The visual system of teleost fish grows continuously,which is a useful model for studying regeneration of the central nervous system.Glial cells are key for this process,but their contribution is still not well define...The visual system of teleost fish grows continuously,which is a useful model for studying regeneration of the central nervous system.Glial cells are key for this process,but their contribution is still not well defined.We followed oligodendrocytes in the visual system of adult zebrafish during regeneration of the optic nerve at 6,24,and 72 hours post-lesion and at 7 and 14 days post-lesion via the sox10:tagRFP transgenic line and confocal microscopy.To understand the changes that these oligodendrocytes undergo during regeneration,we used Sox2 immunohistochemistry,a stem cell marker involved in oligodendrocyte differentiation.We also used the Click-iT™ Plus TUNEL assay to study cell death and a BrdU assay to determine cell proliferation.Before optic nerve crush,sox10:tagRFP oligodendrocytes are located in the retina,in the optic nerve head,and through all the entire optic nerve.Sox2-positive cells are present in the peripheral germinal zone,the mature retina,and the optic nerve.After optic nerve crush,sox10:tagRFP cells disappeared from the optic nerve crush zone,suggesting that they died,although they were not TUNEL positive.Concomitantly,the number of Sox2-positive cells increased around the crushed area,the optic nerve head,and the retina.Then,between 24 hours post-lesion and 14 days post-lesion,double sox10:tagRFP/Sox2-positive cells were detected in the retina,optic nerve head,and whole optic nerve,together with a proliferation response at 72 hours post-lesion.Our results confirm that a degenerating process may occur prior to regeneration.First,sox10:tagRFP oligodendrocytes that surround the degenerated axons stop wrapping them,change their“myelinating oligodendrocyte”morphology to a“nonmyelinating oligodendrocyte”morphology,and die.Then,residual oligodendrocyte progenitor cells in the optic nerve and retina proliferate and differentiate for the purpose of remyelination.As new axons arise from the surviving retinal ganglion cells,new sox10:tagRFP oligodendrocytes arise from residual oligodendrocyte progenitor cells to guide,nourish and myelinate them.Thus,oligodendrocytes play an active role in zebrafish axon regeneration and remyelination.展开更多
Neural stem cells(NSCs)have the potential for self-renewal and multidirectional differentiation,and their transplantation has achieved good efficacy in a variety of diseases.However,only 1%-10%of transplanted NSCs sur...Neural stem cells(NSCs)have the potential for self-renewal and multidirectional differentiation,and their transplantation has achieved good efficacy in a variety of diseases.However,only 1%-10%of transplanted NSCs survive in the ischemic and hypoxic microenvironment of posthemorrhagic hydrocephalus.^(Sox2)is an important factor for NSCs to maintain proliferation.Therefore,^(Sox2)-overexpressing NSCs(NSC^(Sox2))may be more successful in improving neurological dysfunction after posthemorrhagic hydrocephalus.In this study,human NSC^(Sox2)was transplanted into a posthemorrhagic hydrocephalus mouse model,and retinoic acid was administered to further promote NSC differentiation.The results showed that NSC^(Sox2)attenuated the ventricular enlargement caused by posthemorrhagic hydrocephalus and improved neurological function.NSC^(Sox2)also promoted nerve regeneration,inhibited neuroinflammation and promoted M2 polarization(anti-inflammatory phenotype),thereby reducing cerebrospinal fluid secretion in choroid plexus.These findings suggest that NSC^(Sox2)rescued ventricular enlargement and neurological dysfunction induced by posthemorrhagic hydrocephalus through neural regeneration and modulation of inflammation.展开更多
有性生殖是多细胞动物物种繁衍的普遍方式。本研究以大型淡水贝类中国圆田螺(Cipangopaludina chinensis)为例,从全基因组层面鉴定与中国圆田螺性别分化相关的Dmrt(double-sex and mab-3 relatated transcription factor)、Sox(sry-rela...有性生殖是多细胞动物物种繁衍的普遍方式。本研究以大型淡水贝类中国圆田螺(Cipangopaludina chinensis)为例,从全基因组层面鉴定与中国圆田螺性别分化相关的Dmrt(double-sex and mab-3 relatated transcription factor)、Sox(sry-related high mobility group box)和Wnt(wingless-type mmtv integration site family)基因家族并分析其特点、演化及在性腺中的表达模式。结果显示中国圆田螺CchDmrt、CchSox和CchWnt基因家族分别含有3个、5个和10个成员,主要分布于2号、3号、5号、7号和9号染色体上。选取腹足纲代表性物种构建系统进化树,结果显示Dmrt、Sox和Wnt各亚家族分布聚类在一起,其中中国圆田螺与铜锈环棱螺(Bellamya aeruginosa)亲缘关系最近。3个基因家族中,CchSox10、CchWnt5和CchWnt7受到正选择作用,相对进化速率较快。三维结构分析显示CchDmrt和CchWnt基因家族编码的蛋白质较为保守,而CchSox基因家族编码的蛋白质之间则存在一定差异性,提示其存在功能差异。雌雄性腺转录组表达分析显示,CchSox5、CchWnt2和CchWnt16在精巢中的表达量高于卵巢的,并形成差异表达基因,而CchDmrt4、CchSox2、CchSox10、CchSox12和CchFoxl2基因则相反,说明不同家族成员在性腺发育中的作用不同。选取差异表达基因进行实时定量PCR(real-time quantitative PCR,RT-qPCR)验证,结果与转录组结果一致。本研究鉴定了中国圆田螺Dmrt、Sox和Wnt等性别决定相关基因,为其功能研究和中国圆田螺遗传育种研究提供基础资料。展开更多
基金supported by the Lanzadera TCUE and C2 program(Universidad de Salamanca)(to ASL)the Spanish National Research Council(CSIC)funded by the Junta de Castilla y León and co-financed by the European Regional Development Fund(ERDF“Europe drives our growth”):Internationalization Project“CL-EI-2021-08-IBFG Unit of Excellence”,Grant(PID2022-138478OA-100)funded by MICIU/AEI/10.13039/501100011033 and,by FEDER,UE(to MGM)+3 种基金Junta de Castilla y León(SA225P23)Gerencia Regional de Salud(2701/A1/2023)(to AV)the Plan Especial Grado Medicina(USAL)(to CPM)a Ramón y Cajal researcher:Grant RYC2021-033684-I funded by MICIU/AEI/10.13039/501100011033 and,by European Union NextGenerationEU/PRTR.
文摘The visual system of teleost fish grows continuously,which is a useful model for studying regeneration of the central nervous system.Glial cells are key for this process,but their contribution is still not well defined.We followed oligodendrocytes in the visual system of adult zebrafish during regeneration of the optic nerve at 6,24,and 72 hours post-lesion and at 7 and 14 days post-lesion via the sox10:tagRFP transgenic line and confocal microscopy.To understand the changes that these oligodendrocytes undergo during regeneration,we used Sox2 immunohistochemistry,a stem cell marker involved in oligodendrocyte differentiation.We also used the Click-iT™ Plus TUNEL assay to study cell death and a BrdU assay to determine cell proliferation.Before optic nerve crush,sox10:tagRFP oligodendrocytes are located in the retina,in the optic nerve head,and through all the entire optic nerve.Sox2-positive cells are present in the peripheral germinal zone,the mature retina,and the optic nerve.After optic nerve crush,sox10:tagRFP cells disappeared from the optic nerve crush zone,suggesting that they died,although they were not TUNEL positive.Concomitantly,the number of Sox2-positive cells increased around the crushed area,the optic nerve head,and the retina.Then,between 24 hours post-lesion and 14 days post-lesion,double sox10:tagRFP/Sox2-positive cells were detected in the retina,optic nerve head,and whole optic nerve,together with a proliferation response at 72 hours post-lesion.Our results confirm that a degenerating process may occur prior to regeneration.First,sox10:tagRFP oligodendrocytes that surround the degenerated axons stop wrapping them,change their“myelinating oligodendrocyte”morphology to a“nonmyelinating oligodendrocyte”morphology,and die.Then,residual oligodendrocyte progenitor cells in the optic nerve and retina proliferate and differentiate for the purpose of remyelination.As new axons arise from the surviving retinal ganglion cells,new sox10:tagRFP oligodendrocytes arise from residual oligodendrocyte progenitor cells to guide,nourish and myelinate them.Thus,oligodendrocytes play an active role in zebrafish axon regeneration and remyelination.
基金supported by the National Natural Science Foundation of China,Nos.82473334(to LZ),82401629(to XL)the Major Scientific and Technological Achievements Transformation Project of Ningxia Hui Autonomous Region,No.2022CJE09013(to LZ)+4 种基金Mianyang Science and Technology Bureau(Mianyang Science and Technology Program),No.2023ZYDF097(to LZ)NHC Key Laboratory of Nuclear Technology Medical Transformation(Mianyang Central Hospital),No.2023HYX001(to LZ)Spinal Cord Diseases Clinical Medical Center of Yunnan Province,No.2024JSKFKT-16(to BG)the Natural Science Foundation of Sichuan Province,No.2024NSFSC1646(to XL)the China Postdoctoral Science Foundation,Nos.GZC20231811(to XL),2024T170601(to XL)and 2024M76228(to XL).
文摘Neural stem cells(NSCs)have the potential for self-renewal and multidirectional differentiation,and their transplantation has achieved good efficacy in a variety of diseases.However,only 1%-10%of transplanted NSCs survive in the ischemic and hypoxic microenvironment of posthemorrhagic hydrocephalus.^(Sox2)is an important factor for NSCs to maintain proliferation.Therefore,^(Sox2)-overexpressing NSCs(NSC^(Sox2))may be more successful in improving neurological dysfunction after posthemorrhagic hydrocephalus.In this study,human NSC^(Sox2)was transplanted into a posthemorrhagic hydrocephalus mouse model,and retinoic acid was administered to further promote NSC differentiation.The results showed that NSC^(Sox2)attenuated the ventricular enlargement caused by posthemorrhagic hydrocephalus and improved neurological function.NSC^(Sox2)also promoted nerve regeneration,inhibited neuroinflammation and promoted M2 polarization(anti-inflammatory phenotype),thereby reducing cerebrospinal fluid secretion in choroid plexus.These findings suggest that NSC^(Sox2)rescued ventricular enlargement and neurological dysfunction induced by posthemorrhagic hydrocephalus through neural regeneration and modulation of inflammation.
文摘有性生殖是多细胞动物物种繁衍的普遍方式。本研究以大型淡水贝类中国圆田螺(Cipangopaludina chinensis)为例,从全基因组层面鉴定与中国圆田螺性别分化相关的Dmrt(double-sex and mab-3 relatated transcription factor)、Sox(sry-related high mobility group box)和Wnt(wingless-type mmtv integration site family)基因家族并分析其特点、演化及在性腺中的表达模式。结果显示中国圆田螺CchDmrt、CchSox和CchWnt基因家族分别含有3个、5个和10个成员,主要分布于2号、3号、5号、7号和9号染色体上。选取腹足纲代表性物种构建系统进化树,结果显示Dmrt、Sox和Wnt各亚家族分布聚类在一起,其中中国圆田螺与铜锈环棱螺(Bellamya aeruginosa)亲缘关系最近。3个基因家族中,CchSox10、CchWnt5和CchWnt7受到正选择作用,相对进化速率较快。三维结构分析显示CchDmrt和CchWnt基因家族编码的蛋白质较为保守,而CchSox基因家族编码的蛋白质之间则存在一定差异性,提示其存在功能差异。雌雄性腺转录组表达分析显示,CchSox5、CchWnt2和CchWnt16在精巢中的表达量高于卵巢的,并形成差异表达基因,而CchDmrt4、CchSox2、CchSox10、CchSox12和CchFoxl2基因则相反,说明不同家族成员在性腺发育中的作用不同。选取差异表达基因进行实时定量PCR(real-time quantitative PCR,RT-qPCR)验证,结果与转录组结果一致。本研究鉴定了中国圆田螺Dmrt、Sox和Wnt等性别决定相关基因,为其功能研究和中国圆田螺遗传育种研究提供基础资料。
