Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different p...Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different phenotypes can guide the migration of Schwann cells to the same sensory or motor phenotype.In this study,we analyzed the different effects of peripheral nerve-derived fibroblasts and cardiac fibroblasts on motoneurons.Compared with cardiac fibroblasts,peripheral nerve fibroblasts greatly promoted motoneuron neurite outgrowth.Transcriptome analysis results identified 491 genes that were differentially expressed in peripheral nerve fibroblasts and cardiac fibroblasts.Among these,130 were significantly upregulated in peripheral nerve fibroblasts compared with cardiac fibroblasts.These genes may be involved in axon guidance and neuron projection.Three days after sciatic nerve transection in rats,peripheral nerve fibroblasts accumulated in the proximal and distal nerve stumps,and most expressed brain-derived neurotrophic factor.In vitro,brain-derived neurotrophic factor secreted from peripheral nerve fibroblasts increased the expression ofβ-actin and F-actin through the extracellular regulated protein kinase and serine/threonine kinase pathways,and enhanced motoneuron neurite outgrowth.These findings suggest that peripheral nerve fibroblasts and cardiac fibroblasts exhibit different patterns of gene expression.Peripheral nerve fibroblasts can promote motoneuron neurite outgrowth.展开更多
The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that ...The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that are unrelated in sequence but share conserved function in catalyzing the reversible conversion of CO_(2) and HCO_(3)^(-).Here we have characterized the CA members in the transcriptome of S.japonica using Single-molecule real-time(SMRT)sequencing technology.Approximately 9830.4 megabases from 5,028,003 quality subreads were generated,and they were assembled into 326,512 full-length non-chimeric(FLNC)reads,with an average flnc read length of 2181 bp.After removing redundant sequences,79,010 unique transcripts were obtained of which 38,039 transcripts were successfully annotated.From the full-length transcriptome,we have identified 7 full-length cDNA sequences for CA genes(4α-CAs,1β-CAs and 2γ-CAs)and assessed for their potential functions based on phylogenetic analysis.Characterizations of CAs will provide the ground for future studies to determine the involvement of CAs in inorganic carbon absorption and transportation in S.japonica.展开更多
Vogt-Koyanagi-Harada disease(VKH)is a rare autoimmune disease characterized by diffuse and bilateral uveitis,alopecia,tinnitus,hearing loss,vitiligo and headache.The transcriptional expression pattern of peripheral bl...Vogt-Koyanagi-Harada disease(VKH)is a rare autoimmune disease characterized by diffuse and bilateral uveitis,alopecia,tinnitus,hearing loss,vitiligo and headache.The transcriptional expression pattern of peripheral blood mononuclear cells(PBMC)in VKH remains largely unknown.In this study,mRNA sequencing was conducted in PBMC from VKH patients with active uveitis before treatment(n=7),the same patients after prednisone combined with cyclosporine treatment(n=7)and healthy control subjects strictly matched with gender and age(n=7).We found 118 differentially expressed genes(DEGs)between VKH patients and healthy control subjects,and 21 DEGs between VKH patients before and after treatment.TRIB1 was selected as a potential biomarker to monitor the development of VKH according to the mRNA sequencing.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed to predict the possible biological functions and signaling pathways of DEGs.Neutrophil degranulation,peptidase regulator activity,secretory granule membrane,cellular response to peptide,growth factor binding and cell projection membrane were enriched as GO annotations of DEGs.Arachidonic acid metabolism and mitogen-activated protein kinase(MAPK)signaling pathway were potential signaling pathways involved in pathogenesis and drug response of VKH.A protein–protein interaction(PPI)network was constructed by STRING,and colony stimulating factor 1 receptor(CSF1R)was identified as the hubgene of all DEGs by Cytoscape.The cell type presumed to contribute to the aberrant expression of DEGs was analyzed with the use of publicly available single-cell sequencing data of PBMC from a healthy donor and single-cell sequencing dataset of monocytes from VKH patients.Our findings may help to decipher the underlying cellular and molecular pathogenesis of VKH and may lead novel therapeutic applications.展开更多
Background:Hypertrophic scars cause impaired skin appearance and function,seriously affecting physical and mental health.Due to medical ethics and clinical accessibility,the collection of human scar specimens is frequ...Background:Hypertrophic scars cause impaired skin appearance and function,seriously affecting physical and mental health.Due to medical ethics and clinical accessibility,the collection of human scar specimens is frequently restricted,and the establishment of scar experimental animal models for scientific research is urgently needed.The four most commonly used animal models of hypertrophic scars have the following drawbacks:the rabbit ear model takes a long time to construct;the immunodeficient mouse hypertrophic scar model necessitates careful feeding and experimental operations;female Duroc pigs are expensive to purchase and maintain,and their large size makes it difficult to produce a significant number of models;and mouse scar models that rely on tension require special skin stretch devices,which are often damaged and shed,resulting in unstable model establishment.Our group overcame the shortcomings of previous scar animal models and created a new mouse model of hypertrophic scarring induced by suture anchoring at the wound edge.Methods:We utilized suture anchoring of incisional wounds to impose directional tension throughout the healing process,restrain wound contraction,and generate granulation tissue,thus inducing scar formation.Dorsal paired incisions were generated in mice,with wound edges on the upper back sutured to the rib cage and the wound edges on the lower back relaxed as a control.Macroscopic manifestation,microscopic histological analysis,mRNA sequencing,bioinformatics,and in vitro cell assays were also conducted to verify the reliability of this method.Results:Compared with those in relaxed controls,the fibrotic changes in stretched wounds were more profound.Histologically,the stretched scars were hypercellular,hypervascular,and hyperproliferative with disorganized extracellular matrix deposition,and displayed molecular hallmarks of hypertrophic fibrosis.In addition,the stretched scars exhibited transcriptional overlap with mechanically stretched scars,and human hypertrophic and keloid scars.Phosphatidylinositol 3-kinase-serine/threonine-protein kinase B signaling was implicated as a profibrotic mediator of apoptosis resistance under suture-induced tension.Conclusions:This straightforward murine model successfully induces cardinal molecular and histological features of pathological hypertrophic scarring through localized suture tension to inhibit wound contraction.The model enables us to interrogate the mechanisms of tension-induced fibrosis and evaluate anti-scarring therapies.展开更多
The effects of brassinosteroid signaling on shoot and root development have been characterized in great detail but a simple consistent positive or negative impact on a basic cellular parameter was not identified.In th...The effects of brassinosteroid signaling on shoot and root development have been characterized in great detail but a simple consistent positive or negative impact on a basic cellular parameter was not identified.In this study,we combined digital 3D single-cell shape analysis and single-cell mRNA sequencing to charac-terize root meristems and mature root segments of brassinosteroid-blind mutants and wild type.The resul-tant datasets demonstrate that brassinosteroid signaling affects neither cell volume nor cell proliferation capacity.Instead,brassinosteroid signaling is essential for the precise orientation of cell division planes and the extent and timing of anisotropic cell expansion.Moreover,we found that the cell-aligning effects of brassinosteroid signaling can propagate to normalize the anatomy of both adjacent and distant brassinosteroid-blind cells through non-cell-autonomous functions,which are sufficient to restore growth vigor.Finally,single-cell transcriptome data discern directly brassinosteroid-responsive genes from genes that can react non-cell-autonomously and highlight arabinogalactans as sentinels of brassinosteroid-dependent anisotropic cell expansion.展开更多
基金supported by the National Key Research and Development Program of China,No.2017YFA0104703(to FD)the National Natural Science Foundation of China(Major Program),No.92068112(to FD)+2 种基金Science and Technology Program of Nantong of China,No.JC2020035(to QRH)National Natural Science Foundation of China,Nos.31500927(to QRH)and 31870977(to HYS)the Priority Academic Program Development of Jiangsu High Education Institutions(PAPD)(to FD).
文摘Peripheral nerve fibroblasts play a critical role in nerve development and regeneration.Our previous study found that peripheral nerve fibroblasts have different sensory and motor phenotypes.Fibroblasts of different phenotypes can guide the migration of Schwann cells to the same sensory or motor phenotype.In this study,we analyzed the different effects of peripheral nerve-derived fibroblasts and cardiac fibroblasts on motoneurons.Compared with cardiac fibroblasts,peripheral nerve fibroblasts greatly promoted motoneuron neurite outgrowth.Transcriptome analysis results identified 491 genes that were differentially expressed in peripheral nerve fibroblasts and cardiac fibroblasts.Among these,130 were significantly upregulated in peripheral nerve fibroblasts compared with cardiac fibroblasts.These genes may be involved in axon guidance and neuron projection.Three days after sciatic nerve transection in rats,peripheral nerve fibroblasts accumulated in the proximal and distal nerve stumps,and most expressed brain-derived neurotrophic factor.In vitro,brain-derived neurotrophic factor secreted from peripheral nerve fibroblasts increased the expression ofβ-actin and F-actin through the extracellular regulated protein kinase and serine/threonine kinase pathways,and enhanced motoneuron neurite outgrowth.These findings suggest that peripheral nerve fibroblasts and cardiac fibroblasts exhibit different patterns of gene expression.Peripheral nerve fibroblasts can promote motoneuron neurite outgrowth.
基金This research was supported by Shanghai Universities Peak Discipline Project of Aquaculture and“Double First-Class Initiative”program for its First-Class Discipline of aquaculture.
文摘The carbonic anhydrases(CAs)are a group of enzymes that play an important role in the absorption and transportation of CO_(2) in Saccharina japonica.They are encoded by a superfamily of genes with seven subtypes that are unrelated in sequence but share conserved function in catalyzing the reversible conversion of CO_(2) and HCO_(3)^(-).Here we have characterized the CA members in the transcriptome of S.japonica using Single-molecule real-time(SMRT)sequencing technology.Approximately 9830.4 megabases from 5,028,003 quality subreads were generated,and they were assembled into 326,512 full-length non-chimeric(FLNC)reads,with an average flnc read length of 2181 bp.After removing redundant sequences,79,010 unique transcripts were obtained of which 38,039 transcripts were successfully annotated.From the full-length transcriptome,we have identified 7 full-length cDNA sequences for CA genes(4α-CAs,1β-CAs and 2γ-CAs)and assessed for their potential functions based on phylogenetic analysis.Characterizations of CAs will provide the ground for future studies to determine the involvement of CAs in inorganic carbon absorption and transportation in S.japonica.
基金supported by National Natural Science Foundation Key Program,China(No.81930023)Natural Science Foundation Major International(Regional)Joint Research Project,China(No.81720108009)+2 种基金Chongqing Outstanding Scientists Project(2019),Chongqing Key Laboratory of Ophthalmology,China(No.CSTC,2008CA5003)Chongqing Science&Technology Platform and Base Construction Program,China(No.cstc2014ptsy10002)the Chongqing Chief Medical Scientist Project,China(2018).
文摘Vogt-Koyanagi-Harada disease(VKH)is a rare autoimmune disease characterized by diffuse and bilateral uveitis,alopecia,tinnitus,hearing loss,vitiligo and headache.The transcriptional expression pattern of peripheral blood mononuclear cells(PBMC)in VKH remains largely unknown.In this study,mRNA sequencing was conducted in PBMC from VKH patients with active uveitis before treatment(n=7),the same patients after prednisone combined with cyclosporine treatment(n=7)and healthy control subjects strictly matched with gender and age(n=7).We found 118 differentially expressed genes(DEGs)between VKH patients and healthy control subjects,and 21 DEGs between VKH patients before and after treatment.TRIB1 was selected as a potential biomarker to monitor the development of VKH according to the mRNA sequencing.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed to predict the possible biological functions and signaling pathways of DEGs.Neutrophil degranulation,peptidase regulator activity,secretory granule membrane,cellular response to peptide,growth factor binding and cell projection membrane were enriched as GO annotations of DEGs.Arachidonic acid metabolism and mitogen-activated protein kinase(MAPK)signaling pathway were potential signaling pathways involved in pathogenesis and drug response of VKH.A protein–protein interaction(PPI)network was constructed by STRING,and colony stimulating factor 1 receptor(CSF1R)was identified as the hubgene of all DEGs by Cytoscape.The cell type presumed to contribute to the aberrant expression of DEGs was analyzed with the use of publicly available single-cell sequencing data of PBMC from a healthy donor and single-cell sequencing dataset of monocytes from VKH patients.Our findings may help to decipher the underlying cellular and molecular pathogenesis of VKH and may lead novel therapeutic applications.
基金supported by the National Natural Science Foundation of China(81772091)the Clinical Research Plan of Shanghai Hospital Development Center(No.SHDC2020CR3039B).
文摘Background:Hypertrophic scars cause impaired skin appearance and function,seriously affecting physical and mental health.Due to medical ethics and clinical accessibility,the collection of human scar specimens is frequently restricted,and the establishment of scar experimental animal models for scientific research is urgently needed.The four most commonly used animal models of hypertrophic scars have the following drawbacks:the rabbit ear model takes a long time to construct;the immunodeficient mouse hypertrophic scar model necessitates careful feeding and experimental operations;female Duroc pigs are expensive to purchase and maintain,and their large size makes it difficult to produce a significant number of models;and mouse scar models that rely on tension require special skin stretch devices,which are often damaged and shed,resulting in unstable model establishment.Our group overcame the shortcomings of previous scar animal models and created a new mouse model of hypertrophic scarring induced by suture anchoring at the wound edge.Methods:We utilized suture anchoring of incisional wounds to impose directional tension throughout the healing process,restrain wound contraction,and generate granulation tissue,thus inducing scar formation.Dorsal paired incisions were generated in mice,with wound edges on the upper back sutured to the rib cage and the wound edges on the lower back relaxed as a control.Macroscopic manifestation,microscopic histological analysis,mRNA sequencing,bioinformatics,and in vitro cell assays were also conducted to verify the reliability of this method.Results:Compared with those in relaxed controls,the fibrotic changes in stretched wounds were more profound.Histologically,the stretched scars were hypercellular,hypervascular,and hyperproliferative with disorganized extracellular matrix deposition,and displayed molecular hallmarks of hypertrophic fibrosis.In addition,the stretched scars exhibited transcriptional overlap with mechanically stretched scars,and human hypertrophic and keloid scars.Phosphatidylinositol 3-kinase-serine/threonine-protein kinase B signaling was implicated as a profibrotic mediator of apoptosis resistance under suture-induced tension.Conclusions:This straightforward murine model successfully induces cardinal molecular and histological features of pathological hypertrophic scarring through localized suture tension to inhibit wound contraction.The model enables us to interrogate the mechanisms of tension-induced fibrosis and evaluate anti-scarring therapies.
基金funded by core funding from the University of Lausanne,the Swiss National Science Foundation(grant 310030B_185379,awarded to C.S.H.)The Research Foundation-Flanders(FWO,post-doc fellowship 1215820N,awarded to T.E.)+2 种基金the European Research Council(ERC Start ing Grant TORPEDO,714055,awarded to B.D.R.)the BBSRC(grant BB/S002804/1 to G.W.B.)the Deutsche Forschungsgemeinschaft(DFG,post-doctoral fellowship GR 5009/1-1,awarded to M.G.).
文摘The effects of brassinosteroid signaling on shoot and root development have been characterized in great detail but a simple consistent positive or negative impact on a basic cellular parameter was not identified.In this study,we combined digital 3D single-cell shape analysis and single-cell mRNA sequencing to charac-terize root meristems and mature root segments of brassinosteroid-blind mutants and wild type.The resul-tant datasets demonstrate that brassinosteroid signaling affects neither cell volume nor cell proliferation capacity.Instead,brassinosteroid signaling is essential for the precise orientation of cell division planes and the extent and timing of anisotropic cell expansion.Moreover,we found that the cell-aligning effects of brassinosteroid signaling can propagate to normalize the anatomy of both adjacent and distant brassinosteroid-blind cells through non-cell-autonomous functions,which are sufficient to restore growth vigor.Finally,single-cell transcriptome data discern directly brassinosteroid-responsive genes from genes that can react non-cell-autonomously and highlight arabinogalactans as sentinels of brassinosteroid-dependent anisotropic cell expansion.
