[Objective] The aim was to study the effect of bensulfuron-methyl herbicide on acute toxicity and genetics toxicity of Danio redo. [ Method] Median lethal concentration was calculated by acute toxicity test, and analy...[Objective] The aim was to study the effect of bensulfuron-methyl herbicide on acute toxicity and genetics toxicity of Danio redo. [ Method] Median lethal concentration was calculated by acute toxicity test, and analyzing the herbicide whether existing in potential toxicity to aquatic organisms or not. Based on the study of acute toxicity, genetics toxicity was carried out, by calculating the micronucleus rate to judge bensulfuron-methyl herbicide whether existing in potential toxicity or not. [ Result ] The LD5o （24 h and 48 h） of bensulfuron-methyl herbicide are 0.698 ml/L and 0.637 ml/L respectively, the safe concentration was 0.159 ml/L. The results on the effects of micronucleus （MN） in erythrocytes of Danio redo induced by bensulfuron-methyl at different times and different concentrations showed that the MN rate of control group was 0.010 3%, the highest MN rate of experimental group reached to 0. 372%, it also indicated that bensulfuron-methyl herbicide had genetics toxicity to Danio redo. At the same detection time, there was dose-effect relationship of MN rate in erythrocytes between treatment and control groups with different concentrations. In the same treatment group, the MN rate in erythrocytes reached to peak value at 24 h, and decreased at 48 h and 72 h with the infection time was prolonged. [ Conclusion ] The study provides some basis for scientifically selecting and reasonably using herbicide.展开更多
Perfluorononanoic acid（PFNA） is a nine-carbon perfluoroalkyl acid widely used in industrial and domestic products. It is a persistent organic pollutant found in the environment as well as in the tissues of humans an...Perfluorononanoic acid（PFNA） is a nine-carbon perfluoroalkyl acid widely used in industrial and domestic products. It is a persistent organic pollutant found in the environment as well as in the tissues of humans and wildlife. There is a concern that this chemical might be a developmental toxicant and teratogen in various ecosystems. In the present study,the toxic effects of PFNA were evaluated in zebrafish（Danio rerio） embryos. One hour post-fertilization embryos were treated with 0, 25, 50, 100, 200, 300, 350, and 400 μmol/L PFNA for 96 hr in 6-well plates. Developmental phenotypes and hatching rates were observed and recorded. Nineteen genes related to oxidative stress and lipid metabolism were examined using Quantitative RT-PCR and confirmed by whole mount in situ hybridization（WISH）. Results showed that PFNA delayed the development of zebrafish embryos, reduced the hatching rate, and caused ventricular edema and malformation of the spine. In addition, the amount of reactive oxygen species in the embryo bodies increased significantly after exposure to PFNA compared with that of the control group. The Quantitative RT-PCR and WISH experiments demonstrated that m RNA expression of the lfabp and ucp2 genes increased significantly while that of sod1 and mt-nd1 decreased significantly after PFNA exposure. The m RNA expression levels of gpx1 and mt-atp6 decreased significantly in the high concentration group. However, the m RNA expression levels of both ppara and pparg did not show any significant variation after exposure. These findings suggest that PFNA affected the development of zebrafish embryos at relatively low concentrations.展开更多
Ethylene thiourea(ETU)converted from ethylene bisdithiocarbamate(EBDC)fungicides has aroused great concern because of its prevalence and harmful effects.Although ETU-induced neurotoxicity has been reported,the potenti...Ethylene thiourea(ETU)converted from ethylene bisdithiocarbamate(EBDC)fungicides has aroused great concern because of its prevalence and harmful effects.Although ETU-induced neurotoxicity has been reported,the potential mechanisms remain unclear.This study provided insights into its neurotoxic effects at environmentally relevant concentrations in zebrafish.Our findings showed that embryonic exposure to ETU decreased the hatch rate and delayed somite development.Furthermore,ETU treatment significantly reduced the dark velocity in the locomotion assay.The upregulated tendency of the mitogen-activated protein kinases(MAPK)pathway(mknk1,atf4,mapkapk3)screened by transcriptome analysis implied motor neuron degeneration,which was validated by subsequent morphological observation,as axon length and branches were truncated in the 62.5μg/L ETU group.However,although the rescue experiment with a p38 MAPK inhibitor(SB203580)successfully ameliorated axon degeneration,it failed to reverse the locomotion behaviors.Further exploration of transcriptome data revealed the varied expression of presynaptic scaffold protein-related genes(pcloa,pclob,bsna),whose downregulation might impair the neuromuscular junction(NMJ).Therefore,we reasonably suspected that ETU-induced neurobehavioral deficits might result from the combined effects of the MAPK pathway and presynaptic proteins.Considering this,we highlighted the necessity to take precautions and early interventions for susceptible ETU-exposed populations.展开更多
Decabromodiphenylethane （DBDPE） has been widely used as an alternative flame retardant due to the restriction or phase-out of traditional polybrominated diphenyl ethers （PBDEs）, and is of increasing concern regard...Decabromodiphenylethane （DBDPE） has been widely used as an alternative flame retardant due to the restriction or phase-out of traditional polybrominated diphenyl ethers （PBDEs）, and is of increasing concern regarding its ubiquity, persistence, and potential adverse effects. In the present study, the toxicological effects of DBDPE were evaluated using zebrafish as an in vivo model. Upon being exposed to DBDPE-polluted sediments for a short term, it was found that the mortality and malformation of zebrafish （including edema, bent notochord, and bent tail） were not affected even at the highest concentration tested （1000.0μg/kg dry sediment）. Regarding behavioral responses, it was found that zebraflsh larvae of 48 hours post fertilization （hpf） in all groups escaped successfully with a touch to the dorsal fin. However, when exposed to the highest DBDPE concentration, the larvae of 120 hpf exhibited significantly smaller distances as compared to the control. Moreover, the results of the acetylcholinesterase （ACHE） activity, the expression levels of two important nerve-related genes, and the cell apoptosis all indicated that DBDPE posed low neurotoxicity in embryo-larval zebrafish. The results in this study shed some light on the potential risks of DBDPE in the real environment and highlight the application of the sediment exposure route in the future.展开更多
Mepanipyrim,an anilinopyrimidine fungicide,has been extensively used to prevent fungal diseases in fruit culture.Currently,research on mepanipyrim-induced toxicity in organisms is still very scarce,especially visual d...Mepanipyrim,an anilinopyrimidine fungicide,has been extensively used to prevent fungal diseases in fruit culture.Currently,research on mepanipyrim-induced toxicity in organisms is still very scarce,especially visual developmental toxicity.Here,zebrafish larvae were employed to investigate mepanipyrim-induced visual developmental toxicity.Intense light andmonochromatic light stimuli-evoked escape experiments were used to investigate vision-guided behaviors.Meanwhile,transcriptomic sequencing and real-time quantitative PCR assays were applied to assess the potential mechanisms of mepanipyrim-induced visual developmental toxicity and vision-guided behavioral alteration.Our results showed that mepanipyrim exposure could induce retinal impairment and vision-guided behavioral alteration in larval zebrafish.In addition,the grk1b gene of the phototransduction signaling pathway was found to be a potential aryl hydrocarbon receptor(AhR)-regulated gene.Mepanipyrim-induced visual developmental toxicity was potentially related to the AhR signaling pathway.Furthermore,mepanipyrim-induced behavioral alteration was guided by the visual function,and the effects of mepanipyrim on long and middle wavelength light-sensitive opsins may be the main cause of vision-guided behavioral alteration.Our results provide insights into understanding the relationship between visual development and vision-guided behaviors induced by mepanipyrim exposure.展开更多
Lecithotrophic fish embryos rely on finite maternally deposited yolk resources for early development.Toxicant exposure can disrupt the uptake of yolk resources with consequences for development.In this study,we invest...Lecithotrophic fish embryos rely on finite maternally deposited yolk resources for early development.Toxicant exposure can disrupt the uptake of yolk resources with consequences for development.In this study,we investigate the impacts of altered yolk utilization on fish embryos using the cardiotoxic compound phenanthrene.Zebrafish embryos were exposed to a cardiotoxic concentration of phenanthrene beginning at 6 hpf(hours post-fertilization)until a maximum of 72 hpf.Embryos were stained with Oil Red O to visualize neutral lipids.We then used a nontargeted approach to profile lipids in 24 and 72 hpf embryos after phenanthrene treatment.To assess changes in lipid movement within the embryo,the yolk sac was dissected from the body at 24 and 72 hpf and analyzed separately from the body at 72 hpf.Overall,total metabolites were significantly reduced in the yolk sac,and staining for neutral lipids was reduced in the embryo body at 72 hpf.This result is consistent with significant reductions in triglycerides in both the embryo body and yolk,indicating a limited contribution of impaired cardiac function to lipid mobilization at the dose tested.Additionally,lysophosphatidylcholines and lysophosphatidylethanolamines were significantly increased in the 72 hpf embryo body.Bioinformatic pathway analysis indicated that changes to these lysophospholipids could be linked to a disease model associated with inflammation and neuron demyelination consistent with previously observed injuries to neuronal and eye development in fish embryos and larvae.展开更多
Zebrafish an emerging animal model for studies comparability with humans, good accessibility to the hearing utilize this animal model, methodologies need to be used to on auditory system. This model presents high org...Zebrafish an emerging animal model for studies comparability with humans, good accessibility to the hearing utilize this animal model, methodologies need to be used to on auditory system. This model presents high organ, and high throughput capacity. To better quantify the hearing function of the zebrafish. Zebrafish displays a series of innate and robust behavior related to its auditory function. Here, we reviewed the advantage of using zebrafish in auditory research and then introduced three behavioral tests, as follows： the startle response, the vestibular-ocular reflex, and rheotaxis. These tests are discussed in terms of their physiological characteristics, up-to-date technical development, and apparatus description. Test limitation and areas to improve are also introduced. Finally, we revealed the feasibility of these applications in zebrafish behavioral assessment and their potential in the high-throughput screening on hearing-related genes and drugs.展开更多
Zebrafish (Danio rerio) genetic screens allow isolation of a wide array of problems in vertebrate biology. The effects of base analogues 5-bromouracil and 6-aminopurine on the development of zebrafish embryos are repo...Zebrafish (Danio rerio) genetic screens allow isolation of a wide array of problems in vertebrate biology. The effects of base analogues 5-bromouracil and 6-aminopurine on the development of zebrafish embryos are reported for the first time in this study. The early development of the zebrafish embryos was little affected by 5-bromouracil and 6-aminopurine, while the late development (organogenesis) was significantly impaired. Embryos exposed to 5-bromouracil mainly showed curled tail, wavy body, golden pigmentation and the mouth with protruding lower jaw. 6-aminopurine-treated embryos had defective anterior structures, curled tails and wavy body. RAPD analysis showed that the majority of 5-bromouracil- and 6-aminopurine-treated larvae and fish shared banding patterns in common with the control, suggesting that most mutagenesis induced by these agents are point mutations. However, some fish derived from 5-bromouracil-treated embryos had golden (gol) pigmentation; and RAPD analysis revealed that their band patterns differed from those of the control. Possibly, 5-bromouracil can occasionally cause relatively extensive changes in the fish genome. The results of this study may provide valuable help for detailed studies of mutagenesis.展开更多
In this study, a recombinant pET28c-gBTB/POZ was constructed by cloning the sequence of the BTB/POZ domain of the zebrafish gcl （germ cell-less） into the expression vector pET28c, and pET28c-gBTB/POZ was transformed...In this study, a recombinant pET28c-gBTB/POZ was constructed by cloning the sequence of the BTB/POZ domain of the zebrafish gcl （germ cell-less） into the expression vector pET28c, and pET28c-gBTB/POZ was transformed into BL21（DE3） pLysS strain to express the fusion protein for the preparation of antibody. Polyclonal-antibody against the GCL-BTB/POZ domain was prepared by immunizing rabbit with the fusion protein, and the Western Blot and immuno-histochemical analysis were performed to detect the quantity of the polyclonal-antibody. The result indicates that the polyclonal-antibodies were of good quantity and specification. Further studies will be performed to demonstrate the function and expression pattern of the GCL protein during the development process of zebrafish with the polyclonal-antibody.展开更多
Zebrafish（Danio rerio） Z-OTU,containing OTU and TUDOR domains,was predicted to be a member of OTU-related protease,a family of the deubiquitylating enzymes（DUBs）.A previous report from our laboratory clearly descr...Zebrafish（Danio rerio） Z-OTU,containing OTU and TUDOR domains,was predicted to be a member of OTU-related protease,a family of the deubiquitylating enzymes（DUBs）.A previous report from our laboratory clearly describes the expression patterns of z-otu mRNA.Here,we characterized the Z-OTU protein during zebrafish oogenesis and early embryogenesis.After prokaryotic expression,the recombinant protein of the OTU domain and GST was purified and injected into rabbits to obtain the polyclonal antibody-anti-Z-OTU,which was used for immunohistochemistry in zebrafish ovaries and embryos.Interestingly,obvious differences existed between the expression patterns of z-otu mRNA and its protein during oogenesis and early embryogenesis.In stage I oocytes,z-otu mRNA was detected in cytoplasm while its protein existed in the germinal vesicle.In addition,its protein was distributed during entire oogenesis,while mRNA was not detected in oocytes at stage IV or mature oocytes.The z-otu mRNA disappeared after midblastula transition（MBT） and its protein gradually decreased after this stage.We inferred that Z-OTU protein,like other OTU-related protease with DUB activity,was required for germinal vesicle breakdown of oocytes during meiosis,germinal vesicle migration,and embryo cleavage maintenance.展开更多
Spring viremia of carp virus(SVCV)is globally widespread and poses a serious threat to aquatic ecology and aquaculture due to its broad host range.To develop effective agents to control SVCV infection,we selected 16 n...Spring viremia of carp virus(SVCV)is globally widespread and poses a serious threat to aquatic ecology and aquaculture due to its broad host range.To develop effective agents to control SVCV infection,we selected 16 naturally active small molecules to assess their anti-SVCV activity.Notably,dihydroartemisinin(DHA)(100μmol/L)and(S,S)-(+)-tetrandrine(TET)(16μmol/L)exhibited high antiviral effects in epithelioma papulosum cyprinid(EPC)cells,with inhibitory rates of 70.11%and 73.54%,respectively.The possible antiviral mechanisms were determined as follows:1.Preincubation with DHA and TET decreased viral particle infectivity in fish cells,suggesting that horizontal transmission of SVCV in the aquatic environment was disrupted;2.Although neither had an effect on viral adhesion,TET(but not DHA)interfered with SVCV entry into host cells(>80%),suggesting that TET may have an antiviral function in early viral replication.For in vivo study,both agents enhanced the survival rate of SVCV-infected zebrafish by 53.3%,significantly decreased viral load,and modulated the expression of antiviralrelated genes,indicating that DHA and TET may stimulate the host innate immune response to prevent viral infection.Overall,our findings indicated that DHA and TET had positive effects on suppressing SVCV infection by affecting early-stage viral replication,thus holding great potential as immunostimulants to reduce the risk of aquatic rhabdovirus disease outbreaks.展开更多
Shoaling behavior provides numerous fitness benefits for fish, including enhanced access to mates, increased success in foraging and protection from predators. We were interested in determining whether shoaling intens...Shoaling behavior provides numerous fitness benefits for fish, including enhanced access to mates, increased success in foraging and protection from predators. We were interested in determining whether shoaling intensity differed throughout the day. To do this we kept adult zebrafish Danio rerio in different lighting conditions for 10 days： ＂Normal＂ （12：12LD, lights on at 0800 hrs）, ＂Reverse＂ （12：12LD, lights on at 2000 hrs）, DD, or LL, and then observed the shoaling behavior at different times during the day. Our findings suggest that daily variations exist in shoaling behavior, with mean shoaling times for fish from the ＇normal＇ group being the lowest at the mid-point of the dark phase in the fish＇s subjective day （00：00 hrs）, then rising signifi- cantly throughout the day, reaching their highest intensity at 20：00 hrs （lights out）. Fish from the ＂reverse＂ LD cycle （lights on at 20：00 hrs） showed differences in the mean shoaling times at different times of day, but did not show a gradual increase in shoaling throughout their subjective day. Fish from the DD and LL groups did not show significant differences in the mean shoaling values at different times of day, suggesting that the differences observed in LD fish may not represent circadian rhythms. Therefore, these results demonstrate the existence of daily variations in the shoaling behavior of fish and suggest that environmental cues in the form of light/dark cycles play an important role in regulating these variations [Current Zoology 58 （1）： 129-137, 2012].展开更多
Vibrio parahaemolyticus(Vp)is an aquatic zoonotic pathogen that causes vibriosis in marine animals as well as sepsis,gastroenteritis and wound infection in human.In vertebrates,the innate immune system plays a critica...Vibrio parahaemolyticus(Vp)is an aquatic zoonotic pathogen that causes vibriosis in marine animals as well as sepsis,gastroenteritis and wound infection in human.In vertebrates,the innate immune system plays a critical defense mechanism against Vp infection including transmigration of neutrophils.In this study,we have examined the genetic recombination and infectious process of Vp in the zebrafish(Danio rerio),a vertebrate model system extensively used for studying host-pathogen interactions.A pathogenic Vp strain,Vp57,tagged with red fluorescent protein(RFP)(Vp57^(RFP))was introduced into larval zebrafish at 3 days post fertilization(dpf),in which the innate immunity is present whereas the adaptive immunity has not yet developed.Vp57 and Vp57^(RFP)showed similar LD50 and induced similar symptoms and pathological changes in the hosts.We microinjected 579 colony-forming units(CFU)Vp57^(RFP)into the zebrafish caudal vein,and observed that neutrophils were recruited to the injection site and within 3 h post infection(hpi)Vp57^(RFP)were mainly distributed in the tail,eyes,heart and optic vesicle.After 3 hpi,the fish died with slight spine bending.Several Vp57^(RFP)were also detected in somites,the phagocytosis of neutrophils was activated through the progress of bacterial infections.We found that chemotaxis and phagocytosis of neutrophils occurs when zebrafish is infected by Vp57^(RFP),whereas Vp57^(RFP)can escape from neutrophils and colonize other remote regions using the blood circulation system.展开更多
Objective:To evaluate the larval consumption rate of Danio rerio(D.rerio)and Oreochromis mossambicus(O.mossambicus)for the control of filarial vector mosquito Culex quinquefasciatus(Cx.quinquefasciatus).Methods:The fi...Objective:To evaluate the larval consumption rate of Danio rerio(D.rerio)and Oreochromis mossambicus(O.mossambicus)for the control of filarial vector mosquito Culex quinquefasciatus(Cx.quinquefasciatus).Methods:The fishes D.rerio and O.mossambicus were tested by feeding assay against Cx.quinquefasciatus for 24 h.Results:After 24 h of feeding assay the fishes D.rerio and O.mossambicus exhibited potential reduction rate of Cx.quinquefasciatus larvae.Conclusions:The present research work demonstrates that the D.rerio and O.mossambicus can be used for integrated mosquito management,which is simple and safe to non target organisms.展开更多
Estradiol, or 17-β-estradiol （E2）, the most potent naturally occurring estrogen, is involved in the hormone-immune system interaction in both mammals and fish. However, in vivo studies are largely limited, and litt...Estradiol, or 17-β-estradiol （E2）, the most potent naturally occurring estrogen, is involved in the hormone-immune system interaction in both mammals and fish. However, in vivo studies are largely limited, and little is known about whether E2 exerts similar effects on both female and male zebrafish （Danio rerio）. Here, we show exposure of both sexes ofD. rerio to 20 nmol/L E2 resulted in a significant increase in Vgl expression, but caused little damage to the hepatocytes, suggesting that this is the optimum E2 concentration. Also, exposure to 20 nmol/L E2 for 20 days caused a marked increase in plasma IgM levels, but had little influence on the peripheral leukocyte density, providing the first evidence of a hormone-immune system interaction in this species.展开更多
Shoaling intensity in zebrafish Danio rerio is believed to vary throughout subjective day and night hours. This experiment examines long term variations in shoaling behavior. Adult zebrafish Dan io rerio were maintain...Shoaling intensity in zebrafish Danio rerio is believed to vary throughout subjective day and night hours. This experiment examines long term variations in shoaling behavior. Adult zebrafish Dan io rerio were maintained under a 12：12 LD cycle （with dim red light serving as reduced visibility during subjective dark hours）, and their shoaling behavior was monitored every hour for a three-day period of time. Our results show that zebrafish perform shoaling behavior throughout subjective day and under reduced visibility conditions, although mean shoaling times during the light phase were significantly higher than mean shoaling times during the dark phase. However, on the 3^rd day of the experiment, mean shoaling times during the subjective night had increased and mean shoaling times during the subjective day had decreased. This shift in intensity was not seen on the first two days of the study, and may represent the influence of experience on the behavior of the test fish. We believe this study shows that shoaling behavior changes with light/dark cycles and that fish shoal even during reduced visibility conditions.展开更多
文摘[Objective] The aim was to study the effect of bensulfuron-methyl herbicide on acute toxicity and genetics toxicity of Danio redo. [ Method] Median lethal concentration was calculated by acute toxicity test, and analyzing the herbicide whether existing in potential toxicity to aquatic organisms or not. Based on the study of acute toxicity, genetics toxicity was carried out, by calculating the micronucleus rate to judge bensulfuron-methyl herbicide whether existing in potential toxicity or not. [ Result ] The LD5o （24 h and 48 h） of bensulfuron-methyl herbicide are 0.698 ml/L and 0.637 ml/L respectively, the safe concentration was 0.159 ml/L. The results on the effects of micronucleus （MN） in erythrocytes of Danio redo induced by bensulfuron-methyl at different times and different concentrations showed that the MN rate of control group was 0.010 3%, the highest MN rate of experimental group reached to 0. 372%, it also indicated that bensulfuron-methyl herbicide had genetics toxicity to Danio redo. At the same detection time, there was dose-effect relationship of MN rate in erythrocytes between treatment and control groups with different concentrations. In the same treatment group, the MN rate in erythrocytes reached to peak value at 24 h, and decreased at 48 h and 72 h with the infection time was prolonged. [ Conclusion ] The study provides some basis for scientifically selecting and reasonably using herbicide.
基金supported by the National Basic Research Program (973) of China (No. 2013CB945204)the National Natural Science Foundation of China (Nos. 31320103915 and 21377128)
文摘Perfluorononanoic acid（PFNA） is a nine-carbon perfluoroalkyl acid widely used in industrial and domestic products. It is a persistent organic pollutant found in the environment as well as in the tissues of humans and wildlife. There is a concern that this chemical might be a developmental toxicant and teratogen in various ecosystems. In the present study,the toxic effects of PFNA were evaluated in zebrafish（Danio rerio） embryos. One hour post-fertilization embryos were treated with 0, 25, 50, 100, 200, 300, 350, and 400 μmol/L PFNA for 96 hr in 6-well plates. Developmental phenotypes and hatching rates were observed and recorded. Nineteen genes related to oxidative stress and lipid metabolism were examined using Quantitative RT-PCR and confirmed by whole mount in situ hybridization（WISH）. Results showed that PFNA delayed the development of zebrafish embryos, reduced the hatching rate, and caused ventricular edema and malformation of the spine. In addition, the amount of reactive oxygen species in the embryo bodies increased significantly after exposure to PFNA compared with that of the control group. The Quantitative RT-PCR and WISH experiments demonstrated that m RNA expression of the lfabp and ucp2 genes increased significantly while that of sod1 and mt-nd1 decreased significantly after PFNA exposure. The m RNA expression levels of gpx1 and mt-atp6 decreased significantly in the high concentration group. However, the m RNA expression levels of both ppara and pparg did not show any significant variation after exposure. These findings suggest that PFNA affected the development of zebrafish embryos at relatively low concentrations.
基金supported by the National Key R&D Program of China (Nos.2018YFC1004300 and 2018YFC1004304)。
文摘Ethylene thiourea(ETU)converted from ethylene bisdithiocarbamate(EBDC)fungicides has aroused great concern because of its prevalence and harmful effects.Although ETU-induced neurotoxicity has been reported,the potential mechanisms remain unclear.This study provided insights into its neurotoxic effects at environmentally relevant concentrations in zebrafish.Our findings showed that embryonic exposure to ETU decreased the hatch rate and delayed somite development.Furthermore,ETU treatment significantly reduced the dark velocity in the locomotion assay.The upregulated tendency of the mitogen-activated protein kinases(MAPK)pathway(mknk1,atf4,mapkapk3)screened by transcriptome analysis implied motor neuron degeneration,which was validated by subsequent morphological observation,as axon length and branches were truncated in the 62.5μg/L ETU group.However,although the rescue experiment with a p38 MAPK inhibitor(SB203580)successfully ameliorated axon degeneration,it failed to reverse the locomotion behaviors.Further exploration of transcriptome data revealed the varied expression of presynaptic scaffold protein-related genes(pcloa,pclob,bsna),whose downregulation might impair the neuromuscular junction(NMJ).Therefore,we reasonably suspected that ETU-induced neurobehavioral deficits might result from the combined effects of the MAPK pathway and presynaptic proteins.Considering this,we highlighted the necessity to take precautions and early interventions for susceptible ETU-exposed populations.
基金Project supported by the National Natural Science Foundation of China(Nos.21407038,21407037,21407051,21307023,and 21377117)the Zhejiang Provincial Natural Science Foundation of China(No.LY18B070008)
文摘Decabromodiphenylethane （DBDPE） has been widely used as an alternative flame retardant due to the restriction or phase-out of traditional polybrominated diphenyl ethers （PBDEs）, and is of increasing concern regarding its ubiquity, persistence, and potential adverse effects. In the present study, the toxicological effects of DBDPE were evaluated using zebrafish as an in vivo model. Upon being exposed to DBDPE-polluted sediments for a short term, it was found that the mortality and malformation of zebrafish （including edema, bent notochord, and bent tail） were not affected even at the highest concentration tested （1000.0μg/kg dry sediment）. Regarding behavioral responses, it was found that zebraflsh larvae of 48 hours post fertilization （hpf） in all groups escaped successfully with a touch to the dorsal fin. However, when exposed to the highest DBDPE concentration, the larvae of 120 hpf exhibited significantly smaller distances as compared to the control. Moreover, the results of the acetylcholinesterase （ACHE） activity, the expression levels of two important nerve-related genes, and the cell apoptosis all indicated that DBDPE posed low neurotoxicity in embryo-larval zebrafish. The results in this study shed some light on the potential risks of DBDPE in the real environment and highlight the application of the sediment exposure route in the future.
基金supported by the National Natural Science Foundation of China (No.42177411)the Natural Science Foundation of Fujian Province of China (No.2018J01067)
文摘Mepanipyrim,an anilinopyrimidine fungicide,has been extensively used to prevent fungal diseases in fruit culture.Currently,research on mepanipyrim-induced toxicity in organisms is still very scarce,especially visual developmental toxicity.Here,zebrafish larvae were employed to investigate mepanipyrim-induced visual developmental toxicity.Intense light andmonochromatic light stimuli-evoked escape experiments were used to investigate vision-guided behaviors.Meanwhile,transcriptomic sequencing and real-time quantitative PCR assays were applied to assess the potential mechanisms of mepanipyrim-induced visual developmental toxicity and vision-guided behavioral alteration.Our results showed that mepanipyrim exposure could induce retinal impairment and vision-guided behavioral alteration in larval zebrafish.In addition,the grk1b gene of the phototransduction signaling pathway was found to be a potential aryl hydrocarbon receptor(AhR)-regulated gene.Mepanipyrim-induced visual developmental toxicity was potentially related to the AhR signaling pathway.Furthermore,mepanipyrim-induced behavioral alteration was guided by the visual function,and the effects of mepanipyrim on long and middle wavelength light-sensitive opsins may be the main cause of vision-guided behavioral alteration.Our results provide insights into understanding the relationship between visual development and vision-guided behaviors induced by mepanipyrim exposure.
基金Funding was provided by the UCR/AES resource allocation program.
文摘Lecithotrophic fish embryos rely on finite maternally deposited yolk resources for early development.Toxicant exposure can disrupt the uptake of yolk resources with consequences for development.In this study,we investigate the impacts of altered yolk utilization on fish embryos using the cardiotoxic compound phenanthrene.Zebrafish embryos were exposed to a cardiotoxic concentration of phenanthrene beginning at 6 hpf(hours post-fertilization)until a maximum of 72 hpf.Embryos were stained with Oil Red O to visualize neutral lipids.We then used a nontargeted approach to profile lipids in 24 and 72 hpf embryos after phenanthrene treatment.To assess changes in lipid movement within the embryo,the yolk sac was dissected from the body at 24 and 72 hpf and analyzed separately from the body at 72 hpf.Overall,total metabolites were significantly reduced in the yolk sac,and staining for neutral lipids was reduced in the embryo body at 72 hpf.This result is consistent with significant reductions in triglycerides in both the embryo body and yolk,indicating a limited contribution of impaired cardiac function to lipid mobilization at the dose tested.Additionally,lysophosphatidylcholines and lysophosphatidylethanolamines were significantly increased in the 72 hpf embryo body.Bioinformatic pathway analysis indicated that changes to these lysophospholipids could be linked to a disease model associated with inflammation and neuron demyelination consistent with previously observed injuries to neuronal and eye development in fish embryos and larvae.
文摘Zebrafish an emerging animal model for studies comparability with humans, good accessibility to the hearing utilize this animal model, methodologies need to be used to on auditory system. This model presents high organ, and high throughput capacity. To better quantify the hearing function of the zebrafish. Zebrafish displays a series of innate and robust behavior related to its auditory function. Here, we reviewed the advantage of using zebrafish in auditory research and then introduced three behavioral tests, as follows： the startle response, the vestibular-ocular reflex, and rheotaxis. These tests are discussed in terms of their physiological characteristics, up-to-date technical development, and apparatus description. Test limitation and areas to improve are also introduced. Finally, we revealed the feasibility of these applications in zebrafish behavioral assessment and their potential in the high-throughput screening on hearing-related genes and drugs.
基金This work was supported by Ministry of Science and Technology (MOST) of China (G 1999012005).
文摘Zebrafish (Danio rerio) genetic screens allow isolation of a wide array of problems in vertebrate biology. The effects of base analogues 5-bromouracil and 6-aminopurine on the development of zebrafish embryos are reported for the first time in this study. The early development of the zebrafish embryos was little affected by 5-bromouracil and 6-aminopurine, while the late development (organogenesis) was significantly impaired. Embryos exposed to 5-bromouracil mainly showed curled tail, wavy body, golden pigmentation and the mouth with protruding lower jaw. 6-aminopurine-treated embryos had defective anterior structures, curled tails and wavy body. RAPD analysis showed that the majority of 5-bromouracil- and 6-aminopurine-treated larvae and fish shared banding patterns in common with the control, suggesting that most mutagenesis induced by these agents are point mutations. However, some fish derived from 5-bromouracil-treated embryos had golden (gol) pigmentation; and RAPD analysis revealed that their band patterns differed from those of the control. Possibly, 5-bromouracil can occasionally cause relatively extensive changes in the fish genome. The results of this study may provide valuable help for detailed studies of mutagenesis.
基金Supported by the National Natural Science Foundation of China (30570968, 30370744)
文摘In this study, a recombinant pET28c-gBTB/POZ was constructed by cloning the sequence of the BTB/POZ domain of the zebrafish gcl （germ cell-less） into the expression vector pET28c, and pET28c-gBTB/POZ was transformed into BL21（DE3） pLysS strain to express the fusion protein for the preparation of antibody. Polyclonal-antibody against the GCL-BTB/POZ domain was prepared by immunizing rabbit with the fusion protein, and the Western Blot and immuno-histochemical analysis were performed to detect the quantity of the polyclonal-antibody. The result indicates that the polyclonal-antibodies were of good quantity and specification. Further studies will be performed to demonstrate the function and expression pattern of the GCL protein during the development process of zebrafish with the polyclonal-antibody.
基金supported by grants from the ICGEB (International Center for Genetic Engineering and Biotechnology) (CRP/CHN02-01) (SONG Ping)the National Basic Research Program of China (2004CB117400) (SONG Ping)+1 种基金the National Natural Science Foundation of China (30150005 30270675) (SONG Ping)
文摘Zebrafish（Danio rerio） Z-OTU,containing OTU and TUDOR domains,was predicted to be a member of OTU-related protease,a family of the deubiquitylating enzymes（DUBs）.A previous report from our laboratory clearly describes the expression patterns of z-otu mRNA.Here,we characterized the Z-OTU protein during zebrafish oogenesis and early embryogenesis.After prokaryotic expression,the recombinant protein of the OTU domain and GST was purified and injected into rabbits to obtain the polyclonal antibody-anti-Z-OTU,which was used for immunohistochemistry in zebrafish ovaries and embryos.Interestingly,obvious differences existed between the expression patterns of z-otu mRNA and its protein during oogenesis and early embryogenesis.In stage I oocytes,z-otu mRNA was detected in cytoplasm while its protein existed in the germinal vesicle.In addition,its protein was distributed during entire oogenesis,while mRNA was not detected in oocytes at stage IV or mature oocytes.The z-otu mRNA disappeared after midblastula transition（MBT） and its protein gradually decreased after this stage.We inferred that Z-OTU protein,like other OTU-related protease with DUB activity,was required for germinal vesicle breakdown of oocytes during meiosis,germinal vesicle migration,and embryo cleavage maintenance.
基金supported by the National Natural Science Foundation of China(31902410)Natural Science Foundation of Zhejiang Province(LQ22C190002)+3 种基金Natural Science Foundation of Ningbo City(2021J117)Program of Science and Technology Department of Ningbo City(2021S058)One Health Interdisciplinary Research Project of Ningbo University(HZ202201)Zhejiang Xinmiao Talents Programs(2022R405B066)。
文摘Spring viremia of carp virus(SVCV)is globally widespread and poses a serious threat to aquatic ecology and aquaculture due to its broad host range.To develop effective agents to control SVCV infection,we selected 16 naturally active small molecules to assess their anti-SVCV activity.Notably,dihydroartemisinin(DHA)(100μmol/L)and(S,S)-(+)-tetrandrine(TET)(16μmol/L)exhibited high antiviral effects in epithelioma papulosum cyprinid(EPC)cells,with inhibitory rates of 70.11%and 73.54%,respectively.The possible antiviral mechanisms were determined as follows:1.Preincubation with DHA and TET decreased viral particle infectivity in fish cells,suggesting that horizontal transmission of SVCV in the aquatic environment was disrupted;2.Although neither had an effect on viral adhesion,TET(but not DHA)interfered with SVCV entry into host cells(>80%),suggesting that TET may have an antiviral function in early viral replication.For in vivo study,both agents enhanced the survival rate of SVCV-infected zebrafish by 53.3%,significantly decreased viral load,and modulated the expression of antiviralrelated genes,indicating that DHA and TET may stimulate the host innate immune response to prevent viral infection.Overall,our findings indicated that DHA and TET had positive effects on suppressing SVCV infection by affecting early-stage viral replication,thus holding great potential as immunostimulants to reduce the risk of aquatic rhabdovirus disease outbreaks.
文摘Shoaling behavior provides numerous fitness benefits for fish, including enhanced access to mates, increased success in foraging and protection from predators. We were interested in determining whether shoaling intensity differed throughout the day. To do this we kept adult zebrafish Danio rerio in different lighting conditions for 10 days： ＂Normal＂ （12：12LD, lights on at 0800 hrs）, ＂Reverse＂ （12：12LD, lights on at 2000 hrs）, DD, or LL, and then observed the shoaling behavior at different times during the day. Our findings suggest that daily variations exist in shoaling behavior, with mean shoaling times for fish from the ＇normal＇ group being the lowest at the mid-point of the dark phase in the fish＇s subjective day （00：00 hrs）, then rising signifi- cantly throughout the day, reaching their highest intensity at 20：00 hrs （lights out）. Fish from the ＂reverse＂ LD cycle （lights on at 20：00 hrs） showed differences in the mean shoaling times at different times of day, but did not show a gradual increase in shoaling throughout their subjective day. Fish from the DD and LL groups did not show significant differences in the mean shoaling values at different times of day, suggesting that the differences observed in LD fish may not represent circadian rhythms. Therefore, these results demonstrate the existence of daily variations in the shoaling behavior of fish and suggest that environmental cues in the form of light/dark cycles play an important role in regulating these variations [Current Zoology 58 （1）： 129-137, 2012].
基金This work was supported by the Innovation Program of Shanghai Municipal Education Commission(13ZZ127)the Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry(D-8002-15-0042)+1 种基金the Doctor Startup Fund of Shanghai Ocean University(A-0209-13-0105344)the SHOU&MSU Marine Joint Research Center Grant(A1-0209-15-0806).
文摘Vibrio parahaemolyticus(Vp)is an aquatic zoonotic pathogen that causes vibriosis in marine animals as well as sepsis,gastroenteritis and wound infection in human.In vertebrates,the innate immune system plays a critical defense mechanism against Vp infection including transmigration of neutrophils.In this study,we have examined the genetic recombination and infectious process of Vp in the zebrafish(Danio rerio),a vertebrate model system extensively used for studying host-pathogen interactions.A pathogenic Vp strain,Vp57,tagged with red fluorescent protein(RFP)(Vp57^(RFP))was introduced into larval zebrafish at 3 days post fertilization(dpf),in which the innate immunity is present whereas the adaptive immunity has not yet developed.Vp57 and Vp57^(RFP)showed similar LD50 and induced similar symptoms and pathological changes in the hosts.We microinjected 579 colony-forming units(CFU)Vp57^(RFP)into the zebrafish caudal vein,and observed that neutrophils were recruited to the injection site and within 3 h post infection(hpi)Vp57^(RFP)were mainly distributed in the tail,eyes,heart and optic vesicle.After 3 hpi,the fish died with slight spine bending.Several Vp57^(RFP)were also detected in somites,the phagocytosis of neutrophils was activated through the progress of bacterial infections.We found that chemotaxis and phagocytosis of neutrophils occurs when zebrafish is infected by Vp57^(RFP),whereas Vp57^(RFP)can escape from neutrophils and colonize other remote regions using the blood circulation system.
文摘Objective:To evaluate the larval consumption rate of Danio rerio(D.rerio)and Oreochromis mossambicus(O.mossambicus)for the control of filarial vector mosquito Culex quinquefasciatus(Cx.quinquefasciatus).Methods:The fishes D.rerio and O.mossambicus were tested by feeding assay against Cx.quinquefasciatus for 24 h.Results:After 24 h of feeding assay the fishes D.rerio and O.mossambicus exhibited potential reduction rate of Cx.quinquefasciatus larvae.Conclusions:The present research work demonstrates that the D.rerio and O.mossambicus can be used for integrated mosquito management,which is simple and safe to non target organisms.
基金Supported by the Natural Science Foundation of Shandong Province (No. 2007ZRB02238)the Natural Science Foundation of Qingdao Municipality (No. 06-2-2-15-jch)
文摘Estradiol, or 17-β-estradiol （E2）, the most potent naturally occurring estrogen, is involved in the hormone-immune system interaction in both mammals and fish. However, in vivo studies are largely limited, and little is known about whether E2 exerts similar effects on both female and male zebrafish （Danio rerio）. Here, we show exposure of both sexes ofD. rerio to 20 nmol/L E2 resulted in a significant increase in Vgl expression, but caused little damage to the hepatocytes, suggesting that this is the optimum E2 concentration. Also, exposure to 20 nmol/L E2 for 20 days caused a marked increase in plasma IgM levels, but had little influence on the peripheral leukocyte density, providing the first evidence of a hormone-immune system interaction in this species.
文摘Shoaling intensity in zebrafish Danio rerio is believed to vary throughout subjective day and night hours. This experiment examines long term variations in shoaling behavior. Adult zebrafish Dan io rerio were maintained under a 12：12 LD cycle （with dim red light serving as reduced visibility during subjective dark hours）, and their shoaling behavior was monitored every hour for a three-day period of time. Our results show that zebrafish perform shoaling behavior throughout subjective day and under reduced visibility conditions, although mean shoaling times during the light phase were significantly higher than mean shoaling times during the dark phase. However, on the 3^rd day of the experiment, mean shoaling times during the subjective night had increased and mean shoaling times during the subjective day had decreased. This shift in intensity was not seen on the first two days of the study, and may represent the influence of experience on the behavior of the test fish. We believe this study shows that shoaling behavior changes with light/dark cycles and that fish shoal even during reduced visibility conditions.
