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1.
Artigo em Inglês | MEDLINE | ID: mdl-38735623

RESUMO

Aquatic environments are subject to ultraviolet B (UVB) radiation incidence, and its effects on organisms are dose-dependent. Besides DNA, mitochondria are an important target of this radiation that causes structural damage and impairs its functional dynamics. Here, we hypothesize that mitophagy acts as an organelle quality control mechanism to mitigate UVB impacts in embryonic cells. Then, freshwater prawn Macrobrachium olfersii embryos was used as a model to investigate the effects of UVB on genes (Tomm20, Opa1, Pink, Prkn, Sqstm1, and Map1lc3) and proteins (TOM20, PINK1, p62 and LC3B) involved in mitophagy modulation. The choice of genes and proteins was based on the identification of mitochondrial membrane (Tomm20, Opa1 and TOM20), mediation of mitophagy (Pink1, Prkn and PINK1), and recognition of mitochondria by the autophagosome membrane (Sqstm1, Map1lc3, p62 and LC3B). First, the phylogeny of all genes presented bootstrap values >80 and conserved domains among crustacean species. Gene expression was inherently modulated during development, with transcripts (Tomm20, Opa1, Pink, Prkn, Sqstm1, and Map1lc3) overexpressed in the initial and final stages of development. Moreover, UVB radiation induced upregulation of Tomm20, Opa1, Pink, Prkn, Sqstm1, and Map1lc3 genes at 6 h after exposure. Interestingly, after 12 h, the protein content of PINK1, p62, and LC3B increased, while TOM20 was not responsive. Despite UVB radiation's harmful effects on embryonic cells, the chronology of gene expression and protein content indicates rapid activation of mitophagy, serving as an organelle quality control mechanism, given the analyzed cells' integrity.


Assuntos
Mitofagia , Palaemonidae , Raios Ultravioleta , Animais , Raios Ultravioleta/efeitos adversos , Mitofagia/efeitos da radiação , Palaemonidae/efeitos da radiação , Palaemonidae/embriologia , Palaemonidae/genética , Mitocôndrias/metabolismo , Mitocôndrias/efeitos da radiação , Embrião não Mamífero/efeitos da radiação , Embrião não Mamífero/metabolismo , Proteínas de Artrópodes/metabolismo , Proteínas de Artrópodes/genética , Filogenia , Organelas/metabolismo , Organelas/efeitos da radiação
2.
Bull Exp Biol Med ; 176(5): 645-648, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38727954

RESUMO

Using the method of dominant lethal mutations, we assessed the frequency of the death of Drosophila melanogaster embryos under combined exposure to ionizing γ-radiation and non-ionizing pulsed magnetic field at various doses and modes of exposure. Mutagenic effect of combined exposure is antagonistic in nature. The antagonism is more pronounced when the following mode of exposure was used: exposure to non-ionizing pulsed magnetic field for 5 h followed by exposure to γ-radiation at doses of 3, 10, and 60 Gy. In case of reverse sequence of exposures, the antagonistic effect was statistically significant after exposure to γ-radiation at doses of 3 and 10 Gy, whereas at a dose of 20 Gy, a synergistic interaction was noted.


Assuntos
Drosophila melanogaster , Raios gama , Animais , Drosophila melanogaster/efeitos da radiação , Drosophila melanogaster/genética , Raios gama/efeitos adversos , Radiação Eletromagnética , Relação Dose-Resposta à Radiação , Campos Eletromagnéticos/efeitos adversos , Embrião não Mamífero/efeitos da radiação , Radiação Ionizante , Mutação/efeitos da radiação , Mutagênese/efeitos da radiação
3.
J Radiat Res ; 65(3): 315-322, 2024 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-38648785

RESUMO

Ionizing radiation (IR) causes DNA damage, particularly DNA double-strand breaks (DSBs), which have significant implications for genome stability. The major pathways of repairing DSBs are homologous recombination (HR) and nonhomologous end joining (NHEJ). However, the repair mechanism of IR-induced DSBs in embryos is not well understood, despite extensive research in somatic cells. The externally developing aquatic organism, Xenopus tropicalis, serves as a valuable model for studying embryo development. A significant increase in zygotic transcription occurs at the midblastula transition (MBT), resulting in a longer cell cycle and asynchronous cell divisions. This study examines the impact of X-ray irradiation on Xenopus embryos before and after the MBT. The findings reveal a heightened X-ray sensitivity in embryos prior to the MBT, indicating a distinct shift in the DNA repair pathway during embryo development. Importantly, we show a transition in the dominant DSB repair pathway from NHEJ to HR before and after the MBT. These results suggest that the MBT plays a crucial role in altering DSB repair mechanisms, thereby influencing the IR sensitivity of developing embryos.


Assuntos
Blástula , Quebras de DNA de Cadeia Dupla , Reparo do DNA , Animais , Quebras de DNA de Cadeia Dupla/efeitos da radiação , Reparo do DNA/efeitos da radiação , Blástula/efeitos da radiação , Blástula/metabolismo , Xenopus/embriologia , Reparo do DNA por Junção de Extremidades/efeitos da radiação , Embrião não Mamífero/efeitos da radiação , Embrião não Mamífero/metabolismo , Raios X
4.
Radiother Oncol ; 194: 110197, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38447870

RESUMO

PURPOSE: A better characterization of the dependence of the tissue sparing effect at ultra-high dose rate (UHDR) on physical beam parameters (dose, dose rate, radiation quality) would be helpful towards a mechanistic understanding of the FLASH effect and for its broader clinical translation. To address this, a comprehensive study on the normal tissue sparing at UHDR using the zebrafish embryo (ZFE) model was conducted. METHODS: One-day-old ZFE were irradiated over a wide dose range (15-95 Gy) in three different beams (proton entrance channel, proton spread out Bragg peak and 30 MeV electrons) at UHDR and reference dose rate. After irradiation the ZFE were incubated for 4 days and then analyzed for four different biological endpoints (pericardial edema, curved spine, embryo length and eye diameter). RESULTS: Dose-effect curves were obtained and a sparing effect at UHDR was observed for all three beams. It was demonstrated that proton relative biological effectiveness and UHDR sparing are both relevant to predict the resulting dose response. Dose dependent FLASH modifying factors (FMF) for ZFE were found to be compatible with rodent data from the literature. It was found that the UHDR sparing effect saturates at doses above âˆ¼ 50 Gy with an FMF of âˆ¼ 0.7-0.8. A strong dose rate dependence of the tissue sparing effect in ZFE was observed. The magnitude of the maximum sparing effect was comparable for all studied biological endpoints. CONCLUSION: The ZFE model was shown to be a suitable pre-clinical high-throughput model for radiobiological studies on FLASH radiotherapy, providing results comparable to rodent models. This underlines the relevance of ZFE studies for FLASH radiotherapy research.


Assuntos
Relação Dose-Resposta à Radiação , Elétrons , Embrião não Mamífero , Peixe-Zebra , Animais , Peixe-Zebra/embriologia , Elétrons/uso terapêutico , Embrião não Mamífero/efeitos da radiação , Terapia com Prótons/métodos , Dosagem Radioterapêutica , Prótons , Eficiência Biológica Relativa
5.
Environ Sci Technol ; 58(5): 2204-2213, 2024 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-38269402

RESUMO

Ionizing radiation exposure induces significant DNA damage and cell death in aquatic species. Accurate sensing and quantification play pivotal roles in environmental monitoring and surveillance. Zebrafish (Danio rerio) is a well-suited animal model for research into this aspect, especially with recent development of cytogenetic and transgenic tools. In this study, we present time-course studies of chromosome aberrations and cell death in zebrafish embryos exposed to 2 Gy 137Cs total-body irradiation. Using a cytogenetic approach, we quantified chromosome and chromatid aberrations in irradiated embryos at 6, 14, 20, and 24 h postirradiation. Metaphases with aberrations showed rapid declining kinetics, accompanied by incomplete karyotypes and irregular chromatin contents. Using an apoptosis-reporting transgenic zebrafish, we found increasing cell death along these time points, with the embryonic eyes and brain contributing the majority of the cell death volumes. We provide evidence that self-proliferating progenitor cells form the underlying linkage between the two kinetics and their positions define radiosensitive niches in zebrafish embryos. Our results provide detailed chromosome aberration and cell death dynamics in 137Cs-irradiated zebrafish embryos and unveil the appropriate timeline and tissue positions for accurate sensing and quantification of radiation-induced damages in zebrafish embryos.


Assuntos
Aberrações Cromossômicas , Peixe-Zebra , Animais , Peixe-Zebra/genética , Raios gama , Cromossomos , Apoptose , Embrião não Mamífero/efeitos da radiação
6.
Radiat Res ; 199(3): 229-239, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36745564

RESUMO

Among the various types of cell death induced by ionizing radiation, apoptosis is a highly regulated and well-characterized form. Investigating radiation-induced apoptosis in an intact organism offers advantages in capturing the dynamics of apoptosis under preserved physiology, although high resolution imaging remains challenging. Owing to their optical transparency and genetic amenability, zebrafish is an ideal animal model for research into this aspect. In this study, we present a secA5 transgenic zebrafish expressing genetically encoded secreted ANNEXIN V fused with mVenus, a yellow fluorescent protein that enables reporting of radiation-induced apoptosis. Using in vivo imaging approach, we show that after 2 Gy total-body irradiation, apoptosis could be visualized at single-cell resolution in different cell types throughout the embryo. Elevated apoptosis could be imaged and quantified in the neuroepithelium of the embryonic brain, as well as the proliferative zone and parenchyma of the larval brain. In addition, clearance of apoptotic cells by microglia, the professional phagocytes residing in the brain, could be imaged at single-cell resolution in irradiated larvae. These results establish transgenic secA5 zebrafish as a useful and versatile in vivo system for investigating the dynamic process of radiation-induced apoptosis.


Assuntos
Apoptose , Peixe-Zebra , Animais , Peixe-Zebra/genética , Apoptose/genética , Apoptose/efeitos da radiação , Microglia , Diagnóstico por Imagem , Encéfalo , Embrião não Mamífero/metabolismo , Embrião não Mamífero/efeitos da radiação
7.
Ecotoxicol Environ Saf ; 231: 113211, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35051758

RESUMO

Ultraviolet (UV) rays can be both harmful and beneficial to humans. This study aimed to investigate the toxicity and safety of ultraviolet C (UVC) exposure in living organisms and the corresponding biodefense molecular mechanisms. Zebrafish embryos, at an early developmental stage (5-6 h post-fertilization), were irradiated with increasing UVC dosages using high-efficiency deep-ultraviolet light-emitting diodes (278 nm). Morphological phenotypes including survival rate, hatching rate, heart rate, and malformation rate were evaluated. Compared to un-irradiated controls, all zebrafish embryos exposed to 4.5 mJ/cm2 UVC survived and showed no significant difference in hatching and heart rate. However, 7.5 mJ/cm2 of UVC irradiation caused a significantly decreased survival rate (37.5%) and an increased malformation rate (81.8%). Therefore, 4.5 mJ/cm2 was chosen as the limit dosage that the internal biodefense system of zebrafish embryos can protect against UVC radiation. Transcriptome analysis (RNA sequencing) performed on 3 min and 3 days post-irradiation embryos (4.5 mJ/cm2) revealed the molecular mechanisms underlying the response of zebrafish embryos to irradiation. The embryos quickly responded to UVC-induced stress by activating the p53 signaling pathway. In addition, after 3 days of recuperation, the embryos showed activation of signal transducer and activator of transcription (STAT) signaling pathway. To our knowledge, this is the first study to evaluate the toxicological effects and the molecular mechanism of biodefense in zebrafish embryos upon 278 nm UVC irradiation.


Assuntos
Embrião não Mamífero/efeitos da radiação , Transcriptoma , Raios Ultravioleta , Peixe-Zebra , Animais , Perfilação da Expressão Gênica , Peixe-Zebra/genética
8.
Lab Invest ; 102(3): 298-311, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34773069

RESUMO

Pancreatic cancer (PC) is highly resistant to chemo and radiotherapy. Radiation-induced fibrosis (RIF) is a major cause of clinical concern for various malignancies, including PC. In this study, we aimed to evaluate the radiosensitizing and anti-RIF potential of fluvastatin in PC. Short-term viability and clonogenic survival assays were used to evaluate the radiosensitizing potential of fluvastatin in multiple human and murine PC cell lines. The expression of different proteins was analyzed to understand the mechanisms of fluvastatin-mediated radiosensitization of PC cells and its anti-RIF effects in both mouse and human pancreatic stellate cells (PSCs). Finally, these effects of fluvastatin and/or radiation were assessed in an immune-competent syngeneic murine model of PC. Fluvastatin radiosensitized multiple PC cell lines, as well as radioresistant cell lines in vitro, by inhibiting radiation-induced DNA damage repair response. Nonmalignant cells, such as PSCs and NIH3T3 cells, were less sensitive to fluvastatin-mediated radiosensitization than PC cells. Interestingly, fluvastatin suppressed radiation and/or TGF-ß-induced activation of PSCs, as well as the fibrogenic properties of these cells in vitro. Fluvastatin considerably augmented the antitumor effect of external radiation therapy and also suppressed intra-tumor RIF in vivo. These findings suggested that along with radiation, fluvastatin co-treatment may be a potential therapeutic approach against PC.


Assuntos
Fluvastatina/farmacologia , Neoplasias Pancreáticas/patologia , Tolerância a Radiação/efeitos dos fármacos , Fator de Crescimento Transformador beta/farmacologia , Animais , Apoptose/efeitos dos fármacos , Apoptose/efeitos da radiação , Autofagia/efeitos dos fármacos , Autofagia/efeitos da radiação , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Células Cultivadas , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/embriologia , Embrião não Mamífero/efeitos da radiação , Fibrose/prevenção & controle , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Células NIH 3T3 , Neoplasias Experimentais/tratamento farmacológico , Neoplasias Experimentais/patologia , Neoplasias Experimentais/radioterapia , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/radioterapia , Peixe-Zebra/embriologia
9.
Int J Mol Sci ; 22(16)2021 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-34445758

RESUMO

Radiotherapy is still a long way from personalizing cancer treatment plans, and its effectiveness depends on the radiosensitivity of tumor cells. Indeed, therapies that are efficient and successful for some patients may be relatively ineffective for others. Based on this, radiobiological research is focusing on the ability of some reagents to make cancer cells more responsive to ionizing radiation, as well as to protect the surrounding healthy tissues from possible side effects. In this scenario, zebrafish emerged as an effective model system to test for radiation modifiers that can potentially be used for radiotherapeutic purposes in humans. The adoption of this experimental organism is fully justified and supported by the high similarity between fish and humans in both their genome sequences and the effects provoked in them by ionizing radiation. This review aims to provide the literature state of the art of zebrafish in vivo model for radiobiological studies, particularly focusing on the epigenetic and radiomodifying effects produced during fish embryos' and larvae's exposure to radiotherapy treatments.


Assuntos
Epigênese Genética/efeitos da radiação , Radiossensibilizantes/efeitos adversos , Radioterapia/efeitos adversos , Peixe-Zebra , Animais , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/efeitos da radiação , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/efeitos da radiação , Modelos Animais
10.
PLoS Genet ; 17(5): e1009544, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33999957

RESUMO

Here we describe the development and characterization of the photo-N-degron, a peptide tag that can be used in optogenetic studies of protein function in vivo. The photo-N-degron can be expressed as a genetic fusion to the amino termini of other proteins, where it undergoes a blue light-dependent conformational change that exposes a signal for the class of ubiquitin ligases, the N-recognins, which mediate the N-end rule mechanism of proteasomal degradation. We demonstrate that the photo-N-degron can be used to direct light-mediated degradation of proteins in Saccharomyces cerevisiae and Drosophila melanogaster with fine temporal control. In addition, we compare the effectiveness of the photo-N-degron with that of two other light-dependent degrons that have been developed in their abilities to mediate the loss of function of Cactus, a component of the dorsal-ventral patterning system in the Drosophila embryo. We find that like the photo-N-degron, the blue light-inducible degradation (B-LID) domain, a light-activated degron that must be placed at the carboxy terminus of targeted proteins, is also effective in eliciting light-dependent loss of Cactus function, as determined by embryonic dorsal-ventral patterning phenotypes. In contrast, another previously described photosensitive degron (psd), which also must be located at the carboxy terminus of associated proteins, has little effect on Cactus-dependent phenotypes in response to illumination of developing embryos. These and other observations indicate that care must be taken in the selection and application of light-dependent and other inducible degrons for use in studies of protein function in vivo, but importantly demonstrate that N- and C-terminal fusions to the photo-N-degron and the B-LID domain, respectively, support light-dependent degradation in vivo.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Drosophila melanogaster/efeitos da radiação , Optogenética/métodos , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/efeitos da radiação , Animais , Arginina/metabolismo , Avena , Núcleo Celular/metabolismo , Núcleo Celular/efeitos da radiação , Escuridão , Drosophila melanogaster/embriologia , Embrião não Mamífero/metabolismo , Embrião não Mamífero/efeitos da radiação , Feminino , Fluorescência , Lasers , Luz , Mutação com Perda de Função , Masculino , Proteínas de Neoplasias/metabolismo , Fenótipo , Complexo de Endopeptidases do Proteassoma/metabolismo , Domínios Proteicos/efeitos da radiação , Proteínas Serina-Treonina Quinases/química , Proteólise/efeitos da radiação , Ubiquitina-Proteína Ligases/metabolismo
11.
Int J Mol Sci ; 22(3)2021 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-33530555

RESUMO

The number of sources of anthropogenic magnetic and electromagnetic fields generated by various underwater facilities, industrial equipment, and transferring devices in aquatic environment is increasing. These have an effect on an array of fish life processes, but especially the early developmental stages. The magnitude of these effects depends on field strength and time of exposure and is species-specific. We review studies on the effect of magnetic fields on the course of embryogenesis, with special reference to survival, the size of the embryos, embryonic motor function, changes in pigment cells, respiration hatching, and directional reactions. We also describe the effect of magnetic fields on sperm motility and egg activation. Magnetic fields can exert positive effects, as in the case of the considerable extension of sperm capability of activation, or have a negative influence in the form of a disturbance in heart rate or developmental instability in inner ear organs.


Assuntos
Embrião não Mamífero/efeitos da radiação , Desenvolvimento Embrionário/efeitos da radiação , Peixes , Campos Magnéticos , Animais , Permeabilidade da Membrana Celular/efeitos da radiação , Peixes/crescimento & desenvolvimento , Peixes/fisiologia , Larva , Óvulo/efeitos da radiação
12.
Nat Commun ; 11(1): 6214, 2020 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-33277480

RESUMO

Radioprotectors for acute injuries caused by large doses of ionizing radiation are vital to national security, public health and future development of humankind. Here, we develop a strategy to explore safe and efficient radioprotectors by combining Hantzsch's reaction, high-throughput methods and polymer chemistry. A water-soluble polymer with low-cytotoxicity and an excellent anti-radiation capability has been achieved. In in vivo experiments, this polymer is even better than amifostine, which is the only approved radioprotector for clinical applications, in effectively protecting zebrafish embryos from fatally large doses of ionizing radiation (80 Gy X-ray). A mechanistic study also reveals that the radioprotective ability of this polymer originates from its ability to efficiently prevent DNA damage due to high doses of radiation. This is an initial attempt to explore polymer radioprotectors via a multi-component reaction. It allows exploiting functional polymers and provides the underlying insights to guide the design of radioprotective polymers.


Assuntos
Técnicas de Química Sintética/métodos , Embrião não Mamífero/efeitos da radiação , Fibroblastos/efeitos da radiação , Polímeros/síntese química , Protetores contra Radiação/síntese química , Raios X , Amifostina/farmacologia , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Ensaio Cometa , Dano ao DNA/efeitos dos fármacos , Dano ao DNA/efeitos da radiação , Embrião não Mamífero/efeitos dos fármacos , Embrião não Mamífero/embriologia , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Camundongos , Modelos Químicos , Estrutura Molecular , Polímeros/química , Polímeros/farmacologia , Protetores contra Radiação/química , Protetores contra Radiação/farmacologia , Peixe-Zebra/embriologia
13.
Anticancer Res ; 40(11): 6123-6135, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33109550

RESUMO

BACKGROUND/AIM: The importance of hadron therapy in the cancer management is growing. We aimed to refine the biological effect detection using a vertebrate model. MATERIALS AND METHODS: Embryos at 24 and 72 h postfertilization were irradiated at the entrance plateau and the mid spread-out Bragg peak of a 150 MeV proton beam and with reference photons. Radiation-induced DNA double-strand breaks (DSB) and histopathological changes of the eye, muscles and brain were evaluated; deterioration of specific organs (eye, yolk sac, body) was measured. RESULTS: More and longer-lasting DSBs occurred in eye and muscle cells due to proton versus photon beams, albeit in different numbers. Edema, necrosis and tissue disorganization, (especially in the eye) were observed. Dose-dependent morphological deteriorations were detected at ≥10 Gy dose levels, with relative biological effectiveness between 0.99±0.07 (length) and 1.12±0.19 (eye). CONCLUSION: Quantitative assessment of radiation induced changes in zebrafish embryos proved to be beneficial for the radiobiological characterization of proton beams.


Assuntos
Fótons , Prótons , Peixe-Zebra/fisiologia , Animais , Encéfalo/efeitos da radiação , Dano ao DNA , Modelos Animais de Doenças , Relação Dose-Resposta à Radiação , Embrião não Mamífero/efeitos da radiação , Olho/patologia , Olho/efeitos da radiação , Cinética , Tamanho do Órgão/efeitos da radiação , Eficiência Biológica Relativa , Saco Vitelino/patologia , Saco Vitelino/efeitos da radiação , Peixe-Zebra/embriologia
14.
Int J Mol Sci ; 21(16)2020 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-32796742

RESUMO

Continuous overexposure to sunlight increases its harmful effects on the skin. For this reason, there is a growing need to characterize economic models more representative of the negative effects and counteracting responses that irradiation causes on human skin. These models will serve for the screening of protective compounds against damage caused by ultraviolet (UV) and high energy visible light (HEV). Therefore, two common in vitro models employed for sunlight irradiation studies, namely human keratinocyte HaCat culture and reconstructed human epidermis (RHE), were compared with the medaka fish embryo model, traditionally used in other scientific disciplines. Using suberythemal doses of UVA and HEV to determine the level of Reactive Oxygen Species (ROS) generation and thymine dimers formed by UVB, we show that medaka embryo responds with a lower damage level, more comparable to human skin, than the other two models, probably due to the protective mechanisms that work in a complete organism. In the same way, the protective effects of antioxidant compounds have the greatest effect on medaka embryos. Taken together, these findings suggest that medaka embryos would be a good alternative in vitro model for sunlight effect studies, and for the screening of molecules with counteracting capacity against the damage caused by UV and HEV.


Assuntos
Dano ao DNA , Avaliação Pré-Clínica de Medicamentos , Embrião não Mamífero/efeitos da radiação , Modelos Biológicos , Oryzias/embriologia , Raios Ultravioleta , Animais , Antioxidantes/farmacologia , Epiderme/efeitos da radiação , Células HaCaT , Humanos , Espécies Reativas de Oxigênio/metabolismo
15.
Rev Sci Instrum ; 91(6): 063303, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32611048

RESUMO

The development from single shot basic laser plasma interaction research toward experiments in which repetition rated laser-driven ion sources can be applied requires technological improvements. For example, in the case of radio-biological experiments, irradiation duration and reproducible controlled conditions are important for performing studies with a large number of samples. We present important technological advancements of recent years at the ATLAS 300 laser in Garching near Munich since our last radiation biology experiment. Improvements range from target positioning over proton transport and diagnostics to specimen handling. Exemplarily, we show the current capabilities by performing an application oriented experiment employing the zebrafish embryo model as a living vertebrate organism for laser-driven proton irradiation. The size, intensity, and energy of the laser-driven proton bunches resulted in evaluable partial body changes in the small (<1 mm) embryos, confirming the feasibility of the experimental system. The outcomes of this first study show both the appropriateness of the current capabilities and the required improvements of our laser-driven proton source for in vivo biological experiments, in particular the need for accurate, spatially resolved single bunch dosimetry and image guidance.


Assuntos
Aceleração , Embrião não Mamífero/efeitos da radiação , Lasers , Prótons , Radiobiologia/métodos , Peixe-Zebra/embriologia , Animais , Estudos de Viabilidade
16.
Proc Natl Acad Sci U S A ; 117(16): 8973-8979, 2020 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-32245815

RESUMO

The disposable soma theory is a central tenet of the biology of aging where germline immortality comes at the cost of an aging soma [T. B. L. Kirkwood, Nature 270, 301-304 (1977); T. B. L. Kirkwood, Proc. R. Soc. Lond. B Biol. Sci. 205, 531-546 (1979); T. B. L. Kirkwood, S. N. Austad, Nature 408, 233-238 (2000)]. Limited resources and a possible trade-off between the repair and maintenance of the germ cells and growth and maintenance of the soma may explain the deterioration of the soma over time. Here we show that germline removal allows accelerated somatic healing under stress. We tested "the expensive germ line" hypothesis by generating germline-free zebrafish Danio rerio and testing the effect of the presence and absence of the germ line on somatic repair under benign and stressful conditions. We exposed male fish to sublethal low-dose ionizing radiation, a genotoxic stress affecting the soma and the germ line, and tested how fast the soma recovered following partial fin ablation. We found that somatic recovery from ablation occurred substantially faster in irradiated germline-free fish than in the control germline-carrying fish where somatic recovery was stunned. The germ line did show signs of postirradiation recovery in germline-carrying fish in several traits related to offspring number and fitness. These results support the theoretical conjecture that germline maintenance is costly and directly trades off with somatic maintenance.


Assuntos
Envelhecimento/fisiologia , Regeneração/fisiologia , Estresse Fisiológico , Animais , Animais Geneticamente Modificados , Embrião não Mamífero/efeitos da radiação , Feminino , Técnicas de Silenciamento de Genes , Células Germinativas/fisiologia , Células Germinativas/efeitos da radiação , Masculino , Modelos Animais , Proteínas de Ligação a RNA/genética , Fatores Sexuais , Irradiação Corporal Total , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/genética
17.
Aquat Toxicol ; 222: 105468, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32199137

RESUMO

The extracellular matrix (ECM) is a non-cellular and three-dimensional structure, constituted by a macromolecular dynamic network that involves the cells in all animal tissues, including embryonic ones. Several studies with vertebrates and cell cultures have reported deleterious effects of ultraviolet-B (UVB) radiation on the components associated with the ECM. However, studies focusing on the UVB radiation effects on ECM components of crustaceans during embryonic development are very scarce. Thus, the aim of this study was to identify the coding sequences of components associated with the ECM and to evaluate the effect of UVB radiation on embryos of the ecologically-important decapod Macrobrachium olfersii. To evaluate the modulation of these ECM components during embryonic development, the transcript levels of Col4α1, Itgß, Lamα, Mmp1 and Timp in M. olfersii embryos were analyzed at early developmental stages (E1, E3 and E4), intermediate developmental stage (E7) and late developmental stages (E10 and E14). In addition, embryos at E7, which correspond to a landmark of crustacean development, were analyzed after 12 h of UVB exposure to verify UVB effects on the ECM components. The ECM component sequences were similar to other decapods, suggesting conservation of these genes among crustaceans. The results showed modulations of the ECM components of M. olfersii embryos that reflect the need for each component in the cellular mechanisms, necessary for normal embryonic development. After UVB exposure, embryos showed opacity of embryonic tissues and it was found the overexpression of Col4α1, Itgß, Mmp1 and Timp transcript levels (1.82-, 1.52-, 2.34- and 6.27-fold, respectively). These impairments can compromise important events for normal embryonic development, such as growth of optic lobes, caudal papilla, ramification of appendages and differentiation of organic systems. The results presented here, together with the effects on morphology, cell proliferation, differentiation, and apoptosis demonstrated previously, strengthen the knowledge of the complex impacts of UVB radiation on freshwater embryos. Nevertheless, our results encourage further investigations focusing on the assessment of UVB effects on different organisms in order to better understand the myriad of UVB effects on ECM components.


Assuntos
Embrião não Mamífero/efeitos da radiação , Desenvolvimento Embrionário/efeitos da radiação , Matriz Extracelular/efeitos da radiação , Palaemonidae/efeitos da radiação , Transcrição Gênica/efeitos da radiação , Raios Ultravioleta , Animais , Apoptose/efeitos da radiação , Diferenciação Celular/efeitos da radiação , Proliferação de Células/efeitos da radiação , Embrião não Mamífero/metabolismo , Embrião não Mamífero/patologia , Desenvolvimento Embrionário/genética , Matriz Extracelular/genética , Água Doce/química , Palaemonidae/genética , Palaemonidae/crescimento & desenvolvimento
18.
Chemosphere ; 248: 126005, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32032873

RESUMO

The Chernobyl and Fukushima nuclear power plant (NPP) accidents that occurred in 1986 and 2011 respectively have led to many years of chronic radiation exposure of wildlife. However, controversies remain on the dose threshold above which an impact on animal health occurs. Fish have been highly exposed immediately after both accidents in freshwater systems around Chernobyl and in freshwater and marine systems around Fukushima. The dose levels decreased during the years after the accidents, however, little is known about the effects of environmental low doses of radiation on fish health. The present laboratory study assesses the effects of an environmentally relevant dose range of radiation (0.1, 1 and 10 mGy/day) on early life stages of the 3-spined stickleback, Gasterosteus aculeatus. The cardiac physiology and developmental features (head width, diameter, area) of high exposed embryos (10 mGy/day) showed no significant change when compared to controls. Embryos exposed to the medium and high dose were slower to hatch than the controls (between 166 and 195 h post-fertilization). After 10 days of exposure (at 240 h post-fertilization), larvae exposed to the high dose displayed comparable growth to controls. High-throughput sequence analysis of transcriptional changes at this time point revealed no significant changes in gene regulation compared to controls regardless of exposure conditions. Our results suggest that exposure of fish embryos to environmental radiation elicits subtle delays in hatching times, but does not impair the overall growth and physiology, nor the gene expression patterns in the recently hatched larvae.


Assuntos
Desenvolvimento Embrionário/efeitos da radiação , Smegmamorpha/embriologia , Poluentes Radioativos da Água/análise , Animais , Animais Selvagens , Embrião não Mamífero/efeitos da radiação , Peixes , Água Doce , Larva
19.
Radiat Environ Biophys ; 59(2): 211-220, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31927628

RESUMO

The purpose of this investigation was to study the effect of acute γ-irradiation of parent adults on the endoreduplication of giant chromosomes in F1 generation of Drosophila melanogaster Meig. A wild-type Oregon-R strain was used as the material. Virgin females and males of Drosophila adults at the age of 3 days were irradiated with doses of 8, 16 and 25 Gy. Giant chromosomes were studied by cytomorphometry on squashed preparations of Drosophila salivary glands stained with acetoorsein. The preparations were obtained at late third instar larvae. The mean values of the polyteny degree of chromosomes (PDC) in males increased after 8 Gy by 10.6%, after 25 Gy by 7.4%, and did not change after the dose of 16 Gy. In females, the PDC did not differ from the control irrespective of the irradiation dose. An increase in endoreduplication was also evidenced by the accelerated development of offsprings of both sexes after irradiation of parents with 25 Gy, and in males also at a dose of 16 Gy. The statistical impact of power of radiation on polyteny was 26.8%, while the impact of sex was 4.9%. The impact of power of radiation on the developmental rate of offspring was 4.4% in males and 7.5% in females. The enhancement of endoreduplication is considered as a consequence of increasing selection pressure after irradiation. The possible involvement of epigenetic effects in the effect of ionizing radiation on endoreduplication is discussed.


Assuntos
Drosophila melanogaster/efeitos da radiação , Endorreduplicação/efeitos da radiação , Raios gama/efeitos adversos , Animais , Cromossomos de Insetos/efeitos da radiação , Drosophila melanogaster/genética , Embrião não Mamífero/efeitos da radiação , Feminino , Larva/genética , Larva/efeitos da radiação , Masculino , Glândulas Salivares
20.
Nanotoxicology ; 14(2): 232-249, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31738614

RESUMO

The use of nanotechnology to enhance pesticide formulations holds the promise of reduced pesticide use, reduced mobility in soils, and overall improvements in agricultural practices while simultaneously maintaining yields. However, the toxicity of nano-enabled pesticides, including azoxystrobin (Az), has not been well studied compared with their conventional form. This study investigates both lethal and sub-lethal endpoints in zebrafish embryos up to 120 h post-fertilization (hpf) under either laboratory light or simulated UV light. The median lethal concentration (LC50) value of nano-enabled Az (nAz) was significantly lower than the conventional form (Az). Interestingly, artificial UV light significantly increased toxicity (decreased LC50) of both Az and nAz. Malformations were not observed but the remaining yolk sac volume was significantly increased in both types of Az at both light conditions. This decreased yolk consumption is in agreement with reduced oxygen consumption and heart rate. Catalase enzyme activity was only reduced to UV light while superoxide dismutase activity was significantly reduced by co-exposure of UV light, and either type of Az at a nominal concentration of 100 µg L-1. The co-exposure of Az at 100 µg L-1 and UV light significantly upregulated sod1, sod2, and gpx1b expression and both types of Az significantly reduced gpx1a expression. Lipid peroxidation was significantly increased in nAz and Az at 100 µg L-1 under laboratory light, while UV light induced even higher level of lipid peroxidation. The results will provide important information on the toxicity of nAz under ecologically realistic conditions.


Assuntos
Embrião não Mamífero/efeitos dos fármacos , Nanopartículas/toxicidade , Praguicidas/toxicidade , Pirimidinas/toxicidade , Estrobilurinas/toxicidade , Raios Ultravioleta , Poluentes Químicos da Água/toxicidade , Peixe-Zebra , Animais , Catalase/metabolismo , Relação Dose-Resposta a Droga , Embrião não Mamífero/efeitos da radiação , Desenvolvimento Embrionário/efeitos dos fármacos , Nanopartículas/química , Estresse Oxidativo/efeitos dos fármacos , Praguicidas/química , Pirimidinas/química , Estrobilurinas/química , Poluentes Químicos da Água/química , Saco Vitelino/metabolismo
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