RESUMO
Organotin compounds, such as tributyltin (TBT) and triphenyltin (TPT), have been widely used to control marine fouling. Here, we show that organotin stimulation reduces the hormone levels in the plasma of two economically important aquaculture fish. Blood plasma samples were collected from juvenile red seabream and black rockfish exposed to environmentally realistic concentrations of TBT and TPT for 14â¯days. The levels of two plasma biomarkers, namely the yolk protein precursor vitellogenin (VTG) and the sex steroid 17ß-estradiol (E2), were measured to determine the endocrine disrupting potential of the organotin compounds. Both organotin compounds were dose-dependently accumulated in the blood of two fish. Exposure to waterborne TBT and TBT significantly decreased the plasma VTG levels in both the juvenile fish in a dose-dependent manner. In contrast, the treatment with E2, a well-known VTG inducer, significantly increased the plasma VTG levels in both the fish. In addition, the mRNA levels of vtg were also downregulated in the liver tissues of both the fish at 100 and/or 1000â¯ngâ¯L-1 of TBT or TPT exposure. The plasma E2 titers were significantly suppressed at 100 and/or 1000â¯ngâ¯L-1 of TBT or TPT exposure for 14â¯days compared to their titer in the control. Since estrogen directly regulates vtg gene expression and VTG synthesis, our results reveal the endocrine disrupting potential of organotin compounds, and subsequently the endocrine modulation at early stage of fish can trigger further fluctuations in sexual differentiation, maturation, sex ration or egg production. In addition, the results demonstrate their effects on non-target organisms, particularly on animals reared in aquaculture and fisheries.
Assuntos
Disruptores Endócrinos/toxicidade , Estradiol/sangue , Compostos Orgânicos de Estanho/toxicidade , Percas/sangue , Dourada/sangue , Vitelogeninas/sangue , Poluentes Químicos da Água/toxicidade , Animais , Aquicultura , Biomarcadores/sangue , Antagonistas de Estrogênios/química , Antagonistas de Estrogênios/toxicidade , Feminino , Proteínas de Peixes/agonistas , Proteínas de Peixes/antagonistas & inibidores , Proteínas de Peixes/sangue , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Hormese/efeitos dos fármacos , Masculino , Concentração Osmolar , Percas/crescimento & desenvolvimento , Reprodutibilidade dos Testes , República da Coreia , Dourada/crescimento & desenvolvimento , Especificidade da Espécie , Compostos de Trialquitina/toxicidade , Vitelogeninas/agonistas , Vitelogeninas/antagonistas & inibidores , Vitelogeninas/genéticaRESUMO
Tamoxifen is a widely used anticancer drug with both an estrogen agonist and antagonist effect. This study focused on its endocrine disrupting effect, and overall environmental significance. Zebrafish embryos were exposed to different concentrations (0.5, 5, 50 and 500 µg l(-1) ) of tamoxifen for 96 h. The results showed a complex effect of tamoxifen on zebrafish embryo development. For the 500 µg l(-1) exposure group, the heart rate was decreased by 20% and mild defects in caudal fin and skin were observed. Expressions of a series of genes related to endocrine and morphological changes were subsequently tested through quantitative real-time polymerase chain reaction. Bisphenol A as a known estrogen was also tested as an endocrine-related comparison. Among the expression of endocrine-related genes, esr1, ar, cyp19a1b, hsd3b1 and ugt1a1 were all increased by tamoxifen exposure, similar to bisphenol A. The cyp19a1b is a key gene that controls estrogen synthesis. Exposure to 0.5, 5, 50 and 500 µg l(-1) of tamoxifen caused upregulation of cyp19a1b expression to 152%, 568%, 953% and 2024% compared to controls, higher than the effects from the same concentrations of bisphenol A treatment, yet vtg1 was suppressed by 24% from exposure to 500 µg l(-1) tamoxifen. The expression of metabolic-related genes such as cyp1a, cyp1c2, cyp3a65, gpx1a, gstp1, gsr and genes related to observed morphological changes such as krt17 were also found to be upregulated by high concentrations of tamoxifen. These findings indicated the potential environmental effect of tamoxifen on teleost early development. Copyright © 2015 John Wiley & Sons, Ltd.
Assuntos
Embrião não Mamífero/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Tamoxifeno/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra , Nadadeiras de Animais/anormalidades , Nadadeiras de Animais/efeitos dos fármacos , Nadadeiras de Animais/embriologia , Animais , Antineoplásicos Hormonais/toxicidade , Aromatase/genética , Aromatase/metabolismo , Embrião não Mamífero/anormalidades , Embrião não Mamífero/enzimologia , Embrião não Mamífero/metabolismo , Estrogênios não Esteroides/toxicidade , Frequência Cardíaca/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Larva/metabolismo , Concentração Osmolar , Distribuição Aleatória , Moduladores Seletivos de Receptor Estrogênico/toxicidade , Pele/efeitos dos fármacos , Pele/embriologia , Anormalidades da Pele/induzido quimicamente , Anormalidades da Pele/embriologia , Anormalidades da Pele/veterinária , Teratogênicos/toxicidade , Vitelogeninas/antagonistas & inibidores , Vitelogeninas/genética , Vitelogeninas/metabolismo , Peixe-Zebra/anormalidades , Peixe-Zebra/embriologia , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/agonistas , Proteínas de Peixe-Zebra/antagonistas & inibidores , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo , Zigoto/efeitos dos fármacos , Zigoto/crescimento & desenvolvimento , Zigoto/metabolismoRESUMO
BACKGROUND: Protein Tyrosine Phosphatases (PTPs) are enzymes that catalyze phosphotyrosine dephosphorylation and modulate cell differentiation, growth and metabolism. In mammals, PTPs play a key role in the modulation of canonical pathways involved in metabolism and immunity. PTP1B is the prototype member of classical PTPs and a major target for treating human diseases, such as cancer, obesity and diabetes. These signaling enzymes are, hence, targets of a wide array of inhibitors. Anautogenous mosquitoes rely on blood meals to lay eggs and are vectors of the most prevalent human diseases. Identifying the mosquito ortholog of PTP1B and determining its involvement in egg production is, therefore, important in the search for a novel and crucial target for vector control. METHODOLOGY/PRINCIPAL FINDINGS: We conducted an analysis to identify the ortholog of mammalian PTP1B in the Aedes aegypti genome. We identified eight genes coding for classical PTPs. In silico structural and functional analyses of proteins coded by such genes revealed that four of these code for catalytically active enzymes. Among the four genes coding for active PTPs, AAEL001919 exhibits the greatest degree of homology with the mammalian PTP1B. Next, we evaluated the role of this enzyme in egg formation. Blood feeding largely affects AAEL001919 expression, especially in the fat body and ovaries. These tissues are critically involved in the synthesis and storage of vitellogenin, the major yolk protein. Including the classical PTP inhibitor sodium orthovanadate or the PTP substrate DiFMUP in the blood meal decreased vitellogenin synthesis and egg production. Similarly, silencing AAEL001919 using RNA interference (RNAi) assays resulted in 30% suppression of egg production. CONCLUSIONS/SIGNIFICANCE: The data reported herein implicate, for the first time, a gene that codes for a classical PTP in mosquito egg formation. These findings raise the possibility that this class of enzymes may be used as novel targets to block egg formation in mosquitoes.
Assuntos
Aedes/enzimologia , Genoma de Inseto , Oviposição/genética , Proteína Tirosina Fosfatase não Receptora Tipo 1/genética , Vitelogeninas/genética , Aedes/efeitos dos fármacos , Aedes/genética , Sequência de Aminoácidos , Animais , Corpo Adiposo/efeitos dos fármacos , Corpo Adiposo/enzimologia , Feminino , Regulação da Expressão Gênica , Himecromona/análogos & derivados , Himecromona/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Ovário/efeitos dos fármacos , Ovário/enzimologia , Oviposição/efeitos dos fármacos , Estrutura Terciária de Proteína , Proteína Tirosina Fosfatase não Receptora Tipo 1/antagonistas & inibidores , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Vanadatos/farmacologia , Vitelogeninas/antagonistas & inibidores , Vitelogeninas/biossínteseRESUMO
In this study, male Chinese loaches in a semistatic waterborne exposure system were used to study the effects of tributyltin (TBT) on vitellogenin (Vtg) production induced by 17beta-estradiol (E2), TBT accumulation and distribution in tissues, and the effects of E2 on the distribution of TBT. The results demonstrate that TBT does not induce the synthesis of Vtg in male Chinese loach and that TBT could significantly inhibit Vtg production induced by E2, after exposure to binary mixtures of E2 and TBT for 14 days. TBT was found to accumulate in the liver, kidney, gills, intestines, and muscles of male Chinese loach, wherein the liver, kidney, gills, and intestines were found to have the most TBT accumulation. The existence of E2 did not significantly affect tissue distribution of TBT.
Assuntos
Cipriniformes/metabolismo , Estradiol/toxicidade , Compostos de Trialquitina/toxicidade , Vitelogeninas/sangue , Poluentes Químicos da Água/toxicidade , Animais , Estradiol/metabolismo , Brânquias/metabolismo , Mucosa Intestinal/metabolismo , Rim/metabolismo , Fígado/metabolismo , Masculino , Músculos/metabolismo , Compostos de Trialquitina/metabolismo , Vitelogeninas/antagonistas & inibidores , Poluentes Químicos da Água/metabolismoRESUMO
Effects of Al and Cd on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 beta (E(2)) were examined in primary hepatocyte cultures of rainbow trout. Hepatocytes were precultured for 2 days and then E(2) (2 x 10(-6) M) and Al (10(-6)-10(-4) M) or Cd (10(-9)-10(-6) M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metals had no appreciable effect on the viability of hepatocytes in culture. However, Al and Cd interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction occurred at Al concentrations greater than 5 x 10(-5) M. VTG mRNA expression also decreased with a negative correlation with Al concentrations (r = -0.98). The inhibition of VTG production by Cd was not concentration-dependent. This metal markedly inhibited VTG production and VTG mRNA expression at 10(-6) M. The Al-induced inhibition of VTG production was restored 7 days after Al removal, but the Cd-induced inhibition was not restored. These results suggest that Al and Cd inhibit VTG production at the transcriptional level to reduce VTG mRNA expression by different mechanisms.