Paraneoplastic neurological syndrome refers to certain malignant tumors that have affected the distant nervous system and caused corresponding dysfunction in the absence of tumor metastasis.Patients with this syndrome...Paraneoplastic neurological syndrome refers to certain malignant tumors that have affected the distant nervous system and caused corresponding dysfunction in the absence of tumor metastasis.Patients with this syndrome produce multiple antibodies,each targeting a different antigen and causing different symptoms and signs.The CV2/collapsin response mediator protein 5(CRMP5)antibody is a major antibody of this type.It damages the nervous system,which often manifests as limbic encephalitis,chorea,ocular manifestation,cerebellar ataxia,myelopathy,and peripheral neuropathy.Detecting CV2/CRMP5 antibody is crucial for the clinical diagnosis of paraneoplastic neurological syndrome,and anti-tumor and immunological therapies can help to alleviate symptoms and improve prognosis.However,because of the low incidence of this disease,few repo rts and no reviews have been published about it so far.This article intends to review the research on CV2/CRMP5antibody-associated paraneoplastic neurological syndrome and summarize its clinical features to help clinicians comprehensively understand the disease.Additionally,this review discusses the curre nt challenges that this disease poses,and the application prospects of new detection and diagnostic techniques in the field of paraneoplastic neurological syndrom e,including CV2/CRMP5-associated paraneoplastic neurological syndrome,in recent years.展开更多
目的探讨CRMP5对大鼠海马神经元突起生长的影响。方法将带FAM标记的CRMP5的特异性干扰片段及阴性对照转染培养成熟的海马神经元,用免疫荧光的方法验证干扰片段对神经元内源性CRMP5的干扰效果,并利用共聚焦显微镜观察神经元突起以及侧枝...目的探讨CRMP5对大鼠海马神经元突起生长的影响。方法将带FAM标记的CRMP5的特异性干扰片段及阴性对照转染培养成熟的海马神经元,用免疫荧光的方法验证干扰片段对神经元内源性CRMP5的干扰效果,并利用共聚焦显微镜观察神经元突起以及侧枝的形成。结果携带FAM的si RNA可以成功的进入细胞,分布于神经元的胞体以及树突;免疫荧光证实CRMP5 si RNA可以有效的沉默CRMP5蛋白的表达;沉默CRMP5基因表达后的海马神经元突起短小,而且缺少分支,而对照细胞突起长,分支多;定量分析显示,导入CRMP5 si RNA的细胞突起的长度较对照细胞缩短,差异显著(P<0.05);突起的数目比较,一级突起数目无显著差异,而二级及其以上突起的数目明显减少,差异显著(P<0.05)。结论沉默CRMP5可抑制海马神经元突起的生长和侧枝形成。展开更多
Recent studies have shown that mutation at Ser522 causes inhibition of collapsin response mediator protein 2(CRMP2) phosphorylation and induces axon elongation and partial recovery of the lost sensorimotor function af...Recent studies have shown that mutation at Ser522 causes inhibition of collapsin response mediator protein 2(CRMP2) phosphorylation and induces axon elongation and partial recovery of the lost sensorimotor function after spinal cord injury(SCI).We aimed to reveal the intracellular mechanism in axotomized neurons in the CRMP2 knock-in(CRMP2KI) mouse model by performing transcriptome analysis in mouse sensorimotor cortex using micro-dissection punching system.Prior to that, we analyzed the structural pathophysiology in axotomized or neighboring neurons after SCI and found that somatic atrophy and dendritic spine reduction in sensorimotor cortex were suppressed in CRMP2KI mice.Further analysis of the transcriptome has aided in the identification of four hemoglobin genes Hba-a1, Hba-a2, Hbb-bs, and Hbb-bt that are significantly upregulated in wild-type mice with concomitant upregulation of genes involved in the oxidative phosphorylation and ribosomal pathways after SCI.However, we observed substantial upregulation in channel activity genes and downregulation of genes regulating vesicles, synaptic function, glial cell differentiation in CRMP2KI mice.Moreover, the transcriptome profile of CRMP2KI mice has been discussed wherein energy metabolism and neuronal pathways were found to be differentially regulated.Our results showed that CRMP2KI mice displayed improved SCI pathophysiology not only via microtubule stabilization in neurons, but also possibly via the whole metabolic system in the central nervous system, response changes in glial cells, and synapses.Taken together, we reveal new insights on SCI pathophysiology and the regenerative mechanism of central nervous system by the inhibition of CRMP2 phosphorylation at Ser522.All these experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee at Waseda University, Japan(2017-A027 approved on March 21, 2017;2018-A003 approved on March 25, 2018;2019-A026 approved on March 25, 2019).展开更多
CRMP2 is one of the well-studied members of the CRMPs family,which has been demonstrated not only play the roles in neurite extension,dendrites and dendritic spine development,but also in the AMPA receptor trafficking...CRMP2 is one of the well-studied members of the CRMPs family,which has been demonstrated not only play the roles in neurite extension,dendrites and dendritic spine development,but also in the AMPA receptor trafficking.However,the role of CRMP2 on the trafficking of AMPA receptor GluA2 subunit especially phosphorylated GluA2 is unknown.GST-Pull down and C-IP assays,immunofluorescence staining and whole cell patch clamp recording technique were performed in this study.The results showed that CRMP2 interacted with wild-type GluA2 and promoted the surface expression of wild-type GluA2.No matter GluA2 S880 was exogenously phosphorylated by phorbol myristate acetate(TPA)or endogenously phosphorylated by PKC5,CRMP2 can promote the surface expression of phosphorylated GluA2 and enhance its amplitude and frequency of mEPSCs,increasing the slope of the I-V curve.展开更多
基金National Natural Science Foundation of China,No.U1604181Henan Province Key R&D and Promotion Special Project (Science and Technology Tackle),No.212102310834+1 种基金Henan Medical Education Research Project,No.Wjlx2020531the Joint project of Medical Science and Technology Research Program of Henan Province,No.LHGJ20190078 (all to JW)。
文摘Paraneoplastic neurological syndrome refers to certain malignant tumors that have affected the distant nervous system and caused corresponding dysfunction in the absence of tumor metastasis.Patients with this syndrome produce multiple antibodies,each targeting a different antigen and causing different symptoms and signs.The CV2/collapsin response mediator protein 5(CRMP5)antibody is a major antibody of this type.It damages the nervous system,which often manifests as limbic encephalitis,chorea,ocular manifestation,cerebellar ataxia,myelopathy,and peripheral neuropathy.Detecting CV2/CRMP5 antibody is crucial for the clinical diagnosis of paraneoplastic neurological syndrome,and anti-tumor and immunological therapies can help to alleviate symptoms and improve prognosis.However,because of the low incidence of this disease,few repo rts and no reviews have been published about it so far.This article intends to review the research on CV2/CRMP5antibody-associated paraneoplastic neurological syndrome and summarize its clinical features to help clinicians comprehensively understand the disease.Additionally,this review discusses the curre nt challenges that this disease poses,and the application prospects of new detection and diagnostic techniques in the field of paraneoplastic neurological syndrom e,including CV2/CRMP5-associated paraneoplastic neurological syndrome,in recent years.
文摘目的探讨CRMP5对大鼠海马神经元突起生长的影响。方法将带FAM标记的CRMP5的特异性干扰片段及阴性对照转染培养成熟的海马神经元,用免疫荧光的方法验证干扰片段对神经元内源性CRMP5的干扰效果,并利用共聚焦显微镜观察神经元突起以及侧枝的形成。结果携带FAM的si RNA可以成功的进入细胞,分布于神经元的胞体以及树突;免疫荧光证实CRMP5 si RNA可以有效的沉默CRMP5蛋白的表达;沉默CRMP5基因表达后的海马神经元突起短小,而且缺少分支,而对照细胞突起长,分支多;定量分析显示,导入CRMP5 si RNA的细胞突起的长度较对照细胞缩短,差异显著(P<0.05);突起的数目比较,一级突起数目无显著差异,而二级及其以上突起的数目明显减少,差异显著(P<0.05)。结论沉默CRMP5可抑制海马神经元突起的生长和侧枝形成。
基金supported by Grants-in-Aid for Scientific Research on Priority Areas from The Ministry of Education, Culture, Sports, Science and Technology(No.26430043to TO)。
文摘Recent studies have shown that mutation at Ser522 causes inhibition of collapsin response mediator protein 2(CRMP2) phosphorylation and induces axon elongation and partial recovery of the lost sensorimotor function after spinal cord injury(SCI).We aimed to reveal the intracellular mechanism in axotomized neurons in the CRMP2 knock-in(CRMP2KI) mouse model by performing transcriptome analysis in mouse sensorimotor cortex using micro-dissection punching system.Prior to that, we analyzed the structural pathophysiology in axotomized or neighboring neurons after SCI and found that somatic atrophy and dendritic spine reduction in sensorimotor cortex were suppressed in CRMP2KI mice.Further analysis of the transcriptome has aided in the identification of four hemoglobin genes Hba-a1, Hba-a2, Hbb-bs, and Hbb-bt that are significantly upregulated in wild-type mice with concomitant upregulation of genes involved in the oxidative phosphorylation and ribosomal pathways after SCI.However, we observed substantial upregulation in channel activity genes and downregulation of genes regulating vesicles, synaptic function, glial cell differentiation in CRMP2KI mice.Moreover, the transcriptome profile of CRMP2KI mice has been discussed wherein energy metabolism and neuronal pathways were found to be differentially regulated.Our results showed that CRMP2KI mice displayed improved SCI pathophysiology not only via microtubule stabilization in neurons, but also possibly via the whole metabolic system in the central nervous system, response changes in glial cells, and synapses.Taken together, we reveal new insights on SCI pathophysiology and the regenerative mechanism of central nervous system by the inhibition of CRMP2 phosphorylation at Ser522.All these experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee at Waseda University, Japan(2017-A027 approved on March 21, 2017;2018-A003 approved on March 25, 2018;2019-A026 approved on March 25, 2019).
文摘CRMP2 is one of the well-studied members of the CRMPs family,which has been demonstrated not only play the roles in neurite extension,dendrites and dendritic spine development,but also in the AMPA receptor trafficking.However,the role of CRMP2 on the trafficking of AMPA receptor GluA2 subunit especially phosphorylated GluA2 is unknown.GST-Pull down and C-IP assays,immunofluorescence staining and whole cell patch clamp recording technique were performed in this study.The results showed that CRMP2 interacted with wild-type GluA2 and promoted the surface expression of wild-type GluA2.No matter GluA2 S880 was exogenously phosphorylated by phorbol myristate acetate(TPA)or endogenously phosphorylated by PKC5,CRMP2 can promote the surface expression of phosphorylated GluA2 and enhance its amplitude and frequency of mEPSCs,increasing the slope of the I-V curve.
