Alzheimer＇s disease, the leading cause of dementia in the elderly, is a complex neurodegenerative disorder which leads to a progressive decline in cognitive functions. A rapid screening model is highly demanded for i...Alzheimer＇s disease, the leading cause of dementia in the elderly, is a complex neurodegenerative disorder which leads to a progressive decline in cognitive functions. A rapid screening model is highly demanded for identification and evaluation of novel anti-Alzheimer＇s disease drugs from a large numbers of compounds. Until now, numerous studies utilized zebrafish model for drug discovery. Since aluminum can induce a similar biological activity in zebrafish as in Alzheimer patients, in this study, we developed a novel animal model using 3 to 5 day post-fertilization larval zebrafish by optimizing the doses and duration of aluminum chloride exposure. Six anti-Alzheimer＇s disease drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model. Importantly, Rivastigmine, ThT, Flurbiprofen and AM-117 could increase the value of Dyskinesia Recovery Rate by 53.4-64%, 169.4-200%, 54.5-96% and 70.9-121%, respectively. Rivastigmine, Memantine, ThT, Flurbiprofen, Rosiglitazone and AM-117 improved the value of Response Efficiency by 86.6-175.1%, 28.2-66.6%, 127.2-236.5%, 118.3-323.7%, 26.6-140.8% and 70.2-161.4%, respectively. Our results suggest that the zebrafish model developed in this study could be a useful tool for high throughput screening of potential novel anti-Alzheimer＇s disease leading compounds targeting acetylcholinesterase, N-methyl-D-aspartic acid receptor, γ-secretase, peroxisome proliferator-activated receptor-γand amyloid-β.展开更多
OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular...OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular mechanism of hypoxia still remains unclear.In this work,we further explore the molecular mechanism of hypoxia and adaption to attenuate the damage in zebrafish model that have potential to resist hypoxic environment.METHODS The hypoxic zebrafish model was established in different concentration of oxygen with 3%,5%,10%,21%in water.The brain tissue was separated and the RNA-seq was used to identify the differentially expressed genes.The related endogenous metabolites profiles were obtained by LC-HDMS,and the multivariate statistics was applied to discover the important metabolites candidates in hypoxic zebrafish.The candidates were searched in HMDB,KEGG and Lipid Maps databases.RESULTS The zebrafish hypoxic model was successfully constructed via the different concentration of oxygen,temperature and hypoxic time.The activities of the related hypoxic metabolic enzymes and factors including HIF-1a,actate dehydrogenase(LDH)and citrate synthase(CS)were evaluated.Significant differences(P<0.05 and fold change>2)in the expression of 422 genes were observed between the normal and 3% hypoxic model.Among them,201 genes increased depended on the lower concentration of oxygen.53 metabolites were identified that had significant difference between the hypoxia and control groups(P<0.05,fold change>1.5 and VIP>1.5).The ten key metabolites were increased gradually while six compounds were decreased.The endogenous hypoxic metabolites of phenylalanine,D-glucosamine-6P and several important lipids with the relevant hub genes had similar change in hypoxic model.In addition,the metabolic pathways of phenylalanine,glutamine and glycolipid were influenced in both the levels of genes and metabolites.CONCLUSION The up-regulation of phenylalanine,D-glucosamine-6P and lipid may have further understanding of protective effect in hypoxia.Our data provided an insight to further reveal the hypoxia and adaptation mechanism.展开更多
AIM: To establish a model of retinal neurodegeneration induced by N-Methyl-D-aspartic acid(NMDA) in adult zebrafish.METHODS: We compared the effects of three different NMDA delivery methods on retinal neurodegeneratio...AIM: To establish a model of retinal neurodegeneration induced by N-Methyl-D-aspartic acid(NMDA) in adult zebrafish.METHODS: We compared the effects of three different NMDA delivery methods on retinal neurodegeneration in adult zebrafish: immersion(I.M.), intravitreal injection(I.V.), and intraperitoneal injection(I.P.), and examined retinal pathology and degeneration by hematoxylin and eosin and TUNEL staining in the treated zebrafish. Effects of the NMDA receptor antagonist MK-801 and the natural product resveratrol on NMDA-induced retinal neurodegeneration were also assessed.RESULTS: The thickened inner retina was seen in histology with 100 μmol/L NMDA by I.M. administration. Significant apoptosis in the retinal ganglion cell layer and retinal thickness reduction occurred in 0.5 mol/L NMDA I.P. administration group.Seizure-like behavioral changes, but no retinal histological alteration occurred in 16 mg/kg NMDA I.P. administration group. Resveratrol and MK-801 prevented NMDA-induced retinal neurodegeneration in the zebrafish. CONCLUSION: Among the three drug administration methods, I.V. injection of NMDA is the most suitable for establishment of an acute retinal damage model inzebrafish. I.M. with NMDA is likely the best for use as a chronic retinal damage model. I.P. treatment with NMDA causes brain damage. Resveratrol and MK801 may be a clinically valuable treatment for retinal neurodegeneration.展开更多
The Editors-in-Chiefs would like to alert readers to the fact that concerns have been raised related to the publication“Nervonic acid suppresses MPTP-induced Parkinson’s disease in an adult zebrafish model by regula...The Editors-in-Chiefs would like to alert readers to the fact that concerns have been raised related to the publication“Nervonic acid suppresses MPTP-induced Parkinson’s disease in an adult zebrafish model by regulating the MAPK/NF-κB signaling pathway,inflammation,apoptosis,and oxidative stress”,which was published in Food Bioscience Volume 59,June 2024,103777,https://doi.org/10.1016/j.fbio.2024.103777.展开更多
Background:Despite the success of tyrosine kinase inhibitors in chronic myeloid leukemia(CML)therapy,CML still faces the challenges of drug resistance and progression to blast crisis.Twenty-five percent of patients ha...Background:Despite the success of tyrosine kinase inhibitors in chronic myeloid leukemia(CML)therapy,CML still faces the challenges of drug resistance and progression to blast crisis.Twenty-five percent of patients have imatinib resistance and treatment difficulties due to heterogeneity after progression,but little is known about the mechanism.A key transcription factor in hematopoiesis,MYB,has been reported to increase abnormally in several types of aggressive blood disorders including CML.Methods:This study used a zebrafish model to explore the relationship between BCR/ABL1 and c-myb in CML progression.A CML zebrafish model was crossed with a c-myb hyperactivity transgenic line.Results:It was found that both exogenous BCR/ABL1 and c-myb could up-regulate the expression of neutrophil-related genes.More seriously,neutrophil accumulation was observed when BCR/ABL1 was combined with c-myb overexpression.Further studies showed that c-myb may be one of the downstream targets of BCR/ABL1 and the effect of BCR/ABL1 on neutrophils was c-myb dependent.Taking advantage of this inheritable in vivo model,it was shown that a combination of imatinib and flavopiridol,a cyclin-dependent kinase inhibitor targeting MYB,could more effectively alleviate the aggressive phenotype of the double transgene line.Conclusion:In summary,this study suggests that c-myb acts downstream of BCR/ABL1 and is involved in CML progression and is therefore a risk factor and a valuable target for the treatment of CML progression.The model used in the study could be helpful in high-throughput drug screening in CML transformation.展开更多
Zebrafish are increasingly being utilized as a laboratory animal species to study various biological processes,both normal and pathological.It is crucial to comprehend the dynamics of zebrafish locomotion and put fort...Zebrafish are increasingly being utilized as a laboratory animal species to study various biological processes,both normal and pathological.It is crucial to comprehend the dynamics of zebrafish locomotion and put forth realistic models since their locomotion characteristics are employed as feedback indicators in diverse experiments.In this study,we conducted experimental research on the locomotion of zebrafish across various spatial sizes,focusing on the analysis of motion step size and motion direction.The results indicated that the motion step exhibits long-range correlations,the motion direction shows unbiased randomness,and the data characteristics are not influenced by spatial size.The dynamic mechanisms are complicated dynamical processes rather than fractional Brownian or Lévy processes motion.Based on the experimental results,we proposed a model for describing the movement of zebrafish in a circular container.Our findings shed light on the locomotion characteristics of zebrafish,and have the potential to benefit both the biological outcomes of animal tests and the welfare of the subjects.展开更多
Because the physiological characteristics and melanin regulation mechanism of zebrafish are highly similar with those of humans,it is of high reference value to use zebrafish model in the evaluation of cosmetic whiten...Because the physiological characteristics and melanin regulation mechanism of zebrafish are highly similar with those of humans,it is of high reference value to use zebrafish model in the evaluation of cosmetic whitening efficacy.In this study,zebrafish embryos are used as biological models to evaluate the whitening efficacy of six kinds of cosmetics raw materials,such as antioxidant,preservative and essence,and the formula of facial cleanser and facial mask products,and the limitations of the zebrafish melanin production grayscale detection method in practical application are discussed.The results show that the selection of different types of components can also reduce the production of melanin and show whitening effect.It can be seen that the gray scale method of melanin production in zebrafish is suitable for the evaluation of the efficacy of raw materials.In practical application,due to the complexity of the formula,the toxic effects of different types of ingredients may interfere with the melanin generation of zebrafish,affecting the judgment and evaluation of whitening efficacy.For the detection of whitening efficacy of products,a comprehensive evaluation system should be built together with other methods to accurately evaluate the whitening efficacy.展开更多
Cilia, microtubule-based structures found on the surface of almost all vertebrate cells, play an array of diverse biological functions. Abnormal ciliary axonemal structure and function can result in a class of genetic...Cilia, microtubule-based structures found on the surface of almost all vertebrate cells, play an array of diverse biological functions. Abnormal ciliary axonemal structure and function can result in a class of genetic disorders that are collectively termed ciliopathies. Model organisms, including Chlamydomonas reinhardtii and Caenorhabditis elegans have been widely used to study the complex genetic basis of ciliopathies. Here, we review the advantages of the zebrafish as a vertebrate model for human ciliopathies. We summarize the features of zebrafish cilia, and the major findings and contributions of the zebrafish model in recent studies of human ciliopathies. We also discuss the new genome editing approaches being efficiently used in zebrafish, and the exciting prospects of these approaches in modeling human ciliopathies.展开更多
Zebrafish（Danio rerio） is an ideal model for studying the mechanism of infectious disease and the interaction between host and pathogen.As a teleost,zebrafish has developed a complete immune system which is similar ...Zebrafish（Danio rerio） is an ideal model for studying the mechanism of infectious disease and the interaction between host and pathogen.As a teleost,zebrafish has developed a complete immune system which is similar to mammals.Moreover,the easy acquirement of large amounts of transparent embryos makes it a good candidate for gene manipulation and drug screening.In a zebrafish infection model,all of the site,timing,and dose of the bacteria microinjection into the embryo are important factors that determine the bacterial infection of host.Here,we established a multi-site infection model in zebrafish larvae of 36 hours post-fertilization（hpf） by micro-injecting wild-type or GFP-expressing Staphylococcus aereus（5.aureus） with gradient burdens into different embryo sites including the pericardial cavity（PC）,eye,the fourth hindbrain ventricle（4V）,yolk circulation valley（YCV）,caudal vein（CV）,yolk body（YB）,and Duct of Cuvier（DC） to resemble human infectious disease.With the combination of GFP-expressing S.aureus and transgenic zebrafish Tg（corola：eGFP;lyz：Dsred） and Tg（lyz：Dsred） lines whose macrophages or neutrophils are fluorescent labeled,we observed the dynamic process of bacterial infection by in vivo multicolored confocal fluorescence imaging.Analyses of zebrafish embryo survival, bacterial proliferation and myeloid cells phagocytosis show that the site- and dose-dependent differences exist in infection of different bacterial entry routes.This work provides a consideration for the future study of pathogenesis and host resistance through selection of multi-site infection model.More interaction mechanisms between pathogenic bacteria virulence factors and the immune responses of zebrafish could be determined through zebrafish multi-site infection model.展开更多
OBJECTIVE To assess the efficancy of Yangxue Qingnao granules on simvastatininduced vascular injury model in zebrafish.METHODS Since statins can inhibit vascular development in zebrafish,in this study,we developed a n...OBJECTIVE To assess the efficancy of Yangxue Qingnao granules on simvastatininduced vascular injury model in zebrafish.METHODS Since statins can inhibit vascular development in zebrafish,in this study,we developed a novel animal model using 1 to 3 day post-fertilization larval zebrafish by optimizing the doses and duration of simvastatin exposure.Five pro-angiogenic drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model.Zebrafish was treated with different concentration of Yangxue Qingnao granules(62.5,125 and 250 μg·mL-1) for 2 d then tested for the area of subintestinal vein vessels.RESULTS Vascular regeneration promoting effect of five pro-angiogenic drugs(calycosin,astragaloside,chlorogenic acid,ferulic acid and Panax Notoginseng Saponins) were 8-48%,24-51%,35-58%,28-75% and 37-69%,respectively.In 62.5,125 and 250 μg·mL-1 Yangxue Qingnao granules group,vascular regeneration promoting effect were 21%(P>0.05),84%(P<0.01) and 53%(P<0.01).CONCLUSION Our results demonstrate that the zebrafish vascular injury model validated in this study could be used for in vivo angiogenesis studies and drug screening and for assessing pro-angiogenic drugs with different mechanisms.Yangxue Qingnao granules could promote the vascular regeneration in zebrafish.展开更多
Crohn’s disease and ulcerative colitis—inflammatory bowel diseases (IBD)—are chronic conditions with an inadequately understood pathogenesis. Employing a set of novel molecular markers in a gene expression assay (q...Crohn’s disease and ulcerative colitis—inflammatory bowel diseases (IBD)—are chronic conditions with an inadequately understood pathogenesis. Employing a set of novel molecular markers in a gene expression assay (qPCR), we have used adult zebrafish to investigate two acute inflammatory models, induced by the haptenizing agents oxazolone and TNBS. In addition, goblet cell dynamics in the scales and intestine and 5-HT (serotonin) in intestinal tissues were investigated through optical projection tomography. Gene expression studies revealed a distinct and significant upregulation of proinflammatory cytokines, acute-phase reactants and metalloprotease 9 in both chemical models, primarily after 72 hours. In comparison, transcription factors and cytokines associated with Th1 and Th17 (Crohn’s) and Th2 (ulcerative colitis) were mainly not affected in this acute setting. However, elevated transcript levels were detected in Foxp3, IL-10 and T-bet, which are linked with tolerance and Tregs in mammals. Goblet cells in scales were depleted in both chemical models and in the intestine of oxazolone-treated fish. A marked 5-HT signal was noted in intestinal tissue of some chemically treated zebrafish. In conclusion, a distinct acute inflammatory reaction was induced in both chemical models. Further, oxazolone and TNBS did not result in clear-cut Th2 and Th1/Th17 pathway signaling at this early timepoint, but the applied molecular tools may provide further insight to the IBD pathogenesis and translational value of the IBD zebrafish model.展开更多
Background:The CUG-binding protein Elav-like family member 2(CELF2)gene has been linked to the pathogenesis of epilepsy,but its precise role remains unclear.This study aimed to investigate the pathogenic mechanisms of...Background:The CUG-binding protein Elav-like family member 2(CELF2)gene has been linked to the pathogenesis of epilepsy,but its precise role remains unclear.This study aimed to investigate the pathogenic mechanisms of CELF2 mutation in epilepsy,utilizing zebrafish models to explore its molecular pathways and biological impact.Methods:Whole-exome sequencing was performed to identify CELF2mutations associated with epilepsy.CELF2 zebrafish model was generated using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-related protein 9technology and morpholinos,followed by behavioral and electroencephalographic analyses to confirm epileptic phenotypes.Proteomic and metabolomic analyses were conducted to examine the impact of CELF2 deficiency on metabolic pathways,and single-cell sequencing was used to assess alterations in neuronal cell populations.Results:An infant with infantile epileptic spasms syndrome associated with a CELF2(p.Pro520Arg)gene mutation was reported.We established zebrafish models with celf2 gene knockout and knockdown and found that zebrafish with celf2mutations exhibited epilepsy-like behaviors,which could be rescued by injection of CELF2 wild-type mRNA.Significant changes were observed in crucial marker genes associated with the nervous system in the celf2^(+/−)group,including FOS,BDNF,NPAS4,GABRA1,GABRG2,and PYYA.Disruptions in lipid metabolism,heat shock protein 90 beta1(Hsp90b1),were identified in proteomic and metabolomic analyses.Single-cell sequencing showed changes in nucleosome localization,nucleosome DNA binding,arginine and proline metabolic pathways,gonadotropin-releasing hormone signaling pathway,and nucleotide-binding oligomerization domain receptor signaling pathway.Conclusions:Our study has revealed a promising association between defects in the CELF2 gene and epilepsy using a zebrafish model,suggesting that CLEF2 is a causative gene in epilepsy.These findings not only indicate the potential impact on the biological process influenced by the CELF2 gene defect but also offer hopeful insights into the pathogenesis of epilepsy and potential therapeutic targets.展开更多
Animal models are necessary to investigate the pathogenic features underlying motor neuron degeneration and for therapeutic development in amyotrophic lateral sclerosis（ALS）. Measures of model validity allow for a c...Animal models are necessary to investigate the pathogenic features underlying motor neuron degeneration and for therapeutic development in amyotrophic lateral sclerosis（ALS）. Measures of model validity allow for a critical interpretation of results from each model and caution from over-interpretation of experimental models. Face and construct validity refer to the similarity in phenotype and the proposed causal factor to the human disease, respectively. More recently developed models are restricted by limited phenotype characterization, yet new models hold promise for novel disease insights, thus highlighting their importance. In this article, we evaluate the features of face and construct validity of our new zebrafish model of environmentally-induced motor neuron degeneration and discuss this in the context of current environmental and genetic ALS models, including C9 orf72, mutant Cu/Zn superoxide dismutase 1 and TAR DNA-binding protein 43 mouse and zebrafish models. In this mini-review, we discuss the pros and cons to validity criteria in each model. Our zebrafish model of environmentally-induced motor neuron degeneration displays convincing features of face validity with many hallmarks of ALS-like features, and weakness in construct validity. However, the value of this model may lie in its potential to be more representative of the pathogenic features underlying sporadic ALS cases, where environmental factors may be more likely to be involved in disease etiology than single dominant gene mutations. It may be necessary to compare findings between different strains and species modeling specific genes or environmental factors to confirm findings from ALS animal models and tease out arbitrary strain-and overexpression-specific effects.展开更多
目的分析斑马鱼模型在药源性肾损伤评价中的应用价值。方法检索PubMed、Web of Science、中国知网(CNKI)数据库中斑马鱼模型在药物及化学品诱导肾毒性研究中的文献,检索时限为各数据库自建库起至2023年12月31日,归纳其方法学、评价指标...目的分析斑马鱼模型在药源性肾损伤评价中的应用价值。方法检索PubMed、Web of Science、中国知网(CNKI)数据库中斑马鱼模型在药物及化学品诱导肾毒性研究中的文献,检索时限为各数据库自建库起至2023年12月31日,归纳其方法学、评价指标、毒性机制及在中医药评价中的具体实践,展望其在新领域的应用潜力。结果斑马鱼因与人类高度的遗传与生理相似性、早期胚胎透明、繁殖快速等特点,在肾毒性评价中展现出独特优势,可作为补充模型。斑马鱼模型肾损伤评价指标包括表型和组织病理形态学指标、滤过功能相关指标、细胞生物学过程和生化指标、肾损伤细胞标志物,涵盖从整体表型到分子标志物的多层次检测体系。化学品诱导斑马鱼模型肾毒性的主要机制包括细胞凋亡、细胞死亡、纤维化、炎性反应、氧化应激等。该模型已成功应用于马兜铃酸等中药毒性物质的评价及大黄酸、大豆苷等潜在肾保护成分的筛选,但未来仍需进一步明确斑马鱼模型在化学品毒性筛选中的地位,考察其对药物毒性动力学、实验操作及结果评价的重现性和标准,并加快高端配套设备的研发。结论斑马鱼模型可作为衔接体外筛选与哺乳动物体内实验、进行药物肾毒性快速评价与机制研究的一种重要补充模型,且在复杂的中医药体系安全性评价中具有潜力。展开更多
基金Acknowledgments The authors thank the National Natural Science Foundation of China （81302646）, Natural Science Foundation of Zhejiang Province （LQ13H300002）, Science Technology Department of Zhejiang Province （2015F50015） and Health and Family Planning commission of Zhejiang Province （XKQ-010-001 and 2013KYB070） for financial support.
文摘Alzheimer＇s disease, the leading cause of dementia in the elderly, is a complex neurodegenerative disorder which leads to a progressive decline in cognitive functions. A rapid screening model is highly demanded for identification and evaluation of novel anti-Alzheimer＇s disease drugs from a large numbers of compounds. Until now, numerous studies utilized zebrafish model for drug discovery. Since aluminum can induce a similar biological activity in zebrafish as in Alzheimer patients, in this study, we developed a novel animal model using 3 to 5 day post-fertilization larval zebrafish by optimizing the doses and duration of aluminum chloride exposure. Six anti-Alzheimer＇s disease drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model. Importantly, Rivastigmine, ThT, Flurbiprofen and AM-117 could increase the value of Dyskinesia Recovery Rate by 53.4-64%, 169.4-200%, 54.5-96% and 70.9-121%, respectively. Rivastigmine, Memantine, ThT, Flurbiprofen, Rosiglitazone and AM-117 improved the value of Response Efficiency by 86.6-175.1%, 28.2-66.6%, 127.2-236.5%, 118.3-323.7%, 26.6-140.8% and 70.2-161.4%, respectively. Our results suggest that the zebrafish model developed in this study could be a useful tool for high throughput screening of potential novel anti-Alzheimer＇s disease leading compounds targeting acetylcholinesterase, N-methyl-D-aspartic acid receptor, γ-secretase, peroxisome proliferator-activated receptor-γand amyloid-β.
基金supported by National Natural Science Foundation of China(81573683 and 81173121)
文摘OBJECTIVE Hypoxia is associated with many complicated pathophysiological and biochemical processes that integrated and regulated via the key gene,protein and endogenous metabolite levels.Up to date,the exact molecular mechanism of hypoxia still remains unclear.In this work,we further explore the molecular mechanism of hypoxia and adaption to attenuate the damage in zebrafish model that have potential to resist hypoxic environment.METHODS The hypoxic zebrafish model was established in different concentration of oxygen with 3%,5%,10%,21%in water.The brain tissue was separated and the RNA-seq was used to identify the differentially expressed genes.The related endogenous metabolites profiles were obtained by LC-HDMS,and the multivariate statistics was applied to discover the important metabolites candidates in hypoxic zebrafish.The candidates were searched in HMDB,KEGG and Lipid Maps databases.RESULTS The zebrafish hypoxic model was successfully constructed via the different concentration of oxygen,temperature and hypoxic time.The activities of the related hypoxic metabolic enzymes and factors including HIF-1a,actate dehydrogenase(LDH)and citrate synthase(CS)were evaluated.Significant differences(P<0.05 and fold change>2)in the expression of 422 genes were observed between the normal and 3% hypoxic model.Among them,201 genes increased depended on the lower concentration of oxygen.53 metabolites were identified that had significant difference between the hypoxia and control groups(P<0.05,fold change>1.5 and VIP>1.5).The ten key metabolites were increased gradually while six compounds were decreased.The endogenous hypoxic metabolites of phenylalanine,D-glucosamine-6P and several important lipids with the relevant hub genes had similar change in hypoxic model.In addition,the metabolic pathways of phenylalanine,glutamine and glycolipid were influenced in both the levels of genes and metabolites.CONCLUSION The up-regulation of phenylalanine,D-glucosamine-6P and lipid may have further understanding of protective effect in hypoxia.Our data provided an insight to further reveal the hypoxia and adaptation mechanism.
基金Supported by the National Natural Science Foundation of China (No.81271425 No.81260148+5 种基金 No.31400988 No.81160144 No.31171044)the Jiangxi Provincial Natural Science Foundation (No.20181ACG70010)the Natural Science Foundation of Jiangxi (No.20151BBG70243 No.20122BCB23007)
文摘AIM: To establish a model of retinal neurodegeneration induced by N-Methyl-D-aspartic acid(NMDA) in adult zebrafish.METHODS: We compared the effects of three different NMDA delivery methods on retinal neurodegeneration in adult zebrafish: immersion(I.M.), intravitreal injection(I.V.), and intraperitoneal injection(I.P.), and examined retinal pathology and degeneration by hematoxylin and eosin and TUNEL staining in the treated zebrafish. Effects of the NMDA receptor antagonist MK-801 and the natural product resveratrol on NMDA-induced retinal neurodegeneration were also assessed.RESULTS: The thickened inner retina was seen in histology with 100 μmol/L NMDA by I.M. administration. Significant apoptosis in the retinal ganglion cell layer and retinal thickness reduction occurred in 0.5 mol/L NMDA I.P. administration group.Seizure-like behavioral changes, but no retinal histological alteration occurred in 16 mg/kg NMDA I.P. administration group. Resveratrol and MK-801 prevented NMDA-induced retinal neurodegeneration in the zebrafish. CONCLUSION: Among the three drug administration methods, I.V. injection of NMDA is the most suitable for establishment of an acute retinal damage model inzebrafish. I.M. with NMDA is likely the best for use as a chronic retinal damage model. I.P. treatment with NMDA causes brain damage. Resveratrol and MK801 may be a clinically valuable treatment for retinal neurodegeneration.
文摘The Editors-in-Chiefs would like to alert readers to the fact that concerns have been raised related to the publication“Nervonic acid suppresses MPTP-induced Parkinson’s disease in an adult zebrafish model by regulating the MAPK/NF-κB signaling pathway,inflammation,apoptosis,and oxidative stress”,which was published in Food Bioscience Volume 59,June 2024,103777,https://doi.org/10.1016/j.fbio.2024.103777.
基金National Key R&D Program of ChinaGrant/Award Number:2018YFA0801000+5 种基金National Natural Science Foundation of ChinaGrant/Award Number:32170830Natural Science Foundation of Guangdong ProvinceChinaGrant/Award Number:2021A1515010422South China University of Technology。
文摘Background:Despite the success of tyrosine kinase inhibitors in chronic myeloid leukemia(CML)therapy,CML still faces the challenges of drug resistance and progression to blast crisis.Twenty-five percent of patients have imatinib resistance and treatment difficulties due to heterogeneity after progression,but little is known about the mechanism.A key transcription factor in hematopoiesis,MYB,has been reported to increase abnormally in several types of aggressive blood disorders including CML.Methods:This study used a zebrafish model to explore the relationship between BCR/ABL1 and c-myb in CML progression.A CML zebrafish model was crossed with a c-myb hyperactivity transgenic line.Results:It was found that both exogenous BCR/ABL1 and c-myb could up-regulate the expression of neutrophil-related genes.More seriously,neutrophil accumulation was observed when BCR/ABL1 was combined with c-myb overexpression.Further studies showed that c-myb may be one of the downstream targets of BCR/ABL1 and the effect of BCR/ABL1 on neutrophils was c-myb dependent.Taking advantage of this inheritable in vivo model,it was shown that a combination of imatinib and flavopiridol,a cyclin-dependent kinase inhibitor targeting MYB,could more effectively alleviate the aggressive phenotype of the double transgene line.Conclusion:In summary,this study suggests that c-myb acts downstream of BCR/ABL1 and is involved in CML progression and is therefore a risk factor and a valuable target for the treatment of CML progression.The model used in the study could be helpful in high-throughput drug screening in CML transformation.
基金Project supported by the National Natural Science Foundation of China(Grant No.12205006)the Excellent Youth Scientific Research Project of Anhui Province,China(Grant No.2022AH030107)。
文摘Zebrafish are increasingly being utilized as a laboratory animal species to study various biological processes,both normal and pathological.It is crucial to comprehend the dynamics of zebrafish locomotion and put forth realistic models since their locomotion characteristics are employed as feedback indicators in diverse experiments.In this study,we conducted experimental research on the locomotion of zebrafish across various spatial sizes,focusing on the analysis of motion step size and motion direction.The results indicated that the motion step exhibits long-range correlations,the motion direction shows unbiased randomness,and the data characteristics are not influenced by spatial size.The dynamic mechanisms are complicated dynamical processes rather than fractional Brownian or Lévy processes motion.Based on the experimental results,we proposed a model for describing the movement of zebrafish in a circular container.Our findings shed light on the locomotion characteristics of zebrafish,and have the potential to benefit both the biological outcomes of animal tests and the welfare of the subjects.
文摘Because the physiological characteristics and melanin regulation mechanism of zebrafish are highly similar with those of humans,it is of high reference value to use zebrafish model in the evaluation of cosmetic whitening efficacy.In this study,zebrafish embryos are used as biological models to evaluate the whitening efficacy of six kinds of cosmetics raw materials,such as antioxidant,preservative and essence,and the formula of facial cleanser and facial mask products,and the limitations of the zebrafish melanin production grayscale detection method in practical application are discussed.The results show that the selection of different types of components can also reduce the production of melanin and show whitening effect.It can be seen that the gray scale method of melanin production in zebrafish is suitable for the evaluation of the efficacy of raw materials.In practical application,due to the complexity of the formula,the toxic effects of different types of ingredients may interfere with the melanin generation of zebrafish,affecting the judgment and evaluation of whitening efficacy.For the detection of whitening efficacy of products,a comprehensive evaluation system should be built together with other methods to accurately evaluate the whitening efficacy.
基金supported by the grants from the National Natural Science Foundation of China (Nos. 31372274 and 31422051)Shandong Provincial Natural Science Foundation, China (No. JQ201506) to C.Z.supported by the grant from NIH/NIDCR (R01DE018043) to P.C.Y.
文摘Cilia, microtubule-based structures found on the surface of almost all vertebrate cells, play an array of diverse biological functions. Abnormal ciliary axonemal structure and function can result in a class of genetic disorders that are collectively termed ciliopathies. Model organisms, including Chlamydomonas reinhardtii and Caenorhabditis elegans have been widely used to study the complex genetic basis of ciliopathies. Here, we review the advantages of the zebrafish as a vertebrate model for human ciliopathies. We summarize the features of zebrafish cilia, and the major findings and contributions of the zebrafish model in recent studies of human ciliopathies. We also discuss the new genome editing approaches being efficiently used in zebrafish, and the exciting prospects of these approaches in modeling human ciliopathies.
基金supported by the grants of"Hundred Talent"of Chinese Academy of Sciencesthe National Natural Science Foundation of China(No.31070950) to B.Hu
文摘Zebrafish（Danio rerio） is an ideal model for studying the mechanism of infectious disease and the interaction between host and pathogen.As a teleost,zebrafish has developed a complete immune system which is similar to mammals.Moreover,the easy acquirement of large amounts of transparent embryos makes it a good candidate for gene manipulation and drug screening.In a zebrafish infection model,all of the site,timing,and dose of the bacteria microinjection into the embryo are important factors that determine the bacterial infection of host.Here,we established a multi-site infection model in zebrafish larvae of 36 hours post-fertilization（hpf） by micro-injecting wild-type or GFP-expressing Staphylococcus aereus（5.aureus） with gradient burdens into different embryo sites including the pericardial cavity（PC）,eye,the fourth hindbrain ventricle（4V）,yolk circulation valley（YCV）,caudal vein（CV）,yolk body（YB）,and Duct of Cuvier（DC） to resemble human infectious disease.With the combination of GFP-expressing S.aureus and transgenic zebrafish Tg（corola：eGFP;lyz：Dsred） and Tg（lyz：Dsred） lines whose macrophages or neutrophils are fluorescent labeled,we observed the dynamic process of bacterial infection by in vivo multicolored confocal fluorescence imaging.Analyses of zebrafish embryo survival, bacterial proliferation and myeloid cells phagocytosis show that the site- and dose-dependent differences exist in infection of different bacterial entry routes.This work provides a consideration for the future study of pathogenesis and host resistance through selection of multi-site infection model.More interaction mechanisms between pathogenic bacteria virulence factors and the immune responses of zebrafish could be determined through zebrafish multi-site infection model.
文摘OBJECTIVE To assess the efficancy of Yangxue Qingnao granules on simvastatininduced vascular injury model in zebrafish.METHODS Since statins can inhibit vascular development in zebrafish,in this study,we developed a novel animal model using 1 to 3 day post-fertilization larval zebrafish by optimizing the doses and duration of simvastatin exposure.Five pro-angiogenic drugs with a variety of mechanisms were tested to validate the newly developed zebrafish model.Zebrafish was treated with different concentration of Yangxue Qingnao granules(62.5,125 and 250 μg·mL-1) for 2 d then tested for the area of subintestinal vein vessels.RESULTS Vascular regeneration promoting effect of five pro-angiogenic drugs(calycosin,astragaloside,chlorogenic acid,ferulic acid and Panax Notoginseng Saponins) were 8-48%,24-51%,35-58%,28-75% and 37-69%,respectively.In 62.5,125 and 250 μg·mL-1 Yangxue Qingnao granules group,vascular regeneration promoting effect were 21%(P>0.05),84%(P<0.01) and 53%(P<0.01).CONCLUSION Our results demonstrate that the zebrafish vascular injury model validated in this study could be used for in vivo angiogenesis studies and drug screening and for assessing pro-angiogenic drugs with different mechanisms.Yangxue Qingnao granules could promote the vascular regeneration in zebrafish.
文摘Crohn’s disease and ulcerative colitis—inflammatory bowel diseases (IBD)—are chronic conditions with an inadequately understood pathogenesis. Employing a set of novel molecular markers in a gene expression assay (qPCR), we have used adult zebrafish to investigate two acute inflammatory models, induced by the haptenizing agents oxazolone and TNBS. In addition, goblet cell dynamics in the scales and intestine and 5-HT (serotonin) in intestinal tissues were investigated through optical projection tomography. Gene expression studies revealed a distinct and significant upregulation of proinflammatory cytokines, acute-phase reactants and metalloprotease 9 in both chemical models, primarily after 72 hours. In comparison, transcription factors and cytokines associated with Th1 and Th17 (Crohn’s) and Th2 (ulcerative colitis) were mainly not affected in this acute setting. However, elevated transcript levels were detected in Foxp3, IL-10 and T-bet, which are linked with tolerance and Tregs in mammals. Goblet cells in scales were depleted in both chemical models and in the intestine of oxazolone-treated fish. A marked 5-HT signal was noted in intestinal tissue of some chemically treated zebrafish. In conclusion, a distinct acute inflammatory reaction was induced in both chemical models. Further, oxazolone and TNBS did not result in clear-cut Th2 and Th1/Th17 pathway signaling at this early timepoint, but the applied molecular tools may provide further insight to the IBD pathogenesis and translational value of the IBD zebrafish model.
基金supported by grants from the National Science Foundation of China(Nos.82071686 and 82271692)the Clinical Research Fund of West China Second University Hospital(Nos.KL115 and KL072)the Natural Science Foundation of Sichuan Province(No.2022NSFSC0782).
文摘Background:The CUG-binding protein Elav-like family member 2(CELF2)gene has been linked to the pathogenesis of epilepsy,but its precise role remains unclear.This study aimed to investigate the pathogenic mechanisms of CELF2 mutation in epilepsy,utilizing zebrafish models to explore its molecular pathways and biological impact.Methods:Whole-exome sequencing was performed to identify CELF2mutations associated with epilepsy.CELF2 zebrafish model was generated using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-related protein 9technology and morpholinos,followed by behavioral and electroencephalographic analyses to confirm epileptic phenotypes.Proteomic and metabolomic analyses were conducted to examine the impact of CELF2 deficiency on metabolic pathways,and single-cell sequencing was used to assess alterations in neuronal cell populations.Results:An infant with infantile epileptic spasms syndrome associated with a CELF2(p.Pro520Arg)gene mutation was reported.We established zebrafish models with celf2 gene knockout and knockdown and found that zebrafish with celf2mutations exhibited epilepsy-like behaviors,which could be rescued by injection of CELF2 wild-type mRNA.Significant changes were observed in crucial marker genes associated with the nervous system in the celf2^(+/−)group,including FOS,BDNF,NPAS4,GABRA1,GABRG2,and PYYA.Disruptions in lipid metabolism,heat shock protein 90 beta1(Hsp90b1),were identified in proteomic and metabolomic analyses.Single-cell sequencing showed changes in nucleosome localization,nucleosome DNA binding,arginine and proline metabolic pathways,gonadotropin-releasing hormone signaling pathway,and nucleotide-binding oligomerization domain receptor signaling pathway.Conclusions:Our study has revealed a promising association between defects in the CELF2 gene and epilepsy using a zebrafish model,suggesting that CLEF2 is a causative gene in epilepsy.These findings not only indicate the potential impact on the biological process influenced by the CELF2 gene defect but also offer hopeful insights into the pathogenesis of epilepsy and potential therapeutic targets.
基金supported by a grant from Estate of Luther Allyn Shourds Dean,No.20R17162(to CAS)
文摘Animal models are necessary to investigate the pathogenic features underlying motor neuron degeneration and for therapeutic development in amyotrophic lateral sclerosis（ALS）. Measures of model validity allow for a critical interpretation of results from each model and caution from over-interpretation of experimental models. Face and construct validity refer to the similarity in phenotype and the proposed causal factor to the human disease, respectively. More recently developed models are restricted by limited phenotype characterization, yet new models hold promise for novel disease insights, thus highlighting their importance. In this article, we evaluate the features of face and construct validity of our new zebrafish model of environmentally-induced motor neuron degeneration and discuss this in the context of current environmental and genetic ALS models, including C9 orf72, mutant Cu/Zn superoxide dismutase 1 and TAR DNA-binding protein 43 mouse and zebrafish models. In this mini-review, we discuss the pros and cons to validity criteria in each model. Our zebrafish model of environmentally-induced motor neuron degeneration displays convincing features of face validity with many hallmarks of ALS-like features, and weakness in construct validity. However, the value of this model may lie in its potential to be more representative of the pathogenic features underlying sporadic ALS cases, where environmental factors may be more likely to be involved in disease etiology than single dominant gene mutations. It may be necessary to compare findings between different strains and species modeling specific genes or environmental factors to confirm findings from ALS animal models and tease out arbitrary strain-and overexpression-specific effects.
文摘目的分析斑马鱼模型在药源性肾损伤评价中的应用价值。方法检索PubMed、Web of Science、中国知网(CNKI)数据库中斑马鱼模型在药物及化学品诱导肾毒性研究中的文献,检索时限为各数据库自建库起至2023年12月31日,归纳其方法学、评价指标、毒性机制及在中医药评价中的具体实践,展望其在新领域的应用潜力。结果斑马鱼因与人类高度的遗传与生理相似性、早期胚胎透明、繁殖快速等特点,在肾毒性评价中展现出独特优势,可作为补充模型。斑马鱼模型肾损伤评价指标包括表型和组织病理形态学指标、滤过功能相关指标、细胞生物学过程和生化指标、肾损伤细胞标志物,涵盖从整体表型到分子标志物的多层次检测体系。化学品诱导斑马鱼模型肾毒性的主要机制包括细胞凋亡、细胞死亡、纤维化、炎性反应、氧化应激等。该模型已成功应用于马兜铃酸等中药毒性物质的评价及大黄酸、大豆苷等潜在肾保护成分的筛选,但未来仍需进一步明确斑马鱼模型在化学品毒性筛选中的地位,考察其对药物毒性动力学、实验操作及结果评价的重现性和标准,并加快高端配套设备的研发。结论斑马鱼模型可作为衔接体外筛选与哺乳动物体内实验、进行药物肾毒性快速评价与机制研究的一种重要补充模型,且在复杂的中医药体系安全性评价中具有潜力。
