratogenic effects of thermal stress were studied in the oviparous Italian wall lizard Podarcis sicula. To this purpose, the eggs were exposed to continuous or temporary cold (15°C) or warm (30°C) stresses and the effects were... more
The teratogenic effects of thermal stress were studied in the oviparous Italian wall lizard Podarcis sicula. To this purpose, the eggs were exposed to continuous or temporary cold (15°C) or warm (30°C) stresses and the effects were analysed at the cytological and molecular levels. The results demonstrated the lethality of the continuous regimes and of the warm temporary regime, no matter if given at early (5 days) or late (15 days) stages of development. Temporary cold stress also resulted in lethality, but only if given in the early stage; later, in fact, it resulted in an abnormal development, with marked alterations in the encephalic vesicles, in the eyes and the trunk organs. By in situ hybridization, it was demonstrated that these alterations were often correlated with changes in HSP70 expression. In conclusion, our data indicate that Podarcis embryos have a limited potential to tolerate thermal changes, especially warm ones. The average predicted temperature increase of 2-4°C ...
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The aim of this study was to verify if the freshwater safety values established from the European Community (1998) and the Italian Ministry of Health (2001) for cadmium (44.5nM/L in drinking water and 178nM/L in sewage waters) were safe... more
The aim of this study was to verify if the freshwater safety values established from the European Community (1998) and the Italian Ministry of Health (2001) for cadmium (44.5nM/L in drinking water and 178nM/L in sewage waters) were safe for amphibians, since at these same concentrations cadmium induced endocrine disruption in the newt Triturus carnifex. Adult male specimens of T. carnifex were exposed daily to cadmium (44.5nM/L and 178nM/L as CdCl2, nominal concentrations), respectively, during 3- and 9-months; at the same time, control newts were exposed to tap water only. The accumulation of cadmium in the skin, liver and kidney, the levels of metallothioneins in the skin and the liver, the expression of metallothionein mRNA in the liver, as well as the presence of histological alterations and of apoptosis in the target organs were evaluated. The 9-months exposure induced cadmium accumulation in all the tissues examined; moreover, histological changes were observed in all the tissues examined, irrespective of the dose or the time of exposure. Apoptosis was only detected in the kidney, whereas metallothioneins and metallothionein mRNA did not increase. This study demonstrates that the existing chronic water quality criterion established for cadmium induces in the newt T. carnifex cadmium accumulation and histological alterations in the target organs examined. Together with our previous results, showing that, at these same concentrations, cadmium induced endocrine disruption, the present results suggest that the existing chronic water quality criterion for cadmium appears to be not protective of amphibians.
Research Interests: Aquatic Toxicology, Apoptosis, Italy, Biological Sciences, Environmental Sciences, and 14 moreKidney, Liver, Endocrine disruptors, Animals, Male, Metallothionein, Skin, Cadmium, CHEMICAL SCIENCES, Endocrine Disruptors, Environmental Exposure, Gene Expression Regulation, Triturus, and Cadmium Chloride
Obesity and dietary fats are well known risk factors for the pathogenesis of neurodegenerative diseases. The analysis of specific markers, whose brain level can be affected by diet, might contribute to unveil the intersection between... more
Obesity and dietary fats are well known risk factors for the pathogenesis of neurodegenerative diseases. The analysis of specific markers, whose brain level can be affected by diet, might contribute to unveil the intersection between inflammation/obesity and neurodegeneration. Haptoglobin (Hpt) is an acute phase protein, which acts as antioxidant by binding free haemoglobin (Hb), thus neutralizing its pro-oxidative action. We previously demonstrated that Hpt plays critical functions in brain, modulating cholesterol trafficking in neuroblastoma cell lines, beta-amyloid (Aβ) uptake by astrocyte, and limiting Aβ toxicity on these cells. A major aim of this study was to evaluate whether a long term (12 or 24 weeks) high-fat diet (HFD) influences Hpt and Hb expression in rat hippocampus. We also assessed the development of obesity-induced inflammation by measuring hippocampal level of TNF-alpha, and the extent of protein oxidation by titrating nitro-tyrosine (N-Tyr). Hpt concentration wa...
Reverse transcriptase-mediated PCR has been used to isolate two distinct metallothionein (MT) cDNA species from RNA extracted from icefish liver, namely MT-I and MT-II. Northern blot analysis with these cDNA species revealed that... more
Reverse transcriptase-mediated PCR has been used to isolate two distinct metallothionein (MT) cDNA species from RNA extracted from icefish liver, namely MT-I and MT-II. Northern blot analysis with these cDNA species revealed that significant endogenous levels of MT mRNA were present in liver tissues of normal animals despite the fact that no MT protein could be found accumulating in the same tissue. However, multiple injections of CdCl2 induced high levels of both MT mRNA and MT protein. Sequence analysis of the cDNA species that were present after cadmium injection revealed the presence of both isoforms. Quantification of the MT-I and MT-II transcripts from normal and heavy-metal-treated fish showed an alteration in the ratio of the MT isoform transcripts. Endogenous transcripts consisted mostly of MT-II, whereas the MT-I transcript was preferentially accumulated only in response to the cadmium salt. The protein encoded by each cDNA isoform was isolated from the heavy-metal-treated...
Research Interests: Capillary electrophoresis, Biological Sciences, Sequence alignment, Liver, Animals, and 13 moreFishes, Polymerase Chain Reaction, Metallothionein, Biochemical, Cadmium, CHEMICAL SCIENCES, Molecular cloning, Amino Acids, Amino Acid Sequence, Protein Biosynthesis, Gene Expression Regulation, Molecular Sequence Data, and Medical and Health Sciences
Icefish (family Channichthyidae, suborder Nothothenioidei) are a group of Antarctic fish that have evolved unique phenotypes in order to adapt to the environment in which they live. Besides the lack of haemoglobin and the drastic... more
Icefish (family Channichthyidae, suborder Nothothenioidei) are a group of Antarctic fish that have evolved unique phenotypes in order to adapt to the environment in which they live. Besides the lack of haemoglobin and the drastic reduction in the number of erythrocyte-like cells, another striking feature of the icefish is that their liver is devoid of metallothionein. These cysteine-rich heavy-metal-binding proteins are usually present in large amounts in a large variety of organisms, from bacteria to mammals. Despite the failure to detect appreciable levels of metallothionein in icefish liver, a cDNA encoding metallothionein was produced from total RNA by reverse transcriptase PCR. The icefish metallothionein showed high percentage identity with metallothionein from Trematomus bernachii, a red-blooded Antarctic fish in which a normal content of hepatic metallothionein was found. Steady-state mRNA levels were assessed in fish liver by high-stringency hybridization of the metallothio...
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Cadmium is a persistent contaminant accumulated in the environment from both anthropogenic and natural sources. Every year, large quantities of this metal are released in the different environmental compartments and may pose a significant... more
Cadmium is a persistent contaminant accumulated in the environment from both anthropogenic and natural sources. Every year, large quantities of this metal are released in the different environmental compartments and may pose a significant threat to the biota exposed. Intracellular damage caused by cadmium exposure includes protein denaturation, lipid peroxidation, generation of reactive oxygen species and DNA strand breaks. Many studies have also demonstrated that this ion has a teratogenic or lethal effect on embryos, related to the dose and exposure time. In spite of the wide number of studies carried out in laboratory mammals, data on cadmium effects on fertility, reproduction and embryonic development of wild terrestrial vertebrates are still limited. In particular, information on the consequences of environmental cadmium exposure on reptiles survival and biodiversity are particularly scanty. Reptiles are presently considered highly susceptible to a number of environmental pollu...
In the present work, we have studied the evolutionary mechanism underlying the fate of duplicate genes in the aspartic proteinase family. Special emphasis has been given to the nothepsin gene. In fish, nothepsin is expressed exclusively... more
In the present work, we have studied the evolutionary mechanism underlying the fate of duplicate genes in the aspartic proteinase family. Special emphasis has been given to the nothepsin gene. In fish, nothepsin is expressed exclusively in female liver, whilst in lizard the expression pattern of nothepsin resembles that of an housekeeping gene, such as cathepsin D. The finding of nothepsin in a reptile is indicative of the presence of this gene in organisms other than fish. However, the failure to find any nothepsin-like gene in murine and human genome suggests that the gene has been lost in certain lineages during evolution. our results, obtained by combining adaptive evolution and functional divergence analyses, indicates that cathepsin D accumulated amino acid substitutions, whereas nothepsin retained most of the ancestral functions. We suggest a mechanism that accounts for the persistence of the two paralogous genes without invoking positive selection or functional divergence. W...
Molecular cloning and sequence determination of a novel aspartic proteinase from Antarctic fish 1 The nucleotide sequences reported in this paper have been submitted to the GenBank/EMBL/DDJB databases with accession no. AJ223836 for C. hamatus and Y08567 for T. bernacchii aspartic proteinases. 1more
In the present report, we describe a novel aspartic proteinase from the liver of two Antarctic fish species. The nucleotide sequences of the cDNA obtained from the two fishes show 90% identity with each other but only 58% identity with... more
In the present report, we describe a novel aspartic proteinase from the liver of two Antarctic fish species. The nucleotide sequences of the cDNA obtained from the two fishes show 90% identity with each other but only 58% identity with aspartic proteinases from other sources. Sequence analysis shows features for the Antarctic enzymes which are not present in related enzymes
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ABSTRACT The changes in metallothionein content during development of embryos have been studied by gel permeation chromatography of the cytosolic proteins. In unfertilized eggs, the maternal metallothionein pool is relatively high, but... more
ABSTRACT The changes in metallothionein content during development of embryos have been studied by gel permeation chromatography of the cytosolic proteins. In unfertilized eggs, the maternal metallothionein pool is relatively high, but decreases through cleavage until the late blastula stage, increasing again at the gastrula stage. metallothionein synthesis in embryos incubated with [35S]-cysteine is higher at the gastrula stage than at the earlier stages. These results suggest that the changes in metallothionein content and synthesis may be somehow correlated to the metabolic activity of the growing embryo.
... ed.) © 1999 Birkhauser Verlag Basel/Switzerland Accumulation of untranslated metallothionein mRNA in antarctic hemoglobinless fish (icefish) Vincenzo Carginale', Rosaria ... 7 Scudiero R, Capasso C, carginale V, Riggio M, Capasso... more
... ed.) © 1999 Birkhauser Verlag Basel/Switzerland Accumulation of untranslated metallothionein mRNA in antarctic hemoglobinless fish (icefish) Vincenzo Carginale', Rosaria ... 7 Scudiero R, Capasso C, carginale V, Riggio M, Capasso A, Ciaramella M, Filosa S, Parisi E (1997 ...
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Pituitary adenylate cyclase-activating peptide (PACAP) is a neuropeptide involved in multiple functions, including vertebrate reproduction. Recently, we reported the presence of PACAP in the testis of Italian wall lizard Podarcis sicula... more
Pituitary adenylate cyclase-activating peptide (PACAP) is a neuropeptide involved in multiple functions, including vertebrate reproduction. Recently, we reported the presence of PACAP in the testis of Italian wall lizard Podarcis sicula during reproductive period (May-June). Herein we investigated the PACAP mRNA expression and the localization of PACAP/PACAP receptor system, in the other periods of the Podarcis reproductive cycle, namely in summer stasis, early autumnal resumption, mid-autumnal resumption, winter stasis, and spring resumption. Using biomolecular and immunohistochemical investigations, we demonstrated that PACAP mRNA was widely expressed in all germ and somatic cells; in summer stasis (July-August) and early autumnal resumption (September) in particular, the mRNA was always found in Sertoli cells while was transiently expressed in germ and in Leydig cells. Differently from the mRNA, the protein was always present in germ and somatic cells independently from the reproductive cycle phase. As PACAP, the PAC1 receptor was always present in the testis, except for the summer stasis (July-August) and the early autumnal resumption (September), when PACAP was lacking in germ and somatic cells (Leydig and Sertoli cells). The present results strongly suggest that PACAP/PAC1 receptor system is widely represented during the reproductive cycle of male lizard. The possible involvement of PACAP/PACAP receptor system in the control of spermatogenesis is discussed.
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The possible role of metallothionein (MT) in metal homeostasis has been investigated in growing oocytes and eggs of the lizard Podarcis sicula. Chromatographic analysis does not reveal the presence of MT in both ovary and eggs, the only... more
The possible role of metallothionein (MT) in metal homeostasis has been investigated in growing oocytes and eggs of the lizard Podarcis sicula. Chromatographic analysis does not reveal the presence of MT in both ovary and eggs, the only metal-binding proteins detected being represented by high molecular mass components. De novo synthesis of MT could be observed in the ovary of cadmium-treated lizards. A cDNA encoding MT was obtained from the liver of P. sicula by RT-PCR followed by a RACE strategy, using primers designed on consensus motifs of vertebrate MT. In spite of the lack of MT in the ovary of untreated animals, Northern blot analysis demonstrates that the maternal untranslated MT transcript is expressed constitutively in the ovary in all the periods of the ovarian cycle. MT mRNA content increases during the oocyte growth, reaching the highest level in ovulated eggs, concomitantly with the accumulation of zinc and copper. Our findings suggest that maternal MT mRNA accumulates in the egg and is translated sometime during development to cope with the future needs of the growing embryo. The appearance of MT after cadmium treatment suggests that the block that makes the oocytarian MT mRNA untranslatable is removed by the metal.
Research Interests: Oogenesis, Sequence alignment, Liver, Copper, Lizards, and 14 moreFemale, Animals, Zinc, Metallothionein, Superoxide Dismutase, Molecular cloning, Amino Acid Sequence, Base Sequence, Ovary, Oocytes, Biochemistry and cell biology, Molecular Sequence Data, ovum, and Paediatrics and reproductive medicine
Organisms living in the Southern Ocean are exposed to strong environmental constraints, especially temperature. The Perciform suborder Notothenioidei, today largely endemic to the Antarctic, has developed numerous cold-adapted characters.... more
Organisms living in the Southern Ocean are exposed to strong environmental constraints, especially temperature. The Perciform suborder Notothenioidei, today largely endemic to the Antarctic, has developed numerous cold-adapted characters. The most striking peculiarity of this group of fish is the drastic reduction of hemoglobin content in their blood. This condition gives rise to a low metabolic demand of iron. Recently, we have undertaken a study to add new insights on iron metabolism in hemoglobinless fish. By taking advantage to our previous studies on transferrins from Antarctic Notothenioids, in this article we compared the evolutionary properties of these sequences to those from other fish groups and mammals. Our findings based on analysis of dN/dS ratio and on the results of the relative rate ratio test suggest that the transferrin gene has undergone positive selection in fish but not in mammals. The results of type I functional divergence provide statistical evidence for shifted evolutionary rate after speciation. Finally, when superimposed onto the three-dimensional structure of transferrin, the sites identified as responsible of the divergence in Notothenioids appear to cluster preferentially at non-iron binding locations, occupying surface locations that might affect the rate of iron binding and/or release in a cold environment.
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Iron is an essential element for metabolic processes intrinsic to life, but the properties that make iron a necessity also make it potentially deleterious. To avoid harm, iron homeostasis is achieved through iron transport, storage and... more
Iron is an essential element for metabolic processes intrinsic to life, but the properties that make iron a necessity also make it potentially deleterious. To avoid harm, iron homeostasis is achieved through iron transport, storage and regulatory proteins. The functions of some of these molecules are well described, for example transferrin and ferritin, whereas the roles of others remain unclear. The past decade has seen the identification of new molecules involved in iron metabolism, such as divalent metal transporter-1, and hepcidin. The present review aims at surveying the studies carried out on some of the most important genes involved in transport and storage of iron in Antarctic Notothenioidei, a dominating fish group endowed of a number of striking adaptive characters, including reduced (or absence of) hematocrit. This unique peculiarity among vertebrates makes this fish group a suitable system to studying the relationship between hemoglobin and iron metabolism and to understanding the adaptive changes occurred in Antarctic fish metabolism during their evolution to avoid the deleterious effects of iron overload in the absence of hemoglobin. The results summarised here indicate that the loss of hemoglobin in the most specialized group of Antarctic notothenioids, belonging to the Channychthyidae family, is accompanied by remodulation of the iron metabolism.
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Two low-molecular-weight zinc-binding proteins were purified from eggs of the sea urchin Strongylocentrotus intermedius (Echinodermata: Echinoidea) by gel-permeation and anion-exchange chromatography followed by high performance liquid... more
Two low-molecular-weight zinc-binding proteins were purified from eggs of the sea urchin Strongylocentrotus intermedius (Echinodermata: Echinoidea) by gel-permeation and anion-exchange chromatography followed by high performance liquid chromatography. The characteristics of these proteins are distinct from those of metallothionein. Amino acid compositions show a low or intermediate content of cysteine and high amounts of acidic amino acids, glycine and histidine; they
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Research Interests: Evolutionary Biology, Genetics, Molecular Evolution, Mammals, Molecular, and 15 morePhylogeny, Humans, Animals, Heavy Metal, Fishes, Detoxification, Metallothionein, Body Temperature, Amino Acids, Genetic variation, Amino Acid Sequence, Low molecular weight, Biochemistry and cell biology, Molecular Sequence Data, and Amino acid composition
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Lizards are soil surface animals that represent an important link between invertebrates and higher predators. Being part of wild fauna, they can be affected by contamination from anthropic activities and in particular, pesticides and... more
Lizards are soil surface animals that represent an important link between invertebrates and higher predators. Being part of wild fauna, they can be affected by contamination from anthropic activities and in particular, pesticides and chemical substances of various nature that reach the soil surface directly or through fall out. Among these substances, heavy metals such as cadmium may exert particularly marked toxic effect on both adult and embryos. In lizards, recent studies show that cadmium may cause developmental defects, including alteration of eye development, with appearance of unilateral microphthalmia and retinal folding. In the present study, the effects of cadmium incubation on retinal development were investigated demonstrating that cadmium interferes with cell cycle regulation by increasing proliferation. An increased expression of Otx2 and Pax6 genes, markers of retinal differentiation, was also found. However, the cellular localization of Pax6 and Otx2 transcripts did not change in treated embryos: in the early stages of retinogenesis, the two genes were expressed in all retinal cells; in the differentiated retina, Otx2 remained in the cellular bodies of retinal cells forming the nuclear and the ganglion layers, whereas Pax6 was expressed only in the cells of the inner nuclear and the ganglion layers. Data suggest that the increased expression of Pax6 and Otx2 could be ascribed to the hyperproliferation of retinal cells rather than to an effective gene overexpression.
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Starting from the knowledge that in the reproductive period the Vasoactive Intestinal Peptide (VIP) is widely distributed in Podarcis sicula testis, we studied VIP expression and the localization of the neuropeptide and its receptors in... more
Starting from the knowledge that in the reproductive period the Vasoactive Intestinal Peptide (VIP) is widely distributed in Podarcis sicula testis, we studied VIP expression and the localization of the neuropeptide and its receptors in the testis of the Italian wall lizard P. sicula in the other phases of its reproductive cycle (summer stasis, autumnal resumption, winter stasis, spring resumption). By Real Time-PCR, we demonstrated that testicular VIP mRNA levels change during the reproductive cycle, showing a cyclic trend with two peaks, one in the mid-autumnal resumption and the other in the reproductive period. By in situ hybridization and immunohistochemistry, we demonstrated that both VIP mRNA and protein were widely distributed in the testis in almost all the phases of the cycle, except in the early autumnal resumption. As regards the receptors, the VPAC1R was localized mainly in Leydig cells, while the VPAC2R showed the same distribution of VIP. Our results demonstrate that, differently from mammals, where VIP is present only in nerve fibres innerving the testis, an endotesticular synthesis takes place in the lizard and the VIP synthesis changes throughout the reproductive cycle. Moreover, the VIP/VPAC receptor system distribution observed in germ and somatic cells in various phases of the cycle, and particularly in the autumnal resumption and the reproductive period, strongly suggests its involvement in both spermatogenesis and steroidogenesis. Finally, the wider distribution of VIP in lizards with respect to mammals leads us to hypothesize that during the evolution the synthesis sites have been transferred from the testis to other districts, such as the brain.
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Research Interests: Genetics, Functional Analysis, Gene regulation, DNA, Protein Structure and Function, and 15 moreHumans, Sequence alignment, Animals, Fishes, Metallothionein, Gene, Transcription Factor, Cadmium, Metals, Molecular cloning, Protein isoforms, Base Sequence, Gene Expression Regulation, Molecular Sequence Data, and Reporter gene
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Research Interests: Genetics, Molecular Evolution, PCR, Phylogeny, Reptiles, and 15 moreAnimals, Metallothionein, Phylogenetic analysis, Evolutionary History, Molecular Phylogenetics and Evolution, Gene, Gene Duplication, Bp, Molecular cloning, cDNA, Protein isoforms, Amino Acid Sequence, Base Sequence, UTR, and Molecular Sequence Data
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Full-length zebrafish cDNAs encoding two aspartic proteinases were cloned and sequenced. One of the two cDNAs was a 1708 bp product with an open reading frame of 398 amino acid residues corresponding to a cathepsin D. The other was a 1383... more
Full-length zebrafish cDNAs encoding two aspartic proteinases were cloned and sequenced. One of the two cDNAs was a 1708 bp product with an open reading frame of 398 amino acid residues corresponding to a cathepsin D. The other was a 1383 bp product encoding a polypeptide chain of 416 amino acids homologous to nothepsin, an aspartic proteinase first identified by us in the liver of Antarctic Notothenioidei. Gene expression assessed by RT-PCR and northern blot hybridization of RNA from different tissues showed that the expression was tissue- and sex-specific. Whereas the cathepsin D gene was expressed in all the tissues examined independently of the sex, the nothepsin gene was expressed exclusively in female livers.
Research Interests: Genetics, RNA, Gene expression, Danio rerio, Phylogeny, and 15 morecathepsin D, Female, Animals, Male, Gene, ribosomal RNA, Sex Factors, Amino Acid Sequence, Base Sequence, Open Reading Frame, Reverse Transcriptase, Tissue Specificity, Molecular Sequence Data, Sodium Dodecyl Sulfate, and Tissue distribution
Research Interests: Genetics, Molecular Biology, Gene expression, Ultraviolet, Animals, and 11 moreGene, Cadmium, Steady state, Perciformes, Heat Shock Protein, Differential Display, Reverse Transcriptase, Gene Expression Regulation, Sodium Dodecyl Sulfate, reverse transcriptase polymerase chain reaction, and Reverse transcription polymerase chain reaction
Research Interests: Genetics, Molecular Evolution, Relative Growth Rate, Phylogeny, Humans, and 14 moreMaximum Likelihood, Animals, Molecular Phylogenetics and Evolution, Gene, Gene Duplication, Gene Conversion, Gene Families, Positive Selection, Evolutionary rate, Adaptive evolution, Genetic variation, SPR, Gene Family, and Nucleotide sequence
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Research Interests: Environmental Toxicology, Reproduction, Apoptosis, Oogenesis, Biological Sciences, and 15 moreEnvironmental Sciences, Lizards, Female, Animals, Metallothionein, CHEMICAL SCIENCES, Ovarian Follicle, Environmental Pollutants, Cell Proliferation, Ovary, Somatic Cell Count, Zona Pellucida, Environmental, Cell count, and Cadmium Chloride
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Research Interests: Gene expression, Biological Sciences, Phylogeny, Sequence alignment, Animals, and 13 moreMolecular Phylogenetics and Evolution, Cichlids, Perciformes, Salmo salar, Zebrafish, Protein isoforms, Species Specificity, Amino Acid Sequence, Base Sequence, Gene Expression Regulation, Molecular Sequence Data, Sequence homology, and conserved sequence
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Research Interests: Oogenesis, Danio rerio, Biological Sciences, Environmental Sciences, Copper, and 15 moreComparative Biochemistry, Female, Animals, Male, DNA methylation, Early development, Metallothionein, Cadmium, Oocyte Maturation, Embryonic Development, Zebrafish, Oocytes, Embryos, Embryogenesis, and Medical and Health Sciences
This study examined the cytological and molecular effects of cadmium, a toxic heavy metal, in the liver of the Italian wall lizard Podarcis sicula. Cadmium was administered in single dose, by diet, to induce a concentration comparable... more
This study examined the cytological and molecular effects of cadmium, a toxic heavy metal, in the liver of the Italian wall lizard Podarcis sicula. Cadmium was administered in single dose, by diet, to induce a concentration comparable with that measured in animals living in contaminated sites. For comparison, cadmium was also administered in multiple doses by food (chronic) or in a single dose intraperitoneally (i.p.); the effects were followed at regular time intervals up to 30 days post treatments. Atomic absorption spectrometry analysis demonstrated cadmium ion uptake and accumulation in the parenchyma with an estimated half-life of approximately 8 days. Cytological analyses revealed that the metal induced oedema, activated metallothionein expression in Kupffer cells and extracellular matrix production in fat storing cells. It also caused swelling and alteration in lipid and sugar metabolism in hepatocytes. In conclusion, in the wall lizard cadmium is toxic to the liver even at very low concentrations, the response is not strictly dose and time dependent and almost no recovery occurs in short (30 days) time periods.
Research Interests: Immunohistochemistry, Transmission Electron Microscopy, Apoptosis, Gene expression, Extracellular Matrix, and 15 moreIn Situ Hybridization, Biological Sciences, Environmental Sciences, Liver, Lizards, Animals, Heavy Metal, Metallothionein, Atomic Absorption Spectrometry, Cadmium, Time Dependent, Hepatocytes, Cell Proliferation, Kupffer cells, and Medical and Health Sciences
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The present report is an attempt to investigate the influence of intraperitoneal and dietary cadmium exposure on the distribution of cadmium accumulation and induction of metallothionein gene expression in different tissues of the lizard... more
The present report is an attempt to investigate the influence of intraperitoneal and dietary cadmium exposure on the distribution of cadmium accumulation and induction of metallothionein gene expression in different tissues of the lizard Podarcis sicula. Cadmium accumulation in liver, kidney, ovary, brain and intestine was measured by atomic absorption spectrometry. Metallothionein gene induction was determined by dot blot analyses on the total RNA extracted from the same organs. Our data indicate that cadmium exposure results in significant cadmium uptake, but the patterns of this uptake varies with organ and exposure route. After a single intraperitoneal treatment, concentrations of cadmium and metallothionein transcript are positively correlated in kidney, liver and ovary. Following a dietary cadmium treatment, a positive correlation between the increase of metallothionein mRNA and cadmium accumulation is found in intestine, ovary and kidney, while no correlation is present in liver and brain.
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Screening for differentially expressed genes is a straightforward approach to study the molecular basis of contaminant toxicity. In this paper, the mRNA differential display technique was applied to analyze transcriptional regulation in... more
Screening for differentially expressed genes is a straightforward approach to study the molecular basis of contaminant toxicity. In this paper, the mRNA differential display technique was applied to analyze transcriptional regulation in response to cadmium exposure in the lizard embryos. Lizard eggs may be particularly susceptible to soil contamination and in ovo exposure may interfere or disrupt normal physiological function in the developing embryo, including regulation of gene expression. Fertilized eggs of the lizard Podarcis sicula were incubated in cadmium-contaminated soil at 25 degrees C for 20 days. Gene expression profiling showed 5 down- and 9 up-regulated genes. Four cDNAs had no homology to known gene sequences, thus suggesting that may either encode not yet identified proteins, or correspond to untranslated regions of mRNA molecules. Four fragments exhibited significant sequence similarity with genes encoding novel proteins or ESTs derived from other vertebrates. The remaining genes are mainly involved in molecular pathways associated with processes such as membrane trafficking, signal transduction, cytoskeletal organization, cell proliferation and differentiation. Cadmium also affected the expression of factors actively involved in the regulation of the transcription machinery. Down-regulated genes are mainly associated with cellular metabolism and cell-cycle regulation and apoptosis. All of these differentially expressed genes may represent candidates that function in cadmium responses. The present study leads to an increased understanding of genes and/or the biochemical pathways involved in perturbation of embryo development following cadmium exposure.
Research Interests: Membrane Trafficking, Gene expression, Biological Sciences, Environmental Sciences, Regulation of Gene Expression, and 15 moreLizards, Animals, Gene Regulatory Networks, Contaminated soil, Cell Cycle regulation, Reproducibility of Results, Cell Proliferation, Differential Display, Embryos, Differential Gene Expression, Gene expression profiling, Embryo Development, reverse transcriptase polymerase chain reaction, Medical and Health Sciences, and Cadmium Chloride
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Metallothionein levels were determined in the eggs of two sea urchin species, the Mediterranean Sphaerechinus granularis and the Antarctic Sterechinus neumayeri. While appreciable levels of metallothionein were found in S. granularis... more
Metallothionein levels were determined in the eggs of two sea urchin species, the Mediterranean Sphaerechinus granularis and the Antarctic Sterechinus neumayeri. While appreciable levels of metallothionein were found in S. granularis eggs, a negligible amount was detected in S. neumayeri. Two metallothionein isoforms were purified from S. granularis, and metallothionein cDNAs were obtained by means of reverse transcriptase-polymerase chain reaction
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Copper (Cu) is an essential element required in many biological processes including cellular growth and development. The molecular mechanisms involved in copper homeostasis include proteins that play a role in Cu uptake. Genes encoding... more
Copper (Cu) is an essential element required in many biological processes including cellular growth and development. The molecular mechanisms involved in copper homeostasis include proteins that play a role in Cu uptake. Genes encoding high affinity copper transporters (Ctr) have been identified in yeast, plant and mammalian cells. Analysis of copper and zinc content in growing ovarian follicles and ovulated eggs of the reptilian Podarcis sicula demonstrated that the levels of both metals rise during oocyte growth, reaching the maximum in ovulated eggs. By exploiting the remarkable evolutionary conservation of the primary structure of Ctr proteins, cDNA encoding a Ctr was isolated from the liver of the lizard P. sicula by reverse transcriptase PCR and RACE strategy by using primers designed based on consensus motifs present in mammalian Ctr. The predicted protein sequence contains three transmembrane domains and a putative hydrophilic extracellular amino-terminal domain. Besides complementing the respiratory deficiency of yeast cells defective in high affinity Cu transport, expression of lizard Ctr1(1) in Hek293 cells stimulates Cu uptake.Gene expression assessed by Northern blot hybridization of RNA from different tissues of P. sicula shows the highest levels of transcript in both intestine and liver. The profile of Ctr1 mRNA in growing ovarian follicles and eggs demonstrates that the transcript accumulates during the oocyte growth and reaches the highest levels in ovulated eggs. These results suggest that lizard Ctr1 protein may function in Cu acquisition in growing oocytes and eggs.
Research Interests: Genetics, Gene expression, Oogenesis, Molecular Mechanics, Biological Sciences, and 15 moreCell line, Humans, Copper, Lizards, Animals, Growth and development, Physical sciences, Membrane transport proteins, Molecular cloning, Biological Process, Amino Acid Sequence, Base Sequence, Oocytes, Molecular Sequence Data, and ovum
Research Interests: Italy, Biological Sciences, Sequence alignment, Liver, cathepsin D, and 15 moreAnimals, Fishes, Physical sciences, Temperature, High Temperature, Affinity chromatography, Enzyme, Molecular cloning, Molecular Mass, Amino Acid Sequence, Base Sequence, Open Reading Frame, Kinetic Parameter, Molecular Sequence Data, and reverse transcriptase polymerase chain reaction
ABSTRACT The concentration of cadmium and lead ions was determined in digestive gland and mantle of mussels (Mytilus galloprovincialis) from five aquaculture stations located in the Gulf of Naples (Southern Italy, Tyrrhenian Sea).... more
ABSTRACT The concentration of cadmium and lead ions was determined in digestive gland and mantle of mussels (Mytilus galloprovincialis) from five aquaculture stations located in the Gulf of Naples (Southern Italy, Tyrrhenian Sea). Metallothionein (MT) levels in the same tissues were also evaluated. This gulf represents a greatly urbanized area, characterized by an important commercial port and past industrial activities, terminated in the last decade of the 20th century. The results were compared with those obtained from mussels cultivated in a more pristine neighbouring area. Data demonstrate that the amount of both metals found in mussels harvested at the highly anthropogenic sites in the Gulf of Naples was comparable with that found in mussels from the more naturalistic site. The content of both cadmium and lead was not related with that of MT. Indeed, the amount of both metals in the digestive gland was greater than in the mantle, whereas the MT preferentially accumulated in the mantle. Possibly, the MT content in mantle was associated to the physiological function of the tissue, rather than heavy metals exposure.