The autotetraploid Carassius auratus(4nRR,4n=200,RRRR)is derived from whole-genome duplication of Carassius auratus red var.(RCC,2n=100,RR).In the current study,we demonstrated that chromatophores and pigment changes ...The autotetraploid Carassius auratus(4nRR,4n=200,RRRR)is derived from whole-genome duplication of Carassius auratus red var.(RCC,2n=100,RR).In the current study,we demonstrated that chromatophores and pigment changes directly caused the coloration and variation of 4nRR skin(red in RCC,brownish-yellow in4nRR).To further explore the molecular mechanisms underlying coloration formation and variation in 4nRR,we performed transcriptome profiling and molecular functional verification in RCC and 4nRR.Results revealed that scarb1,associated with carotenoid metabolism,underwent significant down-regulation in 4nRR.Efficient editing of this candidate pigment gene provided clear evidence of its significant role in RCC coloration.Subsequently,we identified four divergent scarb1 homeologs in 4nRR:two original scarb1 homeologs from RCC and two duplicated ones.Notably,three of these homeologs possessed two highly conserved alleles,exhibiting biased and allelespecific expression in the skin.Remarkably,after precise editing of both the original and duplicated scarb1homeologs and/or alleles,4nRR individuals,whether singly or multiply mutated,displayed a transition from brownishyellow skin to a cyan-gray phenotype.Concurrently,the proportional areas of the cyan-gray regions displayed a gene-dose correlation.These findings illustrate the subfunctionalization of duplicated scarb1,with all scarb1genes synergistically and equally contributing to the pigmentation of 4nRR.This is the first report concerning the functional differentiation of duplicated homeologs in an autopolyploidfish,substantiallyenrichingour understanding of coloration formation and change within this group of organisms.展开更多
The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency.However, the proof of chondrogenic potential of the cells is scarce.Therefore,we investigated ...The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency.However, the proof of chondrogenic potential of the cells is scarce.Therefore,we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor(TGF)-p3 and bone morphogenetic protein(BMP)-6.After isolation of periodontal ligament stem cells(PDLSCs) from human periodontal ligament,the cells were cultured in Dulbecco’s modified Eagle’s medium(DMEM) with 20%fetal bovine serum(FBS).A mechanical force initiated chondrogenic differentiation of the cells.For chondrogenic differentiation,10μg·LTGF-β3 or 100μg·LBMP-6 and the combination treating group for synergistic effect of the growth factors.We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay,histology,immunohistochemistry and genetic analysis.PDLSCs showed mesenchymal stem cell properties proved by FACS analysis.Glycosaminoglycans contents were increased 217%by TGF-β3 and 220%by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281%compared to control.The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls,but also TGF-P3 or BMP-6 single treatment dramatically.The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions.The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis,which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.展开更多
Aim: To identify proteins induced by androgen in Sertoli cells during spermatogenesis. Methods: We analyzed protein profiles in TM4 Sertoli cells treated with dihydrotestosterone (DHT) using surface enhanced laser...Aim: To identify proteins induced by androgen in Sertoli cells during spermatogenesis. Methods: We analyzed protein profiles in TM4 Sertoli cells treated with dihydrotestosterone (DHT) using surface enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Results: We found increases in the expression of a 5.0-kDa protein at 15 min, an 11.3-kDa protein at 24 h and 4.3 kDa, 5.7 kDa, 5.8 kDa, 9.95 kDa and 9.98 kDa proteins at 48 h after the treatment. In contrast, the expression of 6.3 kDa and 8.6 kDa proteins decreased at 30 min, and 4.9 kDa, 5.0 kDa, 12.4 kDa and 19.8 kDa proteins at 48 h after the treatment. The ll.3-kDa protein was identified as macrophage migration inhibitory factor (MIF) known to having various functions. The 9.98-kDa protein was identified as calgizzarin related to calcium channels. The timing of their expression suggests that MIF and calgizzarin are involved in late regulation of spermatogenesis in Sertoli cells by androgen. Conclusion: MIF and calgizzarin are two important androgen-responsive proteins produced by Sertoli cells and they might play a role in regulating spermatogenesis.展开更多
Genotoxicity of dichlorvos and butachlor on erythrocytes of Rana zhenhaiensis tadpoles was investigated by the alkaline single-cell gel electrophoresis assay or comet assay.Tadpoles were treated for 24h in the laborat...Genotoxicity of dichlorvos and butachlor on erythrocytes of Rana zhenhaiensis tadpoles was investigated by the alkaline single-cell gel electrophoresis assay or comet assay.Tadpoles were treated for 24h in the laboratory with different concentrations of the testing agents,2.256,4.512,6.768,9.024,11.280mg/L for dichlorvos and 0.292,0.584,0.876,1.168,1.460mg/L for butachlor,to use the comet Assay to test for the significance of dosage responsiveness to an increase in DNA damage,asmeasured by themean DNA tail length-to-width ratio.The concentrations of 4.512mg/L dichlorvos and 0.876mg/L butachlor resulted in highly significant increases in DNA damage of the tadpoles.There were linear correlations between themean DNA tail length-to-width ratio and the concentrations of the two test substances.Our results showed that the two commonly used agricultural chemicals caused dose dependent DNA damage of amphibians,and that comet assaymight be a useful tool formeasuring DNA damage of tadpoles exposed in the field.展开更多
The basic mechanism of heterosis has not been systematically and completely characterized.In previous studies,we obtained three economically important fishes that exhibit rapid growth,WR(WCC♀×RCC♂),WR-Ⅱ(WR♀...The basic mechanism of heterosis has not been systematically and completely characterized.In previous studies,we obtained three economically important fishes that exhibit rapid growth,WR(WCC♀×RCC♂),WR-Ⅱ(WR♀×WCC♂),and WR-Ⅲ(WR-Ⅱ♀×4nAU♂),through distant hybridization.However,the mechanism underlying this rapid growth remains unclear.In this study,we found that WR,WR-Ⅱ,and WR-Ⅲshowed muscle hypertrophy and higher muscle protein and fat contents compared with their parent species(RCC and WCC).Candidate genes responsible for this rapid growth were then obtained through an analysis of 12 muscle transcriptomes.Notably,the mRNA level of mstnb(myostatin b),which is a negative regulator of myogenesis,was significantly reduced in WR,WR-Ⅱ,and WR-Ⅲcompared with the parent species.To verify the function of mstnb,a mstnb-deficient mutant RCC line was generated using the CRISPR-Cas9 technique.The average body weight of mstnb-deficient RCC at 12 months of age was significantly increased by 29.57%compared with that in wild-type siblings.Moreover,the area and number of muscle fibers were significantly increased in mstnb-deficient RCC,indicating hypertrophy and hyperplasia.Furthermore,the muscle protein and fat contents were significantly increased in mstnb-deficient RCC.The molecular regulatory mechanism of mstnb was then revealed by transcription profiling,which showed that genes related to myogenesis(myod,myog,and myf5),protein synthesis(PI3K-AKT-mTOR),and lipogenesis(pparγand fabp3)were highly activated in hybrid fishes and mstnb-deficient RCC.This study revealed that low expression or deficiency of mstnb regulates somatic growth by promoting myogenesis,protein synthesis,and lipogenesis in hybrid fishes and mstnb-deficient RCC,which provides evidence for the molecular mechanism of heterosis via distant hybridization.展开更多
Objective To investigate the mechanistic basis for the anti-proliferation and anti-invasion effect of tumor necrosis factor-related apoptosis-induced ligand(TRAIL)and celastrol combination treatment(TCCT)in glioblasto...Objective To investigate the mechanistic basis for the anti-proliferation and anti-invasion effect of tumor necrosis factor-related apoptosis-induced ligand(TRAIL)and celastrol combination treatment(TCCT)in glioblastoma cells.Methods Cell counting kit-8 was used to detect the effects of different concentrations of celastrol(0-16µmol/L)and TRAIL(0-500 ng/mL)on the cell viability of glioblastoma cells.U87 cells were randomly divided into 4 groups,namely control,TRAIL(TRAIL 100 ng/mL),Cel(celastrol 0.5µmol/L)and TCCT(TRAIL 100 ng/mL+celastrol 0.5µmol/L).Cell proliferation,migration,and invasion were detected by colony formation,wound healing,and Transwell assays,respectively.Quantitative reverse transcription polymerase chain reaction and Western blotting were performed to assess the levels of epithelial-mesenchymal transition(EMT)markers(zona occludens,N-cadherin,vimentin,zinc finger E-box-binding homeobox,Slug,and β-catenin).Wnt pathway was activated by lithium chloride(LiCl,20 mol/L)and the mechanism for action of TCCT was explored.Results Celastrol and TRAIL synergistically inhibited the proliferation,migration,invasion,and EMT of U87 cells(P<0.01).TCCT up-regulated the expression of GSK-3β and down-regulated the expression of β-catenin and its associated proteins(P<0.05 or P<0.01),including c-Myc,Cyclin-D1,and matrix metalloproteinase(MMP)-2.In addition,LiCl,an activator of the Wnt signaling pathway,restored the inhibitory effects of TCCT on the expression of β-catenin and its downstream genes,as well as the migration and invasion of glioblastoma cells(P<0.05 or P<0.01).Conclusions Celastrol and TRAIL can synergistically suppress glioblastoma cell migration,invasion,and EMT,potentially through inhibition of Wnt/β-catenin pathway.This underlies a novel mechanism of action for TCCT as an effective therapy for glioblastoma.展开更多
Adrenomedullin,a peptide with multiple physiological functions in nervous system injury and disease,has aroused the interest of researchers.This review summarizes the role of adrenomedullin in neuropathological disord...Adrenomedullin,a peptide with multiple physiological functions in nervous system injury and disease,has aroused the interest of researchers.This review summarizes the role of adrenomedullin in neuropathological disorders,including pathological pain,brain injury and nerve regeneration,and their treatment.As a newly characterized pronociceptive mediator,adrenomedullin has been shown to act as an upstream factor in the transmission of noxious information for various types of pathological pain including acute and chronic inflammatory pain,cancer pain,neuropathic pain induced by spinal nerve injury and diabetic neuropathy.Initiation of glia-neuron signaling networks in the peripheral and central nervous system by adrenomedullin is involved in the formation and maintenance of morphine tolerance.Adrenomedullin has been shown to exert a facilitated or neuroprotective effect against brain injury including hemorrhagic or ischemic stroke and traumatic brain injury.Additionally,adrenomedullin can serve as a regulator to promote nerve regeneration in pathological conditions.Therefore,adrenomedullin is an important participant in nervous system diseases.展开更多
This study aims to improve control schemes for COVID-19 by a numerical model with estimation of parameters.We established a multi-level and multi-objective nonlinear SEIDR model to simulate the virus transmission.The ...This study aims to improve control schemes for COVID-19 by a numerical model with estimation of parameters.We established a multi-level and multi-objective nonlinear SEIDR model to simulate the virus transmission.The early spread in Japan was adopted as a case study.The first 96 days since the infection were divided into five stages with parameters estimated.Then,we analyzed the trend of the parameter value,age structure ratio,and the defined PCR test index(standardization of the scale of PCR tests).It was discovered that the self-healing rate and confirmed rate were linear with the age structure ratio and the PCR test index using the stepwise regression method.The transmission rates were related to the age structure ratio,PCR test index,and isolation efficiency.Both isolation measures and PCR test medical screening can effectively reduce the number of infected cases based on the simulation results.However,the strategy of increasing PCR test medical screening would encountered a bottleneck effect on the virus control when the index reached 0.3.The effectiveness of the policy would decrease and the basic reproduction number reached the extreme value at 0.6.This study gave a feasible combination for isolation and PCR test by simulation.The isolation intensity could be adjusted to compensate the insufficiency of PCR test to control the pandemic.展开更多
Parkinson’s disease is the second most common neurodegenerative disease and has currently no effective treatment,one that would be able to stop or reverse the loss of dopaminergic neurons in the substantia nigra pars...Parkinson’s disease is the second most common neurodegenerative disease and has currently no effective treatment,one that would be able to stop or reverse the loss of dopaminergic neurons in the substantia nigra pars compacta.In addition,Parkinson’s disease diagnosis is typically done when a significant percentage of the dopaminergic neurons is already lost.In neurodegenerative disorders,some therapeutic strategies could be effective only at inhibiting further degeneration;on the other hand,cell replacement therapies aim at replacing lost neurons,an approach that would be ideal for the treatment of Parkinson’s disease.Many cell replacement therapies have been tested since the 1970s in the field of Parkinson’s disease;however,there are still significant limitations prohibiting a successful clinical application.From the first fetal midbrain intrastriatal graft to the most recent conversion of astrocytes into dopaminergic neurons,we have gained equally,significant insights and questions still looking for an answer.This review aims to summarize the main milestones in cell replacement approaches against Parkinson’s disease.By focusing on achievements and failures,as well as on the additional research steps needed,we aim to provide perspective on how future cell replacement therapies treats Parkinson’s disease.展开更多
Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenes...Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenesis at both transcriptional and posttranscriptional levels,little is currently known regarding the regulation of sperm flagella assembly by microRNAs(miRNAs).The role of miRNAs in the development of sperm abnormalities in sterile triploid fish has not been studied.Results:In this study,we found that miR-199-5p was widely expressed in all detected tissues of different-ploidy crucian carp.As one of the testis-specific candidate markers,Tekt1 was predominantly expressed in the testis.Quantitative real-time PCR(qRT-PCR)analyses showed that the expression trend of miR-199-5p was exactly opposite to that of Tekt1.Through bioinformatics analysis,we identified a putative miR-199-5p binding site in the Tekt1 mRNA.We further identified Tekt1 as a target of miR-199-5p using luciferase reporter assay.Finally,we confirmed that miR-199-5p was necessary for sperm flagellar assembly and spermatogenesis in vivo via intraperitoneal injection of miR-199-5p antagomir or agomir in diploid red crucian carp.Moreover,miR-199-5p gainof-function could lead to spermatids apoptosis and abnormal spermatozoa structure,which is similar to that of allotriploid crucian carp.Conclusions:Our studies suggested that abnormally elevated miR-199-5p inhibited the sperm flagella formation in spermiogenesis by negatively regulating the expression of Tekt1,thereby causing sperm abnormalities of male allotriploid crucian carp.展开更多
Recently,cilia defects have been proposed to contribute to scoliosis.Here,we demonstrate that coiled-coil domain-containing 57(Ccdc57)plays an essential role in straightening the body axis of zebrafish by regulating c...Recently,cilia defects have been proposed to contribute to scoliosis.Here,we demonstrate that coiled-coil domain-containing 57(Ccdc57)plays an essential role in straightening the body axis of zebrafish by regulating ciliary beating in the brain ventricle(BV).Zygotic ccdc57(Zccdc57)mutant zebrafish developes scoliosis without significant changes in their bone density and calcification,and the maternal-zygotic ccdc57(MZccdc57)mutant embryos display curved bodies since the long-pec stage.The expression of ccdc57 is enriched in ciliated tissues and immunofluorescence analysis reveals colocalization of Ccdc57-HA with acetylated a-tubulin,implicating it in having a role in ciliary function.Further examination reveals that it is the coordinated cilia beating of multiple cilia bundles(MCB)in the MZccdc57 mutant embryos that is affected at 48 hours post fertilization,when the compromised cerebrospinal fluid flow and curved body axis have already occurred.Either ccdc57 m RNA injection or epinephrine treatment reverses the spinal curvature in MZccdc57 mutant larvae from ventrally curly to straight or even dorsally curly and significantly upregulates urotensin signaling.This study reveals the role of ccdc57 in maintaining coordinated cilia beating of MCB in the BV.展开更多
Cartilage development is controlled by the highly synergistic proliferation and differentiation of growth plate chondrocytes,in which the Indian hedgehog(IHH)and parathyroid hormone-related protein-parathyroid hormone...Cartilage development is controlled by the highly synergistic proliferation and differentiation of growth plate chondrocytes,in which the Indian hedgehog(IHH)and parathyroid hormone-related protein-parathyroid hormone-1 receptor(PTHrP-PTH1R)feedback loop is crucial.The inositol-requiring enzyme 1a/X-box-binding protein-1 spliced(IRE1α/XBP1s)branch of the unfolded protein response(UPR)is essential for normal cartilage development.However,the precise role of ER stress effector IRE1α,encoded by endoplasmic reticulum to nucleus signaling 1(ERN1),in skeletal development remains unknown.Herein,we reported that loss of IRE1α accelerates chondrocyte hypertrophy and promotes endochondral bone growth.ERN1 acts as a negative regulator of chondrocyte proliferation and differentiation in postnatal growth plates.Its deficiency interrupted PTHrP/PTH1R and IHH homeostasis leading to impaired chondrocyte hypertrophy and differentiation.XBP1s,produced by p-IRE1α-mediated splicing,binds and up-regulates PTH1R and IHH,which coordinate cartilage development.Meanwhile,ER stress cannot be activated normally in ERN1-deficient chondrocytes.In conclusion,ERN1 deficiency accelerates chondrocyte hypertrophy and cartilage mineralization by impairing the homeostasis of the IHH and PTHrP/PTH1R feedback loop and ER stress.ERN1 may have a potential role as a new target for cartilage growth and maturation.展开更多
As an increasingly used alternative to perfluorooctanoic acid(PFOA),hexafluoropropylene oxide trimer acid(HFPO-TA)has been widely detected in global water environments.However,little is known regarding its toxic effec...As an increasingly used alternative to perfluorooctanoic acid(PFOA),hexafluoropropylene oxide trimer acid(HFPO-TA)has been widely detected in global water environments.However,little is known regarding its toxic effects on cardiovascular development.Here,zebrafish embryos were treated with egg water containing 0,60,120,or 240 mg/L HFPO-TA.Results showed that HFPO-TA treatment led to a significant reduction in both larval survival percentage and heart rate.Furthermore,HFPO-TA exposure caused severe pericardial edema and elongation of the sinus venous to bulbus arteriosus distance(SV-BA)in Tg(myl7:GFP)transgenic larvae,disrupting the expression of genes involved in heart development and thus causing abnormal heart looping.Obvious sprouting angiogenesis was observed in the 120 and 240 mg/L exposed Tg(fli:GFP)transgenic larvae.HFPO-TA treatment also impacted the mRNA levels of genes involved in the vascular endothelial growth factor(VEGF)pathway and embryonic vascular development.HFPO-TA exposure significantly decreased erythrocyte number in Tg(gata1:DsRed)transgenic embryos and influenced gene expression associated with the heme metabolism pathway.HFPO-TA also induced oxidative stress and altered the transcriptional levels of genes related to cell cycle and apoptosis,inhibiting cell proliferation while promoting apoptosis.Therefore,HFPO-TA exposure may induce abnormal development of the cardiovascular and hematopoietic systems in zebrafish embryos,suggesting it may not be a suitable or safe alternative for PFOA.展开更多
Viral infectious diseases significantly threaten the sustainability of freshwater fish aquaculture.The lack of studies on epidemic transmission patterns and mechanisms inhibits the development of containment strategie...Viral infectious diseases significantly threaten the sustainability of freshwater fish aquaculture.The lack of studies on epidemic transmission patterns and mechanisms inhibits the development of containment strategies from the viewpoint of veterinary public health.This study raises an epidemic mathematical model considering water transmission with the aim of analyzing the transmission process more accurately.The basic reproduction number R0 was derived by the model parameter including the water transmission coefficient and was used for the analysis of the virus transmission.Spring viremia of carp virus(SVCV)and zebrafish were used as model viruses and animals,respectively,to conduct the transmission experiment.Transmission through water was achieved by connecting two aquarium tanks with a water channel but blocking the fish movement between the tanks.With the collected experimental data,we determined the optimal hybrid machine learning algorithm to analyze the transmission process using an established mathematical model.In addition,future transmission was predicted and validated using the epidemic model and an optimal algorithm.Finally,the sensitivity of model parameters and the simulations of R0 variation were performed based on the modified complex epidemic model.This study is of significance in providing theoretical guidance for minimizing R0 by manipulating model parameters with containment measures.More importantly,since the modified model and algorithm demonstrated better performance in handling freshwater fish transmission problems,this study advances the future application of transmissible disease modeling with larger datasets in freshwater fish aquaculture.展开更多
Hybridization is widely used. However, for a long time, systematic theories and technologies related to hybridization in fish have been lacking. In this study, through long-term systematic research, we investigated an...Hybridization is widely used. However, for a long time, systematic theories and technologies related to hybridization in fish have been lacking. In this study, through long-term systematic research, we investigated and obtained the main rules regarding inheritance and reproduction related to fish distant hybridization. Furthermore, we established one-step and multistep breeding technologies that were suitable for interspecific hybridization and intraspecific hybridization. Simultaneously, we used these two breeding technologies to produce a batch of diploid fish lineages and tetraploid fish lineages and improved fishes. In addition, we widely discuss the methods, technologies and results of hybridization breeding, referring to the domestic and foreign literature on fish hybridization. We hope that this paper will be beneficial for the research and application of fish hybrid breeding.展开更多
Hybridization can combine the genomes of different strains or species, which leads to changes of genotype and phenotype in the hybrids. In this study, we aimed to investigate the genetic variations of hybrids(WR-F1 an...Hybridization can combine the genomes of different strains or species, which leads to changes of genotype and phenotype in the hybrids. In this study, we aimed to investigate the genetic variations of hybrids(WR-F1 and WR-F2) derived from the intraspecific hybridization of white crucian carp(Carassius auratus cuvieri, WCC, ♀) and red crucian carp(Carassius auratus red var., RCC, ♂). Here, we compared the orthologous genes in the liver transcriptomes of hybrids with those of WCC and RCC, and classified the orthologous genes into eight gene patterns within three categories(chimera, mutant, and biparental origin genes).The results revealed 19.04%, 4.17% chimeric genes and 6.90%, 5.05% mutations of orthologous genes in WR-F1 and WR-F2 respectively. Seventeen of twenty-three characterized genes(77%) were confirmed to be the chimeras at the genomic DNA level.The GO classification discovered that some chimeric and mutant genes were related to metabolic process, immune system and developmental process in WR-F1. Our results provide the new evidence that hybridization can combine the parental genomes,leading to changes in the genotype of the resultant hybrids. This is the first report on the formation of chimeric genes from fish intraspecific hybridization, which is potentially interesting from the context of both evolution and the genetic breeding of fish.展开更多
Distant hybridization refers to crosses between two different species, genera, or higher-ranking taxa, which can break species limits, increase genetic variation, and combine the biological characteristics of existing...Distant hybridization refers to crosses between two different species, genera, or higher-ranking taxa, which can break species limits, increase genetic variation, and combine the biological characteristics of existing species. It is an important way of creating genetic variation, fertile strains, and excellent characteristics in new strains and populations. Combining analyses and summaries from many inter-related documents in plants and animals, both domestic and international, including examples and long-standing research on distant hybridization in fish from our laboratory, we summarize and compare the similarities and differences in plant and animal distant hybridization. In addition, we analyze and review the biological characteristics of their different ploidy progenies and the possible causes of disparity in survival rates. Mechanisms of sterility in animal and plant distant hybrids are also discussed, and research methods for the study of biological characteristics of hybrids, including morphology, cytology,and molecular cytogenetics are presented. This paper aims to provide comprehensive research materials and to systematically compare the general and specific characteristics of plant and animal hybrids with regards to reproduction, genetics, growth traits,and other biological characteristics. It is hoped that this paper will have great theoretical and practical significance for the study of genetic breeding and biological evolution of plant and animal distant hybridization.展开更多
The white crucian carp(Carassius auratus cuvieri,WCC) not only is one of the most economically important fish in Asia,characterized by strong reproductive ability and rapid growth rates,but also represents a good germ...The white crucian carp(Carassius auratus cuvieri,WCC) not only is one of the most economically important fish in Asia,characterized by strong reproductive ability and rapid growth rates,but also represents a good germplasm to produce hybrid progenies with heterosis.Gene knockout technique provides a safe and acceptant way for fish breeding.Achieving gene knockout in WCC and its hybrid progeny will be of great importance for both genetic studies and hybridization breeding.Tyrosinase(TYR) is a key enzyme in melanin synthesis.Depletion of tyr in zebrafish and mice results in mosaic pigmentation or total albinism.Here,we successfully used CRISPR-Cas9 to target tyr in WCC and its hybrid progeny(WR) derived from the cross of WCC(♀) and red crucian carp(Carassius auratus red var.,RCC,♂).The level of TYR protein was significantly reduced in mutant WCC.Both the mutant WCC and the mutant WR showed different degrees of melanin reduction compared with the wild-type sibling control fish,resulting from different mutation efficiency ranging from 60% to 90%.In addition,the transcriptional expression profiles of a series of pivotal pigment synthesis genes,i.e.tyrp1,mitfa,mitfb,dct and sox10,were down-regulated in tyr-CRISPR WCC,which ultimately caused a reduction in melanin synthesis.These results demonstrated that tyr plays a key role in melanin synthesis in WCC and WR,and CRISPR-Cas9 is an effective tool for modifying the genome of economical fish.Furthermore,the tyr-CRISPR models could be valuable in understanding fundamental mechanisms of pigment formation in non-model fish.展开更多
Histone post-modifications are important epigenetic markers involved in multiple cellular processes via regulation of gene transcription or remodeling of chromatin structure. Oocyte development is a critical process u...Histone post-modifications are important epigenetic markers involved in multiple cellular processes via regulation of gene transcription or remodeling of chromatin structure. Oocyte development is a critical process under rigorous control to prevent the generation of aberrant gametes. However, the regulatory mechanism of oocyte early development is not well-understood due to the tiny size and poor distinguishability of the gonad in juvenile stages. Here, two cyprinid hybrid fishes, a sterile allotriploid fish and a gynogenetic hybrid fish with delayed oocyte development, provided research models to investigate the mechanisms involved. We used cytogenetic and molecular methods to confirm the pachytene arrest of oocytes in allotriploid fish and gynogenetic hybrid fish. On the basis of these developmental differences, we screened 21 different histone H3 modifications by ELISA and found that four modifications(H3 K4 me3, H3 K9 me3, H3 K79 me, and H3 K79 me3) differed significantly in the two cyprinid hybrid fishes. Changes in histone methylation at the three residues(H3 K4, K9, K79) were caused by specific methyltransferases and demethylases. Our results provide new insights into the epigenetic regulation of oocyte early development in fish, a process critical for understanding of reproductive biology and with practical applications in the aquacultural breeding industry.展开更多
Focusing on adaptation of aquatic organisms, especially fish, can help elucidate complex dynamics in freshwater ecology. The differences in genetic and epigenetic regulation between diploid and triploid Carassius aura...Focusing on adaptation of aquatic organisms, especially fish, can help elucidate complex dynamics in freshwater ecology. The differences in genetic and epigenetic regulation between diploid and triploid Carassius auratus affect survival under eutrophication. To identify the underlying mechanisms that lead to better adaption of triploids than diploids, we compared mRNA and microRNA(miRNA) expressions in liver tissue of diploid and triploid individuals obtained from the Dongting lake water system in central China. Differential expression analysis revealed that 566 transcripts were significantly up-regulated, whereas758 were down-regulated in triploids; of these differentially expressed transcripts, 33 transcripts including cacna1 d, nfkb2, hspa1 and fgfr4 were involved in the MAPK signaling pathway, and eight transcripts were determined to be regulated by seven miRNAs. Additionally, four of 25 differential expressed(DE) transcripts(mhc1, irf7, nfkb2 and pik3 c) involving the viral carcinogenesis pathway were regulated by four miRNAs. Furthermore, genetic polymorphisms analysis showed that more heterozygous mutations were detected in triploids than diploids. The d N/d S results revealed that 21 genes were under positive selection(d N/d S>1) in C. auratus complex. We hypothesize that these changes related to genetic and epigenetic regulation may be caused by abiotic stresses, and facilitate adaptation to environmental changes.展开更多
基金supported by the National Natural Science Foundation of China (32172972,U19A2040)Science and Technology Innovation Program of Hunan Province (2021RC4028)+4 种基金Earmarked Fund for China Agriculture Research System (CARS-45)Hunan Provincial Science and Technology Department (2019RS5001)Special Funds for Construction of Innovative Provinces in Hunan Province (2021NK1010)Special Science Found of Nansha-South China Agricultural University Fishery Research Institute,Guangzhou (NSYYKY202305,NSYYKY202306)Aid Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province。
文摘The autotetraploid Carassius auratus(4nRR,4n=200,RRRR)is derived from whole-genome duplication of Carassius auratus red var.(RCC,2n=100,RR).In the current study,we demonstrated that chromatophores and pigment changes directly caused the coloration and variation of 4nRR skin(red in RCC,brownish-yellow in4nRR).To further explore the molecular mechanisms underlying coloration formation and variation in 4nRR,we performed transcriptome profiling and molecular functional verification in RCC and 4nRR.Results revealed that scarb1,associated with carotenoid metabolism,underwent significant down-regulation in 4nRR.Efficient editing of this candidate pigment gene provided clear evidence of its significant role in RCC coloration.Subsequently,we identified four divergent scarb1 homeologs in 4nRR:two original scarb1 homeologs from RCC and two duplicated ones.Notably,three of these homeologs possessed two highly conserved alleles,exhibiting biased and allelespecific expression in the skin.Remarkably,after precise editing of both the original and duplicated scarb1homeologs and/or alleles,4nRR individuals,whether singly or multiply mutated,displayed a transition from brownishyellow skin to a cyan-gray phenotype.Concurrently,the proportional areas of the cyan-gray regions displayed a gene-dose correlation.These findings illustrate the subfunctionalization of duplicated scarb1,with all scarb1genes synergistically and equally contributing to the pigmentation of 4nRR.This is the first report concerning the functional differentiation of duplicated homeologs in an autopolyploidfish,substantiallyenrichingour understanding of coloration formation and change within this group of organisms.
基金supported by the Bio & Medical Technology Development Program of the National Research Foundation(NRF) funded by the Korean government(MEST)(No.860-20110087)
文摘The periodontal ligament-derived mesenchymal stem cell is regarded as a source of adult stem cells due to its multipotency.However, the proof of chondrogenic potential of the cells is scarce.Therefore,we investigated the chondrogenic differentiation capacity of periodontal ligament derived mesenchymal stem cells induced by transforming growth factor(TGF)-p3 and bone morphogenetic protein(BMP)-6.After isolation of periodontal ligament stem cells(PDLSCs) from human periodontal ligament,the cells were cultured in Dulbecco’s modified Eagle’s medium(DMEM) with 20%fetal bovine serum(FBS).A mechanical force initiated chondrogenic differentiation of the cells.For chondrogenic differentiation,10μg·LTGF-β3 or 100μg·LBMP-6 and the combination treating group for synergistic effect of the growth factors.We analyzed the PDLSCs by fluorescence-activated cell sorting and chondrogenesis were evaluated by glycosaminoglycans assay,histology,immunohistochemistry and genetic analysis.PDLSCs showed mesenchymal stem cell properties proved by FACS analysis.Glycosaminoglycans contents were increased 217%by TGF-β3 and 220%by BMP-6. The synergetic effect of TGF-β3 and BMP-6 were shown up to 281%compared to control.The combination treatment increased Sox9, aggrecan and collagen II expression compared with not only controls,but also TGF-P3 or BMP-6 single treatment dramatically.The histological analysis also indicated the chondrogenic differentiation of PDLSCs in our conditions.The results of the present study demonstrate the potential of the dental stem cell as a valuable cell source for chondrogenesis,which may be applicable for regeneration of cartilage and bone fracture in the field of cell therapy.
文摘Aim: To identify proteins induced by androgen in Sertoli cells during spermatogenesis. Methods: We analyzed protein profiles in TM4 Sertoli cells treated with dihydrotestosterone (DHT) using surface enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Results: We found increases in the expression of a 5.0-kDa protein at 15 min, an 11.3-kDa protein at 24 h and 4.3 kDa, 5.7 kDa, 5.8 kDa, 9.95 kDa and 9.98 kDa proteins at 48 h after the treatment. In contrast, the expression of 6.3 kDa and 8.6 kDa proteins decreased at 30 min, and 4.9 kDa, 5.0 kDa, 12.4 kDa and 19.8 kDa proteins at 48 h after the treatment. The ll.3-kDa protein was identified as macrophage migration inhibitory factor (MIF) known to having various functions. The 9.98-kDa protein was identified as calgizzarin related to calcium channels. The timing of their expression suggests that MIF and calgizzarin are involved in late regulation of spermatogenesis in Sertoli cells by androgen. Conclusion: MIF and calgizzarin are two important androgen-responsive proteins produced by Sertoli cells and they might play a role in regulating spermatogenesis.
基金granted by the Natural Science Foundation of Fujian,China(2009J01129)the Education Department Foundation of Fujian,China(2007F5033)
文摘Genotoxicity of dichlorvos and butachlor on erythrocytes of Rana zhenhaiensis tadpoles was investigated by the alkaline single-cell gel electrophoresis assay or comet assay.Tadpoles were treated for 24h in the laboratory with different concentrations of the testing agents,2.256,4.512,6.768,9.024,11.280mg/L for dichlorvos and 0.292,0.584,0.876,1.168,1.460mg/L for butachlor,to use the comet Assay to test for the significance of dosage responsiveness to an increase in DNA damage,asmeasured by themean DNA tail length-to-width ratio.The concentrations of 4.512mg/L dichlorvos and 0.876mg/L butachlor resulted in highly significant increases in DNA damage of the tadpoles.There were linear correlations between themean DNA tail length-to-width ratio and the concentrations of the two test substances.Our results showed that the two commonly used agricultural chemicals caused dose dependent DNA damage of amphibians,and that comet assaymight be a useful tool formeasuring DNA damage of tadpoles exposed in the field.
基金supported by the National Natural Science Foundation of China(32002382,32293252,U19A2040,32293254)the National Key Research and Development Program of China(2023YFD2400202)+5 种基金the Natural Science Foundation of Hunan Province(2021JJ40339)the Training Program for Excellent Young Innovators of Changsha(kq2209013)the Earmarked Fund for Agriculture Research System of China(CARS-45)the Laboratory of Lingnan Modern Agriculture Project(NT2021008)the 111 Project(D20007)Special Science Found of Nansha-South China Agricultural University Fishery Research Institute,Guangzhou。
文摘The basic mechanism of heterosis has not been systematically and completely characterized.In previous studies,we obtained three economically important fishes that exhibit rapid growth,WR(WCC♀×RCC♂),WR-Ⅱ(WR♀×WCC♂),and WR-Ⅲ(WR-Ⅱ♀×4nAU♂),through distant hybridization.However,the mechanism underlying this rapid growth remains unclear.In this study,we found that WR,WR-Ⅱ,and WR-Ⅲshowed muscle hypertrophy and higher muscle protein and fat contents compared with their parent species(RCC and WCC).Candidate genes responsible for this rapid growth were then obtained through an analysis of 12 muscle transcriptomes.Notably,the mRNA level of mstnb(myostatin b),which is a negative regulator of myogenesis,was significantly reduced in WR,WR-Ⅱ,and WR-Ⅲcompared with the parent species.To verify the function of mstnb,a mstnb-deficient mutant RCC line was generated using the CRISPR-Cas9 technique.The average body weight of mstnb-deficient RCC at 12 months of age was significantly increased by 29.57%compared with that in wild-type siblings.Moreover,the area and number of muscle fibers were significantly increased in mstnb-deficient RCC,indicating hypertrophy and hyperplasia.Furthermore,the muscle protein and fat contents were significantly increased in mstnb-deficient RCC.The molecular regulatory mechanism of mstnb was then revealed by transcription profiling,which showed that genes related to myogenesis(myod,myog,and myf5),protein synthesis(PI3K-AKT-mTOR),and lipogenesis(pparγand fabp3)were highly activated in hybrid fishes and mstnb-deficient RCC.This study revealed that low expression or deficiency of mstnb regulates somatic growth by promoting myogenesis,protein synthesis,and lipogenesis in hybrid fishes and mstnb-deficient RCC,which provides evidence for the molecular mechanism of heterosis via distant hybridization.
基金Supported by Scientific and Technological Research Programme of Chongqing Municipal Education Commission,China(No.KJ130320)。
文摘Objective To investigate the mechanistic basis for the anti-proliferation and anti-invasion effect of tumor necrosis factor-related apoptosis-induced ligand(TRAIL)and celastrol combination treatment(TCCT)in glioblastoma cells.Methods Cell counting kit-8 was used to detect the effects of different concentrations of celastrol(0-16µmol/L)and TRAIL(0-500 ng/mL)on the cell viability of glioblastoma cells.U87 cells were randomly divided into 4 groups,namely control,TRAIL(TRAIL 100 ng/mL),Cel(celastrol 0.5µmol/L)and TCCT(TRAIL 100 ng/mL+celastrol 0.5µmol/L).Cell proliferation,migration,and invasion were detected by colony formation,wound healing,and Transwell assays,respectively.Quantitative reverse transcription polymerase chain reaction and Western blotting were performed to assess the levels of epithelial-mesenchymal transition(EMT)markers(zona occludens,N-cadherin,vimentin,zinc finger E-box-binding homeobox,Slug,and β-catenin).Wnt pathway was activated by lithium chloride(LiCl,20 mol/L)and the mechanism for action of TCCT was explored.Results Celastrol and TRAIL synergistically inhibited the proliferation,migration,invasion,and EMT of U87 cells(P<0.01).TCCT up-regulated the expression of GSK-3β and down-regulated the expression of β-catenin and its associated proteins(P<0.05 or P<0.01),including c-Myc,Cyclin-D1,and matrix metalloproteinase(MMP)-2.In addition,LiCl,an activator of the Wnt signaling pathway,restored the inhibitory effects of TCCT on the expression of β-catenin and its downstream genes,as well as the migration and invasion of glioblastoma cells(P<0.05 or P<0.01).Conclusions Celastrol and TRAIL can synergistically suppress glioblastoma cell migration,invasion,and EMT,potentially through inhibition of Wnt/β-catenin pathway.This underlies a novel mechanism of action for TCCT as an effective therapy for glioblastoma.
基金supported by the National Natural Science Foundation of China,No.81400922(to DMW),81571084the Natural Science Foundation of Fujian Province of China,No.2018J01813(to DMW)the College of Life Sciences of Fujian Normal University of China,No.FZSKG2018016(to DMW)
文摘Adrenomedullin,a peptide with multiple physiological functions in nervous system injury and disease,has aroused the interest of researchers.This review summarizes the role of adrenomedullin in neuropathological disorders,including pathological pain,brain injury and nerve regeneration,and their treatment.As a newly characterized pronociceptive mediator,adrenomedullin has been shown to act as an upstream factor in the transmission of noxious information for various types of pathological pain including acute and chronic inflammatory pain,cancer pain,neuropathic pain induced by spinal nerve injury and diabetic neuropathy.Initiation of glia-neuron signaling networks in the peripheral and central nervous system by adrenomedullin is involved in the formation and maintenance of morphine tolerance.Adrenomedullin has been shown to exert a facilitated or neuroprotective effect against brain injury including hemorrhagic or ischemic stroke and traumatic brain injury.Additionally,adrenomedullin can serve as a regulator to promote nerve regeneration in pathological conditions.Therefore,adrenomedullin is an important participant in nervous system diseases.
基金National Natural Science Foundation of China under Grant Nos.61803152,31920103016,and 11871475Doctoral Start-Up Foundation of Hunan Normal University under Grant No.0531120-3827Hunan Provincial Education Department under Grant No.HNKCSZ-2020-0813.
文摘This study aims to improve control schemes for COVID-19 by a numerical model with estimation of parameters.We established a multi-level and multi-objective nonlinear SEIDR model to simulate the virus transmission.The early spread in Japan was adopted as a case study.The first 96 days since the infection were divided into five stages with parameters estimated.Then,we analyzed the trend of the parameter value,age structure ratio,and the defined PCR test index(standardization of the scale of PCR tests).It was discovered that the self-healing rate and confirmed rate were linear with the age structure ratio and the PCR test index using the stepwise regression method.The transmission rates were related to the age structure ratio,PCR test index,and isolation efficiency.Both isolation measures and PCR test medical screening can effectively reduce the number of infected cases based on the simulation results.However,the strategy of increasing PCR test medical screening would encountered a bottleneck effect on the virus control when the index reached 0.3.The effectiveness of the policy would decrease and the basic reproduction number reached the extreme value at 0.6.This study gave a feasible combination for isolation and PCR test by simulation.The isolation intensity could be adjusted to compensate the insufficiency of PCR test to control the pandemic.
文摘Parkinson’s disease is the second most common neurodegenerative disease and has currently no effective treatment,one that would be able to stop or reverse the loss of dopaminergic neurons in the substantia nigra pars compacta.In addition,Parkinson’s disease diagnosis is typically done when a significant percentage of the dopaminergic neurons is already lost.In neurodegenerative disorders,some therapeutic strategies could be effective only at inhibiting further degeneration;on the other hand,cell replacement therapies aim at replacing lost neurons,an approach that would be ideal for the treatment of Parkinson’s disease.Many cell replacement therapies have been tested since the 1970s in the field of Parkinson’s disease;however,there are still significant limitations prohibiting a successful clinical application.From the first fetal midbrain intrastriatal graft to the most recent conversion of astrocytes into dopaminergic neurons,we have gained equally,significant insights and questions still looking for an answer.This review aims to summarize the main milestones in cell replacement approaches against Parkinson’s disease.By focusing on achievements and failures,as well as on the additional research steps needed,we aim to provide perspective on how future cell replacement therapies treats Parkinson’s disease.
基金supported by the National Natural Science Foundation of China(Grant No.31872551)Laboratory of Lingnan Modern Agriculture Project(Grant No.NT2021008)+4 种基金the Natural Science Foundation of Hunan Province for Distinguished Young Scholars(Grant No.2020JJ2022)Hunan Provincial Innovation Foundation for Postgraduate(Grant No.CX20200480)the earmarked fund for China Agriculture Research System(Grant No.CARS-45)the Key Research and Development Program of Hunan Province(Grant No.2020NK2016)111 Project(D20007).
文摘Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenesis at both transcriptional and posttranscriptional levels,little is currently known regarding the regulation of sperm flagella assembly by microRNAs(miRNAs).The role of miRNAs in the development of sperm abnormalities in sterile triploid fish has not been studied.Results:In this study,we found that miR-199-5p was widely expressed in all detected tissues of different-ploidy crucian carp.As one of the testis-specific candidate markers,Tekt1 was predominantly expressed in the testis.Quantitative real-time PCR(qRT-PCR)analyses showed that the expression trend of miR-199-5p was exactly opposite to that of Tekt1.Through bioinformatics analysis,we identified a putative miR-199-5p binding site in the Tekt1 mRNA.We further identified Tekt1 as a target of miR-199-5p using luciferase reporter assay.Finally,we confirmed that miR-199-5p was necessary for sperm flagellar assembly and spermatogenesis in vivo via intraperitoneal injection of miR-199-5p antagomir or agomir in diploid red crucian carp.Moreover,miR-199-5p gainof-function could lead to spermatids apoptosis and abnormal spermatozoa structure,which is similar to that of allotriploid crucian carp.Conclusions:Our studies suggested that abnormally elevated miR-199-5p inhibited the sperm flagella formation in spermiogenesis by negatively regulating the expression of Tekt1,thereby causing sperm abnormalities of male allotriploid crucian carp.
基金supported by the National Key Research and Development Program,China(2018YFD0900406)the National Natural Science Foundation,China(31802291)the Natural Science Foundation of Hunan Province(2021JJ40342)。
文摘Recently,cilia defects have been proposed to contribute to scoliosis.Here,we demonstrate that coiled-coil domain-containing 57(Ccdc57)plays an essential role in straightening the body axis of zebrafish by regulating ciliary beating in the brain ventricle(BV).Zygotic ccdc57(Zccdc57)mutant zebrafish developes scoliosis without significant changes in their bone density and calcification,and the maternal-zygotic ccdc57(MZccdc57)mutant embryos display curved bodies since the long-pec stage.The expression of ccdc57 is enriched in ciliated tissues and immunofluorescence analysis reveals colocalization of Ccdc57-HA with acetylated a-tubulin,implicating it in having a role in ciliary function.Further examination reveals that it is the coordinated cilia beating of multiple cilia bundles(MCB)in the MZccdc57 mutant embryos that is affected at 48 hours post fertilization,when the compromised cerebrospinal fluid flow and curved body axis have already occurred.Either ccdc57 m RNA injection or epinephrine treatment reverses the spinal curvature in MZccdc57 mutant larvae from ventrally curly to straight or even dorsally curly and significantly upregulates urotensin signaling.This study reveals the role of ccdc57 in maintaining coordinated cilia beating of MCB in the BV.
基金supported by the National Natural Science Foundation of China(No.81672209,81871769,82272550)the Chongqing Science and Technology Bureau(China)(No.cstc2021jcyj-bshX0214).
文摘Cartilage development is controlled by the highly synergistic proliferation and differentiation of growth plate chondrocytes,in which the Indian hedgehog(IHH)and parathyroid hormone-related protein-parathyroid hormone-1 receptor(PTHrP-PTH1R)feedback loop is crucial.The inositol-requiring enzyme 1a/X-box-binding protein-1 spliced(IRE1α/XBP1s)branch of the unfolded protein response(UPR)is essential for normal cartilage development.However,the precise role of ER stress effector IRE1α,encoded by endoplasmic reticulum to nucleus signaling 1(ERN1),in skeletal development remains unknown.Herein,we reported that loss of IRE1α accelerates chondrocyte hypertrophy and promotes endochondral bone growth.ERN1 acts as a negative regulator of chondrocyte proliferation and differentiation in postnatal growth plates.Its deficiency interrupted PTHrP/PTH1R and IHH homeostasis leading to impaired chondrocyte hypertrophy and differentiation.XBP1s,produced by p-IRE1α-mediated splicing,binds and up-regulates PTH1R and IHH,which coordinate cartilage development.Meanwhile,ER stress cannot be activated normally in ERN1-deficient chondrocytes.In conclusion,ERN1 deficiency accelerates chondrocyte hypertrophy and cartilage mineralization by impairing the homeostasis of the IHH and PTHrP/PTH1R feedback loop and ER stress.ERN1 may have a potential role as a new target for cartilage growth and maturation.
基金supported by the Ministry of Science and Technology of China(Nos.2021YFA1101300 and 2020YFA0112500)the National Key R&D Program of China(2018YFA0801000)the National Natural Science Foundation of China(No.32170853)。
文摘As an increasingly used alternative to perfluorooctanoic acid(PFOA),hexafluoropropylene oxide trimer acid(HFPO-TA)has been widely detected in global water environments.However,little is known regarding its toxic effects on cardiovascular development.Here,zebrafish embryos were treated with egg water containing 0,60,120,or 240 mg/L HFPO-TA.Results showed that HFPO-TA treatment led to a significant reduction in both larval survival percentage and heart rate.Furthermore,HFPO-TA exposure caused severe pericardial edema and elongation of the sinus venous to bulbus arteriosus distance(SV-BA)in Tg(myl7:GFP)transgenic larvae,disrupting the expression of genes involved in heart development and thus causing abnormal heart looping.Obvious sprouting angiogenesis was observed in the 120 and 240 mg/L exposed Tg(fli:GFP)transgenic larvae.HFPO-TA treatment also impacted the mRNA levels of genes involved in the vascular endothelial growth factor(VEGF)pathway and embryonic vascular development.HFPO-TA exposure significantly decreased erythrocyte number in Tg(gata1:DsRed)transgenic embryos and influenced gene expression associated with the heme metabolism pathway.HFPO-TA also induced oxidative stress and altered the transcriptional levels of genes related to cell cycle and apoptosis,inhibiting cell proliferation while promoting apoptosis.Therefore,HFPO-TA exposure may induce abnormal development of the cardiovascular and hematopoietic systems in zebrafish embryos,suggesting it may not be a suitable or safe alternative for PFOA.
基金the National Natural Science Foundation of China(U21A20268,31920103016,32173010)the fellowship of China Postdoctoral Science Foundation(No.2022M711128)+2 种基金Hunan Provincial Science and Technology Department(2021RC2076,2021NK2025,2022JJ40276,2022JJ30383)College Student Innovation and Entrepreneurship Training Program(2022123,2023227)the Modern Agricultural Industry Program of Hunan Province,and the Fish Disease and Vaccine Research and Development Platform for Postgraduates in Hunan Province.
文摘Viral infectious diseases significantly threaten the sustainability of freshwater fish aquaculture.The lack of studies on epidemic transmission patterns and mechanisms inhibits the development of containment strategies from the viewpoint of veterinary public health.This study raises an epidemic mathematical model considering water transmission with the aim of analyzing the transmission process more accurately.The basic reproduction number R0 was derived by the model parameter including the water transmission coefficient and was used for the analysis of the virus transmission.Spring viremia of carp virus(SVCV)and zebrafish were used as model viruses and animals,respectively,to conduct the transmission experiment.Transmission through water was achieved by connecting two aquarium tanks with a water channel but blocking the fish movement between the tanks.With the collected experimental data,we determined the optimal hybrid machine learning algorithm to analyze the transmission process using an established mathematical model.In addition,future transmission was predicted and validated using the epidemic model and an optimal algorithm.Finally,the sensitivity of model parameters and the simulations of R0 variation were performed based on the modified complex epidemic model.This study is of significance in providing theoretical guidance for minimizing R0 by manipulating model parameters with containment measures.More importantly,since the modified model and algorithm demonstrated better performance in handling freshwater fish transmission problems,this study advances the future application of transmissible disease modeling with larger datasets in freshwater fish aquaculture.
基金supported by the National Natural Science Foundation of China (31730098, 31430088, and 31802287)the Earmarked Fund for China Agriculture Research System (CARS-45)+2 种基金the Key Research and Development Program of Hunan Province (2018NK2072)Hunan Provincial Natural Science and Technology Major Project (2017NK1031)the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (20134486)
文摘Hybridization is widely used. However, for a long time, systematic theories and technologies related to hybridization in fish have been lacking. In this study, through long-term systematic research, we investigated and obtained the main rules regarding inheritance and reproduction related to fish distant hybridization. Furthermore, we established one-step and multistep breeding technologies that were suitable for interspecific hybridization and intraspecific hybridization. Simultaneously, we used these two breeding technologies to produce a batch of diploid fish lineages and tetraploid fish lineages and improved fishes. In addition, we widely discuss the methods, technologies and results of hybridization breeding, referring to the domestic and foreign literature on fish hybridization. We hope that this paper will be beneficial for the research and application of fish hybrid breeding.
基金supported by National Natural Science Foundation of China (31210103918, 31430088, 91631305)the earmarked fund for China Agriculture Research System (CARS-45)+1 种基金the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (20134486)the Construction Project of Key Discipline of Hunan Province and China and Natural Science Foundation of Hunan Province (14JJ2062)
文摘Hybridization can combine the genomes of different strains or species, which leads to changes of genotype and phenotype in the hybrids. In this study, we aimed to investigate the genetic variations of hybrids(WR-F1 and WR-F2) derived from the intraspecific hybridization of white crucian carp(Carassius auratus cuvieri, WCC, ♀) and red crucian carp(Carassius auratus red var., RCC, ♂). Here, we compared the orthologous genes in the liver transcriptomes of hybrids with those of WCC and RCC, and classified the orthologous genes into eight gene patterns within three categories(chimera, mutant, and biparental origin genes).The results revealed 19.04%, 4.17% chimeric genes and 6.90%, 5.05% mutations of orthologous genes in WR-F1 and WR-F2 respectively. Seventeen of twenty-three characterized genes(77%) were confirmed to be the chimeras at the genomic DNA level.The GO classification discovered that some chimeric and mutant genes were related to metabolic process, immune system and developmental process in WR-F1. Our results provide the new evidence that hybridization can combine the parental genomes,leading to changes in the genotype of the resultant hybrids. This is the first report on the formation of chimeric genes from fish intraspecific hybridization, which is potentially interesting from the context of both evolution and the genetic breeding of fish.
基金supported by National Natural Science Foundation of China(31430088,31210103918,91331105)National High Technology Research and Development Program of China(2011AA100403)+3 种基金the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province(20134486)Project supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China(20114306130001)Educational Commission of Hunan Province(14B115)National Key Disciplines of Developmental Biology and Key Discipline Construction Project Funding for Developmental Biology of Hunan Province
文摘Distant hybridization refers to crosses between two different species, genera, or higher-ranking taxa, which can break species limits, increase genetic variation, and combine the biological characteristics of existing species. It is an important way of creating genetic variation, fertile strains, and excellent characteristics in new strains and populations. Combining analyses and summaries from many inter-related documents in plants and animals, both domestic and international, including examples and long-standing research on distant hybridization in fish from our laboratory, we summarize and compare the similarities and differences in plant and animal distant hybridization. In addition, we analyze and review the biological characteristics of their different ploidy progenies and the possible causes of disparity in survival rates. Mechanisms of sterility in animal and plant distant hybrids are also discussed, and research methods for the study of biological characteristics of hybrids, including morphology, cytology,and molecular cytogenetics are presented. This paper aims to provide comprehensive research materials and to systematically compare the general and specific characteristics of plant and animal hybrids with regards to reproduction, genetics, growth traits,and other biological characteristics. It is hoped that this paper will have great theoretical and practical significance for the study of genetic breeding and biological evolution of plant and animal distant hybridization.
基金supported by the National Natural Science Foundation of China (31430088, 31730098)the earmarked fund for China Agriculture Research System (CARS-45)+2 种基金the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (20134486)the Natural Science Foundation of Hunan Province (14JJ2062)the Research Foundation of Education Bureau of Hunan Province, China (16B160)
文摘The white crucian carp(Carassius auratus cuvieri,WCC) not only is one of the most economically important fish in Asia,characterized by strong reproductive ability and rapid growth rates,but also represents a good germplasm to produce hybrid progenies with heterosis.Gene knockout technique provides a safe and acceptant way for fish breeding.Achieving gene knockout in WCC and its hybrid progeny will be of great importance for both genetic studies and hybridization breeding.Tyrosinase(TYR) is a key enzyme in melanin synthesis.Depletion of tyr in zebrafish and mice results in mosaic pigmentation or total albinism.Here,we successfully used CRISPR-Cas9 to target tyr in WCC and its hybrid progeny(WR) derived from the cross of WCC(♀) and red crucian carp(Carassius auratus red var.,RCC,♂).The level of TYR protein was significantly reduced in mutant WCC.Both the mutant WCC and the mutant WR showed different degrees of melanin reduction compared with the wild-type sibling control fish,resulting from different mutation efficiency ranging from 60% to 90%.In addition,the transcriptional expression profiles of a series of pivotal pigment synthesis genes,i.e.tyrp1,mitfa,mitfb,dct and sox10,were down-regulated in tyr-CRISPR WCC,which ultimately caused a reduction in melanin synthesis.These results demonstrated that tyr plays a key role in melanin synthesis in WCC and WR,and CRISPR-Cas9 is an effective tool for modifying the genome of economical fish.Furthermore,the tyr-CRISPR models could be valuable in understanding fundamental mechanisms of pigment formation in non-model fish.
基金supported by the National Natural Science Foundation of China (31402297, 31730098)the earmarked fund for China Agriculture Research System (CARS-45)the Natural Science Foundation of Hunan Province (2018JJ3338)
文摘Histone post-modifications are important epigenetic markers involved in multiple cellular processes via regulation of gene transcription or remodeling of chromatin structure. Oocyte development is a critical process under rigorous control to prevent the generation of aberrant gametes. However, the regulatory mechanism of oocyte early development is not well-understood due to the tiny size and poor distinguishability of the gonad in juvenile stages. Here, two cyprinid hybrid fishes, a sterile allotriploid fish and a gynogenetic hybrid fish with delayed oocyte development, provided research models to investigate the mechanisms involved. We used cytogenetic and molecular methods to confirm the pachytene arrest of oocytes in allotriploid fish and gynogenetic hybrid fish. On the basis of these developmental differences, we screened 21 different histone H3 modifications by ELISA and found that four modifications(H3 K4 me3, H3 K9 me3, H3 K79 me, and H3 K79 me3) differed significantly in the two cyprinid hybrid fishes. Changes in histone methylation at the three residues(H3 K4, K9, K79) were caused by specific methyltransferases and demethylases. Our results provide new insights into the epigenetic regulation of oocyte early development in fish, a process critical for understanding of reproductive biology and with practical applications in the aquacultural breeding industry.
基金supported by the National Natural Science Foundation of China (317023343, 1430088, 31730098)the earmarked fund for China Agriculture Research System (CARS-45)+5 种基金the Key Research and Development Project of Hunan Province (2016NK2128)the Key Research and Development Project of Hunan Province (2016NK2128)Hunan Provincial Natural Science and Technology Major Project (2017NK1031)the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province (20134486)the Construction Project of Key Discipline of Hunan Province and China, Natural Science Foundation of Hunan Province (14JJ2148)the Scientific Research Fund of Hunan Provincial Education Department (16C0974)
文摘Focusing on adaptation of aquatic organisms, especially fish, can help elucidate complex dynamics in freshwater ecology. The differences in genetic and epigenetic regulation between diploid and triploid Carassius auratus affect survival under eutrophication. To identify the underlying mechanisms that lead to better adaption of triploids than diploids, we compared mRNA and microRNA(miRNA) expressions in liver tissue of diploid and triploid individuals obtained from the Dongting lake water system in central China. Differential expression analysis revealed that 566 transcripts were significantly up-regulated, whereas758 were down-regulated in triploids; of these differentially expressed transcripts, 33 transcripts including cacna1 d, nfkb2, hspa1 and fgfr4 were involved in the MAPK signaling pathway, and eight transcripts were determined to be regulated by seven miRNAs. Additionally, four of 25 differential expressed(DE) transcripts(mhc1, irf7, nfkb2 and pik3 c) involving the viral carcinogenesis pathway were regulated by four miRNAs. Furthermore, genetic polymorphisms analysis showed that more heterozygous mutations were detected in triploids than diploids. The d N/d S results revealed that 21 genes were under positive selection(d N/d S>1) in C. auratus complex. We hypothesize that these changes related to genetic and epigenetic regulation may be caused by abiotic stresses, and facilitate adaptation to environmental changes.