Zebrafish Offspring Research Articles

DNA methyltransferase 3a mediates developmental thermal plasticity

BackgroundThermal plasticity is pivotal for evolution in changing climates and in mediating resilience to its potentially negative effects. The efficacy to respond to environmental change depends on underlying mechanisms. DNA methylation induced by DNA methyltransferase 3 enzymes in the germline or during early embryonic development may be correlated with responses to environmental change. This developmental plasticity can interact with reversible acclimation within adult organisms, which would increase the speed of response and could alleviate potential mismatches between parental or early embryonic environments and those experienced at later life stages. Our aim was to determine whether there is a causative relationship between DNMT3 enzyme and developmental thermal plasticity and whether either or both interact with short-term acclimation to alter fitness and thermal responses in zebrafish (Danio rerio).ResultsWe developed a novel DNMT3a knock-out model to show that sequential knock-out of DNA methyltransferase 3a isoforms (DNMT3aa−/− and DNMT3aa−/−ab−/−) additively decreased survival and increased deformities when cold developmental temperatures in zebrafish offspring mismatched warm temperatures experienced by parents. Interestingly, short-term cold acclimation of parents before breeding rescued DNMT3a knock-out offspring by restoring survival at cold temperatures. DNMT3a knock-out genotype interacted with developmental temperatures to modify thermal performance curves in offspring, where at least one DNMT3a isoform was necessary to buffer locomotion from increasing temperatures. The thermal sensitivity of citrate synthase activity, an indicator of mitochondrial density, was less severely affected by DNMT3a knock-out, but there was nonetheless a significant interaction between genotype and developmental temperatures.ConclusionsOur results show that DNMT3a regulates developmental thermal plasticity and that the phenotypic effects of different DNMT3a isoforms are additive. However, DNMT3a interacts with other mechanisms, such as histone (de)acetylation, induced during short-term acclimation to buffer phenotypes from environmental change. Interactions between these mechanisms make phenotypic compensation for climate change more efficient and make it less likely that thermal plasticity incurs a cost resulting from environmental mismatches.

Effects of waterborne exposure to environmentally relevant concentrations of selenite on reproductive function of female zebrafish: A life cycle assessment.

Recently, bioaccumulation of dietary organic selenium (Se) in the ovaries and inhibition of reproduction in female aquatic animals have been reported. However, there is limited data on the subtle reproductive impacts of waterborne exposure to inorganic Se in fish. Here, zebrafish embryos (2h post-fertilization) were exposed to solutions with environmentally relevant levels of Na2SeO3 with concentrations of 0 (control), 7.98±0.31, 25.14±0.15, and 79.60±0.81μg Se/L for 120d until they reached sexual maturity. Female zebrafish were selected for reproductive toxicity assessment. In the early embryonic stage, whole-mount in situ hybridization of zebrafish embryos showed that waterborne Na2SeO3 exposure did not affect the observed location of vasa expression in primordial germ cells at 24, 48, and 72h post-fertilization. Life-cycle exposure to 25.14±0.15 and 79.60±0.81μg Se/L Na2SeO3 did not change the testosterone and 17β-estradiol contents in female zebrafish at the endpoint of exposure, but significantly reduced the proportion of early vitellogenic oocytes and mature oocytes. Follicle maturity retardation was accompanied by changes in transcriptional levels of the genes related to the hypothalamus-pituitary-gonad-liver (HPGL) axis. Transcriptional levels of cyp19a and lhr in the ovary were down-regulated, while the transcriptional level of fshr in the ovaries was up-regulated. In the 21-day cumulative spawning experiment, Na2SeO3 (25.14±0.15 and 79.60±0.81μg Se/L) caused fewer eggs to be produced. Additionally, the malformation of zebrafish offspring significantly increased in the group exposed to 79.60±0.81μg Se/L. In conclusion, for the first time, this study shows that life-cycle exposure to environmentally relevant concentrations of waterborne Na2SeO3 significantly delays ovarian maturation and reduces the fertility of the female zebrafish.

The joint effect of parental exposure to microcystin-LR and polystyrene nanoplastics on the growth of zebrafish offspring

The coexistence of nanoplastics (NPs) and various pollutants in the environment has become a problem that cannot be ignored. In order to identify the microcystin-LR (MCLR) bioaccumulation and the potential impacts on the early growth of F1 zebrafish (Danio rerio) offspring in the presence of polystyrene nanoplastics (PSNPs), PSNPs and MCLR were used to expose adult zebrafish for 21days. The exposure groups divided into MCLR (0, 0.9, 4.5 and 22.5μgL−1) alone groups and PSNP (100μgL−1) and MCLR co-exposure groups. F1 embryos were collected and developed to 120 h post-fertilization (hpf) in clear water. Compared with the exposure to MCLR only, the combined exposure increased the parental transfer of MCLR to the offspring and subsequently exacerbated the growth inhibition of F1 larvae. Further research clarified that combined exposure of PSNPs and MCLR could reduce the levels of thyroxine (T4) and 3, 5, 3′-triiodothyronine (T3) by altering the expression of hypothalamus–pituitary–thyroid (HPT) axis-related genes, eventually leading to growth inhibition of F1 larvae. Our results also exhibited combined exposure of PSNPs and MCLR could change the transcription of key genes of the GH/IGF axis compared with MCLR single exposure, suggesting the GH/IGF axis was a potential target for the growth inhibition of F1 larvae in PSNPs and MCLR co-exposure groups. The present study highlights the potential risks of coexistence of MCLR and PSNPs on development of fish offspring, and the environmental risks to aquatic ecosystems.

Exposure of parents to alcohol alters behavior of offspring in zebrafish

Alcoholism and alcohol abuse represent a significant medical and societal problem, and have been thoroughly investigated in humans as well as using animal models. A less well understood aspect of alcohol related disorders is the possible effect of this drug on offspring whose parents were exposed prior to conception. The zebrafish has been successfully employed in alcohol research, however, the effect of exposing the parents to alcohol before fertilization of the eggs on offspring has not been demonstrated in this species. In this proof of concept study, we attempt to address this hiatus. We exposed both adult male and female zebrafish to 0.0% (control) or 0.5% (vol/vol) alcohol chronically for 7 days, subsequently bred the fish within their respective treatment group, collected the fertilized eggs, allowed them to develop, and tested the behavior of free-swimming offspring at their age of 7–9 days post-fertilization. We conducted the analysis in two genetically distinct quasi-inbred strains of zebrafish, AB and TL. Although gross morphology and general activity of the fish appeared unaffected, we found significant behavioral alterations in offspring of alcohol exposed parents compared to offspring of control parents in both strains. These alterations included robustly increased duration and reduced frequency of immobility, increased turn angle, and increased intra-individual variance of turn angle in offspring of alcohol exposed parents in both strains. The mechanisms underlying these behavioral effects or whether the effects are due to exposure of the father, the mother, or both to alcohol are unknown. Nevertheless, our results now set the stage for future studies with zebrafish that will address these questions.

Paternal exposure to di-n-butyl-phthalate induced developmental toxicity in zebrafish (Danio rerio).

Dibutyl phthalate (DBP) is an environmental endocrine disruptor detected in water, soil, and other environmental media frequently. Growing concerns regarding DBP exposure focus on toxicity to male reproduction. Reports about the developmental toxicity of paternal DBP exposure are rare. In this study, we investigated the developmental toxicity of paternal exposure to DBP on offspring in zebrafish. Adult male zebrafish with normal reproductive function were exposed to 0.2, 0.6, 1.8 mg/L of DBP or acetone solvent control for 30 days, and then mated with females. Thirty embryos per group were randomly selected to be observed, and malformations were recorded and photographed. The mating and observations were repeated three times, for a total of 90 embryos per group. The results showed that the percentage of malformations, such as edema and a bent trunk, was increased in the 0.6 and 1.8 mg/L DBP exposure groups, the heart rate and spontaneous contraction decreased in the 0.6 and 1.8 mg/L DBP exposure groups and migration of primordial germ cells was disrupted in some F1 embryos in all DBP exposure group after paternal exposure. The axial skeleton was affected in some F1 adults in the 1.8 mg/L DBP exposure group. Our findings demonstrate the developmental toxicity of paternal DBP exposure in zebrafish.

Maternal exposure to environmental antibiotic mixture during gravid period predicts gastrointestinal effects in zebrafish offspring

Due to overuse, misuse, and poor absorption during treatment, antibiotics are consistently released into the environment, raising concerns about their impacts on ecological sustainability and health. In this study we performed transcriptome profiling to assess potential reproductive effects of an antibiotic mixture in gravid female zebrafish. Gravid fish (150 dpf) were exposed to a mixture of 15 commonly detected antibiotics at 0, 1, and 100 μg/L for 4 weeks. Concentrations of all the 15 antibiotics, especially chlortetracycline, were detected in the F0 ovary and F1 eggs after treatment, indicating maternal transfer of antibiotics. Impaired F0 growth (average 2.2 % and 24.3 % inductions in body length and ovary weight, respectively), and reduced F1 offspring survival (average 4.2 % reductions in survival at 120 hpf) was observed after maternal exposure to the 100 μg/L treatment. Pathway analyses of whole-transcriptome expression profiles from F0 ovaries predicted colorectal disorders. Similarly, pathways of F1 larval transcriptomes from treated females also predicted colorectal disorders along with intestinal apoptosis and oxidative stress, which may be related to growth impairment. These results show that maternal transfer of antibiotics occurs in zebrafish, resulting in transgenerational changes in F1 offspring survival and transcription that predict adverse gastrointestinal effects in offspring.

Nano-TiO2 enhanced bioaccumulation and developmental neurotoxicity of bisphenol a in zebrafish larvae

The titanium dioxide nanoparticles (n-TiO2) could enhance the bioavailability and toxicity of the coexisted organic toxicants in aquatic phase. Parental co-exposure to n-TiO2 and bisphenol A (BPA) could generate developmental neurotoxicity in unexposed zebrafish offspring. However, it remains unexplored regarding the developmental neurotoxicity in larvae fish after co-exposure during the early developmental stage. In present study, fertilized zebrafish eggs were exposed to TiO2 nanoparticles (100 μg/L), BPA (1, 4 and 20 μg/L) or their binary mixtures until 6 days post fertilization (dpf). No significant change was observed in hatching, malformation, survival and weight of the larvae among all groups. However, n-TiO2 significantly increased the body burden of BPA in the 4 and 20 μg/L co-exposure groups, depressed expression of neurodevelopment marker genes (α1-tubulin, mbp and syn2a) as well as the locomotor behavior. The current results indicate that n-TiO2 could strengthen the developmental neurotoxicity and inactive locomotion in co-exposed zebrafish larvae by promoting the bioaccumulation and bioavailability of BPA, which highlighted the similar toxic risks of developmental neurotoxicity after co-exposure at early developmental stage to that of the parental co-exposure.

Parental exposure to cadmium chloride causes developmental toxicity and thyroid endocrine disruption in zebrafish offspring

Cadmium is a common heavy metal pollutant. Previous studies have found that long-term cadmium exposure can cause damage to multiple organs/systems in humans and experimental animals; however, there are few studies that elucidate its effects on offspring development, discuss whether it can be transmitted to offspring from the parent, and debate whether it affects the functional development of the thyroid hormone system in offsprings. In this study, sexually mature zebrafish were exposed to different concentrations of cadmium chloride (0.01 μmol/L, 0.1 μmol/L, and 1 μmol/L) to study reproductive toxicity. It was found that parental zebrafish exposed to 1 μmol/L of cadmium chloride produced offsprings with different degrees of malformation. At 5 days post-fertilization (dpf), the levels of 3,5,3′-triiododenosine (T3) and thyroxine (T4) in the zebrafish were decreased. At 10 dpf, the T4 and T3 levels in the zebrafish of the offspring were significantly reduced. At the same time, the expression of thyroid receptor (trα and trβ) genes in five dpf larvae was significantly up-regulated in the 1 μmol/L treatment group relative to the control group. The mRNAs of thyroid hormone synthesis and metabolism-related genes (tshβ, dio1, dio2, ugt1ab, and ttr) were significantly up-regulated in the 0.1 μmol/L and 1 μmol/L treatment groups. This study demonstrates that parental cadmium chloride exposure produces reproductive toxicity in zebrafish and that the effects can be transferred from the parent to the offspring, resulting in developmental toxicity in the thyroid endocrine system.

Paternal exposure to a common herbicide alters the behavior and serotonergic system of zebrafish offspring.

Increasingly, studies are revealing that endocrine disrupting chemicals (EDCs) can alter animal behavior. Early life exposure to EDCs may permanently alter phenotypes through to adulthood. In addition, the effects of EDCs may not be isolated to a single generation − offspring may indirectly be impacted, via non-genetic processes. Here, we analyzed the effects of paternal atrazine exposure on behavioral traits (distance moved, exploration, bottom-dwelling time, latency to enter the top zone, and interaction with a mirror) and whole-brain mRNA of genes involved in the serotonergic system regulation (slc6a4a, slc6a4b, htr1Aa, htr1B, htr2B) of zebrafish (Danio rerio). F0 male zebraFIsh were exposed to atrazine at 0.3, 3 or 30 part per billion (ppb) during early juvenile development, the behavior of F1 progeny was tested at adulthood, and the effect of 0.3 ppb atrazine treatment on mRNA transcription was quantified. Paternal exposure to atrazine significantly reduced interactions with a mirror (a proxy for aggression) and altered the latency to enter the top zone of a tank in unexposed F1 offspring. Bottom-dwelling time (a proxy for anxiety) also appeared to be somewhat affected, and activity (distance moved) was reduced in the context of aggression. slc6a4a and htr1Aa mRNA transcript levels were found to correlate positively with anxiety levels in controls, but we found that this relationship was disrupted in the 0.3 ppb atrazine treatment group. Overall, paternal atrazine exposure resulted in alterations across a variety of behavioral traits and showed signs of serotonergic system dysregulation, demonstrating intergenerational effects. Further research is needed to explore transgenerational effects on behavior and possible mechanisms underpinning behavioral effects.

Parental exposure to bisphenol A affects pharyngeal cartilage development and causes global transcriptomic changes in zebrafish (Danio rerio) offspring

BackgroundAs one of the most common endocrine-disrupting chemicals (EDCs), bisphenol A (BPA) is a threat to aquatic ecosystems. Despite a rich literature addressing the adverse effects of BPA on various systems in fish models, the potential impact of parental BPA exposure on offspring pharyngeal cartilage development is poorly understood. MethodsAdult zebrafish (F0) were exposed to BPA (1.0 μM) or control for 7 days. Eggs (F1) were collected and exposed to BPA (control, 0.05, 0.1, 1, 10 μM) until 120 h post-fertilization. Histomorphometrical essay was used to quantitatively and qualitatively assess the effects of BPA on pharyngeal cartilage development. RNA sequencing (RNA-seq) was used to discover differentially expressed genes (DEGs), and KEGG pathway and GO enrichment analysis were performed to interpret functional ontology. ResultsParental BPA exposure affected hatchability and heart rates of F1 progeny. By pathology analysis, parental BPA exposure caused craniofacial deformity, characterized by wider angles of cartilage elements, disrupted pharyngeal chondrocytes and promoted apoptosis and elongation of head length. RNA-seq suggested that many DEGs were involved in multiple biological processes and signaling pathways; defense responses, reactive oxygen species metabolic process, apoptosis, p53 signaling pathway and MAPK signaling pathway were closely associated with the toxicity of parental BPA exposure. ConclusionsParental BPA exposure affected chondrogenesis in the viscerocranium of zebrafish offspring and led to global transcriptomic changes involved in apoptosis, hyperplasia and oxidative stress. These newly identified gene expression patterns, pathways and gene networks of zebrafish eleutheroembryos after early-life waterborne BPA exposure, may lead to severe and permanent morphological and functional consequences.

Microcystin-LR triggers different endoplasmic reticulum stress pathways in the liver, ovary, and offspring of zebrafish (Danio rerio)

The existence of microcystins (MCs), the secondary metabolite of cyanobacteria, has become a growing public health concern. Previous researches have proved that MCs can trigger endoplasmic reticulum stress (ERS), but the underlying mechanisms remain unclear. In the present study, adult female zebrafish were exposed to MC-LR (0, 1, 5 and 20 μg/L) for 30 d, and the offspring derived from the treated females and healthy males were cultured in water without MC-LR until 96 h post fertilization (hpf). Our data suggested that MC-LR causes a significant increase in the eif2s1a, atf4, and eif2ak3 transcription levels in the liver and ovary. The mRNA levels of atf4, atf6, bcl-2, hspa5, eif2s1a and eif2ak3 upregulated notably in the offspring. JNK phosphorylation level and cleaved-caspase3 protein expression elevated obviously in the liver and ovary, but had no remarkable change in the offspring. Furthermore, TUNEL results showed that MC-LR significantly induced apoptosis in the liver and ovary, while acridine orange (AO) staining indicated that MC-LR did not cause abnormal apoptosis in offspring. Concisely, the present study indicated that MC-LR leads to apoptosis through different ERS pathways in the liver, ovary and offspring, and also provides a new perspective for understanding the apoptosis caused by MC-LR.

Cryoprotectants synergy improve zebrafish sperm cryopreservation and offspring skeletogenesis

The synergy obtained by the combination of cryoprotectants is a successful strategy that can be beneficial on the optimization of zebrafish sperm cryopreservation. Recently, a protocol was established for this species using an electric ultrafreezer (−150 °C) performing cooling rate (−66 °C/min) and storage within one step. The ultimate objective of sperm cryopreservation is to generate healthy offspring. Therefore, the objective of this study was to select the most adequate cryoprotectant combination, for the previously established protocol, that generate high quality offspring with normal skeletogenesis. Among the permeating cryoprotectant concentrations studied 12.5% and 15% of N,N-dimethylformamide (DMF) yielded high post-thaw sperm quality and hatching rates. For these two concentrations, the presence of bovine serum albumin (10 mg/mL), egg yolk (10%), glycine (30 mM) and bicine (50 mM) was evaluated for post-thaw sperm motility, viability, in vitro fertilization success and offspring skeletal development (30 days post fertilization). Higher concentration of permeating cryoprotectant (15%) decreased the incidence of deformed arches and severe skeletal malformations, which suggests higher capacity to protect the cell against cold stress and DNA damage. Extender containing 15% DMF with Ctrl, Bicine and egg yolk were the non-permeating cryoprotectants with higher post-thaw quality. The use of these compounds results in a reduction in vertebral fusions, compressions and severity of skeletal malformations in the offspring. Therefore, these extender compositions are beneficial for the quality of zebrafish offspring sired by cryopreserved sperm with −66 °C/min freezing rate. To the best of our knowledge, this is the first report on skeletal development of the offspring sired by cryopreserved sperm performed with different freezing media compositions in zebrafish.

Parental exposure to triphenyltin inhibits growth and disrupts thyroid function in zebrafish larvae

Triphenyltin (TPT) is widely used and commonly found in a water environment, so its effects on aquatic systems are of great concern. This study aimed to reveal the effects of chronic parental exposure of TPT on thyroid disruption and growth inhibition in zebrafish. Adult zebrafish (F0 generation) were exposed to environmentally relevant concentrations (1, 10, and 100 ng/L) of TPT for 60 days, and the larvae (F1 generation) were tested without TPT treatment. Results demonstrated that parental exposure to TPT disrupts thyroid function in zebrafish offspring: serum thyroxine (T4) significantly decreased, while serum 3,5,3′-triiodothyronine (T3) increased, and several genes involved in the hypothalamic–pituitary–thyroid (HPT) axis were down-regulated. In addition, we observed developmental abnormalities in the larvae, demonstrated by a significantly altered hatching rate, malformation rate, body length, heart rate, and survival rate, as well as down-regulation of genes involved in the growth hormone/insulin-like growth factor (GH/IGF) axis. Therefore, parental exposure to TPT induces toxicity in fish offspring through perturbation of the HPT and GH/IGF axes.

Maternal ethanol consumption before paternal fertilization: Stimulation of hypocretin neurogenesis and ethanol intake in zebrafish offspring

There are numerous clinical and pre-clinical studies showing that exposure of the embryo to ethanol markedly affects neuronal development and stimulates alcohol drinking and related behaviors. In rodents and zebrafish, our studies show that embryonic exposure to low-dose ethanol, in addition to increasing voluntary ethanol intake during adolescence, increases the density of hypothalamic hypocretin (hcrt) neurons, a neuropeptide known to regulate reward-related behaviors. The question addressed here in zebrafish is whether maternal ethanol intake before conception also affects neuronal and behavioral development, phenomena suggested by clinical reports but seldom investigated. To determine if preconception maternal ethanol consumption also affects these hcrt neurons and behavior in the offspring, we first standardized a method of measuring voluntary ethanol consumption in AB strain adult and larval zebrafish given gelatin meals containing 10% or 0.1% ethanol, respectively. We found the number of bites of gelatin to be an accurate measure of intake in adults and a strong predictor of blood ethanol levels, and also to be a reliable indicator of intake in larval zebrafish. We then used this feeding paradigm and live imaging to examine the effects of preconception maternal intake of 10% ethanol-gelatin compared to plain-gelatin for 14 days on neuronal development in the offspring. Whereas ethanol consumption by adult female HuC:GFP transgenic zebrafish had no impact on the number of differentiated HuC+ neurons at 28 h post-fertilization (hpf), preconception ethanol consumption by adult female hcrt:EGFP zebrafish significantly increased the number of hcrt neurons in the offspring, an effect observed at 28 hpf and confirmed at 6 and 12 days post-fertilization (dpf). This increase in hcrt neurons was primarily present on the left side of the brain, indicating asymmetry in ethanol's actions, and it was accompanied by behavioral changes in the offspring, including a significant increase in novelty-induced locomotor activity but not thigmotaxis measured at 6 dpf and also in voluntary consumption of 0.1% ethanol-gelatin at 12 dpf. Notably, these measures of ethanol intake and locomotor activity stimulated by preconception ethanol were strongly, positively correlated with the number of hcrt neurons. These findings demonstrate that preconception maternal ethanol consumption affects the brain and behavior of the offspring, producing effects similar to those caused by embryonic ethanol exposure, and they provide further evidence that the ethanol-induced increase in hcrt neurogenesis contributes to the behavioral disturbances caused by ethanol.

Parental exposure to tris(1,3-dichloro-2-propyl) phosphate results in thyroid endocrine disruption and inhibition of growth in zebrafish offspring

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is a re-emerging environmental contaminant used as a suitable substitute for brominated flame retardants. The objective of this study was to evaluate the effects of TDCIPP on thyroid disruption and growth inhibition in zebrafish (Danio rerio) offspring after chronic parental exposure, and to examine the possible molecular mechanisms involved. When adult zebrafish (4 months old) were exposed to 5.66, 25.55, or 92.8 μg TDCIPP/L for 90 days, bioconcentration of TDCIPP and its metabolic product [bis(1,3-dichloro-2-propyl) phosphate, BDCIPP] was observed in 7-day postfertilization (dpf) F1 larvae, which suggests the transfer of this compound from adult fish to their offspring. Our results demonstrated that parental exposure to TDCIPP induced thyroid disruption in the offspring, demonstrated by significantly decreased thyroxine (T4) and increased 3,5,3′-triiodothyronine (T3) levels, and disruption of the transcription of several genes and expression of proteins involved in the hypothalamic-pituitary-thyroid (HPT) axis in F1 larvae. Parental exposure to TDCIPP resulted in developmental abnormalities in offspring; the smaller body length that was recorded might be partly the result of the perturbation of the HPT axis. In addition, the results revealed that growth inhibition also resulted from the downregulation of the transcription of genes and expression of proteins involved in the growth hormone/insulin-like growth factor (GH/IGF) axis. Our study provides a new set of evidence showing that parental exposure to TDCIPP can induce thyroid disruption and inhibition of growth in offspring, and that perturbation of the HPT axis and GH/IGF axis contribute to these adverse effects.

Maternal exposure to fipronil results in sulfone metabolite enrichment and transgenerational toxicity in zebrafish offspring: Indication for an overlooked risk in maternal transfer?

Ecotoxicological studies show the association between pesticide pollution and transgenerational toxicity in aquatic organisms. However, a less considered risk is that many pesticides can be metabolized and transferred to offspring as new toxicants. In this study, we used zebrafish to evaluate the maternal transfer risk of fipronil (FIP), which is a great threat to aquatic organisms with toxic metabolite formation. After 28-day exposure to environmentally relevant concentrations (1.0, 5.0 and 10.0 μg/L) of FIP in adult female zebrafish (F0), the toxicants off-loading and transgenerational toxicity in offspring were studied. High burdens of FIP and its sulfone metabolite were found in both F0 and the embryos (F1), resulting in increased CYP450 activity. The residual levels of the metabolite were higher than those of the parent compound. Chiral analysis further showed a preferential accumulation of S-enantiomer of FIP in both F0 and F1. Maternal exposure to FIP increased the malformation rate and decreased the swim speed in larvae. Additionally, after exposure, the levels of thyroid hormones (THs), including triiodothyronine (T3) and thyroxine (T4), decreased in both generations, particularly in the F1. Gene transcription expression along the hypothalamic-pituitary-thyroid (HPT) axis was also significantly affected. Maternal exposure to FIP increased sulfone metabolite enrichment and cause multiple toxic effects in F1. Findings from this study highlight the key role of biologically active product formation in the maternal transfer of pollutants and associated risk assessment.

Maternal Exposure to Dietary Selenium Causes Dopaminergic Hyperfunction and Cognitive Impairment in Zebrafish Offspring.

Maternal exposure to environmental contaminants is a predisposing factor for neurodevelopmental disorders with associated cognitive and social deficits in offspring. In this study, we investigated the effects of maternal exposure to selenium (Se), a contaminant of potential environmental concern in aquatic ecosystems, on cognitive performance and the underlying mechanisms in F1-generation adult zebrafish. Adult female zebrafish were exposed to different concentrations of dietary Se (3.5, 11.1, or 27.4 μg Se/g dry weight) for a period of 60 days. Fish were subsequently bred, and their offspring were collected and raised for 6 months on a normal diet. Maternal exposure to all concentrations of dietary Se induced learning impairment in F1-zebrafish tested in a latent learning task. The results also showed a hyperfunctioning dopaminergic system in fish exhibiting the learning deficit. The hyperfunction of the dopaminergic system was associated with enhanced oxidative stress and alterations in the mRNA abundance of several immediate early and late response genes in the zebrafish brain. Taken together, these results suggest that maternal exposure to dietary Se via alterations in the dopaminergic system leads to persistent neurobehavioral deficits in F1-generation adult zebrafish.

Ionizing radiation induces transgenerational effects of DNA methylation in zebrafish

Ionizing radiation is known to cause DNA damage, yet the mechanisms underlying potential transgenerational effects of exposure have been scarcely studied. Previously, we observed effects in offspring of zebrafish exposed to gamma radiation during gametogenesis. Here, we hypothesize that these effects are accompanied by changes of DNA methylation possibly inherited by subsequent generations. We assessed DNA methylation in F1 embryos (5.5 hours post fertilization) with whole genome bisulfite sequencing following parental exposure to 8.7 mGy/h for 27 days and found 5658 differentially methylated regions (DMRs). DMRs were predominantly located at known regulatory regions, such as gene promoters and enhancers. Pathway analysis indicated the involvement of DMRs related to similar pathways found with gene expression analysis, such as development, apoptosis and cancers, which could be linked to previous observed developmental defects and genomic instability in the offspring. Follow up of 19 F1 DMRs in F2 and F3 embryos revealed persistent effects up to the F3 generation at 5 regions. These results indicate that ionizing radiation related effects in offspring can be linked to DNA methylation changes that partly can persist over generations. Monitoring DNA methylation could serve as a biomarker to provide an indication of ancestral exposures to ionizing radiation.

Transgenerational thyroid endocrine disruption induced by bisphenol S affects the early development of zebrafish offspring

Maternal thyroid hormones (THs) play an essential role in the embryonic and larval development of fish. Previous studies in fish have reported that parental exposure to thyroid disrupting chemicals (TDCs) changed maternal TH levels in the offspring; however, whether this transgenerational thyroid endocrine disruption can further disturb the early development of the offspring still remains largely unknown. Bisphenol S (BPS), a substitute of bisphenol A, has been reported to be a potential TDC. In this study, zebrafish (F0) were exposed to environmentally relevant concentrations (1, 10, and 100 μg/L) of BPS from 2 h post-fertilization to 120 days post-fertilization and then paired to spawn. Plasma levels of thyroxine (T4) were significantly decreased in F0 females while 3,5,3′-triiodothyronine (T3) plasma levels were significantly increased in F0 females and males; moreover, TH content in eggs (F1) spawned by exposed F0 generation exhibited similar changes as the F0 females, with significant decreases in T4 and increases in T3, demonstrating BPS-induced maternal transfer of thyroid endocrine disruption. Further, excessive levels of maternal T3 in the offspring resulted in delayed embryonic development and hatching, swim bladder inflation defect, reduction in motility, developmental neurotoxicity, and lateral stripe hypopigmentation in non-exposed F1 embryos and larvae. These results highlight the adverse effects on the early development of offspring induced by transgenerational thyroid endocrine disruption, which have been ignored by previous studies. Therefore, these results can further improve our understanding of the ecological risks of TDCs.

Parental co-exposure to bisphenol A and nano-TiO2 causes thyroid endocrine disruption and developmental neurotoxicity in zebrafish offspring

The coexistence of organic toxicants and nanoparticles in the environment influences pollutant bioavailability and toxicity. Using chronic co-exposure to an adult zebrafish model, this study investigated the transfer kinetics and transgenerational effects of bisphenol A (BPA) and titanium dioxide nanoparticles (n-TiO2) exposure in F1 offspring. When single and combined exposure to BPA (0, 2, and 20 μg/L) and n-TiO2 (100 μg/L) were compared, combined exposure was found to reciprocally facilitate bioaccumulation in adult fish while enhancing maternal transfer to offspring. Thyroid endocrine disruption and developmental neurotoxicity were observed in larval offspring by parental exposure to BPA alone or in combination with n-TiO2. Exposure to 20 μg/L BPA significantly decreased the thyroxine (T4) concentration in adult plasma, leading to less transfer into the eggs. The presence of 20 μg/L BPA with n-TiO2 further decreased the level of T4 compared to BPA exposure alone. Additionally, offspring larvae derived from exposed parents exhibited lethargic swimming behavior. Overall, this study examined the interactions of BPA and n-TiO2 with regard to their bioaccumulation, maternal transfer, and developmental effects, which highlighted that co-exposure dynamics are important and need to be considered for accurate environmental risk assessment.