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Developmental pruning of early-stage myelin segments during CNS myelination in vivo

细胞研究（英文版） 2013年第7期23卷 962-964页

作者： Peng Liu Jiu-lin Du Cheng He Department of Neurobiology and Key Laboratory of Molecular Neurobiology of Ministry of Education Second Military Medical University 800 Xiangyin Road Shanghai 200433 China Institute of Neuroscience and State Key Laboratory of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China

Dear Editor,Axon-wrapping myelin sheath is essential for proper functioning of the nervous *** is formed from membranous sheets that are elaborated by oligodendrocytes (OLs) in the central nervous system (CNS).Although much is known about the development and migration of OL and its progenitor cell in vitro and in vivo [1-3],how CNS myelin is dynamically formed through OL processes is still unclear [1,3].To examine the process of CNS myelination in vivo,we performed longterm time-lapse confocal imaging of the same OLs in the spinal cord of intact zebrafish larvae during 2-7 days post-fertilization (dpf) with intervals of 2-12 *** OLs were mosaically labeled by membrane-tethered red fluorescent protein (mRFP) under the control of zebrafish Sox10 regulatory DNA [2].We find that,besides extension of some early-stage axon-wrapping myelin segments,28％ segments undergo pruning in an axon diameter-correlated manner during early *** neuregulin-ErbB signaling impairs the pruning but not ***,our study reveals earlystage myelin pruning as a novel process during myelination.

关键词：髓鞘/超微结构中枢神经系统/细胞学体外研究人类

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Production of human CNS neurons from embryonal carcinoma cells using a cell aggregation method

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BIOTECHNIQUES 1999年第5期26卷 946-+页

作者： Cheung, WMW Fu, WY Hui, WS Ip, NY Hong Kong Univ Sci & Technol Dept Biol Clear Water Bay Hong Kong Peoples R China Hong Kong Univ Sci & Technol Biotechnol Res Inst Hong Kong Peoples R China

When treated with retinoic acid (RA), a human embryonal carcinoma (EC) cell line, NTera2 cl.D/1 (NT2), differentiates into several morphologically distinct cell types, which include terminally differentiated postmitotic central nervous system (CNS) neurons. Accumulating evidence has demonstrated the significant potential of NT2 cells in studies related to cancer therapy and neurodegenerative diseases However, preparation of enriched NT2 neurons often requires a lengthy period (co. five,weeks) and depends largely on tedious techniques similar to those used for primary neuronal cultures. Here, we report a rapid protocol for the preparation of these human CNS neurons. Using the method of cell aggregation, enriched NT2 neurons can he obtained in approximately two weeks. We also demonstrated that cell aggregation reduced the time normally required for the induction of neuronal differentiation as revealed by the early expression of neuronal markers. The period of RA treatment could also be reduced if NT2 cells were maintained cls aggregates for a sufficient period of time. Taken together our findings demonstrated that cell aggregation promoted RA-induced neuronal differentiation of NT2 cells and provided a rapid protocol for the efficient production of NT2 neurons. The ability to produce large quantities of human CNS neurons should facilitate future rise of these neurons for basic research and applications in cell therapy.

关键词：生物技术癌胚胎性/病理学细胞集合细胞分化/药物作用中枢神经系统/细胞学中枢神经系统/药物作用中枢神经系统/代谢神经组织蛋白质类/代谢神经突/超微结构神经元/细胞学神经元/药物作用神经元/代谢时间因素维甲酸/药理学肿瘤细胞培养的人类

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Genetic ablation reveals that the roof plate is essential for dorsal interneuron specification

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NATURE 2000年第6771期403卷 734-740页

作者： Lee, KJ Dietrich, P Jessell, TM Columbia Univ Ctr Neurobiol & Behav Dept Biochem & Mol Biophys Howard Hughes Med Inst New York NY 10032 USA Columbia Univ Dept Genet & Dev New York NY 10032 USA

During neural development in vertebrates, a spatially ordered array of neurons is generated in response to inductive signals derived from localized organizing centres, One organizing centre that has been proposed to have a role in the control of neural patterning is the roof plate, To define the contribution of signals derived from the roof plate to the specification of neuronal cell types in the dorsal neural tube, we devised a genetic strategy to ablate the roof plate selectively in mouse embryos, Embryos without a roof plate lack all the interneuron subtypes that are normally generated in the dorsal third of the neural tube. Using a genetically based lineage analysis and in vitro assays, we show that the loss of these neurons results from the elimination of nonautonomous signals provided by the roof plate, These results reveal that the roof plate is essential for specifying multiple classes of neurons in the mammalian central nervous system.

关键词：躯体发育模式/遗传学躯体发育模式/生理学骨形态发生蛋白质类细胞谱系中枢神经系统/细胞学中枢神经系统/胚胎学培养技术白喉毒素/遗传学外胚层/生理学胚胎诱导基因打靶生长物质/遗传学生长物质/生理学同源盒结构域蛋白质类中间神经元/生理学肌蛋白质类/代谢神经组织蛋白质类/代谢 PAX7转录因子信号传导动物小鼠

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Expression of the antiproliferative gene TIS21 at the onset of neurogenesis identifies single neuroepithelial cells that switch from proliferative to neuron-generating division

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 1999年第8期96卷 4639-4644页

作者： Iacopetti, P Michelini, M Stuckmann, I Oback, B Aaku-Saraste, E Huttner, WB Univ Heidelberg Dept Neurobiol D-69120 Heidelberg Germany

At the onset of mammalian neurogenesis, neuroepithelial (NE) cells switch from proliferative to neuron-generating divisions, Understanding the molecular basis of this switch requires the ability to distinguish between these two types of division. Here we show that in the mouse ventricular zone, expression of the mRNA of the antiproliferative gene TIS21 (PC3, BTG2) (i) starts at the onset of neurogenesis, (ii) is confined to a subpopulation of NE cells that increases in correlation with the progression of neurogenesis, and (iii) is not detected in newborn neurons. Expression of the TIS21 mRNA in the NE cells occurs transiently during the cell cycle, i.e., in the G(1) phase. In contrast to the TIS21 mRNA, the TIS21 protein persists through the division of NE cells and is inherited by the neurons, where it remains detectable during neuronal migration and the initial phase of differentiation. Our observations indicate that the TIS21 gene is specifically expressed in those NE cells that, at their next division, will generate postmitotic neurons, but not in proliferating NE cells. Using TIS21 as a marker, we find that the switch from proliferative to neuron-generating divisions is initiated in single NE cells rather than in synchronized neighboring cells.

关键词：动物新生 COS细胞细胞周期蛋白质类/生物合成细胞周期蛋白质类/遗传学细胞分裂中枢神经系统/细胞学中枢神经系统/胚胎学中枢神经系统/代谢上皮细胞/细胞学上皮细胞/生理学表位/分析基因表达调控发育期基因肿瘤抑制即早蛋白质类/生物合成即早蛋白质类/遗传学神经元/细胞学神经元/生理学 RNA 信使/分析 RNA 信使/遗传学重组蛋白质类/生物合成转录遗传转染肿瘤抑制蛋白质类动物小鼠

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Distribution and retrograde transport of trophic factors in the central nervous system: Functional implications for the treatment of neurodegenerative diseases

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PROGRESS IN NEUROBIOLOGY 1999年第4期57卷 451-484页

作者： Mufson, EJ Kroin, JS Sendera, TJ Sobreviela, T Rush Presbyterian St Lukes Med Ctr Rush Alzheimers Dis Ctr Res Ctr Brain Repair Dept Neurol Sci Chicago IL 60612 USA Rush Presbyterian St Lukes Med Ctr Rush Alzheimers Dis Ctr Res Ctr Brain Repair Dept Neurosurg Chicago IL 60612 USA

Neurotrophins play a crucial role in the maintenance, survival and selective vulnerability of various neuronal populations within the normal and diseased brain. Several families of growth promoting substances have been identified within the central nervous system (CNS) including the superfamily of nerve growth factor related neurotrophin factors, glial derived neurotrophic factor (GDNF) and ciliary neurotrophic factor (CNTF). In addition, other non-neuronal growth factors such as fibroblast growth factor (FGF) have also been identified. This article reviews the trophic anatomy of these factors within the CNS. Intraventricular and intra-parenchymal injections of exogenous nerve growth factor result in retrograde labeling mainly within the cholinergic basal forebrain. Distribution of brain derived neurotrophic factor (BDNF) following intraventricular injection is minimal due to the binding to the trkB receptor along the ventricular wall. In contrast, intraparenchymal injections of BDNF results in widespread retrograde transport throughout the CNS. BDNF has also been shown to be transported anterogradely within the CNS. Infusion of GDNF into the CNS results in retrograde transport limited to the nigrostriatal pathway. Hippocampal injections of NT-3 retrogradely label mainly basal forebrain neurons. Retrograde trans port of radiolabeled CNTF has only been observed in sensory neurons of the sciatic nerve. Following intraventricular and intraparenchymal infusion of radiolabeled bFGF, retrograde neuronal labeling was found in the telecephalon, diencephalon, mesencephalon and pens. In contrast retrograde labeling for aFGF was found only in the hypothalamus and midbrain. Since select neurotrophins traffic anterogradely and retrogradely within the nervous system, these proteins could be used to treat neurological diseases such as Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis. (C) 1999 Elsevier Science Ltd. All rights reserved.

关键词：轴突运输中枢神经系统/细胞学中枢神经系统/代谢中枢神经系统/病理学成纤维细胞生长因子/投药和剂量成纤维细胞生长因子/代谢神经生长因子类/投药和剂量神经生长因子类/代谢神经生长因子类/药理学神经组织蛋白质类/投药和剂量神经组织蛋白质类/代谢神经组织蛋白质类/药理学神经变性疾病/代谢神经变性疾病/治疗受体神经生长因子类/代谢动物人类

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Vnd/nkx, ind/gsh, and msh/msx:: conserved regulators of dorsoventral neural patterning?

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CURRENT OPINION IN NEUROBIOLOGY 2000年第1期10卷 63-71页

作者： Cornell, RA Von Ohlen, T Univ Oregon Inst Neurosci Eugene OR 97403 USA

Expression of vnd in ventral, ind in intermediate, and msh in dorsal columns of fly neurectoderm, and of homologous gene families in corresponding domains of vertebrate neurectoderm, suggests that elements of dorsoventral neural patterning have been evolutionarily conserved. However, upstream signaling pathways regulating this columnar gene expression pattern appear to have diverged significantly throughout evolution. In addition, while recent loss-of-function studies in flies and mice indicate that these three genes may have a conserved role in regional specification, there is no obvious conservation of the particular cell fates deriving from corresponding domains. The three-column expression pattern may thus represent a developmental mechanism that is more resistant to evolutionary changes than genetic events upstream or downstream of it.

关键词：躯体发育模式中枢神经系统/细胞学中枢神经系统/胚胎学中枢神经系统/代谢保守序列/遗传学保守序列/生理学果蝇蛋白质类黑腹果蝇基因表达调控发育期同源盒结构域蛋白质类/遗传学同源盒结构域蛋白质类/生理学信号传导转录因子脊椎动物动物

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Characterization of MARCKS (myristoylated alanine-rich C kinase substrate) identified by a monoclonal antibody generated against chick embryo neural retina

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 1999年第2期257卷 480-487页

作者： Zolessi, FR Hellman, U Baz, A Arruti, C Univ Republica Fac Ciencias Lab Cultivo Tejidos Secc Biol Celular Montevideo 11000 Uruguay Univ Republica Fac Ciencias Catedra Inmunol UA Montevideo 11000 Uruguay Univ Republica Fac Med Dept Histol & Embriol Montevideo 11000 Uruguay Ludwig Inst Canc Res S-75124 Uppsala Sweden

To identify molecular markers of cell differentiation in developing nervous tissue, monoclonal antibodies against chick embryo neural retina were made. One of them, 3C3mAb, recognized a developmentally regulated antigen present in several organs of the CNS. Data from MALDI-TOF mass spectrometry and peptide sequencing of the immuno-affinity purified protein indicated identity of the antigen with MARCKS. The immunoreactive material was always found as a unique polypeptide (M-r 71 kDa) in SDS-PAGE, however isoelectrofocusing revealed the existence of several bands (pI ranging from 4.0 to 4.5). Interestingly some retinal cell types, as photoreceptors, exhibited an extremely significant decrease in the intensity of the immunoreactive material during the final phases of terminal differentiation while others, as some retinal neurons, maintained the immunoreactivity when fully differentiated. Taken together these results indicate that MARCKS, a protein susceptible of several posttranslational modifications as myristoylation and phosphorylation at variable extent, may act differently in neural retina cell types. (C) 1999 Academic Press.

关键词：氨基酸序列抗体单克隆/免疫学印迹法蛋白质细胞分化细胞培养的中枢神经系统/细胞学中枢神经系统/生长和发育中枢神经系统/代谢杂交瘤细胞内信号肽和蛋白质类等电聚焦膜蛋白质类分子序列数据分子量辛苯昔醇肽碎片/化学肽碎片/分离和提纯光感受器/细胞学光感受器/生长和发育光感受器/代谢蛋白质类/化学蛋白质类/免疫学蛋白质类/分离和提纯蛋白质类/代谢视网膜神经节细胞/细胞学视网膜神经节细胞/代谢序列分析光谱法质量基质辅助激光解吸电离动物鸡胚小鼠

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Neuromedin B

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PROGRESS IN NEUROBIOLOGY 2000年第3期62卷 297-312页

作者： Ohki-Hamazaki, H Tokyo Med & Dent Univ Med Res Inst Dept Mol Neurosci Bunkyo Ku Tokyo 1138519 Japan

Neuromedin B (NMB) is one of the bombesin (BN)-related peptides in mammals. It was originally purified from pig spinal cords, and it has been shown to be present in central nervous system as well as in gastrointestinal tract. BN and its related peptides have various physiological effects. These include regulation of exocrine and endocrine secretions, smooth muscle contraction, feeding, blood pressure, blood glucose, body temperature and cell growth. NMB exerts its effect by binding to the cell surface receptor. A high affinity receptor, NMB receptor (NMB-R) has been identified. This is a G-protein coupled receptor with seven membrane-spanning regions. Upon agonist binding, several intracellular signaling cascades including phospholipase activation, calcium mobilization and protein kinase C (PKC) activation lead to expression of several genes, DNA synthesis or cellular effects such as secretion. Existence of NMB-R has been demonstrated in several brain regions, notably in olfactory and thalamic regions, and in gastrointestinal tracts. Recent analysis using NMB-R-deficient mice, generated by gene-targeting technique, enables to distinguish functional properties of NMB-R from GRP-R. In this review, molecular characterization, anatomical distribution and pharmacological properties of NMB and NMB-R will be presented. Moreover, physiological roles of NMB and its receptor demonstrated by the analysis of NMB-R-deficient mice will be reported. Comparison with GRP/GRP-R system will provide important information about BN-like peptide systems in mammals. (C) 2000 Elsevier Science Ltd. All rights reserved.

关键词：中枢神经系统/细胞学中枢神经系统/代谢消化系统/细胞学消化系统/代谢神经激肽B/类似物和衍生物神经激肽B/代谢动物人类

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Embryogenesis of the central nervous system of the pond snail Lymnaea stagnalis L.: An ultrastructural study

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JOURNAL OF NEUROCYTOLOGY 2000年第1期29卷 43-60页

作者： Nagy, T Elekes, K Hungarian Acad Sci Balaton Limnol Res Inst Dept Expt Zool H-8237 Tihany Hungary

The ultrastructural characteristics of the developing CNS of the pond snail, Lymnaea stagnalis, were investigated, with special attention paid to three specific stages of embryonic development, representing distinctly different phases of both the body morphogenesis and gangliogenesis. These were the 35% (veliger), the 50-55% (metamorphic), and the 75% (post-metamorphic, adult-like) stages of embryonic development. Also, a brief comparison was done with the CNS of hatchlings (100% of embryonic development). During embryogenesis specialized axo-axonic synapses and elements of the glial system, including the ganglionic (neural) sheath, were rarely observed, whereas the frequent occurrence of unspecialized axo-somatic contacts could be demonstrated. Synapse-like axo-axonic connections could be found first in 75% embryos, showing asymmetric vesicle clustering on the "presynaptic" side and increased electron density of the apposed membranes. These phenomena may reflect the dominance of modulatory processes in the CNS during embryogenesis, and the absence of the neural sheath may facilitate trophic and/or hormonal influences within the developing ganglia. The gradual increase in the size of ganglia and the diameter of their neuropils was not accompanied by any widening of the cell body layer or increasing diameter of the nerve cell bodies until the very end of embryogenesis. With respect to the ultrastructural organization of the neuropil, and possibly the entire CNS, a determining stage seems to be that of metamorphosis. Two types of neuropil could be observed at this time;the metamorphosing neuropil with an irregular organization of wavy axon profiles, and well-structured neuropil with a regular organization of axon profiles. Ganglia with irregular or regular neuropil occurred simultaneously in the developing CNS.

关键词：中枢神经系统/细胞学中枢神经系统/胚胎学神经节无脊椎动物/细胞学神经节无脊椎动物/胚胎学椎实螺属/胚胎学显微镜检查电子神经元/超微结构神经毡/超微结构突触/超微结构动物

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Atm and Bax cooperate in ionizing radiation-induced apoptosis in the central nervous system

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第2期97卷 889-894页

作者： Chong, MJ Murray, MR Gosink, EC Russell, HRC Srinivasan, A Kapsetaki, M Korsmeyer, SJ McKinnon, PJ St Jude Childrens Res Hosp Dept Genet Memphis TN 38101 USA IDUN Pharmaceut Inc La Jolla CA 92037 USA Harvard Univ Sch Med Dana Farber Canc Inst Dept Pathol & Med Boston MA 02115 USA

Ataxia-telangiectasia is a hereditary multisystemic disease resulting from mutations of ataxia telangiectasia, mutated (ATM) and is characterized by neurodegeneration, cancer, immune defects, and hypersensitivity to ionizing radiation. The molecular details of ATM function in the nervous system are unclear, although the neurological lesion in ataxia-telangiectasia becomes apparent early in life, suggesting a developmental origin. The central nervous system (CNS) of Atm-null mice shows a pronounced defect in apoptosis induced by genetoxic stress, suggesting ATM functions to eliminate neurons with excessive genomic damage. Here, we report that the death effector Bax is required for a large proportion of Atm-dependent apoptosis in the developing CNS after ionizing radiation (IR). Although many of the same regions of the CNS in both Bax-/- and Atm-/- mice were radioresistant, mice nullizygous for both Bax and Atm showed additional reduction in IR-induced apoptosis in the CNS. Therefore, although the major IR-induced apoptotic pathway in the CNS requires Atm and Bax, a p53-dependent collateral pathway exists that has both Atm- and Bax-independent branches. Further, Atm- and Bax-dependent apoptosis in the CNS also required caspase-3 activation. These data implicate Bax and caspase-3 as death effecters in neurodegenerative pathways.

关键词：细胞凋亡/遗传学细胞凋亡/辐射效应印迹法蛋白质半胱氨酸天冬氨酸蛋白酶3 半胱氨酸天冬氨酸蛋白酶/代谢细胞周期蛋白质类中枢神经系统/细胞学中枢神经系统/代谢中枢神经系统/辐射效应小脑/代谢小脑/辐射效应 DNA结合蛋白质类齿状回/代谢齿状回/辐射效应酶激活/辐射效应基因型小鼠基因敲除蛋白质丝氨酸苏氨酸激酶/遗传学蛋白质丝氨酸苏氨酸激酶/代谢蛋白质丝氨酸苏氨酸激酶/生理学原癌基因蛋白质类/遗传学原癌基因蛋白质类/代谢原癌基因蛋白质类/生理学原癌基因蛋白质c-bcl-2 辐射电离视网膜/代谢视网膜/辐射效应肿瘤抑制蛋白质p53/遗传学肿瘤抑制蛋白质p53/代谢肿瘤抑制蛋白质p53/生理学肿瘤抑制蛋白质类 bcl-2相关X蛋白质动物小鼠

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