RESUMEN
Because of widespread environmental contamination, there is growing concern that nanoplastics may pose a risk to humans and the environment. Due to their small particle size, nanoplastics may cross the blood-nerve barrier and distribute within the nervous system. The present study systematically investigated the uptake/distribution and developmental/neurobehavioral toxicities of different sizes (80, 200, and 500 nm) of polystyrene nanoplastics (PS) in embryonic and juvenile zebrafish. The results indicate that all three sizes of PS could cross the chorion, adsorb by the yolk, and distribute into the intestinal tract, eye, brain, and dorsal trunk of zebrafish, but with different patterns. The organ distribution and observed developmental and neurobehavioral effects varied as a function of PS size. Although all PS exposures induced cell death and inflammation at the cellular level, only exposures to the larger PS resulted in oxidative stress. Meanwhile, exposure to the 80 nm PS increased the expression of neural and optical-specific mRNAs. Collectively, these studies indicate that early life-stage exposures to PS adversely affect zebrafish neurodevelopment and that the observed toxicities are influenced by particle size.
Asunto(s)
Nanopartículas , Contaminantes Químicos del Agua , Humanos , Animales , Poliestirenos/toxicidad , Poliestirenos/metabolismo , Pez Cebra/metabolismo , Microplásticos/toxicidad , Microplásticos/metabolismo , Contaminantes Químicos del Agua/toxicidad , Nanopartículas/toxicidad , Nanopartículas/metabolismoRESUMEN
Glioblastoma as the most common and aggressive central nervous system tumor in adults. Its prognosis and therapeutic outcome are poor due to the limited understanding of its molecular mechanism. Apolipoprotein C-1 (APOC1) as a member of the apolipoprotein family that acts as a tumor promoter in various cancers. MicroRNA (miRNA) can silence gene expression and suppress tumor progression. However, the role of APOC1 and its upstream miRNA has not been explored in glioblastoma. Two glioblastoma cell lines (U87 and U251) were used to explore the role of APOC1 and its upstream miRNA-660-3p in glioblastoma tumorigenesis in vitro. Cells with APOC1/miRNA-660-3p overexpression or knockdown were assessed for their proliferation, migration, and invasion in vitro, and tumorigenesis in vivo. Gene and protein expression was assessed by qRT-PCR and western blot, respectively. Cell proliferation was assessed by the MTT assay and the EdU and Ki67 staining. Cell migration and invasion were assessed by the transwell assay. Tumorigenesis in vivo was assessed in U87 cells with a xenograft mouse model. APOC1 was overexpressed in glioblastoma compared with normal peritumoral tissue and was inversely related to patient prognosis. APOC1 overexpression promotes cell proliferation, migration, and invasion in vitro. APOC1 inhibition reduced tumor growth in vivo. miRNA-660-3p inhibits tumorigenesis by directly targeting APOC1. Mechanistically, APOC1 drives the malignancy of glioblastoma by activating the TGFß2 signaling pathway. miRNA-660-3p suppresses tumorigenesis by targeting APOC1. Therefore, miRNA-660-3p/APOC1 axis can serve as potential intervention targets in managing glioblastoma progression.
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Glioblastoma , MicroARNs , Adulto , Humanos , Animales , Ratones , Glioblastoma/genética , Carcinogénesis , Transducción de Señal , Transformación Celular Neoplásica , MicroARNs/genética , Modelos Animales de Enfermedad , ApolipoproteínasRESUMEN
Autism spectrum disorder (ASD) is a neurodevelopmental disorder that has become more prevalent in recent years. Environmental endocrine disruptor bisphenol A (BPA) has been linked to ASD. BPA analogues (BPs) are structure-modified substitutes widely used as safer alternatives in consumer products, yet few studies have explored the developmental neurotoxicity (DNT) of BPA analogues. In the present study, we used the larval zebrafish model to assess the DNT effects of BPA and its analogues. Our results showed that many BPA analogues are more toxic than BPA in the embryonic zebrafish assay regarding teratogenic effect and mortality, which may partially due to differences in lipophilicity and/or different substitutes of structural function groups such as CF3, benzene, or cyclohexane. At sublethal concentrations, zebrafish embryos exposed to BPA or BPs also displayed reduced prosocial behavior in later larval development, evidenced by increased nearest neighbor distance (NND) and the interindividual distance (IID) in shoaling, which appears to be structurally independent. An in-depth analysis of BPA, bisphenol F (BPF), and bisphenol S (BPS) revealed macrocephaly and ASD-like behavioral deficits resulting from exposures to sublethal concentrations of these chemicals. The ASD-like behavioral deficits were characterized by hyperactivity, increased anxiety-like behavior, and decreased social contact. Mechanistically, accelerated neurogenesis that manifested by increased cell proliferation, the proportion of newborn mature neurons, and the number of neural stem cells in proliferation, as well as upregulated genes related to the K+ channels, may have contributed to the observed ASD-like morphological and behavioral alterations. Our findings indicate that BPF and BPS may also pose significant risks to ASD development in humans and highlight the importance of a comprehensive assessment of DNT effects for all BPA analogues in the future.
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Trastorno del Espectro Autista , Pez Cebra , Humanos , Animales , Recién Nacido , Compuestos de Bencidrilo/análisis , FenotipoRESUMEN
Benzophenone-3 (BP3) is an organic UV filter widely used in the commercial formulations of various personal care products. It has been detected ubiquitously in the environment and human tissues. Recently, BP3-induced neurotoxicity has been identified as the main health risk to humans and aquatic organisms. However, most research has been focused on embryonic development, and few studies explore chronic lifetime exposure. In the present study, we evaluated the neurotoxicity of lifetime exposure to an environmentally relevant concentration of BP3 in zebrafish. Our findings revealed that continuous BP3 exposure at 10 µg/L (0.04 µM) from 6 h post fertilization (hpf) to adulthood at 5 months led to female-biased social behavioral deficits and learning and memory impairment. These neurobehavioral effects were characterized by decreased prosocial activities in the social preference test and mirror biting assay, and reduced learning and memory in a T-maze test. Furthermore, these effects were accompanied by female-specific decreases in brain weight and brain dopamine concentration, female-biased decrease of neurogenesis in the telencephalon as well as female-specific increases in apoptotic cells and expression levels of genes and proteins related to the apoptosis pathway in the brain. Our results suggest that BP3-induced social behavior and learning/memory deficits are correlated to the cell loss in the telencephalon region of the zebrafish brain.
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Benzofenonas , Pez Cebra , Animales , Humanos , Femenino , Adulto , Benzofenonas/toxicidad , Benzofenonas/metabolismo , Conducta Social , CogniciónRESUMEN
Zebrafish is an economical alternative model for developmental neurotoxicity (DNT) testing. DNT studies in zebrafish have been focused on acute effects; few studies explore enduring neurotoxicity in adults. More recently, gut microbiome has emerged as an important modulator between chemical exposure and neurotoxicity, rendering its necessity to be included in DNT testing. The present study used a well-known dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) as a model chemical to explore long-lasting neurotoxicity in adults after transient exposure during early development. We demonstrated that transient MPTP exposure at 1 µM during a sensitive developmental window of 48-96 h post-fertilization (hpf) altered gut microbiome and led to male-biased locomotion and behavioral deficits in adult fish. The locomotion deficit was manifested as hypoactivity observed in adult males under light conditions or specifically the reduction of fast swim bouts. The social behavioral deficits were characterized by the reduced number of times fish crossed the mirror zone in the mirror response assay and the reduced percent time fish spent at the area proximal to conspecific fish shoal in the social preference test. Gut microbiome analysis revealed that transient MPTP exposure during early development might render fish more susceptible to the colonization of the pathogenic Vibrio. In conclusion, our study revealed that transient MPTP exposure during early development could lead to long-lasting neurotoxicity in adult fish and cause altered gut microbiome composition in both larval and adult fish.
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Microbioma Gastrointestinal , Síndromes de Neurotoxicidad , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/farmacología , Animales , Dopamina/farmacología , Larva , Masculino , Pez CebraRESUMEN
Zebrafish represent an economical alternative to rodents for developmental neurotoxicity (DNT) testing. Mechanistic understanding is the key to successfully translating zebrafish findings to humans. In the present study, we used a well-known dopaminergic (DA) neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) as a model chemical to uncover the molecular pathways for observed DNT effects. To enhance the specificity of potential molecular targets, we restricted our exposure to a concentration that is nonteratogenic yet exhibits high DNT effects and an exposure window sensitive to MPTP. Our DNT assessment based on a battery of motor and social behavioral tests revealed an effective concentration of 1 µM and a sensitive window of 48-96 h postfertilization (hpf) for MPTP-induced hypoactivity. It is worth noting that this hypoactivity persisted into later larval development until 28 dpf. We observed increased cell apoptosis, oxidative stress, and decreased ATP levels in larvae immediately after exposure at 96 hpf. Significant reductions of DA neurons were found in the retina at 72, 96, and 120 hpf. No visible deformity was found in motoneurons at 72, 96, and 120 hpf. Transcriptome analysis uncovered a novel pathway manifested by significant upregulation of genes enriched with erythropoiesis. Sensitive window exposure of MPTP and other DA neurotoxins rotenone and paraquat exhibited a concentration-dependent effect on transcriptional changes of embryonic hemoglobins and anemia. Given that anemia is a significant risk factor for Parkinson's disease and MPTP is known to cause parkinsonism in humans, we concluded that anemia resulting from dysregulation of primitive erythropoiesis during embryonic development might serve as a common mechanism underlying DA neurotoxin-induced DNT effects between zebrafish and humans.
Asunto(s)
Anemia , Intoxicación por MPTP , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/farmacología , Animales , Dopamina/metabolismo , Intoxicación por MPTP/metabolismo , Ratones , Ratones Endogámicos C57BL , Neurotoxinas/metabolismo , Neurotoxinas/toxicidad , Pez Cebra/metabolismoRESUMEN
The widespread commercial application of titanium dioxide nanoparticles (TiO2 NPs) leads to ubiquitous presence of TiO2 NPs in the aquatic environment, which highlights the necessity to determine their potential adverse effects on aquatic organisms. The developing nerve system is particularly susceptible to environment perturbation. However, few studies have explored the developmental neurobehavioral toxicity of TiO2 NPs, especially at smaller particle size ranges (≤20 nm) that have relatively longer retention time in the water column. In this study, zebrafish embryos were exposed to non-teratogenic concentrations of 0.1 and 1 mg/L TiO2 NPs (average size of 14-20 nm) from 8 to 108 h post-fertilization (hpf) followed by various assessments at different time points up to 12 days post-fertilization (dpf). Our findings revealed that 1 mg/L TiO2 NPs perturbed the motor and social behaviors in larval zebrafish. These behavioral changes were characterized by decreased swimming speed in a locomotor response test at 5 dpf, increased travel distance in a flash stimulus test at 5 dpf, increased preference to the light zone in a light/dark preference test at 10 dpf, and increased mirror attack and percent time spent in the mirror zone in a mirror stimulus response assay at 12 dpf. Mechanistic examinations at 5 dpf revealed elevated cell apoptosis and oxidative stress. Cell apoptosis was characterized by increased acridine orange (AO) positive cells in the olfactory region and neuromasts of the lateral line system. Oxidative stress was characterized by increased lipid peroxidation, increased ROS production, and upregulated catalase (cat) gene expression. In addition, TiO2 NP exposure also upregulated genes associated with the developmental nervous system such as the growth associated protein 43 (gap43) and proliferating cell nuclear antigen (pcna). Our results suggest that the neurobehavioral changes in larvae exposed to 1 mg/L TiO2 NPs during early development may result from cell apoptosis and oxidative stress induced neuronal damages.
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Nanopartículas , Contaminantes Químicos del Agua , Animales , Nanopartículas/toxicidad , Estrés Oxidativo , Titanio/toxicidad , Contaminantes Químicos del Agua/toxicidad , Pez CebraRESUMEN
Aristolochic acids (AA) are nitrophenanthrene carboxylic acids found in plants of the Aristolochiaceae family. Humans are exposed to AA by deliberately taking herbal medicines or unintentionally as a result of environmental contamination. AA is notorious for its nephrotoxicity, however, fewer studies explore potential neurotoxicity associated with AA exposure. The developing nervous system is vulnerable to xenobiotics, and pregnant women exposed to AA may put their fetuses at risk. In the present study, we used the embryonic zebrafish model to evaluate the developmental neurotoxicity associated with AA exposure. At non-teratogenic concentrations (≤ 4 µM), continuous AA exposure from 8 to 120 hours post fertilization (hpf) resulted in larval hyperactivity that was characterized by increased moving distance, elevated activity and faster swimming speeds in several behavioral assays. Further analysis revealed that 8-24 hpf is the most sensitive exposure window for AA-induced hyperactivity. AA exposures specifically increased motor neuron proliferation, increased apoptosis in the eye, and resulted in cellular oxidative stress. In addition, AA exposures increased larval eye size and perturbed the expression of vision genes. Our study, for the first time, demonstrates that AA is neurotoxic to the developmental zebrafish with a sensitive window distinct from its well-documented nephrotoxicity.
RESUMEN
In recent years, an increasing number of studies have shown that fibroblast growth factor 12 (FGF12) plays important roles in regulating neural development and function. Importantly, changes of FGF12 expression are thought to be related to the pathophysiology of many neurological diseases. However, little research has been performed to explore the protective effect of FGF12 on nerve damage. This study aims to explore its neuroprotective effects using our recombinant humanized FGF12 (rhFGF12). The hFGF12 gene was cloned and ligated into an expression vector to construct a recombinant plasmid pET-3a-hFGF12. Single colonies were screened to obtain high expression engineering strains, and fermentation and purification protocols for rhFGF12 were designed and optimized. The biological activities and related mechanisms of rhFGF12 were investigated by MTT assay using NIH3T3 and PC12 cell lines. The in vitro neurotoxicity model of H2O2-induced oxidative injury in PC12 cells was established to explore the protective effects of rhFGF12. The results indicate that the beneficial effects of rhFGF12 were most likely achieved by promoting cell proliferation and reducing apoptosis. Moreover, a transgenic zebrafish (islet) with strong GFP fluorescence in the motor neurons of the hindbrain was used to establish a central injury model caused by mycophenolate mofetil (MMF). The results suggested that rhFGF12 could ameliorate central injury induced by MMF in zebrafish. In conclusion, we have established an efficient method to express and purify active rhFGF12 using an Escherichia coli expression system. Besides, rhFGF12 plays a protective effect of on nerve damage, and it provides a promising therapeutic approach for nerve injury. KEY POINTS: ⢠Effective expression and purification of bioactive rhFGF12 protein in E. coli. ⢠ERK/MAPK pathway is involved in rhFGF12-stimulated proliferation on PC12 cells. ⢠The rhFGF12 has the neuroprotective effects by inhibiting apoptosis.
Asunto(s)
Fármacos Neuroprotectores , Animales , Escherichia coli/genética , Factores de Crecimiento de Fibroblastos/genética , Humanos , Peróxido de Hidrógeno , Ratones , Células 3T3 NIH , Fármacos Neuroprotectores/farmacología , Ratas , Pez CebraRESUMEN
Aging plays an important role in many diseases, including breast cancer. Aged mammary stem/progenitor cells are perceived to be the cells of origin in breast tumorigenesis; however, the extensive use of mice who have aged naturally for research is hampered by cost, time, disease complications, and high mortality. In this study, we characterized murine mammary stem/progenitor cells in a D-galactose-induced accelerated aging model and compared them with findings from our earlier study on mice from natural aging. Our results showed that mammary glands in the D-galactose-induced aging model mimic natural aging in terms of pathological changes, epithelial cell composition, and mammary stem/progenitor cell function. These changes are accompanied by elevated inflammatory responses both systemically in the blood and locally in the mammary glands, which is similar in mice who age naturally. Our study for the first time evaluated the mammary glands and mammary stem/progenitor function in a D-galactose-induced aging model in rodents, and our findings suggest that D-galactose treatment can be used as a surrogate to study the role aged stem/progenitor cells play in breast tumorigenesis.
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Envejecimiento/efectos de los fármacos , Células Epiteliales/patología , Glándulas Mamarias Animales/patología , Neoplasias Mamarias Experimentales/patología , Células Madre/patología , Envejecimiento/patología , Animales , Diferenciación Celular/efectos de los fármacos , Transformación Celular Neoplásica/efectos de los fármacos , Transformación Celular Neoplásica/patología , Células Epiteliales/efectos de los fármacos , Femenino , Galactosa/administración & dosificación , Galactosa/toxicidad , Glándulas Mamarias Animales/citología , Glándulas Mamarias Animales/efectos de los fármacos , Neoplasias Mamarias Experimentales/inducido químicamente , Ratones , Células Madre/efectos de los fármacosRESUMEN
Both tetrabromobisphenol A (TBBPA) and titanium dioxide nanoparticle (TiO2 NP) have widespread commercial applications, resulting in their ubiquitous co-presence in the environment and biota. Although environmental chemicals exist as mixtures, toxicity studies are nearly always conducted with single chemicals. Few studies explore potential interactions of different chemical mixtures. In this study, we employ the sensitive developing nerve system in zebrafish to assess the neurotoxicity of TBBPA/TiO2 NP mixtures. Specifically, zebrafish embryos were exposed to solvent control (0.1% DMSO), 2 µM TBBPA, 0.1 mg/L TiO2 NP, and their mixture from 8 to 120 h post fertilization (hpf), and motor/social behavioral assessments were conducted on embryos/larvae at different developmental stages. Our results showed that TBBPA/TiO2 NP single or co-exposures increased spontaneous movement, decreased touch response and swim speed, and affected social behaviors of light/dark preference, shoaling, mirror attack and social contact. In particular, many of these phenotypes were manifested with higher magnitude of changes from the mixture exposure. These behavioral deficits were also accompanied with increased cell death in olfactory region and neuromasts in the lateral line system, increased ROS in gallbladder, pancreas, liver, and intestine, as well as increased lipid peroxidation and decreased ATP levels in whole larval tissue homogenates. Further, genes coding for key cell apoptosis marker and antioxidant enzyme were significantly upregulated by these two chemicals, in particular to their mixture. Interestingly, the co-presence of TBBPA also increased the mean particle size of TiO2 NP in the exposure solutions and the TiO2 NP content in larval tissue. Together, our analysis suggests that TBBPA/TiO2 NP induced behavioral changes may be due to physical accumulation of these two chemicals in the target organs, and TiO2 NP may serve as carriers for increased accumulation of TBBPA. To conclude, we demonstrated that TBBPA/TiO2 NP together cause increased bioaccumulation of TiO2, and heightened responses in behavior, cell apoptosis and oxidative stress. Our findings also highlight the importance of toxicity assessment using chemical mixtures.
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Nanopartículas/toxicidad , Bifenilos Polibrominados/toxicidad , Titanio/toxicidad , Contaminantes Químicos del Agua/toxicidad , Pez Cebra/fisiología , Animales , Antioxidantes/metabolismo , Apoptosis , Bioacumulación , Larva/metabolismo , Peroxidación de Lípido , Hígado/metabolismo , Estrés Oxidativo , Conducta SocialRESUMEN
Behavior phenotypes are a powerful means of uncovering subtle xenobiotic chemical impacts on vertebrate nervous system development. Rodents manifest complex and informative behavior phenotypes but are generally not practical models in which to screen large numbers of chemicals. Zebrafish recapitulate much of the behavioral complexity of higher vertebrates, develop externally and are amenable to assay automation. Short duration automated assays can be leveraged to screen large numbers of chemicals or comprehensive dose-response for fewer chemicals. Here we describe a series of mostly automated assays including larval photomotor response, strobe light response, blue color avoidance, shoaling and mirror stimulus-response performed on the ZebraBox (ViewPoint Behavior Technologies) instrument platform. To explore the sensitivity and uniqueness of each assay endpoint, larval cohorts from 5 to 28 days post fertilization were acutely exposed to several chemicals broadly understood to impact different neuro-activities. We highlight the throughput advantages of using the same instrument platform for multiple assays and the ability of different assays to detect unique phenotypes among different chemicals.
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Conducta Animal/fisiología , Ensayos Analíticos de Alto Rendimiento/métodos , Actividad Motora/efectos de los fármacos , Animales , Automatización de Laboratorios/métodos , Embrión no Mamífero , Larva/metabolismo , Actividad Motora/fisiología , Fenotipo , Conducta Social , Pez Cebra/metabolismo , Proteínas de Pez Cebra/metabolismoRESUMEN
Benzophenone-3 (BP3 or oxybenzone) is an organic UV filter that has been widely used in personal care products. Its frequent detection in the environment and humans as well as its structural similarity to estradiol have prompted most research focus on its endocrine effect. However, these effects are usually associated with concentrations 10-100 fold higher than its environmental relevant concentrations. Few studies explore its adverse effects at environmental relevant concentrations. In the present study, we evaluated the developmental neurotoxic (DNT) effects of low concentration BP3 exposure during a sensitive developmental window in zebrafish. Our findings revealed that BP3 exposure at 10 µg/L (0.04 µM) during 6-24 h post fertilization (hpf) led to various DNT effects such as increased spontaneous movement at 21 and 24 hpf, decreased touch response at 27 hpf, heightened hyperactivity in locomotor response at 5 day post fertilization (dpf), decreased shoaling behavior at 11 dpf and decreased mirror attacks at 12 dpf. These effects were accompanied with decreased axonal growth at 27 hpf, decreased cell proliferation and increased cell apoptosis in the head region of larval zebrafish immediately after BP3 exposure at 24 hpf, and increased expression of retinoid X receptor gene rxrgb at 5 dpf. Interestingly, rxrgb knockdown through morpholino injection largely restored most of BP3-induced DNT effects, axonal growth delay, cell proliferation and cell apoptosis, suggesting that BP3-induced DNT effects are likely mediated through the Rxrgb receptor. In considering with recent findings on the endocrine effects of BP3, we conclude that BP3 at environmental relevant concentrations has limited estrogenic effect, but is neurotoxic to developing embryos in zebrafish.
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Embrión no Mamífero , Pez Cebra , Animales , Benzofenonas , Relación Dosis-Respuesta a Droga , LarvaRESUMEN
Alkyl phenanthrene (A-Phen) and Dechlorane Plus (DP) are ubiquitous environmental pollutants that widely co-exist in the environment. It has been established that both A-Phen and DP elicit neurotoxicity, but the potential interactive toxicity of these contaminants is not well-known. To determine whether a mixture of A-Phen and DP would exhibit interactive effects on neurodevelopment, we co-exposed 3-methylphenanthrene (3-MP), a representative of A-Phen, with DP. Our results illustrated that exposure to 5 or 20⯵g/L 3-MP alone or in combination with 60⯵g/L DP caused neurobehavioral anomalies in zebrafish. In accordance with the behavioral deficits, 3-MP alone or co-exposed with DP significantly decreased axonal growth of secondary motoneurons, altered intracellular Ca2+ homeostasis and induced cell apoptosis in the muscle of zebrafish. Additionally, 3-MP alone or co-exposed with DP significantly increased reactive oxygen species (ROS) and the mRNA levels of apoptosis-related genes. These findings indicate that 3-MP alone or co-exposed with DP induces neurobehavioral deficits through the combined effects on neuronal connectivity and muscle function. Chemical analysis revealed significant increases in 3-MP and DP bioaccumulation in zebrafish co-exposed with 3-MP and DP. Elevated bioaccumulation resulting from mixture exposure may represent a significant contribution of the synergistic effects observed in combined chemical exposure.
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Hidrocarburos Clorados/toxicidad , Sistema Nervioso/efectos de los fármacos , Fenantrenos/toxicidad , Compuestos Policíclicos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Pez Cebra/fisiología , Animales , Proteínas Reguladoras de la Apoptosis/genética , Sinergismo Farmacológico , Sistema Nervioso/crecimiento & desarrollo , Fenantrenos/síntesis química , ARN Mensajero/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Pez Cebra/crecimiento & desarrolloRESUMEN
Lipid rafts and associated membrane proteins (flotillin, caveolin) play important roles in cell signaling and sperm fertilization while heat shock proteins (Hsp) ensure properly protein folding to fulfill their physiological functions. The markedly reduced fertility in thawed sperm after cryopreservation could result from disrupted membrane lipid rafts and these proteins. To explore the effect of sperm cryopreservation on lipid rafts and heat shock proteins, we compared lipid raft integrity, and the expression levels of lipid raft associated proteins (Flot-1, Flot-2, Cav-1) as well as heat shock proteins (Hsp90, Hsp70) in fresh and thawed sperm cryopreserved under different scenarios in yellow catfish. We found higher lipid raft integrity, higher protein expression levels of Flot-1, Flot-2, Cav-1, Hsp90, and Hsp70 in fresh sperm samples than in thawed sperm samples, in thawed sperm samples cryopreserved with optimal cooling rate than those cryopreserved with sub-optimal cooling rate, and in thawed sperm samples cryopreserved with extenders supplemented with cholesterol than those supplemented with methyl-ß-cyclodextrin (for cholesterol removal). Our findings indicate that lipid raft integrity, and expression levels of Flot-1, Flot-2, Cav-1, Hsp90, and Hsp70 are clearly associated with sperm quality, and together they may play a cumulative role in reduced fertility associated with thawed sperm in aquatic species.
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Caveolinas/metabolismo , Criopreservación/métodos , Proteínas HSP70 de Choque Térmico/metabolismo , Proteínas HSP90 de Choque Térmico/metabolismo , Microdominios de Membrana/metabolismo , Proteínas de la Membrana/metabolismo , Animales , Bagres/fisiología , Colesterol/farmacología , Masculino , Análisis de Semen , Transducción de Señal , Motilidad Espermática/efectos de los fármacos , Espermatozoides/efectos de los fármacosRESUMEN
Polybrominated diphenyl ethers (PBDEs) and lead (Pb) are common pollutants that co-exist in the environment. These chemicals may be associated with autism spectrum disorder (ASD), yet direct evidence is lacking. More importantly, how co-exposure of these chemicals might affect ASD has never been explored. For assessing the relationship between PBDE/Pb exposure and ASD, pregnant C57BL/6 J female mice were exposed to BDE209 (0.12 ng/day), Pb (1.2 ng/day), or a BDE209/Pb mixture from gestational day (GD) 9.5 to postnatal day (PND) 21 using ALZET osmotic pumps. Polyinosinic-polycytidylic acid (poly I:C) was included as a positive control, as its single dose injection (20 mg/kg.bw; i.p.) at mid-pregnancy (GD 12.5) produces ASD-like behaviors in mouse offspring. These ASD-like phenotypes include decreased preference for social novelty, increased marble burying behavior, and learning impairment. Similar to the poly I:C control, perinatal exposure to Pb or BDE209/Pb mixture elicited increased marble burying and learning impairment, but it had no effect on sociability. Consistent with these behavioral anomalies, Pb and BDE209/Pb co-exposure as well as poly I:C exposure increased the production of pro-inflammation cytokines interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 10 (IL-10), tumor necrosis factor α (TNFα), interferon γ (IFNγ), and interleukin 17 A (IL-17 A) in the serum, and decreased neuronal cells in the CA1 and CA3 subregions of the hippocampus. The majority of these changes in the BDE209/Pb mixture group were due to the effect of Pb rather than BDE209. However, BDE209/Pb co-exposure elicited a synergistic increase in the production of IL-4, IL-6, TNFα, IFNγ, and IL-17A in the serum. BDE209 exposure alone also significantly affected spatial learning and increased the production of IL-10, TNFα, and IL-17 A in the serum of male offspring. Our work demonstrates that perinatal exposure to a low dose of Pb or the BDE209/Pb mixture, although it did not induce typical ASD-like symptoms, elicited restricted, repetitive patterns of behavior and affected learning in male offspring. In addition, the synergistic increase in the systemic inflammatory response in the BDE209/Pb co-exposure group underscores the importance of evaluating chemical mixtures in disease onset.
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Trastorno del Espectro Autista/inducido químicamente , Conducta Animal/efectos de los fármacos , Éteres Difenilos Halogenados/toxicidad , Hipocampo/efectos de los fármacos , Plomo/toxicidad , Efectos Tardíos de la Exposición Prenatal , Animales , Trastorno del Espectro Autista/sangre , Trastorno del Espectro Autista/fisiopatología , Trastorno del Espectro Autista/psicología , Muerte Celular/efectos de los fármacos , Citocinas/sangre , Femenino , Edad Gestacional , Aseo Animal/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/patología , Hipocampo/fisiopatología , Mediadores de Inflamación/sangre , Masculino , Exposición Materna/efectos adversos , Aprendizaje por Laberinto/efectos de los fármacos , Ratones Endogámicos C57BL , Embarazo , Conducta Estereotipada/efectos de los fármacosRESUMEN
Polyhalogenated carbazoles (PHCZs), which have the similar structure of dioxin, have been reported ubiquitous in the environments and drawn wide concerns. However, their potential ecological and health risks are still poorly understood. Here, wildtype zebrafish embryos were used to evaluate the environmental risks of 2,7-dibromocarbazole (2,7-DBCZ), 3,6-dibromocarbazole (3,6-DBCZ), and 3,6-dichlorocarbazole (3,6-DCCZ). 2,7-DBCZ was the most toxic compound with the 96-h LC50 value of 581.8 ± 29.3 µg·L-1 and the EC50 value of 201.5 ± 6.5 µg·L-1 for pericardial edema. The teratogenic effects of 2,7-DBCZ were tested using transgenic zebrafish larvae. The transcriptomic analysis revealed that 90 genes in zebrafish expressed differently after exposure to 2,7-DBCZ, and many pathways were related to aryl hydrocarbon receptor (AhR) activation. The qRT-PCR also showed that expression levels of AhR1 and CYP1 A in zebrafish were significantly up-regulated after exposure to 2,7-DBCZ. In conclusion, 2,7-DBCZ exhibited more potent toxicity and cardiac teratogenic effects, and presented developmental toxicity partially consistent with AhR activation. Our results will be of great help to the risk assessment and regulation-making of PHCZs. Meanwhile, further studies should be promoted to illustrate the potential mechanism between PHCZs and AhR in the near future.
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Carbazoles/toxicidad , Teratógenos/toxicidad , Pez Cebra , Animales , Animales Modificados Genéticamente , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Corazón/efectos de los fármacos , Corazón/crecimiento & desarrollo , Larva/efectos de los fármacos , Larva/genética , Larva/crecimiento & desarrollo , Transcriptoma/efectos de los fármacos , Pez Cebra/anomalías , Pez Cebra/genética , Proteínas de Pez Cebra/genéticaRESUMEN
Perfluorooctane sulfonic acid (PFOS), as a potential endocrine disrupting chemical, is widely detected in the environment, wildlife and human. Currently few studies have documented the effects of chronic PFOS exposure on thyroid in aquatic organisms and the underlying mechanisms are largely unknown. The present study assessed the effect of chronic PFOS exposure on thyroid structure and function using zebrafish model. Zebrafish at 8 h post fertilization (hpf) were exposed to PFOS (250 µg/l) until 120 d post fertilization (dpf). Thyroid hormone (T3 and T4) level, thyroid morphology and thyroid function related gene expression were evaluated in zebrafish at 120 dpf. Our findings demonstrated that chronic PFOS exposure altered thyroid hormone level, thyroid follicular cell structure and thyroid hormone related gene expression, suggesting the validity of zebrafish as an alternative model for PFOS chronic toxicity screening.
Asunto(s)
Ácidos Alcanesulfónicos/toxicidad , Disruptores Endocrinos/toxicidad , Fluorocarburos/toxicidad , Glándula Tiroides/efectos de los fármacos , Pez Cebra , Animales , Embrión no Mamífero/efectos de los fármacos , Embrión no Mamífero/fisiología , Glándula Tiroides/fisiologíaRESUMEN
Autism spectrum disorder (ASD) has complex neurodevelopmental impairments and origins that are linked to both genetic and environmental factors. Hence, there is an urgency to establish animal models with ASD-like characteristics to understand the underlying mechanisms of ASD. Prenatal exposure to valproic acid (VPA) produced ASD-like symptoms in humans, rats, and recently zebrafish. The present study investigated the use of VPA exposure to generate an ASD model in zebrafish. Early life stage exposures produced ASD-like phenotypes in the developing brain development and behavioral changes in embryonic and larval zebrafish. Our findings revealed that treating zebrafish embryos with VPA starting at 8h post fertilization (hpf) resulted in significant: increase in the ASD macrocephalic phenotype; hyperactivity of embryo/larvae movement behaviors; and increases of ASD-like larval social behaviors. Further analysis showed increases in cell proliferation, the proportion of mature newborn neurons, and neural stem cell proliferation in the brain region, which may contribute to the brain overgrowth and macrocephaly observed following VPA exposure. Our study demonstrated that VPA exposure generates ASD-like phenotypes and behaviors, indicating that zebrafish is an alternative model to investigate underlying ASD mechanisms.
Asunto(s)
Trastorno del Espectro Autista/inducido químicamente , Modelos Animales de Enfermedad , Embrión no Mamífero/efectos de los fármacos , Desarrollo Embrionario/efectos de los fármacos , Ácido Valproico/toxicidad , Pez Cebra , Animales , Conducta Animal/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Pez Cebra/embriologíaRESUMEN
One-monomer molecularly imprinted magnetic nanoparticles were prepared as adsorbents for selective extraction of bisphenol A from water in this study. A single bi-functional monomer was adopted for preparation of the molecularly imprinted polymer, avoiding the tedious trial-and-error optimizations as traditional strategy. Moreover, bisphenol F was used as the dummy template for bisphenol A to avoid the interference from residual template molecules. These nanoparticles showed not only large adsorption capacity and good selectivity to the bisphenol A but also outstanding magnetic response performance. Furthermore, they were successfully used as magnetic solid-phase extraction adsorbents of bisphenol A from various water samples, including tap water, river water, and seawater. The developed method was found to be much more efficient, convenient, and economical for selective extraction of bisphenol A compared with the traditional solid-phase extraction. Separation of these nanoparticles can be easily achieved with an external magnetic field, and the optimized adsorption time was only 15 min. The recoveries of bisphenol A in different water samples ranged from 85.38 to 93.75%, with relative standard deviation lower than 7.47%. These results showed that one-monomer molecularly imprinted magnetic nanoparticles had the potential to be popular adsorbents for selective extraction of pollutants from water.