RESUMO
BACKGROUND: The etiology of the metabolic syndrome is complex, and is determined by the interplay of both genetic and environmental factors. The present study was designed to identify genes and proteins in the adipose tissues with altered expression in the spontaneously hypertensive/NIH -corpulent rat, SHR/NDmcr-cp (CP) and to find possible molecular targets associated with the pathogenesis or progression of obesity related to the metabolic syndrome. METHODS: We extracted RNAs and proteins from the epididymal adipose tissues in CP, SHR/Lean (Lean), and Wistar Kyoto (WKY) rats and performed microarray analysis and two-dimensional difference in gel electrophoresis (2D-DIGE) linked to a matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS). RESULTS: The results showed different mRNA and protein expression levels in the adipose tissue: oligo DNA microarray identified 33 genes that were significantly (P < 0.01) up-regulated and 17 genes significantly down-regulated in CP compared with WKY and Lean rats at both 6 and 25 weeks of age. The affected genes-proteins were associated with lipolytic enzymes stimulated by peroxisome proliferator-activated receptor (PPAR) signaling. Further analysis using the 2D-DIGE connected with MALDI-TOF/TOF analysis, the expression of monoglyceride lipase (MGLL) was significantly up-regulated and that of carboxylesterase 3 (CES3) was significantly down-regulated in 6- and 25-week-old CP compared with age-matched control (WKY and Lean rats). CONCLUSIONS: Our results suggest the possible involvement of proteins associated with adipocyte lipolysis in obesity related to the metabolic syndrome.
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BACKGROUND: It is difficult to study the mechanisms of the metabolic syndrome in humans due to the heterogeneous genetic background and lifestyle. The present study investigated changes in the gene and protein profiles in an animal model of the metabolic syndrome to identify the molecular targets associated with the pathogenesis and progression of obesity related to the metabolic syndrome. METHODS: We extracted mRNAs and proteins from the liver tissues of 6- and 25-week-old spontaneously hypertensive/NIH -corpulent rat SHR/NDmcr-cp (CP), SHR/Lean (Lean) and Wistar Kyoto rats (WKY) and performed microarray analysis and two-dimensional difference in gel electrophoresis (2D-DIGE) linked to a matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS). RESULTS: The microarray analysis identified 25 significantly up-regulated genes (P < 0.01; log10 > 1) and 31 significantly down-regulated genes (P < 0.01; log10 < -1) in 6- and 25-week-old CP compared with WKY and Lean. Several of these genes are known to be involved in important biological processes such as electron transporter activity, electron transport, lipid metabolism, ion transport, transferase, and ion channel activity. MALDI-TOF/TOF MS identified 31 proteins with ±1.2 fold change (P < 0.05) in 6- and 25-week-old CP, compared with age-matched WKY and Lean. The up-regulated proteins are involved in metabolic processes, biological regulation, catalytic activity, and binding, while the down-regulated proteins are involved in endoplasmic reticulum stress-related unfolded protein response. CONCLUSION: Genes with significant changes in their expression in transcriptomic analysis matched very few of the proteins identified in proteomics analysis. However, annotated functional classifications might provide an important reference resource to understand the pathogenesis of obesity associated with the metabolic syndrome.
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Zinc pyrithione has been used in anti-dandruff shampoos and in anti-fouling paint on ships. However, little is known of its mode of action. We characterized the effects of sub-lethal concentrations of zinc pyrithione (Zpt) on Saccharomyces cerevisiae using DNA microarrays. The majority of the strongly upregulated genes are related to iron transport, and many of the strongly downregulated genes are related to the biosynthesis of cytochrome (heme). These data suggest that Zpt induces severe iron starvation. To confirm the DNA microarray data, we supplemented cultures containing Zpt with iron, and the growth of the yeast was restored significantly. From these results, we propose that the principal toxicity of zinc pyrithione arises from iron starvation.
Assuntos
Perfilação da Expressão Gênica/métodos , Ferro/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Compostos Organometálicos/administração & dosagem , Piridinas/administração & dosagem , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/fisiologia , Antifúngicos/administração & dosagem , Relação Dose-Resposta a Droga , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Regulação Fúngica da Expressão Gênica/fisiologia , Saccharomyces cerevisiae/efeitos dos fármacosRESUMO
To assess the toxicity of the C1 compounds methanol and formaldehyde, gene expression profiles of treated baker's yeast were analyzed using DNA microarrays. Among approximately 6,000 open reading frames (ORFs), 314 were repressed and 375 were induced in response to methanol. The gene process category "energy" comprised the greatest number of induced genes while "protein synthesis" comprised the greatest number of repressed genes. Products of genes induced by methanol were mainly integral membrane proteins or were localized to the plasma membrane. A total of 622 and 610 ORFs were induced or repressed by formaldehyde, respectively. More than one-third of the genes found to be strongly repressed by formaldehyde belonged to the "protein synthesis" functional category. Conversely, genes in the subcategory of "nitrogen, sulfur, and selenium metabolism" within "metabolism" and in the category of "cell rescue, defense, and virulence" were up-regulated by exposure to formaldehyde. Our data suggest that membrane structure is a major target of methanol toxicity, while proteins were major targets of formaldehyde toxicity.
Assuntos
Formaldeído/toxicidade , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Metanol/toxicidade , Análise de Sequência com Séries de Oligonucleotídeos , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Testes de Toxicidade , Análise por Conglomerados , Reparo do DNA/efeitos dos fármacos , Reparo do DNA/genética , Perfilação da Expressão Gênica , Inativação Metabólica/genética , Fases de Leitura Aberta/genética , Saccharomyces cerevisiae/citologia , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMO
The fish acute toxicity test in the Organisation for Economic CoOperation and Development "OECD Guidelines for Testing of Chemicals" is an essential test for environmental toxicity. Here, we have tried to evaluate the physiology of medaka during these test procedures by using genomics. Genomics technology can provide genome-wide expression profiles of mRNA, and these profiles correspond to the physiology of organisms. Thus, a comparison of mRNA expression profiles gives information on the reproducibility of experimental conditions. Expression profiles of mRNA were measured for medakas maintained within the allowable range of the test conditions and also under extreme conditions beyond the guideline limits. We confirmed the high physiological reproducibility of medaka kept in the recommended conditions of the fish acute toxicity test in the "OECD Guidelines for Testing of Chemicals" from the expression profiles obtained under all experimental conditions, except for the types of feeding.
Assuntos
Poluentes Ambientais/toxicidade , Perfilação da Expressão Gênica , Modelos Animais , Oryzias/genética , Oryzias/fisiologia , Testes de Toxicidade , Animais , Comportamento Alimentar/efeitos dos fármacos , Regulação da Expressão Gênica , Guias como Assunto , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Oxigênio , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Padrões de Referência , Reprodutibilidade dos Testes , Solubilidade/efeitos dos fármacos , Testes de Toxicidade AgudaRESUMO
The effect of the heavy metal copper on the expression of a wide spectrum of genes was analyzed by using a DNA microarray. The gene expression profile of baker's yeast Saccharomyces cerevisiae grown in a medium containing a sublethal concentration of cupric sulfate was compared with that of yeast grown in a normal medium. Among approximately 6000 yeast ORFs, 143 ORFs were induced more than twofold to resist copper toxicity after exposure to copper. Copper metallothionein CUP1-1 and CUP1-2 were induced more than 20-fold. Some genes related to sulfur metabolism and oxidative stress response were also up-regulated. This DNA microarray analysis identified several molecular targets of copper toxicity.
Assuntos
Sulfato de Cobre/toxicidade , Cobre/toxicidade , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Saccharomyces cerevisiae/efeitos dos fármacos , Proteínas de Transporte/genética , Proteínas de Ligação a DNA/genética , Perfilação da Expressão Gênica , Metalotioneína , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Oxidativo/genética , RNA Mensageiro/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Enxofre/metabolismo , Fatores de Transcrição/genéticaRESUMO
T-2 toxin is a mycotoxin that belongs to a group of type A tricothecenes found in agricultural products. The cytotoxicity of T-2 toxin was characterized by analysis of the yeast transcriptome upon challenge with T-2 toxin. Interestingly, T-2 toxin-induced yeast gene expression profiles were found to be similar to profiles obtained following cycloheximide treatment. Moreover, T-2 toxin treatment was found to activate facilitators, gluconeogenesis and cell arrest related genes such as mitogen-activated protein kinase genes (FUS3). T-2 toxin attacks the membrane and as a result the membrane transport system was disturbed. A large number of genes are induced to restore the toxicity caused by T-2 toxin. However, the data did not suggest that DNA damage by alkylation (Mag1, a gene 3-methyl-adenine DNA glycosylase, 0.46-fold down regulated), no induction of DNA repair mechanisms such as recombination (RAD26, RAD52 and etc.) and excision repair (RAD7, RAD14, RAD16, RAD23 and etc.). These results suggested that the toxicity of the T-2 toxin was due to the disturbance of the cell membrane of the yeast cell and that T-2 toxin caused mild mutagenesis.
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A bioassay is a system for monitoring toxicity of chemicals in the environment via the biological responses of experimental organisms. These responses can be detected by analysis of genome-wide changes in mRNA expression levels using DNA microarrays. We applied this system for evaluation of synergistic toxicity by cadmium and thiuram, as this combination showed mutual growth inhibition in yeast. Hierarchical cluster analysis using mRNA expression profiles suggested the response of yeast to this combination is similar to that seen with cadmium treatment alone. Functional characterization of genes induced by this combinational treatment also suggests that the toxicity of cadmium was enhanced. This toxicity was observed as the damage to mitochondrial functions, which were not observed with either cadmium or thiuram treatments alone. The potential toxicity to mitochondria by this combinational treatment was confirmed as the result of deletion of mitochondria with petite colony formation frequency. We could evaluate synergistic toxicity by cadmium and thiuram and show the possible use of transcriptome bioassay for synergistic toxicity.
Assuntos
Cádmio/análise , Perfilação da Expressão Gênica , Mitocôndrias/metabolismo , Mutação , Tiram/análise , Bioensaio , Cádmio/química , Análise por Conglomerados , DNA Mitocondrial/química , Relação Dose-Resposta a Droga , Análise de Sequência com Séries de Oligonucleotídeos , Fases de Leitura Aberta , Filogenia , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saccharomyces cerevisiae/genéticaRESUMO
BACKGROUND: Mycotoxins are fungal secondary metabolites commonly present in feed and food, and are widely regarded as hazardous contaminants. Citrinin, one of the very well known mycotoxins that was first isolated from Penicillium citrinum, is produced by more than 10 kinds of fungi, and is possibly spread all over the world. However, the information on the action mechanism of the toxin is limited. Thus, we investigated the citrinin-induced genomic response for evaluating its toxicity. RESULTS: Citrinin inhibited growth of yeast cells at a concentration higher than 100 ppm. We monitored the citrinin-induced mRNA expression profiles in yeast using the ORF DNA microarray and Oligo DNA microarray, and the expression profiles were compared with those of the other stress-inducing agents. Results obtained from both microarray experiments clustered together, but were different from those of the mycotoxin patulin. The oxidative stress response genes--AADs, FLR1, OYE3, GRE2, and MET17--were significantly induced. In the functional category, expression of genes involved in "metabolism", "cell rescue, defense and virulence", and "energy" were significantly activated. In the category of "metabolism", genes involved in the glutathione synthesis pathway were activated, and in the category of "cell rescue, defense and virulence", the ABC transporter genes were induced. To alleviate the induced stress, these cells might pump out the citrinin after modification with glutathione. While, the citrinin treatment did not induce the genes involved in the DNA repair. CONCLUSION: Results from both microarray studies suggest that citrinin treatment induced oxidative stress in yeast cells. The genotoxicity was less severe than the patulin, suggesting that citrinin is less toxic than patulin. The reproducibility of the expression profiles was much better with the Oligo DNA microarray. However, the Oligo DNA microarray did not completely overcome cross hybridization.
Assuntos
Citrinina/toxicidade , Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Oligonucleotídeos , Fases de Leitura Aberta , Saccharomyces cerevisiae/efeitos dos fármacos , Testes de Toxicidade/métodos , Análise por Conglomerados , Genes Fúngicos , Glutationa/metabolismo , Redes e Vias Metabólicas/efeitos dos fármacos , Redes e Vias Metabólicas/genética , Metionina/metabolismo , Modelos Biológicos , Oligonucleotídeos/análise , Patulina/toxicidade , Saccharomyces cerevisiae/genéticaRESUMO
Throughout Central and South Kalimantan, Indonesia, strongly acidic soil (pH 2.1-3.7) is widely distributed, and the local acidic soil-tolerant plants, including local rice varieties, often possess sphingomonads in their rhizosphere and rhizoplane. To investigate the behavior of sphingomonads inhabiting the rhizosphere of such acid-tolerant plants, we designed 13 different DNA array probes (each of 72 mer) specific to a group of sphingomonads, using a hypervariable V6 region of the 16S rRNA gene. This DNA array system was used preliminarily for an analysis of microfloral dynamisms, particularly of sphingomonads, in acidic paddock ecosystems, and the results suggest that the acid-tolerant local rice shares rhizospherous sphingomonads with wild Juncus sp., a predominant weed that thrives in acidic paddocks during the off-season for rice farming. This tentative conclusion supports the bio-rationality of the traditional rice farming system with respect to functional rhizobacteria.
Assuntos
Sondas de DNA/química , Microbiologia do Solo , Sphingomonas/genética , Burkholderia cepacia/química , Burkholderia cepacia/genética , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Concentração de Íons de Hidrogênio , Indonésia , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos , Oryza/microbiologia , RNA Ribossômico 16S/genética , Solo/análise , Sulfatos/químicaAssuntos
Enterotoxinas/genética , Intoxicação Alimentar Estafilocócica/microbiologia , Staphylococcus aureus/genética , Surtos de Doenças , Eletroforese em Gel de Campo Pulsado/métodos , Enterotoxinas/deficiência , Humanos , Japão/epidemiologia , Reação em Cadeia da Polimerase/métodos , Intoxicação Alimentar Estafilocócica/epidemiologia , Staphylococcus aureus/isolamento & purificaçãoRESUMO
Gene profiling of Japanese medaka (Oryzias latipes) was performed using an oligonucleotide DNA microarray representing 22,587 TIGR O. latipes gene indices (OLGIs). The average correlation coefficients for gene expression between individual mature fish were high (>0.95) for both female and male, indicating that the physiological status of medaka is highly reproducible under prescribed growth conditions. Of the 22,587 OLGIs, 2575 showed significant differences in expression between female and male. Exposure to 17beta-estradiol (E2) revealed 381 E2-responsive OLGIs in male medaka. Feminization and male-dysfunction factors of the E2-treated males calculated using the combination of Pearson correlation coefficient and Euclidean distances indicate that E2 treatment "weakly feminized" male medaka, while male physiological functions were not significantly disrupted. This study demonstrates the possibility of using medaka microarrays to estimate the overall effects of hormonally active chemicals.
Assuntos
Estradiol/farmacologia , Proteínas de Peixes/genética , Perfilação da Expressão Gênica , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Oryzias/genética , Animais , Feminino , Feminização/induzido quimicamente , Feminização/metabolismo , Proteínas de Peixes/metabolismo , Expressão Gênica/efeitos dos fármacos , Masculino , Oryzias/fisiologia , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores SexuaisRESUMO
Patulin, 4-hydroxy-4H-furo[3,2c]pyran-2(6H)-one, is one of the best characterized and most widely disseminated mycotoxins found in agricultural products. Nonetheless, the mechanisms by which patulin causes toxicity are not well understood. Thus, the cytotoxicity of patulin was characterized by analysis of the yeast transcriptome upon challenge with patulin. Interestingly, patulin-induced yeast gene expression profiles were found to be similar to gene expression patterns obtained after treatment with the antifungal agricultural chemicals thiuram, maneb, and zineb. Moreover, patulin treatment was found to activate protein degradation, especially proteasome activities, sulfur amino acid metabolism, and the defense system for oxidative stress. Damage to DNA by alkylation was also suggested, and this seemed to be repaired by recombinational and excision repair mechanisms. Furthermore, the results provide potential biomarker genes for the detection of patulin in agricultural products. The results suggest the possibility of applying the yeast transcriptome system for the evaluation of chemicals, especially for natural chemicals that are difficult to get by organic synthesis.
Assuntos
Patulina/toxicidade , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/genética , Aminoácidos Sulfúricos/metabolismo , Proteínas Fúngicas/metabolismo , Expressão Gênica/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Oxidativo , Saccharomyces cerevisiae/efeitos dos fármacosRESUMO
Adaptation to temperature fluctuation is essential for the survival of all living organisms. Although extensive research has been done on heat and cold shock responses, there have been no reports on global responses to cold shock below 10 degrees C or near-freezing. We examined the genome-wide expression in Saccharomyces cerevisiae, following exposure to 4 degrees C. Hierarchical cluster analysis showed that the gene expression profile following 4 degrees C exposure from 6 to 48 h was different from that at continuous 4 degrees C culture. Under 4 degrees C exposure, the genes involved in trehalose and glycogen synthesis were induced, suggesting that biosynthesis and accumulation of those reserve carbohydrates might be necessary for cold tolerance and energy preservation. The observed increased expression of phospholipids, mannoproteins, and cold shock proteins (e.g., TIP1) is consistent with membrane maintenance and increased permeability of the cell wall at 4 degrees C. The induction of heat shock proteins and glutathione at 4 degrees C may be required for revitalization of enzyme activity, and for detoxification of active oxygen species, respectively. The genes with these functions may provide the ability of cold tolerance and adaptation to yeast cells.
Assuntos
Temperatura Baixa , Regulação Fúngica da Expressão Gênica , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/fisiologia , Adaptação Fisiológica , Regulação para Baixo , Família Multigênica , Análise de Sequência com Séries de Oligonucleotídeos , Saccharomyces cerevisiae/crescimento & desenvolvimento , Regulação para CimaRESUMO
It is well documented that iodine kills microorganisms with a broad spectrum, but a systematic study of its mechanism of action has not yet been reported. Here we found the action of iodine on gene expression level, using the yeast Saccharomyces cerevisiae with a DNA microarray. It was found that, like antimicrobial activity, iodine causes an immediate and dose-dependent (0.5 mM, 0.75 mM and 1 mM) transcriptional alteration in yeast cells. The effects of iodine continued after the first immediate response. Genes for c-compound and carbohydrate metabolism, for energy, and for cell rescue were continuously up-regulated. On the other hand, genes related to protein fate were induced especially at 0.5 h. The gene expression profile at 0.5 h was significantly different from that of a longer iodine exposed condition. The main reaction at 0.5 h after iodine addition might be due to oxidative toxicity, and the profile at 0.5 h was similar to that of an agricultural bactericide.
Assuntos
Regulação Fúngica da Expressão Gênica/efeitos dos fármacos , Iodo/farmacologia , Saccharomyces cerevisiae/genética , Metabolismo dos Carboidratos/fisiologia , Análise por Conglomerados , Meios de Cultura , Primers do DNA , DNA Fúngico/biossíntese , DNA Fúngico/genética , Metabolismo Energético/efeitos dos fármacos , Genes Fúngicos/genética , Família Multigênica/genética , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saccharomyces cerevisiae/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacosRESUMO
Genome-wide mRNA expression profiles of Saccharomyces cerevisiae growing under hydrostatic pressure were characterized. We selected a hydrostatic pressure of 30 MPa at 25 degrees C because yeast cells were able to grow under these conditions, while cell size and complexity were increased after decompression. Functional characterization of pressure-induced genes suggests that genes involved in protein metabolism and membrane metabolism were induced. The response to 30 MPa was significantly different from that observed under lethal conditions because protein degradation was not activated under 30 MPa pressure. Strongly induced genes those that contribute to membrane metabolism and which are also induced by detergents, oils, and membrane stabilizers.
Assuntos
Adaptação Fisiológica , Saccharomyces cerevisiae/fisiologia , Sequência de Bases , Primers do DNA , Citometria de Fluxo , Perfilação da Expressão Gênica , Genes Fúngicos , Pressão , RNA Fúngico/genética , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimentoRESUMO
Plant-derived essential oils with monoterpenoids have been used as antifungal drugs since ancient times, but the mode of action of these natural hydrocarbons at the molecular level is not understood. In order to understand the mechanisms of toxicity of alpha-terpinene (a cyclic monoterpene), a culture of Saccharomyces cerevisiae was exposed to 0.02% alpha-terpinene for 2 h and transcript profiles were obtained using yeast DNA arrays. These profiles, when compared with transcript profiles of untreated cultures, revealed that the expression of 793 genes was affected. For 435 genes, mRNA levels in treated cells compared with control cells differed by more than two-fold, whereas for 358 genes, it was <0.5-fold. Northern blots were performed for selected genes to verify the microarray results. Functional analysis of the up-regulated genes indicates that, similar to commonly used antifungal drugs, alpha-terpinene exposure affected genes involved in ergosterol biosynthesis and sterol uptake. In addition, transcriptional induction of genes related to lipid metabolism, cell wall structure and function, detoxification and cellular transport was observed in response to terpinene toxicity. Notably, the functions of 192 up-regulated genes are still unknown, but their characterization will probably shed light on the mechanisms of drug resistance and sensitivity. Taken together, this study showed that alpha-terpinene has strong antifungal activities and its modes of action resemble those of presently used antifungal drugs.
Assuntos
Antifúngicos/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Terpenos/farmacologia , Northern Blotting , Parede Celular/genética , Parede Celular/fisiologia , Sondas de DNA , DNA Complementar/biossíntese , DNA Fúngico/biossíntese , Ergosterol/farmacologia , Testes de Sensibilidade Microbiana , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos , Fosfolipídeos/biossíntese , RNA Fúngico/biossíntese , RNA Mensageiro/biossíntese , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMO
Sodium n-dodecyl benzene sulfonate (LAS) and sodium dodecyl sulfate (SDS) are popular anionic detergents (surfactants) that are used worldwide and the toxicities of these chemicals have been characterized. We applied these chemicals in a DNA microarray bioassay and determined that the microarray data reflects previous findings and also provides some new information about anionic detergent toxicity. The mRNA expression profiles suggest that LAS and SDS cause damage to membranes and alterations in carbon metabolism, and induce the oxidative stress response. We also found that LAS and SDS induce the pleiotropic drug-resistance network, and that LAS and SDS may be pumped out of yeast cells by this network. Hierarchical clustering of the expression profiles showed that LAS and SDS cause similar features of toxicity and that the toxicity is similar to that of capsaicin but different from that of cadmium and mercury.
Assuntos
Benzenossulfonatos/toxicidade , Análise de Sequência com Séries de Oligonucleotídeos , Dodecilsulfato de Sódio/toxicidade , Tensoativos/toxicidade , Poluentes Químicos da Água/toxicidade , Bioensaio/métodos , Perfilação da Expressão Gênica , RNA Mensageiro/biossíntese , Saccharomyces cerevisiae/genéticaRESUMO
The herbicide Roundup, which contains glyphosate as the active ingredient, was first introduced in 1974 and has enjoyed widespread use in Japan and elsewhere in the world. Roundup-induced reactions occurring in the yeast Saccharomyces cerevisiae may have a predictive value for understanding responses in higher eukaryotes, and we applied yeast DNA microarray analysis for this purpose. Functional characterization of up-regulated open reading frames (ORFs) following Roundup treatment suggests that Roundup affects membrane structures and cellular organelles. Expression profiles induced by treatments with detergents, oils and hydrostatic pressure were similar to those following Roundup treatment based on cluster analysis. Glyphosate alone was not found to inhibit yeast growth at the concentration contained in the Roundup treatment used for microarray analysis. The toxicity of Roundup appeared to be due to detergent in the product.
Assuntos
DNA Fúngico/genética , Glicina/análogos & derivados , Glicina/toxicidade , Herbicidas/toxicidade , Polietilenoglicóis/toxicidade , Saccharomyces cerevisiae/efeitos dos fármacos , Dano ao DNA , Combinação de Medicamentos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Concentração Inibidora 50 , Análise de Sequência com Séries de Oligonucleotídeos , Fases de Leitura Aberta/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , GlifosatoRESUMO
DNA microarrays are becoming increasingly popular in toxicogenomic studies. It is common knowledge that multiple repeats of microarray experiments have to be performed with multiple samples to get reliable data, due to experimental variations (both technical and biological) contained in microarray experiments. However, the use of multiple samples and replicate experiments is not practical in the field of environmental toxicology, since environmental samples are limited and microarray experiments are expensive. Thus it is desirable to obtain reliable data from a minimum number of repeats of microarray experiments. To further establish and extend gene expression profiling for toxicogenomics using microarrays, it is necessary to establish an analytical method of microarrays for toxicogenomics. In this study, we attempted to construct a "Data Dependent Analysis (DDA)" yeast cDNA microarray experiment from 100 microarray datasets in order to get reliable data from one microarray experiment.