Signal transducer and activator of transcription 5b（STAT5b） has been identified as a key downstream mediator of growth hormone（GH） signaling in somatic growth of mammalian. However, the corresponding homologue gen...Signal transducer and activator of transcription 5b（STAT5b） has been identified as a key downstream mediator of growth hormone（GH） signaling in somatic growth of mammalian. However, the corresponding homologue gene of Stat5b is unknown in fish species. In this study, we generated loss-offunction mutants in stat5.1 and stat5.2, two stat5 homologues existing in zebrafish. In stat5.1-deficient zebrafish, a significant reduction of body length and body weight was detected in the embryos/larvae and adults compared with the wild-type control fish, and sexual size dimorphism in adult zebrafish was also eliminated. However, the stat5.2-deficient zebrafish displayed a normal developmental phenotype during all lifespan. Chromatin immunoprecipitation combined with deep sequencing（Ch IP-seq） method was adopted to further investigate the potential transcriptional targets of Stat5 protein and cast much light upon the biological function of Stat5. We identified more than 800 genes as transcriptional targets of Stat5 during zebrafish embryogenesis. KEGG analysis indicated that the Stat5 target gene network is predominantly linked to the metabolic pathways, neuroactive ligand-receptor interaction and JAK-STAT signaling pathways. Further validation studies suggested that Stat5.1 protein could directly regulate the expression of gh1, and stat5.1-mutated zebrafish showed a reduction of gh1 mRNA level. In the present study, stat5.1 was revealed as the corresponding homologue gene of Stat5b in fish species. Additionally,we found a novel molecular interaction between Stat5.1/Stat5b and GH, and unraveled a positive feedback loop Stat5.1-GH-Stat5.1 which is necessary for somatic growth and body development in zebrafish.展开更多
Bioactive ceramics have been used in bone tissue repair and regeneration.However,because of the complex in vivo osteogenesis process,long cycle,and difficulty of accurately tracking,the mechanism of interaction betwee...Bioactive ceramics have been used in bone tissue repair and regeneration.However,because of the complex in vivo osteogenesis process,long cycle,and difficulty of accurately tracking,the mechanism of interaction between materials and cells has yet to be fully understood,hindering its development.The ceramic microbridge micro-fluidic chip system may solve the problem and provide an in vitro method to simulate the microenvironment in vivo.Nevertheless,the complex microenvironment parameters of the chip system need to be studied in detail.Computer simulation bionics can provide clues for the setting of microenvironment parameters.This study used a computational bionic model to simulate the bone growth process in the presence of immune-related factors.The osteoblast differentiation of mesenchymal stem cells of calcium phosphate ceramics in a macrophage-dominated immune microenvironment was studied using a microfluidic chip system.The computational biomimetic model and microfluidic chip findings were basically consistent with the reported results of the animal experiments.These findings suggest that studying the osteogenic behavior of calcium phosphate ceramics using a microfluidic chip model is feasible.The method model provided in this study can be extended to other biomaterials,providing a viable path for their research and evaluation.展开更多
The sex-determining genes are found to be variable among different fish species. Yellow catfish(Pelteobagrus fulvidraco) is an important aquaculture fish species in China with XX/XY sex-determining type. Recently, YY ...The sex-determining genes are found to be variable among different fish species. Yellow catfish(Pelteobagrus fulvidraco) is an important aquaculture fish species in China with XX/XY sex-determining type. Recently, YY super-male yellow catfish has been successfully produced by combining hormonalinduced sex reversal method with sex chromosome-linked markers. Here, we identified a novel PDZ domain-containing gene in yellow catfish designated as pfpdz1, in whose intron the sex-linked marker was located. The coding sequence of pfpdz1 in Y chromosome was identical to that in X chromosome except a missense SNP(A/T) that changes an amino acid(E8V) in the N-terminal region. Pfpdz1 displayed male-specific expression during sex differentiation. Overexpression of pfpdz1 using additive transgenesis induces XX ovary to differentiate into testis-like tissue, while the targeted inactivation of pfpdz1 in Y chromosome using CRISPR/Cas9-mediated mutagenesis triggers ovarian differentiation. Furthermore,we demonstrated that pfpdz1 initiates testicular differentiation through upregulating expression of amh, dmrt1 and sox9a1, as well as downregulating expression of cyp19a1, foxl2 and wnt4. Our data provide functional evidence that pfpdz1 is significant for male differentiation and maintenance in yellow catfish.展开更多
STAT （Signal Transducers and Activators of Transcription） gene family members have been revealed to be involved in cell growth and differentiation in vertebrates. Despite their physiological importance, their functi...STAT （Signal Transducers and Activators of Transcription） gene family members have been revealed to be involved in cell growth and differentiation in vertebrates. Despite their physiological importance, their functions are poorly studied at organ and systemic levels. In this study, we performed a genome-wide analysis using data from invertebrates to vertebrates to identify STAT genes and analyze their evolutionary history. Interestingly, the STAT gene family undergoes genome duplications during the evolutionary history with STAT3 homologues firstly appearing in the basal extant vertebrate, sea lamprey, suggesting its possible roles in spine formation. To investigate the functions of stat3 in fish species, TALEN technology was performed to generate mutant zebrafish lines, star3 mutant zebrafish showed no obvious defects at early developmental stage but displayed severe lateral and vertical curvature of the spine （scoliosis）, spine fracture and the incomplete bone joints with narrower junction between vertebrae at early juvenile stage, as indicated by Alizarin red and Alcian blue staining, radiography and micro-computed tomography （MicroCT） analysis. Transcriptome analysis reveals dramatic alterations in a number of genes involved in immune and infection response, skeletal development and somatic growth, especially downregulated expression of collagen gene family, in the juvenile stat3 mutant zebrafish. Moreover, most of the collagen genes were detected to have abnormal expression pattern during the formation of spine deformities in stat3 mutants. Our data reveal that stat3 is specially expressed in vertebrates and required for normal spine development and immune function in zebrafish.展开更多
基金supported by the Fundamental Research Funds for the Central Universities(2662017PY013 and 2662015PY101)the Autonomous Project of State Key Laboratory of Freshwater Ecology and Biotechnology(2011FBZ22)the Autonomous Projects of the Institute of Hydrobiology,Chinese Academy of Sciences(Y25A17 and Y45A171301)
文摘Signal transducer and activator of transcription 5b（STAT5b） has been identified as a key downstream mediator of growth hormone（GH） signaling in somatic growth of mammalian. However, the corresponding homologue gene of Stat5b is unknown in fish species. In this study, we generated loss-offunction mutants in stat5.1 and stat5.2, two stat5 homologues existing in zebrafish. In stat5.1-deficient zebrafish, a significant reduction of body length and body weight was detected in the embryos/larvae and adults compared with the wild-type control fish, and sexual size dimorphism in adult zebrafish was also eliminated. However, the stat5.2-deficient zebrafish displayed a normal developmental phenotype during all lifespan. Chromatin immunoprecipitation combined with deep sequencing（Ch IP-seq） method was adopted to further investigate the potential transcriptional targets of Stat5 protein and cast much light upon the biological function of Stat5. We identified more than 800 genes as transcriptional targets of Stat5 during zebrafish embryogenesis. KEGG analysis indicated that the Stat5 target gene network is predominantly linked to the metabolic pathways, neuroactive ligand-receptor interaction and JAK-STAT signaling pathways. Further validation studies suggested that Stat5.1 protein could directly regulate the expression of gh1, and stat5.1-mutated zebrafish showed a reduction of gh1 mRNA level. In the present study, stat5.1 was revealed as the corresponding homologue gene of Stat5b in fish species. Additionally,we found a novel molecular interaction between Stat5.1/Stat5b and GH, and unraveled a positive feedback loop Stat5.1-GH-Stat5.1 which is necessary for somatic growth and body development in zebrafish.
基金supported by the National key research and development programs of China:2022YFC2405800Beijing Natural Science Foundation:L234022supported by China NMPA Research on performance and safety evaluation of innovative biomaterial medical devices:RS2024X004.
文摘Bioactive ceramics have been used in bone tissue repair and regeneration.However,because of the complex in vivo osteogenesis process,long cycle,and difficulty of accurately tracking,the mechanism of interaction between materials and cells has yet to be fully understood,hindering its development.The ceramic microbridge micro-fluidic chip system may solve the problem and provide an in vitro method to simulate the microenvironment in vivo.Nevertheless,the complex microenvironment parameters of the chip system need to be studied in detail.Computer simulation bionics can provide clues for the setting of microenvironment parameters.This study used a computational bionic model to simulate the bone growth process in the presence of immune-related factors.The osteoblast differentiation of mesenchymal stem cells of calcium phosphate ceramics in a macrophage-dominated immune microenvironment was studied using a microfluidic chip system.The computational biomimetic model and microfluidic chip findings were basically consistent with the reported results of the animal experiments.These findings suggest that studying the osteogenic behavior of calcium phosphate ceramics using a microfluidic chip model is feasible.The method model provided in this study can be extended to other biomaterials,providing a viable path for their research and evaluation.
基金supported by the National Natural Science Foundation of China (31672635)China Agricultural Research System (CARS-46)
文摘The sex-determining genes are found to be variable among different fish species. Yellow catfish(Pelteobagrus fulvidraco) is an important aquaculture fish species in China with XX/XY sex-determining type. Recently, YY super-male yellow catfish has been successfully produced by combining hormonalinduced sex reversal method with sex chromosome-linked markers. Here, we identified a novel PDZ domain-containing gene in yellow catfish designated as pfpdz1, in whose intron the sex-linked marker was located. The coding sequence of pfpdz1 in Y chromosome was identical to that in X chromosome except a missense SNP(A/T) that changes an amino acid(E8V) in the N-terminal region. Pfpdz1 displayed male-specific expression during sex differentiation. Overexpression of pfpdz1 using additive transgenesis induces XX ovary to differentiate into testis-like tissue, while the targeted inactivation of pfpdz1 in Y chromosome using CRISPR/Cas9-mediated mutagenesis triggers ovarian differentiation. Furthermore,we demonstrated that pfpdz1 initiates testicular differentiation through upregulating expression of amh, dmrt1 and sox9a1, as well as downregulating expression of cyp19a1, foxl2 and wnt4. Our data provide functional evidence that pfpdz1 is significant for male differentiation and maintenance in yellow catfish.
基金supported by the Fundamental Research Funds for the Central Universities (2662015PY101)the Autonomous Project of State Key Laboratory of Freshwater Ecology and Biotechnology (2011FBZ22)the Autonomous Projects of the Institute of Hydrobiology,Chinese Academy of Sciences (Y25A17, Y45A171301)
文摘STAT （Signal Transducers and Activators of Transcription） gene family members have been revealed to be involved in cell growth and differentiation in vertebrates. Despite their physiological importance, their functions are poorly studied at organ and systemic levels. In this study, we performed a genome-wide analysis using data from invertebrates to vertebrates to identify STAT genes and analyze their evolutionary history. Interestingly, the STAT gene family undergoes genome duplications during the evolutionary history with STAT3 homologues firstly appearing in the basal extant vertebrate, sea lamprey, suggesting its possible roles in spine formation. To investigate the functions of stat3 in fish species, TALEN technology was performed to generate mutant zebrafish lines, star3 mutant zebrafish showed no obvious defects at early developmental stage but displayed severe lateral and vertical curvature of the spine （scoliosis）, spine fracture and the incomplete bone joints with narrower junction between vertebrae at early juvenile stage, as indicated by Alizarin red and Alcian blue staining, radiography and micro-computed tomography （MicroCT） analysis. Transcriptome analysis reveals dramatic alterations in a number of genes involved in immune and infection response, skeletal development and somatic growth, especially downregulated expression of collagen gene family, in the juvenile stat3 mutant zebrafish. Moreover, most of the collagen genes were detected to have abnormal expression pattern during the formation of spine deformities in stat3 mutants. Our data reveal that stat3 is specially expressed in vertebrates and required for normal spine development and immune function in zebrafish.
