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1.
J Environ Sci (China) ; 85: 1-8, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31471016

RESUMO

Metalaxyl is an anilide pesticide that is widely used to control plant diseases caused by Peronosporales species. In order to study the toxic effects, zebrafish embryos were exposed to metalaxyl at nominal concentrations of 5, 50 and 500 ng/L for 72 hr, and the cardiac development and functioning of larvae were observed. The results showed that metalaxyl exposure resulted in increased rates of pericardial edema, heart hemorrhage and cardiac malformation. The distance between the sinus venosus and bulbus arteriosus, stroke volume, cardiac output and heart rate were significantly increased in larvae exposed to 50 and 500 ng/L metalaxyl compared to solvent control larvae. Significant upregulation in the transcription of tbx5, gata4 and myh6 was observed in the 50 and 500 ng/L treatments, and that of nkx2.5 and myl7 was observed in the 5, 50 and 500 ng/L groups. These disturbances may be related to cardiac developmental and functional defects in the larvae. The activity of Na+/K+-ATPase and Ca2+-ATPase was significantly increased in zebrafish embryos exposed to 500 ng/L metalaxyl, and the mRNA levels of genes related to ATPase (atp2a11, atp1b2b, and atp1a3b) (in the 50 and 500 ng/L groups) and calcium channels (cacna1ab) (in the 500 ng/L group) were significantly downregulated; these changes might be associated with heart arrhythmia and functional failure.


Assuntos
Alanina/análogos & derivados , Coração/crescimento & desenvolvimento , Poluentes Químicos da Água/toxicidade , Alanina/toxicidade , Animais , Embrião não Mamífero , Coração/efeitos dos fármacos , Peixe-Zebra/embriologia
2.
Nihon Yakurigaku Zasshi ; 154(2): 78-83, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31406047

RESUMO

Even after entering the era of genomic drug discovery in the 21st century, development of a breakthrough therapeutic drug (first-in-class) for intractable diseases (unmet medical needs) has been extremely difficult, but to the US FDA 62% of the approved first-in-class drugs are found by phenotypic screening. The next-generation zebrafish drug discovery enables high-throughput quantitative live in vivo phenotypic screening, and has been impacting global drug discovery strategies now. Compared to severe immunodeficient mice, zebrafish is expected to become a true individualized medical tool as a clinical ex vivo diagnostic system because of the high efficiency and speed of engraftment of patient-derived cancer xenotransplantation. Phenomics-based personalized medicine with the patient-derived cancer xenograft zebrafish in addition to conventional omics platform of individualized medicine is a true next-generation precision medicine to utilize for selection of therapeutic drugs and decision of their doses for the patient, and emerging paradigm shift is realizing in this century.


Assuntos
Descoberta de Drogas/tendências , Medicina de Precisão/tendências , Animais , Genômica , Humanos , Camundongos , Neoplasias , Estados Unidos , Ensaios Antitumorais Modelo de Xenoenxerto , Peixe-Zebra
3.
Adv Exp Med Biol ; 1155: 471-482, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468424

RESUMO

Endothelial cell dysfunction (ECD) is a broad term, which implies dysregulation of endothelial cell functions. Several factors contribute to ECD including high blood pressure, high cholesterol levels, diabetes, obesity, hyperglycemia, and advanced glycation end products (AGEs). The highly reactive dicarbonyl methylglyoxal (MGO) is mainly formed as byproduct of glycolysis. Therefore, high blood glucose levels result in increased MGO accumulation. Taurine-rich foods are considered to protect against various diseases including vasculopathy and to exert anti-aging effects. Here, we investigated the protective effect of hot water extract of Octopus ocellatus meat (OOM), which contains high amounts of taurine, on MGO-induced cell damage in human umbilical vein endothelial cells and zebrafish embryos. Hot water extract of OOMinhibited MGO-induced cytotoxicity and DNA damage, as well as AGEs accumulation. In addition, hot water extract of OOM protected against vascular damage in zebrafish embryos. These results suggest that hot water extract of OOM possesses protective activity against MGO-induced cytotoxicity in both umbilical vein endothelial cells and zebrafish embryos. Therefore, it could be used as a dietary source of an agent for the prevention of vascular diseases.


Assuntos
Extratos Celulares/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Octopodiformes/química , Aldeído Pirúvico/toxicidade , Taurina/farmacologia , Animais , Células Cultivadas , Embrião não Mamífero/efeitos dos fármacos , Humanos , Carne , Peixe-Zebra
4.
Adv Exp Med Biol ; 1155: 611-626, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31468435

RESUMO

In this study, the antioxidant properties of Viviparus contectus (V. contectus) extract were evaluated for various radical scavenging activities, ferric reducing antioxidant power (FRAP), ABTS radical scavenging activity and oxygen radical absorbance capacity (ORAC). In addition, inhibition effect of the V. contectus extract against DNA scission induced by hydroxyl radical was measured. We also studied the protective effect of V. contectus extract against oxidative damage through measurements of intracellular reactive oxygen species (ROS) in Chang cells and zebrafish embryo. We found that V. contectus extract contains strong radical scavenging activities and antioxidant properties, which prevent tert-butylhydroperoxide (t-BHP)-induced oxidative stress, enhance cell viability, reduce ROS production, inhibit oxidative damage and improve mitochondrial function in Chang cells. Also, we determined that the V. contectus extract reduced ROS production mediated by t-BHP induced oxidative stress on zebrafish embryo.


Assuntos
Antioxidantes/farmacologia , Extratos Celulares/farmacologia , Gastrópodes/química , Estresse Oxidativo , Animais , Linhagem Celular , Embrião não Mamífero , Humanos , Espécies Reativas de Oxigênio/metabolismo , Peixe-Zebra , terc-Butil Hidroperóxido
5.
Ecotoxicol Environ Saf ; 182: 109449, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31398778

RESUMO

The flame retardant, tris(1,3-dichloro-2-propyl) phosphate (TDCIPP), is one of the most developmentally toxic organophosphate flame retardants (OPFRs). However, few mechanistic studies on phenotypic malformation caused by TDCIPP have been conducted. This study investigates the molecular mechanism underlying abnormal tail fin development consistently observed in zebrafish embryos exposed to TDCIPP. The results show that the defects in the tail fin (e.g., bent spine, defective caudal fin, and damaged tip) were associated with altered expression of transcription factors. The significant up-regulation of mmp9 and, among insulin-growth factor (IGF) families, igfbp-1a and igfbp1b was observed, whereas alterations in the expression of cdx4, igf1a, ifg1b, igf2b, and vegaa regulating tail development were dependent on time points. In accordance with changes in mRNA gene expression, TDCIPP impaired vessel formation and disorganized muscle in transgenic Tg(fli-GFP) zebrafish larvae. Furthermore, we found that the overexpression of mmp9 caused by TDCIPP was not linked to igfbp-1. Overall, these findings demonstrate that TDCIPP disrupts the progression of tail fin development, accompanied by defects in vessel and muscle formation in developing zebrafish embryos.


Assuntos
Desenvolvimento Embrionário/efeitos dos fármacos , Retardadores de Chama/toxicidade , Compostos Organofosforados/toxicidade , Animais , Animais Geneticamente Modificados , Retardadores de Chama/metabolismo , Larva , Organofosfatos/metabolismo , Fosfatos/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismo
6.
Hum Genet ; 138(8-9): 993-1000, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31422478

RESUMO

In this brief commentary, we provide some of our thoughts and opinions on the current and future use of zebrafish to model human eye disease, dissect pathological progression and advance in our understanding of the genetic bases of microphthalmia, andophthalmia and coloboma (MAC) in humans. We provide some background on eye formation in fish and conservation and divergence across vertebrates in this process, discuss different approaches for manipulating gene function and speculate on future research areas where we think research using fish may prove to be particularly effective.


Assuntos
Oftalmopatias/genética , Peixe-Zebra/genética , Animais , Coloboma/genética , Humanos , Microftalmia/genética
7.
Results Probl Cell Differ ; 67: 253-276, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31435799

RESUMO

'Does the geometric design of centrioles imply their function? Several principles of construction of a microscopically small device for locating the directions of signal sources in microscopic dimensions: it appears that the simplest and smallest device that is compatible with the scrambling influence of thermal fluctuations, as are demonstrated by Brownian motion, is a pair of cylinders oriented at right angles to each other. Centrioles locate the direction of hypothetical signals inside cells' (Albrecht-Buehler G, Cell Motil, 1:237-245; 1981).Despite a century of devoted efforts (articles on the centrosome always begin like this) its role remains vague and nebulous: does the centrosome suffer from bad press? Likely it does, it has an unfair image problem. It is dispensable in mitosis, but a fly zygote, artificially deprived of centrosomes, cannot start its development; its sophisticated architecture (200 protein types, highly conserved during evolution) constitutes an enigmatic puzzle; centrosome reduction in gametogenesis is a challenging brainteaser; its duplication cycle (only one centrosome per cell) is more complicated than chromosomes. Its striking geometric design (two ninefold symmetric orthogonal centrioles) shows an interesting correspondence with the requirements of a cellular compass: a reference system organizer based on a pair of orthogonal goniometers; through its two orthogonal centrioles, the centrosome may play the role of a cell geometry organizer: it can establish a finely tuned geometry, inherited and shared by all cells. Indeed, a geometrical and informational primary role for the centrosome has been ascertained in Caenorhabditis elegans zygote: the sperm centrosome locates its polarity factors. The centrosome, through its aster of microtubules, possesses all the characteristics necessary to operate as a biophysical geometric compass: it could recognize cargoes equipped with topogenic sequences and drive them precisely to where they are addressed (as hypothesized by Albrecht-Buehler nearly 40 years ago). Recently, this geometric role of the centrosome has been rediscovered by two important findings; in the Kupffer's vesicle (the laterality organ of zebrafish), chiral cilia orientation and rotational movement have been described: primary cilia, in left and right halves of the Kupffer's vesicle, are symmetrically oriented relative to the midline and rotate in reverse direction. In mice node (laterality organ) left and right perinodal cells can distinguish flow directionality through their primary cilia: primary cilium, ninefold symmetric, is strictly connected to the centrosome that is located immediately under it (basal body). Kupffer's vesicle histology and mirror behaviour of mice perinodal cells suggest primary cilia are enantiomeric geometric organelles. What is the meaning of the geometric design of centrioles and centrosomes? Does it imply their function?


Assuntos
Centrossomo/metabolismo , Animais , Centríolos/metabolismo , Cílios , Masculino , Espermatozoides/citologia , Peixe-Zebra
8.
Gene ; 718: 144049, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31430520

RESUMO

The role of epigenetics in development has garnered attention in recent years due to their ability to modulate the embryonic developmental gene expression in response to the environmental cues. The epigenetic mechanisms - DNA methylation, histone modification, and non-coding RNAs have a unique impact on vertebrate development. Zebrafish, a model vertebrate organism is being used widely in developmental studies due to their high fecundability and rapid organogenesis. With increased studies on various aspects of epigenetics in development, this review gives a glimpse of the major epigenetic modifications and their role in zebrafish development. In this review, the basic mechanism behind each modification followed by their status in zebrafish has been reviewed. Further, recent advancements in the epigenetic aspect of zebrafish development have been discussed.


Assuntos
Metilação de DNA/fisiologia , Embrião não Mamífero/embriologia , Desenvolvimento Embrionário/fisiologia , Epigênese Genética/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Peixe-Zebra/embriologia , Animais , Histonas/genética , Histonas/metabolismo , Processamento de Proteína Pós-Traducional/fisiologia , Peixe-Zebra/genética
9.
Ying Yong Sheng Tai Xue Bao ; 30(8): 2845-2853, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31418211

RESUMO

The safety of feed derived from genetically modified (GM) crops is one of the focuses of attention. To evaluate the ecotoxicological effects of transgenic mCry1Ac maize (BT799) on fish, zebrafish (Danio rerio) were fed extruded feeds containing either 20% GM maize (GMF) or its parental control maize (PF), GM maize meal (GMM) or its parental control maize meal (PMM), and a control commercial feed (CF), respectively. The growth performance, histopathology, reproduction, antioxidant enzyme activity and mRNA expression levels of sensitive protein in the liver were investigated over the course of a 98-day feeding trial. The results showed that transgenic mCry1Ac maize had no significant effect on growth, histopathology of the liver, brain and intestinal tract, fecundity, hatching rate of fertilized eggs, superoxide dismutase (SOD), catalase (CAT) activity, mRNA expression levels of SOD and CAT, or heat shock protein 70 (HSP70) and vitellogenin (VTG) in the liver. However, zebrafish fed the commercial feed exhibited significantly greater weight, longer length, and higher specific growth rate than those fed feeds (GMF and PF) and maize meals (GMM and PMM). The hatching rate of zebrafish in the feed groups was significantly lower than that of the maize meal groups and the commercial feed group. The mRNA transcriptional levels of VTG were significantly higher in the liver for the feed groups (3.85±0.76) than that for the maize meal groups (1.60±0.56). These results suggest that transgenic mCry1Ac maize has no ecotoxicological effects on zebrafish. However, the differences in nutrient composition and palatability between the extruded experimental feeds and the commercial feed would lead to significant diffe-rences in some parameters.


Assuntos
Alimentos Geneticamente Modificados , Zea mays/genética , Ração Animal , Animais , Plantas Geneticamente Modificadas , Testes de Toxicidade , Peixe-Zebra/fisiologia
10.
Artigo em Inglês | MEDLINE | ID: mdl-31255699

RESUMO

Organic anion transporters (OATs) are membrane proteins within the Solute carrier family 22 (SLC22). They play important roles in cellular uptake of various organic compounds, and due to their expression in barrier tissues of major excretory and non-excretory organs are considered as crucial elements in absorption and distribution of a wide range of endobiotic and xenobiotic compounds. Based on our previous work and initial insights on SLC22 members in zebrafish (Danio rerio), in this study we aimed at in vitro characterization of Oat1 and Oat3 transporters and understanding of their interaction with potential physiological substrates. We first performed synteny analysis to describe in more detail orthological relationship of zebrafish oat1 and oat3 genes. We then developed stable cell lines overexpressing Oat1 and Oat3, and identified Lucifer yellow as Oat1 model fluorescent substrate (Km = 11.4 µM) and 6-carboxyfluorescein as Oat3 model substrate (Km = 5.8 µM). Initial identification performed using the developed assays revealed Kreb's cycle intermediates, bilirubin, bile salts and steroid hormones as the most potent of Oat1 and Oat3 interactors, with IC50 values in micromolar range. Finally, we showed that bilirubin, deoxycholic acid, α-ketoglutarate, pregnenolone, estrone-3-sulfate and corticosterone are in vitro substrates of zebrafish Oat1, and bilirubin and deoxycholic acid are Oat3 substrates. In conclusion, using the approach described, structural and functional similarities of both transporters to human and mammalian orthologs are revealed, their broad ligand selectivity confirmed, potent interactors among endobiotic compounds identified, and first indications of their potential physiological role(s) in zebrafish obtained.


Assuntos
Proteína 1 Transportadora de Ânions Orgânicos/metabolismo , Transportadores de Ânions Orgânicos Sódio-Independentes/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Animais , Avaliação Pré-Clínica de Medicamentos , Proteína 1 Transportadora de Ânions Orgânicos/antagonistas & inibidores , Transportadores de Ânions Orgânicos Sódio-Independentes/antagonistas & inibidores , Ligação Proteica , Transporte Proteico , Proteínas de Peixe-Zebra/antagonistas & inibidores
11.
Sci Total Environ ; 685: 1294-1305, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31272786

RESUMO

Bisphenol A (BPA) is an abundant environmental contaminant and studies have shown the presence of BPA in the urine of over 90% of population tested in Canada and USA. In addition to its reported harmful effects, there is concern for its transgenerational effects. For a compound to induce transgenerational effect, an epigenetic mark should be mitotically and meiotically stable without reprogramming in primordial germ cells and post fertilization embryos. In the present study, female zebrafish were treated with an environmental dose (20 µg/L) of BPA and then crossed with untreated males. To assess epigenetic effects, transcript levels of several genes involved in female reproduction were measured in adult and in 24 hpf embryos up to F3 generation. Exposure to BPA affected adult female fertility up to F2 generation. In F0, F1 and F2 ovaries transcript levels for several genes involved in reproduction, including esr, star, lhcgr and fshr were affected. To investigate epigenetic mechanisms of gene expression modulation, we studied promoter DNA methylation. Among genes involved in gonadal differentiation, amh transcript level was reduced in 24 hpf embryos, up to the F3 generation. Variation in amh transcript level was associated with hyper-methylation of its promoter and changes in H3K4me3/H3K27me3 enrichment, coherent with gene silencing. The findings provide evidence for transgenerational effects of BPA in zebrafish and demonstrate that amh is susceptible to stable epigenetic alterations. CAPSULE: Transgenerational effects of BPA on female reproductive physiology.


Assuntos
Compostos Benzidrílicos/toxicidade , Fenóis/toxicidade , Reprodução/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Metilação de DNA , Epigênese Genética , Feminino , Histonas , Regiões Promotoras Genéticas , Peixe-Zebra
12.
Environ Monit Assess ; 191(8): 478, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31263965

RESUMO

Carwash effluents contain potentially toxic chemical and microbiological pollutants which may pose public health and ecotoxicological threats if directly discharged into surface waters. This work was aimed at determining the microbiological, physicochemical, and toxicological parameters of carwash effluents. Toxicity assays were determined using whole effluent toxicity (WET) using Danio rerio and Daphnia pulex. For microbiological analysis, sample aliquots were spread plated onto R2A Agar for the isolation of heterotrophic bacteria followed by DNA extraction from axenic cultures for sequencing analysis. The pH of effluent samples lay in the alkaline range, and ranged from pH 7 to pH 10. Sample salinity ranged from 0.2 to 0.3 g/Kg. Electrical conductivity values ranged from 274 to 554 µS/cm. Concentrations of Co, Pb, and Ni were < 1 mg/L in all samples while the concentrations of Cu ranged from 0.94 to 3.8 mg/L and Zn from 1.15 to 3 mg/L. Oil and grease concentrations ranged from 5 to 24 mg/L. The concentrations of TPH-GRO were low at < 1 mg/L in all samples. All the carwash effluents were categorised as acutely toxic, with ≥ 75% mortality recorded for both test organisms within the first 24 h of exposure to the test solutions. Heterotrophic bacteria counts ranged from 2800 to 4600 CFU/100 ml. Sequencing analysis revealed that 57% of the isolates were closely related to Aeromonas species, with 43% closely related to Pseudomonas species. We conclude that carwash effluents are veritable sources of microbiological contaminants and potentially toxic chemical pollutants of public health and ecotoxicological concern.


Assuntos
Monitoramento Ambiental , Veículos Automotores , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade , Animais , Bactérias , Daphnia , Ecotoxicologia , Testes de Toxicidade , Microbiologia da Água , Peixe-Zebra
13.
Chem Commun (Camb) ; 55(65): 9629-9632, 2019 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-31353368

RESUMO

Excessive accumulation of reducing agents in the ER leads to a constitutively high UPR. And the co-function of GSH, Cys and HOCl in biological processes is not well understood. To address this, a TP probe, NPCC, was developed for monitoring reductive stress in the ER. It can also distinguish cancer cells from normal cells.


Assuntos
Cumarínicos/química , Estresse do Retículo Endoplasmático/fisiologia , Retículo Endoplasmático/metabolismo , Corantes Fluorescentes/química , Pirazóis/química , Animais , Cumarínicos/síntese química , Cisteína/química , Cisteína/metabolismo , Corantes Fluorescentes/síntese química , Glutationa/química , Glutationa/metabolismo , Cabras , Células HeLa , Humanos , Ácido Hipocloroso/química , Ácido Hipocloroso/metabolismo , Camundongos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Oxirredução , Pirazóis/síntese química , Peixe-Zebra
14.
Chem Commun (Camb) ; 55(58): 8494-8497, 2019 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-31268095

RESUMO

A rational strategy was reported to construct boranil complexes (DPFB derivatives) with unique aggregation-induced emission effects by installing phenyl rings in the anil ligand as the intramolecular rotors. In view of the good biocompatibility and suitable lipophilicity, DPFB derivatives can serve as excellent fluorescent probes for specific imaging of lipid droplets in living cells and yolk lipids in zebrafish.


Assuntos
Compostos de Anilina/química , Compostos de Boro/química , Complexos de Coordenação/química , Corantes Fluorescentes/química , Gotículas Lipídicas/metabolismo , Compostos de Anilina/síntese química , Animais , Compostos de Boro/síntese química , Complexos de Coordenação/síntese química , Fluorescência , Corantes Fluorescentes/síntese química , Células HeLa , Humanos , Ligantes , Microscopia Confocal/métodos , Estrutura Molecular , Peixe-Zebra
15.
J Agric Food Chem ; 67(28): 7783-7792, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31267752

RESUMO

The increasing use of pesticides in agriculture and gardening has caused severe deterioration to both the ecosystem and the health of users (human beings), so there is an urgent need for eco- and user-friendly pesticides. Among a variety of herbicides, paraquat (PQ), frequently used as an effective herbicidal agent worldwide, is well-known for its serious toxicity that has killed, and harmed, thousands of people and countless wildlife such as fish. Herein, we present a facile supramolecular formulation of PQ@cucurbit[7]uril (PQ@CB[7]), prepared by simply mixing PQ with equivalent (molar) CB[7] in water. With addition of CB[7], PQ's cellular uptake was dramatically inhibited. The reactive oxygen species (ROS) generation and the associated apoptosis otherwise induced by PQ in cellular models were both reduced, resulting in increased cellular viability. In a wildtype zebrafish model that is a typical fragile wildlife species in the ecosystem, the supramolecular formulation exhibited significantly reduced hepatotoxicity and increased survival rate, in comparison with those of the fish exposed to free PQ. In a mouse model that is clinically relevant to human being, the administration of PQ@CB[7] significantly alleviated major organ injuries and unusual hematological parameters that were otherwise induced by free PQ, resulting in a significantly increased survival rate. Meanwhile, this formulation maintained effective herbicidal activity that was equivalent to that of free PQ. Taken together, this facile supramolecular PQ formulation is providing not only an extremely rare example of an eco- and user-friendly herbicide that has been desired for decades but also a practical solution for green agriculture.


Assuntos
Herbicidas/farmacologia , Paraquat/farmacologia , Animais , Apoptose/efeitos dos fármacos , Química Verde , Herbicidas/síntese química , Herbicidas/química , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Paraquat/síntese química , Paraquat/química , Poaceae/efeitos dos fármacos , Poaceae/crescimento & desenvolvimento , Peixe-Zebra/metabolismo
16.
BMC Evol Biol ; 19(1): 137, 2019 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-31269894

RESUMO

BACKGROUND: Previously, we have demonstrated that genes involved in ovarian function are highly conserved throughout evolution. In this study, we aimed to document the conservation of genes involved in spermatogenesis from flies to vertebrates and their expression profiles in vertebrates. RESULTS: We retrieved 379 Drosophila melanogaster genes that are functionally involved in male reproduction according to their mutant phenotypes and listed their vertebrate orthologs. 83% of the fly genes have at least one vertebrate ortholog for a total of 625 mouse orthologs. This conservation percentage is almost twice as high as the 42% rate for the whole fly genome and is similar to that previously found for genes preferentially expressed in ovaries. Of the 625 mouse orthologs, we selected 68 mouse genes of interest, 42 of which exhibited a predominant relative expression in testes and 26 were their paralogs. These 68 mouse genes exhibited 144 and 60 orthologs in chicken and zebrafish, respectively, gathered in 28 groups of paralogs. Almost two thirds of the chicken orthologs and half of the zebrafish orthologs exhibited a relative expression ≥50% in testis. Finally, our focus on functional in silico data demonstrated that most of these genes were involved in the germ cell process, primarily in structure elaboration/maintenance and in acid nucleic metabolism. CONCLUSION: Our work confirms that the genes involved in germ cell development are highly conserved across evolution in vertebrates and invertebrates and display a high rate of conservation of preferential testicular expression among vertebrates. Among the genes highlighted in this study, three mouse genes (Lrrc46, Pabpc6 and Pkd2l1) have not previously been described in the testes, neither their zebrafish nor chicken orthologs. The phylogenetic approach developed in this study finally allows considering new testicular genes for further fundamental studies in vertebrates, including model species (mouse and zebrafish).


Assuntos
Galinhas/genética , Evolução Molecular , Testículo/metabolismo , Peixe-Zebra/genética , Animais , Drosophila melanogaster/genética , Masculino , Camundongos , Filogenia , Espermatogênese/genética , Testículo/citologia
17.
Nat Commun ; 10(1): 2947, 2019 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-31270320

RESUMO

To expand the toolbox of imaging in living cells, we have engineered a single-chain variable fragment binding the linear HA epitope with high affinity and specificity in vivo. The resulting probe, called the HA frankenbody, can light up in multiple colors HA-tagged nuclear, cytoplasmic, membrane, and mitochondrial proteins in diverse cell types. The HA frankenbody also enables state-of-the-art single-molecule experiments in living cells, which we demonstrate by tracking single HA-tagged histones in U2OS cells and single mRNA translation dynamics in both U2OS cells and neurons. Together with the SunTag, we also track two mRNA species simultaneously to demonstrate comparative single-molecule studies of translation can now be done with genetically encoded tools alone. Finally, we use the HA frankenbody to precisely quantify the expression of HA-tagged proteins in developing zebrafish embryos. The versatility of the HA frankenbody makes it a powerful tool for imaging protein dynamics in vivo.


Assuntos
Epitopos/metabolismo , Sondas Moleculares/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Imagem Individual de Molécula , Animais , Linhagem Celular Tumoral , Embrião não Mamífero/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Neurônios/metabolismo , Biossíntese de Proteínas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/isolamento & purificação , Anticorpos de Cadeia Única/metabolismo , Coloração e Rotulagem , Peixe-Zebra/embriologia
18.
Nat Commun ; 10(1): 3053, 2019 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-31311924

RESUMO

The germline is the only cellular lineage capable of transferring genetic information from one generation to the next. Intergenerational transmission of epigenetic memory through the germline, in the form of DNA methylation, has been proposed; however, in mammals this is largely prevented by extensive epigenetic erasure during germline definition. Here we report that, unlike mammals, the continuously-defined 'preformed' germline of zebrafish does not undergo genome-wide erasure of DNA methylation during development. Our analysis also uncovers oocyte-specific germline amplification and demethylation of an 11.5-kb repeat region encoding 45S ribosomal RNA (fem-rDNA). The peak of fem-rDNA amplification coincides with the initial expansion of stage IB oocytes, the poly-nucleolar cell type responsible for zebrafish feminisation. Given that fem-rDNA overlaps with the only zebrafish locus identified thus far as sex-linked, we hypothesise fem-rDNA expansion could be intrinsic to sex determination in this species.


Assuntos
Metilação de DNA/fisiologia , DNA Ribossômico/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Oócitos/metabolismo , Peixe-Zebra/fisiologia , Animais , Desmetilação , Epigênese Genética/fisiologia , Feminino , Masculino , RNA Ribossômico/genética , Caracteres Sexuais
19.
Toxicol Lett ; 314: 43-52, 2019 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-31310794

RESUMO

Thioredoxin is an evolutionarily conserved antioxidant protein that plays a crucial role for fundamental cellular processes and embryonic development. Growing evidence support that Thioredoxin influences cellular response to chemicals insults, particularly those accompanying oxidative stress. The mechanisms underlying the functions of Thioredoxin1 in the embryonic development under the environmental toxicant exposure remain, however, largely unexplored. We report here that thioredoxin1 becomes differentially expressed in zebrafish embryos after exposure to 9 out of 11 environmental chemicals. In situ gene expression analysis show that thioredoxin1 is expressed in neurons, olfactory epithelia, liver and swim bladder under normal conditions. After MeHg exposure, however, thioredoxin1 is ectopically induced in the hair cells of the lateral line and in epithelia cells of the pharynx. Knockdown of Thioredoxin1 induces hydrocephalus and increases cell apoptosis in the brain ventricular epithelia cells. In comparison with 5% malformation in embryos injected with control morpholino, MeHg induces more than 77% defects in Thioredoxin1 knockdown embryos. Our data suggest that there is an association between hydrocephalus and Thioredoxin1 malfunction in embryonic development, and provide valuable information to elucidate the protective role of Thioredoxin1 against chemicals disruption.


Assuntos
Encéfalo/efeitos dos fármacos , Poluentes Ambientais/toxicidade , Hidrocefalia/induzido quimicamente , Tiorredoxinas/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados , Apoptose/efeitos dos fármacos , Encéfalo/embriologia , Encéfalo/metabolismo , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/metabolismo , Embrião não Mamífero/patologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Regulação da Expressão Gênica no Desenvolvimento , Hidrocefalia/embriologia , Hidrocefalia/genética , Hidrocefalia/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Estresse Oxidativo/efeitos dos fármacos , Tiorredoxinas/genética , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
20.
Nature ; 571(7764): 198-204, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31292557

RESUMO

Slow-wave sleep and rapid eye movement (or paradoxical) sleep have been found in mammals, birds and lizards, but it is unclear whether these neuronal signatures are found in non-amniotic vertebrates. Here we develop non-invasive fluorescence-based polysomnography for zebrafish, and show-using unbiased, brain-wide activity recording coupled with assessment of eye movement, muscle dynamics and heart rate-that there are at least two major sleep signatures in zebrafish. These signatures, which we term slow bursting sleep and propagating wave sleep, share commonalities with those of slow-wave sleep and paradoxical or rapid eye movement sleep, respectively. Further, we find that melanin-concentrating hormone signalling (which is involved in mammalian sleep) also regulates propagating wave sleep signatures and the overall amount of sleep in zebrafish, probably via activation of ependymal cells. These observations suggest that common neural signatures of sleep may have emerged in the vertebrate brain over 450 million years ago.


Assuntos
Neurônios/fisiologia , Sono/fisiologia , Peixe-Zebra/fisiologia , Animais , Evolução Biológica , Encéfalo/citologia , Encéfalo/efeitos dos fármacos , Encéfalo/fisiologia , Encéfalo/fisiopatologia , Epêndima/citologia , Movimentos Oculares , Fluorescência , Frequência Cardíaca , Hipnóticos e Sedativos/farmacologia , Hormônios Hipotalâmicos/metabolismo , Melaninas/metabolismo , Neurônios/efeitos dos fármacos , Pigmentação/fisiologia , Hormônios Hipofisários/metabolismo , Polissonografia/métodos , Sono/efeitos dos fármacos , Privação do Sono/fisiopatologia , Sono REM/efeitos dos fármacos , Sono REM/fisiologia , Sono de Ondas Lentas/efeitos dos fármacos , Sono de Ondas Lentas/fisiologia
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