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Identification of transition factors in myotube formation from proteome and transcriptome analyses

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Journal of Integrative Agriculture 2023年第10期22卷 3135-3147页

作者： ZHENG Qi HU Rong-cui ZHU Cui-yun JING Jing LOU Meng-yu ZHANG Si-huan LI Shuang CAO Hong-guo ZHANG Xiao-rong LING Ying-hui College of Animal Science and Technology Anhui Agricultural UniversityHefei 230036P.R.China key laboratory of Embryo Development and Reproductive Regulation of anhui province Fuyang Normal UniversityFuyang 236041P.R.China anhui province key laboratory of local livestock and poultry genetic resource conservation and bio-breeding Anhui Agricultural UniversityHefei 230036P.R.China

Muscle fibers are the main component of skeletal muscle and undergo maturation through the formation of myotubes.During early development,a population of skeletal muscle satellite cells(SSCs)proliferate into myoblasts.The myoblasts then undergo further differentiation and fusion events,leading to the development of myotubes.However,the mechanisms involved in the transition from SSCs to myotube formation remain unclear.In this study,we characterized changes in the proteomic and transcriptomic expression profiles of SSCs,myoblasts(differentiation for 2 d)and myotubes(differentiation for 10 d).Proteomic analysis identified SLMAP and STOM as potentially associated with myotube formation.In addition,some different changes in MyoD,MyoG,Myosin7 and Desmin occurred after silencing SLMAP and STOM,suggesting that they may affect changes in the myogenic marker.GO analysis indicated that the differentiation and migration factors SVIL,ENSCHIG00000026624(AQP1)and SERPINE1 enhanced the transition from SSCs to myoblasts,accompanied by changes in the apoptotic balance.In the myoblast vs.myotube group,candidates related to cell adhesion and signal transduction were highly expressed in the myotubes.Additionally,CCN2,TGFB1,MYL2 and MYL4 were identified as hub-candidates in this group.These data enhance our existing understanding of myotube formation during early development and repair.

关键词： proteome transcriptome skeletal muscle satellite cells myoblast myotube

MicroRNA-370-5p inhibits pigmentation and cell proliferation by downregulating mitogen-activated protein kinase kinase kinase 8 expression in sheep melanocytes

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Journal of Integrative Agriculture 2023年第4期22卷 1131-1141页

作者： JI Kai-yuan WEN Ru-jun WANG Zheng-zhou TIAN Qian-qian ZHANG Wei ZHANG Yun-hai anhui province key laboratory of Veterinary Pathobiology and Disease Control College of Animal Science and TechnologyAnhui Agricultural UniversityHefei 230036P.R.China anhui province key laboratory of local livestock and poultry genetic resources conservation and bio-breeding College of Animal Science and TechnologyAnhui Agricultural UniversityHefei 230036P.R.China Linquan Comprehensive Experimental Station Anhui Agricultural UniversityLinquan 236400P.R.China

In mammals,microRNAs(miRNAs)play key roles in multiple biological processes by regulating the expression of target genes.Studies have found that the levels of miR-370-5p expression differ significantly in the skins of sheep with different hair colors;however,its function remains unclear.In this study,we investigated the roles of miR-370-5p in sheep melanocytes and found that the overexpression of miR-370-5p significantly inhibited cell proliferation(P<0.01),tyrosinase activity(P=0.001)and significantly reduced(P<0.001)melanin production.Functional prediction revealed that the 3′-untranslated region(UTR)of MAP3K8 has a putative miR-370-5p binding site,and the interaction between these two molecules was confirmed using luciferase reporter assays.In situ hybridization assays revealed that MAP3K8 is expressed in the cytoplasm of melanocytes.The results of quantitative RT-PCR and Western blotting analyses revealed that overexpression of miR-370-5p in melanocytes significantly inhibits(P<0.01)MAP3K8 expression via direct targeting of its 3′UTR.Inhibition of MAP3K8 expression by siRNA-MAP3K8 transfection induced a significant inhibition(P<0.01)of melanocyte proliferation and significant reduction(P<0.001)in melanin production,which is consistent with our observations for miR-370-5p.Target gene rescue experiments indicated that the expression of MAP3K8 in melanocytes co-transfected with miR-370-5p and MAP3K8-cDNA(containing sites for the targeted binding to miR-370-5p)was significantly rescued(P≤0.001),which subsequently promoted significant increases in cell proliferation(P<0.001)and melanin production(P<0.01).Collectively,these findings indicate that miR-370-5p plays a functional role in inhibiting sheep melanocyte proliferation and melanogenesis by downregulating the expression of MAP3K8.

关键词： microRNA mitogen-activated protein kinase kinase kinase 8 melanogenesis sheep melanocytes cell proliferation

Chromatin remodeler INO80 mediates trophectoderm permeability barrier to modulate morula-to-blastocyst transition

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Zoological Research 2021年第5期42卷 562-573页

作者： Zu-Bing Cao Di Gao Hui-Qun Yin Hui Li Teng-Teng Xu Meng-Ya Zhang Xin Wang Qiu-Chen Liu Ye-Lian Yan Yang-Yang Ma Tong Yu Yun-Sheng Li Yun-Hai Zhang anhui province key laboratory of local livestock and poultry Genetical Resource Conservation and BreedingCollege of Animal Science and TechnologyAnhui Agricultural UniversityHefeiAnhui 230036China Reproductive Medicine Center 901st HospitalHefeiAnhui 230031China

Inositol requiring mutant 80(INO80)is a chromatin remodeler that regulates pluripotency maintenance of embryonic stem cells and reprogramming of somatic cells into pluripotent stem cells.However,the roles and mechanisms of INO80 in porcine preimplantation embryonic development remain largely unknown.Here,we show that INO80 modulates trophectoderm epithelium permeability to promote porcine blastocyst development.The INO80 protein is highly expressed in the nuclei during morula-toblastocyst transition.Functional studies revealed that RNA interference(RNAi)-mediated knockdown of INO80 severely blocks blastocyst formation and disrupts lineage allocation between the inner cell mass and trophectoderm.Mechanistically,singleembryo RNA sequencing revealed that INO80 regulates multiple genes,which are important for lineage specification,tight junction assembly,and fluid accumulation.Consistent with the altered expression of key genes required for tight junction assembly,a permeability assay showed that paracellular sealing is defective in the trophectoderm epithelium of INO80 knockdown blastocysts.Importantly,aggregation of 8-cell embryos from the control and INO80 knockdown groups restores blastocyst development and lineage allocation via direct complementation of the defective trophectoderm epithelium.Taken together,these results demonstrate that INO80 promotes blastocyst development by regulating the expression of key genes required for lineage specification,tight junction assembly,and fluid accumulation.

关键词： INO80 Blastocyst Trophectoderm Tight junction Permeability

Revisiting the evolutionary history of domestic and wild ducks based on genomic analyses

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Zoological Research 2021年第1期42卷 43-50页

作者： Xing Guo Xin-Xin He Hong Chen Zhi-Cheng Wang Hui-Fang Li Jiang-Xian Wang Ming-Shan Wang Run-Shen Jiang anhui province key laboratory of local livestock and poultry genetic resource conservation and bio-breeding Anhui Agricultural UniversityHefeiAnhui 230036China Jiangsu Institute of poultry Science Chinese Academy of Agriculture ScienceYangzhouJiangsu 225125China Howard Hughes Medical Institute University of California Santa CruzSanta CruzCA 95064USA Department of Ecology and Evolutionary biology University of California Santa CruzCA 95064USA

Although domestic ducks have been important poultry species throughout human history,their origin remains enigmatic,with mallards and/or Chinese spot-billed ducks being proposed as the direct wild ancestor(s)of domestic ducks.Here,we analyzed 118 whole genomes from mallard,Chinese spotbilled,and domestic ducks to reconstruct their evolutionary history.We found pervasive introgression patterns among these duck populations.Furthermore,we showed that domestic ducks separated from mallard and Chinese spotbilled ducks nearly 38 thousand years ago(kya)and 54 kya,respectively,which is considerably outside the time period of presumed duck domestication.Thus,our results suggest that domestic ducks may have originated from another wild duck population that is currently undefined or unsampled,rather than from present-day mallard and/or Chinese spot-billed ducks,as previously thought.Overall,this study provides new insight into the complex evolution of ducks.

关键词： Duck Population structure Demographic histories Domestication

Dual Oxidase Mutant Retards Mauthner-Cell Axon Regeneration at an Early Stage via Modulating Mitochondrial Dynamics in Zebrafish

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Neuroscience Bulletin 2020年第12期36卷 1500-1512页

作者： Lei-Qing Yang Min Chen Da-Long Ren Bing Hu Eye Center The First Affiliated Hospital of USTCHefei National Laboratory for Physical Sciences at the MicroscaleChinese Academy of Sciences Key Laboratory of Brain Function and DiseaseSchool of Life SciencesDivision of Biomedical SciencesUniversity of Science and Technology of ChinaHefei 230026China anhui province key laboratory of local livestock and poultry genetic resource conservation and bio-breeding College of Animal Science and TechnologyAnhui Agricultural UniversityHefei 230036China

Dual oxidase(duox)-deriyed reactive oxygen species(ROS)have been correlated with neuronal polarity,cerebellar development,and neuroplasticity.However,there have not been many comprehensive studies of the effect of individual duox isoforms on central-axon regenerationin vivo.Here,we explored this question in zebrafish,an excellent model organism for central-axon regeneration studies.In our research,mutation of the duox gene with CRISPR/Cas9 significantly retarded the singleaxon regeneration of the zebrafish Mauthner cell in vivo.Using deep transcriptome sequencing,we found that the expression levels of related functional enzymes in mitochondria were down-regulated in duox mutant fish.In vivo imaging showed that duox mutants had significantly disrupted mitochondrial transport and redox state in single Mauthner-cell axon.Our research data provide insights into how duox is involved in central-axon regeneration by changing mitochondrial transport.

关键词： duox Zebrafish Mauthner cell Axon regeneration Mitochondrial dynamics

Switches in transcriptome functions during seven skeletal muscle development stages from fetus to kid in Capra hircus

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Journal of Integrative Agriculture 2021年第1期20卷 212-226页

作者： LING Ying-hui ZHENG Qi JING Jing SUI Meng-hua ZHU Lu LI Yun-sheng ZHANG Yunhai LIU Ya FANG Fu-gui ZHANG Xiao-rong College of Animal Science and Technology Anhui Agricultural UniversityHefei 230036P.R.China School of Natural and Environmental Sciences Newcastle UniversityNewcastle upon TyneNE17RUUK anhui province key laboratory of local livestock and poultry genetic resource conservation and bio-breeding Anhui Agricultural UniversityHefei 230036P.R.China

Skeletal muscle accounts for about 40% of mammalian body weight, the development of which is a dynamic, complex and precisely regulated process that is critical for meat production. We here described the transcriptome expression profile in 21 goat samples collected at 7 growth stages from fetus to kid, including fetal 45(F45), 65(F65), 90(F90), 120(F120), and 135(F135) days, and birth 1(B1) day and 90(B90) days kids. Paraffin sections combined with RNA-seq data of the 7 stages divided the transcriptomic functions of skeletal muscle into 4 states: before F90, F120, F135 and B1, and B90. And the dynamic expression of all 4 793 differentially expressed genes(DEGs) was identified. Furthermore, DEGs were clustered by weighted gene correlation network analysis into 4 modules(turquoise, grey, blue and brown) that corresponded to these 4 states. Functional and pathway analysis indicated that the active genes in the stages before F90(turquoise) were closely related to skeletal muscle proliferation. The DEGs in the F120-related module(grey) were found to participate in the regulation of skeletal muscle structure and skeletal muscle development by regulating t RNA. The brown module(F135 and B1) regulated fatty acid biological processes to maintain the normal development of muscle cells. The DEGs of B90 high correlation module(blue) were involved the strengthening and power of skeletal muscle through the regulation of actin filaments and tropomyosin. Our current data thus revealed the internal functional conversion of the goat skeletal muscle in the growth from fetus to kid. The results provided a theoretical basis for analyzing the involvement of m RNA in skeletal muscle development.

关键词： mRNA goat(Capra hircus) transcriptome skeletal muscle

Genome-wide identification of functional enhancers and their potential roles in pig breeding

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Journal of Animal Science and biotechnology 2022年第6期13卷 1690-1702页

作者： Yinqiao Wu Yuedong Zhang Hang Liu Yun Gao Yuyan Liu Ling Chen Lu Liu David M.Irwin Chunhui Hou Zhongyin Zhou Yaping Zhang State key laboratory of genetic resources and Evolution and Yunnan Laboratory of Molecular Biology of Domestic AnimalsKunming Institute of ZoologyChinese Academy of SciencesKunming 650223YunnanChina Kunming College of Life Science University of Chinese Academy of SciencesKunming 650204YunnanChina State key laboratory for conservation and Utilization of bio-resource in Yunnan Yunnan UniversityKunming 650091YunnanChina School of Life Science Yunnan UniversityKunming 650091YunnanChina Institute of Physical Science and Information Technology Anhui UniversityHefei 230601AnhuiChina Department of laboratory Medicine and Pathobiology University of TorontoTorontoCanada Department of biology School of Life SciencesSouthern University of Science and TechnologyShenzhen 518055China Center for Excellence in Animal Evolution and genetics Chinese Academy of SciencesKunming 650223YunnanChina

Background:The pig is an economically important livestock species and is a widely applied large animal model in medical research.Enhancers are critical regulatory elements that have fundamental functions in evolution,development and disease.Genome-wide quantification of functional enhancers in the pig is needed.Results:We performed self-transcribing active regulatory region sequencing(STARR-seq)in the porcine kidney epithelial PK15 and testicular ST cell lines,and reliably identified 2576 functional enhancers.Most of these enhancers were located in repetitive sequences and were enriched within silent and lowly expressed genes.Enhancers poorly overlapped with chromatin accessibility regions and were highly enriched in chromatin with the repressive histone modification H3K9me3,which is different from predicted pig enhancers detected using ChIP-seq for H3K27ac or/and H3K4me1 modified histones.This suggests that most pig enhancers identified with STARR-seq are endogenously repressed at the chromatin level and may function during cell type-specific development or at specific developmental stages.Additionally,the PPP3CA gene is associated with the loin muscle area trait and the QKI gene is associated with alkaline phosphatase activity that may be regulated by distal functional enhancers.Conclusions:In summary,we generated the first functional enhancer map in PK15 and ST cells for the pig genome and highlight its potential roles in pig breeding.

关键词： breeding Functional enhancers Pig STARR-seq

CLAUDIN7 modulates trophectoderm barrier function to maintain blastocyst development in pigs

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CLAUDIN7 modulates trophectoderm barrier function to maintai...

Histone arginine methyltransferase CARM1-mediated H3R26me2 is essential for morula-to-blastocyst transition in pigs

中国畜牧兽医学会动物繁殖学分会第二十次学术研讨会

作者：刘秋晨 Xiangdong Zhang 张运海曹祖兵 anhui province key laboratory of local livestock and poultry Genetical Resource Conservation and BreedingCollege of Animal Science and TechnologyAnhui Agricultural University

Trophectoderm(TE) barrier function is an essential prerequisite for blastocyst development.CLAUDIN7(CLDN7),a member of CLAUDINS family,is involved in regulating intercellular exchange and cell polarity in epithelium cells.However,the role of CLDN7 in porcine early embryo development is yet to be explored.Here,we found that CLDN7 was highly conserved in different species and was widely expressed in different tissues.Remarkably,CLDN7 expression maintained a low level from GV oocyte to 4-cell stage whereas its expression exhibited a higher level from 8-cell stage onwards.Microinjection of siRNA into cytoplasm effectively knocked down expression of CLDN7 mRNA and protein in porcine embryos.CLDN7 knockdown not only significantly reduced blastocyst rates of embryos derived from parthenogenetic activation and in vitro fertilization,but also reduced number of total cells and TE cells in the resulting blastocysts.Furthermore,CLDN7 knockdown led to a significant reduction in expression of multiple genes associated with tight junction assembly and fluid accumulation.A permeability assay revealed that CLDN7 knockdown disrupted tight junction assembly and paracellular sealing in the TE epithelium.Taken together,these results demonstrate that CLDN7 regulates porcine blastocyst development via modulating trophectoderm barrier function.Materials and Methods Three siRNAs were designed for different sites in the pig CLDN7 coding area,and the embryo,embryo development,embryo development to the blastocar,and collected embryos,and then collected by QPCR,immunofluorescence technology and Western Blotting technology.Blastocysts in each group were incubated in modified PZM-3 medium containing 1 mg/mL 4 kDa FITC-dextran.Discussion In our study,we analyzed the homologous of CLDN7 gene in different species and demonstrated that CLDN7 was highly conserved among the species firstly.Next,we described the expression level of CLDN7 in main tissues of pigs and the dynamic expression patterns during porcine preimplantation development.We found th

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Histone arginine methyltransferase CARM1-mediated H3R26me2 i...