RESUMEN
Acrylamide is a commonly used industrial chemical that is known to be neurotoxic to mammals. However, its developmental toxicity is rarely assessed in mammalian models because of the cost and complexity involved. We used zebrafish to assess the neurotoxicity, developmental and behavioral toxicity of acrylamide. At 6 h post fertilization, zebrafish embryos were exposed to four concentrations of acrylamide (10, 30, 100, or 300 mg/L) in a medium for 114 h. Acrylamide caused developmental toxicity characterized by yolk retention, scoliosis, swim bladder deficiency, and curvature of the body. Acrylamide also impaired locomotor activity, which was measured as swimming speed and distance traveled. In addition, treatment with 100 mg/L acrylamide shortened the width of the brain and spinal cord, indicating neuronal toxicity. In summary, acrylamide induces developmental toxicity and neurotoxicity in zebrafish. This can be used to study acrylamide neurotoxicity in a rapid and cost-efficient manner.
Asunto(s)
Acrilamida/toxicidad , Embrión no Mamífero/efectos de los fármacos , Síndromes de Neurotoxicidad/fisiopatología , Pez Cebra/crecimiento & desarrollo , Acrilamida/farmacología , Sacos Aéreos/patología , Animales , Animales Modificados Genéticamente , Modelos Animales de Enfermedad , Embrión no Mamífero/fisiopatología , Desarrollo Embrionario/efectos de los fármacos , Síndromes de Neurotoxicidad/etiología , Escoliosis/etiología , Natación , Pez Cebra/fisiologíaRESUMEN
To assess the developmental toxicity of trichloroacetate (TCA), zebrafish embryos were exposed to 8 to 48 mM of TCA and evaluated for developmental milestones from 8- to 144-hour postfertilization (hpf). All developmental toxicities are reported in this paper. Embryos were found to have developed edema in response to 16 to 48 mM of TCA exposure at 32- to 80-hpf, experienced delay in hatching success in response to 24 to 48 mM at 80-hpf. Lordosis was observed in developing embryos exposed to 40 to 48 mM at 55- to 144-hpf. The observed toxic effects of TCA exposure were found to be concentration and exposure period independent. Effects were found to be associated with increases in superoxide anion production, but these increases were also found to be concentration and time independent. TCA resulted in concentration-dependent increases in embryonic lethality at 144-hpf, with an LC50 determined to be 29.7 mM.
Asunto(s)
Embrión no Mamífero/efectos de los fármacos , Desarrollo Embrionario , Superóxidos/metabolismo , Ácido Tricloroacético/toxicidad , Pez Cebra/fisiología , Animales , Embrión no Mamífero/metabolismo , Embrión no Mamífero/fisiopatología , Lordosis/etiología , Ácido Tricloroacético/farmacología , Pez Cebra/metabolismoRESUMEN
Embryonic turtles have four distinct vascular beds that separately perfuse the developing embryo's body and the extra-embryonic yolk sac, amnion and chorioallantoic membrane (CAM). The mechanisms enabling differential regulation of blood flow through these separate beds, in order to meet the varying demands of the embryo during development, is of current interest. The present investigation followed the changes in blood flow distribution during an acute exposure to hypoxia and after α-adrenergic blockade. We monitored heart rate (fH), mean arterial pressure (Pm), and determined relative blood flow distribution (%QÌsys) using colored microspheres. At 70% and 90% of the incubation period hypoxia elicited a bradycardia without changing Pm while %QÌsys was altered only at 70%, increasing to the CAM and liver. Blockade of α-adrenergic responses with phentolamine did not change fH or Pm but increased %QÌsys to the shell. These results show the capacity of embryos to redistribute cardiac output during acute hypoxia, however α-adrenergic receptors seemed to play a relatively small role in embryonic cardiovascular regulation.
Asunto(s)
Adrenérgicos/farmacología , Circulación Sanguínea/fisiología , Embrión no Mamífero/fisiopatología , Hipoxia/fisiopatología , Tortugas/embriología , Tortugas/fisiología , Animales , Presión Arterial/efectos de los fármacos , Circulación Sanguínea/efectos de los fármacos , Embrión no Mamífero/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Flujo Sanguíneo Regional/efectos de los fármacosRESUMEN
Reduced oxygen availability (hypoxia) is a potent stressor during embryonic development, altering the trajectory of trait maturation and organismal phenotype. We previously documented that chronic embryonic hypoxia has a lasting impact on the metabolic response to feeding in juvenile snapping turtles (Chelydra serpentina). Turtles exposed to hypoxia as embryos [10% O2 (H10)] exhibited an earlier and increased peak postprandial oxygen consumption rate, compared with control turtles [21% O2 (N21)]. In the current study, we measured central blood flow patterns to determine whether the elevated postprandial metabolic response in H10 turtles is linked to lasting impacts on convective transport. Five years after hatching, turtles were instrumented to quantify systemic ([Formula: see text]) and pulmonary ([Formula: see text]) blood flows and heart rate (fH) before and after a â¼5% body mass meal. In adult N21 and H10 turtles, fH was increased significantly by feeding. Although total stroke volume (VS,tot) remained at fasted values, this tachycardia contributed to an elevation in total cardiac output ([Formula: see text]). However, there was a postprandial reduction in a net left-right (L-R) shunt in N21 snapping turtles only. Relative to N21 turtles, H10 animals exhibited higher [Formula: see text] due to increased blood flow through the right systemic outflow vessels of the heart. This effect of hypoxic embryonic development, reducing a net L-R cardiac shunt, may support the increased postprandial metabolic rate we previously reported in H10 turtles, and is further demonstration of adult reptile cardiovascular physiology being programmed by embryonic hypoxia.
Asunto(s)
Hipoxia/fisiopatología , Pulmón/irrigación sanguínea , Periodo Posprandial/fisiología , Tortugas/embriología , Tortugas/fisiología , Animales , Gasto Cardíaco , Circulación Coronaria , Embrión no Mamífero/fisiopatología , Frecuencia Cardíaca/fisiología , Hipoxia/embriologíaRESUMEN
During embryonic development, environmental perturbations can affect organisms' developing phenotype, a process known as developmental plasticity. Resulting phenotypic changes can occur during discrete, critical windows of development. Critical windows are periods when developing embryos are most susceptible to these perturbations. We have previously documented that hypoxia reduces embryo size and increases relative heart mass in American alligator, and this study identified critical windows when hypoxia altered morphological, cardiovascular function and cardiac gene expression of alligator embryos. We hypothesized that incubation in hypoxia (10% O2) would increase relative cardiac size due to cardiac enlargement rather than suppression of somatic growth. We exposed alligator embryos to hypoxia during discrete incubation periods to target windows where the embryonic phenotype is altered. Hypoxia affected heart growth between 20 and 40% of embryonic incubation, whereas somatic growth was affected between 70 and 90% of incubation. Arterial pressure was depressed by hypoxic exposure during 50-70% of incubation, whereas heart rate was depressed in embryos exposed to hypoxia during a period spanning 70-90% of incubation. Expression of Vegf and PdgfB was increased in certain hypoxia-exposed embryo treatment groups, and hypoxia toward the end of incubation altered ß-adrenergic tone for arterial pressure and heart rate. It is well known that hypoxia exposure can alter embryonic development, and in the present study, we have identified brief, discrete windows that alter the morphology, cardiovascular physiology, and gene expression in embryonic American alligator.
Asunto(s)
Caimanes y Cocodrilos/embriología , Cardiomegalia/embriología , Cardiomegalia/fisiopatología , Embrión no Mamífero/fisiopatología , Hipoxia/embriología , Hipoxia/fisiopatología , Animales , Presión Sanguínea , Embrión no Mamífero/embriología , Frecuencia CardíacaRESUMEN
Studies of embryonic and hatchling reptiles have revealed marked plasticity in morphology, metabolism, and cardiovascular function following chronic hypoxic incubation. However, the long-term effects of chronic hypoxia have not yet been investigated in these animals. The aim of this study was to determine growth and postprandial O2 consumption (VÌo2), heart rate (fH), and mean arterial pressure (Pm, in kPa) of common snapping turtles (Chelydra serpentina) that were incubated as embryos in chronic hypoxia (10% O2, H10) or normoxia (21% O2, N21). We hypothesized that hypoxic development would modify posthatching body mass, metabolic rate, and cardiovascular physiology in juvenile snapping turtles. Yearling H10 turtles were significantly smaller than yearling N21 turtles, both of which were raised posthatching in normoxic, common garden conditions. Measurement of postprandial cardiovascular parameters and O2 consumption were conducted in size-matched three-year-old H10 and N21 turtles. Both before and 12 h after feeding, H10 turtles had a significantly lower fH compared with N21 turtles. In addition, VÌo2 was significantly elevated in H10 animals compared with N21 animals 12 h after feeding, and peak postprandial VÌo2 occurred earlier in H10 animals. Pm of three-year-old turtles was not affected by feeding or hypoxic embryonic incubation. Our findings demonstrate that physiological impacts of developmental hypoxia on embryonic reptiles continue into juvenile life.
Asunto(s)
Sistema Cardiovascular/fisiopatología , Hipoxia/fisiopatología , Tortugas , Adaptación Fisiológica , Factores de Edad , Animales , Presión Arterial , Biomarcadores/sangre , Glucemia/metabolismo , Peso Corporal , Sistema Cardiovascular/embriología , Sistema Cardiovascular/crecimiento & desarrollo , Sistema Cardiovascular/metabolismo , Ingestión de Alimentos , Embrión no Mamífero/fisiopatología , Metabolismo Energético , Frecuencia Cardíaca , Hipoxia/sangre , Hipoxia/embriología , Ácido Láctico/sangre , Consumo de Oxígeno , Fenotipo , Factores de Tiempo , Tortugas/sangre , Tortugas/embriología , Tortugas/crecimiento & desarrolloRESUMEN
New strategies are needed to address the data gap between the bioactivity of chemicals in the environment versus existing hazard information. We address whether a high-throughput screening (HTS) system using a vertebrate organism (embryonic zebrafish) can characterize chemical-elicited behavioral responses at an early, 24 hours post-fertilization (hpf) stage that predict teratogenic consequences at a later developmental stage. The system was used to generate full concentration-response behavioral profiles at 24 hpf across 1060 ToxCast™ chemicals. Detailed, morphological evaluation of all individuals was performed as experimental follow-up at 5 days post-fertilization (dpf). Chemicals eliciting behavioral responses were also mapped against external HTS in vitro results to identify specific molecular targets and neurosignalling pathways. We found that, as an integrative measure of normal development, significant alterations in movement highlighted active chemicals representing several modes of action. These early behavioral responses were predictive for 17 specific developmental abnormalities and mortality measured at 5 dpf, often at lower (i.e., more potent) concentrations than those at which morphological effects were observed. Therefore, this system can provide rapid characterization of chemical-elicited behavioral responses at an early developmental stage that are predictive of observable adverse effects later in life.
Asunto(s)
Conducta Animal/efectos de los fármacos , Embrión no Mamífero/anomalías , Embrión no Mamífero/efectos de los fármacos , Sustancias Peligrosas/toxicidad , Teratógenos/toxicidad , Pez Cebra/embriología , Animales , Relación Dosis-Respuesta a Droga , Embrión no Mamífero/fisiopatología , Ensayos Analíticos de Alto Rendimiento , Valor Predictivo de las Pruebas , Pez Cebra/anomalíasRESUMEN
Objective: To investigate the effect of lmna gene down-regulation on early hematopoietic development of zebrafish. Methods: Phosphorodiamidate morpholino oligomer (PMO) technology was used to downregulate lmna gene expression in Zebrafish. Zebrafish embryos injected phosphorodiamidate morpholino antisense oligonucleotide of lmna gene mRNA by microinjection at unicellular stage were taken as the experimental group, and those injected meaningless phosphorodiamidate morpholino antisense oligonucleotide were taken as the control. The embryos were collected at 18, 24, 30 and 36 hpf after the fertilization. The real-time fluorescent quantitative PCR (RT-PCR) and whole embryo in situ hybridization methods were used to detect the expression of myeloid hematopoietic transcription factor pu. 1 and erythroid hematopoietic transcription factor gata1 in zebrafish. Results: RT-PCR showed that the expressions of pu. 1 and gata1 decreased in the experimental group compared with the control group (all P<0.05). Whole embryo in situ hybridization showed that the blue-black positive hybridization signals of pu. 1 and gata1 in experimental group were shallow than those in the control group. Conclusion: Myeloid hematopoietic and erythroid hematopoietic of zebrafish are blocked with the downregulation of lmna gene.
Asunto(s)
Regulación hacia Abajo/genética , Factor de Transcripción GATA1/genética , Lamina Tipo A/fisiología , Proteínas Proto-Oncogénicas/genética , Transactivadores/genética , Pez Cebra/genética , Animales , Embrión no Mamífero/fisiopatología , Factor de Transcripción GATA1/metabolismo , Técnicas de Silenciamiento del Gen , Hematopoyesis , Hibridación in Situ , Lamina Tipo A/genética , Proteínas Proto-Oncogénicas/metabolismo , Transactivadores/metabolismo , Pez Cebra/embriologíaRESUMEN
Wound repair on the cellular and multicellular levels is essential to the survival of complex organisms. In order to avoid further damage, prevent infection, and restore normal function, cells and tissues must rapidly seal and remodel the wounded area. The cytoskeleton is an important component of wound repair in that it is needed for actomyosin contraction, recruitment of repair machineries, and cell migration. Recent use of model systems and high-resolution microscopy has provided new insight into molecular aspects of the cytoskeletal response during wound repair. Here we discuss the role of the cytoskeleton in single-cell, embryonic, and adult repair, as well as the striking resemblance of these processes to normal developmental events and many diseases.
Asunto(s)
Citoesqueleto/fisiología , Cicatrización de Heridas/fisiología , Actomiosina/fisiología , Animales , Cadherinas/fisiología , Señalización del Calcio/fisiología , Membrana Celular/fisiología , Proteínas del Citoesqueleto/fisiología , Embrión de Mamíferos/fisiopatología , Embrión no Mamífero/lesiones , Embrión no Mamífero/fisiopatología , Humanos , Modelos Biológicos , Morfogénesis/fisiología , Lesiones Prenatales/fisiopatologíaRESUMEN
We used arterial tyramine injections to study development of sympathetic actions on in vivo heart rate and blood pressure in embryonic, hatching and yearling female American alligators. Tyramine is a pharmacological tool for understanding comparative and developmental sympathetic regulation of cardiovascular function, and this indirect sympathomimetic agent causes endogenous neuronal catecholamine release, increasing blood pressure and heart rate. Arterial tyramine injection in hatchling and yearling alligators caused the typical vertebrate response - rise in heart rate and blood pressure. However, in embryonic alligators, tyramine caused a substantial and immediate bradycardia at both 70% and 90% of embryonic development. This embryonic bradycardia was accompanied by hypotension, followed by a sustained hypertension similar to the hatchling and juvenile responses. Pretreatment with atropine injection (cholinergic receptor blocker) eliminated the embryonic hypotensive bradycardia, and phentolamine pretreatment (α-adrenergic receptor blocker) eliminated the embryonic hypotensive and hypertensive responses but not the bradycardia. In addition, hexamethonium pretreatment (nicotinic receptor blocker) significantly blunted embryos' bradycardic tyramine response. However, pretreatment with 6-hydroxydopamine, a neurotoxin that destroys catecholaminergic terminals, did not eliminate the embryonic bradycardia. Tyramine likely stimulated a unique embryonic response - neurotransmitter release from preganglionic nerve terminals (blocked with hexamethonium) and an acetylcholine mediated bradycardia with a secondary norepinephrine-dependent sustained hypertension. In addition, tyramine appears to stimulate sympathetic nerve terminals directly, which contributed to the overall hypertension in the embryonic, hatchling and yearling animals. Data demonstrated that humoral catecholamine control of cardiovascular function was dominant over the immature parasympathetic nervous system in developing alligator embryos, and suggested that sympathetic and parasympathetic nerve terminals were present and developing in ovo but were not tonically active.
Asunto(s)
Caimanes y Cocodrilos/fisiología , Sistema Cardiovascular/inervación , Hemodinámica/fisiología , Sistema Nervioso Simpático/fisiología , Caimanes y Cocodrilos/embriología , Caimanes y Cocodrilos/crecimiento & desarrollo , Análisis de Varianza , Animales , Atropina/administración & dosificación , Atropina/farmacología , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Sistema Cardiovascular/embriología , Sistema Cardiovascular/crecimiento & desarrollo , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/embriología , Embrión no Mamífero/fisiopatología , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Frecuencia Cardíaca/fisiología , Hemodinámica/efectos de los fármacos , Inyecciones Intraarteriales , Parasimpatolíticos/administración & dosificación , Parasimpatolíticos/farmacología , Fentolamina/administración & dosificación , Fentolamina/farmacología , Simpatomiméticos/administración & dosificación , Simpatomiméticos/farmacología , Factores de Tiempo , Tiramina/administración & dosificación , Tiramina/farmacología , Estados UnidosRESUMEN
BACKGROUND: The trajectory to heart defects may start in tubular and looping heart stages when detailed analysis of form and function is difficult by currently available methods. We used a novel method, Doppler optical coherence tomography (OCT), to follow changes in cardiovascular function in quail embryos during acute hypoxic stress. Chronic fetal hypoxia is a known risk factor for congenital heart diseases (CHDs). Decreased fetal heart rates during maternal obstructive sleep apnea suggest that studying fetal heart responses under acute hypoxia is warranted. RESULTS: We captured responses to hypoxia at the critical looping heart stages. Doppler OCT revealed detailed vitelline arterial pulsed Doppler waveforms. Embryos tolerated 1 hr of hypoxia (5%, 10%, or 15% O(2) ), but exhibited changes including decreased systolic and increased diastolic duration in 5 min. After 5 min, slower heart rates, arrhythmic events and an increase in retrograde blood flow were observed. These changes suggested slower filling of the heart, which was confirmed by four-dimensional Doppler imaging of the heart itself. CONCLUSIONS: Doppler OCT is well suited for rapid noninvasive screening for functional changes in avian embryos under near physiological conditions. Analysis of the accessible vitelline artery sensitively reflected changes in heart function and can be used for rapid screening. Acute hypoxia caused rapid hemodynamic changes in looping hearts and may be a concern for increased CHD risk.
Asunto(s)
Sistema Cardiovascular/fisiopatología , Embrión no Mamífero/irrigación sanguínea , Embrión no Mamífero/fisiopatología , Hipoxia Fetal/fisiopatología , Cardiopatías Congénitas/embriología , Hemodinámica , Animales , Cardiopatías Congénitas/fisiopatología , Frecuencia Cardíaca , Codorniz , Tomografía de Coherencia ÓpticaRESUMEN
The first detection of avian nephritis virus (ANV) in goose embryos and of turkey astrovirus-1 (TAstV-1) in duck embryos is described. Intestinal samples from duck and goose embryos from five duck and four goose flocks in Croatia were tested by polymerase chain reaction for the presence of ANV, TAstV-1, turkey astrovirus-2, chicken astrovirus, duck astrovirus and also for the presence of avian reovirus, Derzsy's disease virus and duck enteritis virus. The kidneys from duck and goose embryos were also tested for ANV, while liver samples were tested for duck astrovirus. Duck embryos were also tested to detect duck circovirus and goose embryos for the presence of goose circovirus and goose haemorrhagic polyomavirus. All embryos were in the final stage of incubation and were characterized by moderate to markedly retarded growth. ANV was confirmed in the intestines and kidneys of embryos from two duck and two goose flocks and TAstV-1 was found in embryos from two duck flocks. One duck flock was positive for both ANV and TAstV-1. No other viruses were found in tested flocks. Phylogenetic analysis based on the ANV polymerase gene fragment of ANV sequences detected in duck and goose embryos revealed greatest similarity (88.1 to 97.2%) with ANV isolates from chickens. Further, the existence of at least two types of ANV circulating in Croatian duck and goose flocks was confirmed. Based on the phylogenetic analysis of the portion of TAstV-1 polymerase gene, two detected TAstV-1 nucleotide sequences were 99.5% similar. Compared with six TAstV-1 sequences, Croatian sequences showed one unique nucleotide change. In addition to other possible causes of stunted growth and late embryonic death, these findings suggest that ANV and/or TAstV-1 infection may be a contributing factor in the pre-hatching mortality of ducklings and goslings.
Asunto(s)
Infecciones por Astroviridae/veterinaria , Patos , Embrión no Mamífero/virología , Gansos , Enfermedades de las Aves de Corral/embriología , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/virología , Animales , Infecciones por Astroviridae/embriología , Infecciones por Astroviridae/epidemiología , Avastrovirus/genética , Secuencia de Bases , Croacia/epidemiología , Embrión no Mamífero/fisiopatología , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , Análisis de Secuencia de ADN/veterinaria , Homología de SecuenciaRESUMEN
Skeletal muscle has a remarkable ability to regenerate following injury, which is driven by obligate tissue resident muscle stem cells. Following injury, the muscle stem cell is activated and undergoes cell proliferation to generate a pool of myoblasts, which subsequently differentiate to form new muscle fibers. In many muscle wasting conditions, including muscular dystrophy and ageing, this process is impaired resulting in the inability of muscle to regenerate. The process of muscle regeneration in zebrafish is highly conserved with mammalian systems providing an excellent system to study muscle stem cell function and regeneration, in muscle wasting conditions such as muscular dystrophy. Here, we present a method to examine muscle regeneration in zebrafish models of muscle disease. The first step involves the use of a genotyping platform that allows the determination of the genotype of the larvae prior to eliciting an injury. Having determined the genotype, the muscle is injured using a needle stab, following which polarizing light microscopy is used to determine the extent of muscle regeneration. We therefore provide a high throughput pipeline which allows the examination of muscle regeneration in zebrafish models of muscle disease.
Asunto(s)
Músculo Esquelético/fisiopatología , Enfermedades Musculares/fisiopatología , Regeneración/fisiología , Pez Cebra/fisiología , Animales , Modelos Animales de Enfermedad , Embrión no Mamífero/fisiopatología , Genotipo , Laminina/deficiencia , Laminina/metabolismo , Larva/fisiología , Músculo Esquelético/diagnóstico por imagen , Músculo Esquelético/patología , Enfermedades Musculares/patología , Pez Cebra/embriologíaRESUMEN
The aim of this research was to assess the combined effects of three heavy metals (copper, lead, cadmium) on the fertilization and offspring quality of the sea urchin Paracentrotus lividus at EC50, NOEL, and EC1 concentrations. The observed data were compared with the predictions derived from approaches of Concentration Addition (CA) and Independent Action (IA) in order to evaluate the proper prediction of the observed mixture toxic effect. The P. lividus embryotoxicity of trace metals decreases as follows: Cu > Pb > Cd at all toxicity concentration tested. EC50 mixture revealed less toxic only than Cu; EC50 was 0.80 (± 0.07) mg/l, the offspring malformations were mainly P1 type (skeletal alterations) up to 20% mixture concentration, and P2 type from 70% concentration. The NOEL and EC1 mixtures evidenced that all compounds contribute to the overall toxicity, even if present at low concentrations: the former EC50 was 0.532 (± 0.058) mg/l and the latter was 1.081 (± 0.240) mg/l. The developmental defects observed were mainly P1 type in both mixtures. Both CA and IA models did not accurately predict mixture toxicity for EC50 and NOEL mixtures. Instead, EC1 mixture effects seemed well represented by the IA model. The protective action of the CA model, although quite accurate when applied to simple biological systems like algae and bacteria, but failed to represent the worst-case in this study with more complex organisms. It would be useful to introduce in the models one or more factors that take into account the complexity of these biological systems.
Asunto(s)
Cadmio/toxicidad , Cobre/toxicidad , Plomo/toxicidad , Paracentrotus/embriología , Animales , Embrión no Mamífero/fisiopatología , Desarrollo Embrionario , Fertilización , Modelos Biológicos , Nivel sin Efectos Adversos Observados , Paracentrotus/fisiología , Pruebas de ToxicidadRESUMEN
Mutations in the CFTR gene cause cystic fibrosis (CF) with myocardial dysfunction. However, it remains unknown whether CF-related heart disease is a secondary effect of pulmonary disease, or an intrinsic primary defect in the heart. Here, we used zebrafish, which lack lung tissue, to investigate the role of CFTR in cardiogenesis. Our findings demonstrated that the loss of CFTR impairs cardiac development from the cardiac progenitor stage, resulting in cardiac looping defects, a dilated atrium, pericardial edema, and a decrease in heart rate. Furthermore, we found that cardiac development was perturbed in wild-type embryos treated with a gating-specific CFTR channel inhibitor, CFTRinh-172, at the blastula stage of development, but not at later stages. Gene expression analysis of blastulas indicated that transcript levels, including mRNAs associated with cardiovascular diseases, were significantly altered in embryos derived from cftr mutants relative to controls. To evaluate the role of CFTR in human heart failure, we performed a genetic association study on individuals with dilated cardiomyopathy and found that the I556V mutation in CFTR, which causes a channel defect, was associated with the disease. Similar to other well-studied channel-defective CFTR mutants, CFTR I556V mRNA failed to restore cardiac dysplasia in mutant embryos. The present study revealed an important role for the CFTR ion channel in regulating cardiac development during early embryogenesis, supporting the hypothesis that CF-related heart disease results from an intrinsic primary defect in the heart.
Asunto(s)
Cardiomiopatía Dilatada/genética , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Corazón/crecimiento & desarrollo , Pez Cebra/genética , Animales , Cardiomiopatía Dilatada/fisiopatología , Modelos Animales de Enfermedad , Embrión no Mamífero/metabolismo , Embrión no Mamífero/fisiopatología , Desarrollo Embrionario/genética , Corazón/fisiopatología , Humanos , Mutación/genética , Pez Cebra/crecimiento & desarrolloRESUMEN
BACKGROUND: The cardiotoxicity of isoniazid on zebrafish embryos and its underlying mechanism is unclear. METHODS: Here, we exposed zebrafish embryos at 4 h post-fertilization to different levels of isoniazid and recorded the morphology and number of malformed and dead embryos under the microscope. RESULTS: The high concentration of isoniazid group showed more malformed and dead embryos than the low concentration of isoniazid group and control group. The morphology of the heart and its alteration were visualized using transgenic zebrafish (cmlc2: GFP) and confirmed by in situ hybridization. The negative effects of isoniazid on the developing heart were characterized by lower heart rate and more heart looping disorders. Mechanistically, PCR showed decreased expression of heart-specific transcription factors when exposed to isoniazid. Oxidative stress was induced by isoniazid in cardiomyocytes, mediated by decreased activities of catalase and superoxide dismutase, which were rescued by scavengers of reactive oxygen species. CONCLUSION: In conclusion, this study demonstrated that isoniazid led to heart looping disturbance by the downregulation of cardiac-specific transcription factors and induction of cardiomyocyte apoptosis.
Asunto(s)
Antituberculosos/efectos adversos , Desarrollo Embrionario/efectos de los fármacos , Cardiopatías Congénitas/inducido químicamente , Isoniazida/efectos adversos , Animales , Animales Modificados Genéticamente , Apoptosis/efectos de los fármacos , Regulación hacia Abajo , Embrión no Mamífero/anomalías , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/fisiopatología , Cardiopatías Congénitas/genética , Cardiopatías Congénitas/fisiopatología , Frecuencia Cardíaca/efectos de los fármacos , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Factores de Transcripción/genética , Pez CebraRESUMEN
CHARGE syndrome is an autosomal dominant congenital disorder caused primarily by mutations in the CHD7 gene. Using a small molecule screen in a zebrafish model of CHARGE syndrome, we identified 4 compounds that rescue embryos from disease-like phenotypes. Our screen yielded DAPT, a Notch signaling inhibitor that could ameliorate the craniofacial, cranial neuronal and myelination defects in chd7 morphant zebrafish embryos. We discovered that Procainamide, an inhibitor of DNA methyltransferase 1, was able to recover the pattern of expression of isl2a, a cranial neuronal marker while also reducing the effect on craniofacial cartilage and myelination. M344, an inhibitor of Histone deacetylases had a strong recovery effect on craniofacial cartilage defects and could also modestly revert the myelination defects in zebrafish embryos. CHIC-35, a SIRT1 inhibitor partially restored the expression of isl2a in cranial neurons while causing a partial reversion of myelination and craniofacial cartilage defects. Our results suggest that a modular approach to phenotypic rescue in multi-organ syndromes might be a more successful approach to treat these disorders. Our findings also open up the possibility of using these compounds for other disorders with shared phenotypes.
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Síndrome CHARGE/tratamiento farmacológico , Síndrome CHARGE/fisiopatología , ADN Helicasas/genética , Proteínas de Unión al ADN/genética , Dipéptidos/farmacología , Procainamida/farmacología , Vorinostat/farmacología , Proteínas de Pez Cebra/genética , Pez Cebra/embriología , Animales , Animales Modificados Genéticamente , Síndrome CHARGE/genética , Cartílago/efectos de los fármacos , Cartílago/patología , ADN (Citosina-5-)-Metiltransferasa 1/antagonistas & inhibidores , ADN Helicasas/metabolismo , Proteínas de Unión al ADN/metabolismo , Dipéptidos/uso terapéutico , Modelos Animales de Enfermedad , Embrión no Mamífero/diagnóstico por imagen , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/metabolismo , Embrión no Mamífero/fisiopatología , Técnicas de Silenciamiento del Gen , Inhibidores de Histona Desacetilasas/farmacología , Inhibidores de Histona Desacetilasas/uso terapéutico , Proteínas con Homeodominio LIM/genética , Proteínas con Homeodominio LIM/metabolismo , Fibras Nerviosas Mielínicas/efectos de los fármacos , Fibras Nerviosas Mielínicas/patología , Neuronas/efectos de los fármacos , Neuronas/patología , Procainamida/uso terapéutico , Receptores Notch/antagonistas & inhibidores , Sirtuina 1/antagonistas & inhibidores , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vorinostat/uso terapéutico , Pez Cebra/genética , Proteínas de Pez Cebra/metabolismoRESUMEN
3,3',4,4',5-Pentachlorinated biphenyls 126 (PCB126) is a global environmental contaminant that can induce cellular oxidative stress. We investigated whether vitamin E can protect against toxicity from PCB126 during zebrafish (Danio rerio) development. Zebrafish embryos were exposed to 100nM PCB126 and compared with a second group that was co-exposed with 100muM vitamin E until 5 days post fertilization. PCB126 induced pericardial sac edema, yolk sac edema, and growth retardation in zebrafish embyos. In contrast, vitamin E co-exposure group did not show any gross changes. Real-time PCR results showed that vitamin E co-exposure group were restored to control group for the expression levels of heat shock protein 70 Cognate, aryl hydrocarbon receptor type-2, cytochrome P450 1A, and superoxide dismutase-1. These data give insights into the use of vitamin E to reduce PCB126-mediated toxicity and into the use of zebrafish embryos for exploring mechanisms underlying the oxidative potential of AHR agonists.
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Antioxidantes/farmacología , Embrión no Mamífero/efectos de los fármacos , Antagonistas de Estrógenos/toxicidad , Bifenilos Policlorados/toxicidad , Vitamina E/farmacología , Contaminantes Químicos del Agua/toxicidad , Pez Cebra , Animales , Antagonismo de Drogas , Edema/inducido químicamente , Edema/patología , Embrión no Mamífero/metabolismo , Embrión no Mamífero/fisiopatología , Desarrollo Embrionario/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Pericardio/efectos de los fármacos , Pericardio/patología , ARN Mensajero/metabolismo , Saco Vitelino/efectos de los fármacos , Saco Vitelino/patología , Proteínas de Pez Cebra/genética , Proteínas de Pez Cebra/metabolismoRESUMEN
Freshwater harmful algal blooms produce a broad array of bioactive compounds, with variable polarity. Acute exposure to cyanotoxins can impact the liver, nervous system, gastrointestinal tract, skin, and immune function. Increasing evidence suggests chronic effects from low-level exposures of cyanotoxins and other associated bioactive metabolites of cyanobacterial origin. These sundry compounds persist in drinking and recreational waters and challenge resource managers in detection and removal. A systematic approach to assess the developmental toxicity of cyanobacterial metabolite standards was employed utilizing a robust and high throughput developmental Danio rerio embryo platform that incorporated a neurobehavioral endpoint, photomotor response. Subsequently, we applied the platform to cyanobacterial bloom surface water samples taken from temperate recreational beaches and tropical lake subsistence drinking water sources as a model approach. Dechorionated Danio rerio embryos were statically immersed beginning at four to six hours post fertilization at environmentally relevant concentrations, and then assessed at 24â¯h and 5â¯days for morbidity, morphological changes, and photomotor response. At least one assessed endpoint deviated significantly for exposed embryos for 22 out of 25 metabolites examined. Notably, the alkaloid lyngbyatoxin-a resulted in profound, dose-dependent morbidity and mortality beginning at 5⯵g/L. In addition, hydrophobic components of extracts from beach monitoring resulted in potent morbidity and mortality despite only trace cyanotoxins detected. The hydrophilic extracts with several order of magnitude higher concentrations of microcystins resulted in no morbidity or mortality. Developmental photomotor response was consistently altered in environmental bloom samples, independent of the presence or concentration of toxins detected in extracts. While limited with respect to more polar compounds, this novel screening approach complements specific fingerprinting of acutely toxic metabolites with robust assessment of developmental toxicity, critical for chronic exposure scenarios.
Asunto(s)
Toxinas Bacterianas/toxicidad , Cianobacterias , Floraciones de Algas Nocivas , Microcistinas/toxicidad , Pez Cebra , Animales , Playas , Embrión no Mamífero/fisiopatología , Lagos , Medición de Riesgo , UruguayRESUMEN
As the older class of brominated flame retardants (BFRs) are phased out of commercial use because of findings of neurotoxicity with developmental exposure, a newer class of flame retardants have been introduced, the organophosphate flame retardants (OPFRs). Presently, little is known about the potential for developmental neurotoxicity or the behavioral consequences of OPFR exposure. Our aim was to characterize the life-long neurobehavioral effects of 4 widely used OPFRs using the zebrafish model. Zebrafish embryos were exposed to 0.1% DMSO (vehicle control); or one of the following treatments; isopropylated phenyl phosphate (IPP) (0.01, 0.03, 0.1, 0.3 µM); butylphenyl diphenyl phosphate (BPDP) (0.003, 0.03, 0.3, 3 µM); 2-ethylhexyl diphenyl phosphate (EHDP) (0.03, 0.3, 1 µM); isodecyl diphenyl phosphate (IDDP) (0.1, 0.3, 1, 10 µM) from 0- to 5-days postfertilization. On Day 6, the larvae were tested for motility under alternating dark and light conditions. Finally, at 5-7 months of age the exposed fish and controls were tested on a battery of behavioral tests to assess emotional function, sensorimotor response, social interaction and predator evasion. These tests showed chemical-specific short-term effects of altered motility in larvae in all of the tested compounds, and long-term impairment of anxiety-related behavior in adults following IPP, BPDP, or EHDP exposures. Our results show that OPFRs may not be a safe alternative to the phased-out BFRs and may cause behavioral impacts throughout the lifespan. Further research should evaluate the risk to mammalian experimental models and humans.