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SV2A and SV2B function as redundant Ca²⁺ regulators in neurotransmitter release

NEURON 1999年第4期24卷 1003-1016页

作者： Janz, R Goda, Y Geppert, M Missler, M Südhof, TC Univ Texas SW Med Ctr Howard Hughes Med Inst Ctr Basic Neurosci Dallas TX 75235 USA Univ Texas SW Med Ctr Dept Mol Genet Dallas TX 75235 USA Univ Calif San Diego Dept Biol La Jolla CA 92093 USA Max Planck Inst Expt Med Dept Mol Neurobiol D-37075 Gottingen Germany

SV2 proteins are abundant synaptic vesicle proteins expressed in two major (SV2A and SV2B) and one minor isoform (SV2C) that resemble transporter proteins. We now show that SV2B knockout mice are phenotypically normal while SV2A- and SV2A/SV2B double knockout mice exhibit severe seizures and die postnatally. In electrophysiological recordings from cultured hippocampal neurons, SV2A- or SV2B-deficient cells exhibited no detectable abnormalities. Neurons lacking both SV2 isoforms, however, experienced sustained increases in Ca2+-dependent synaptic transmission when two or more action potentials were triggered in succession. These increases could be reversed by EGTA-AM. Our data suggest that without SV2 proteins, presynaptic Ca2+ accumulation during consecutive action potentials causes abnormal increases in neurotransmitter release that destabilize synaptic circuits and induce epilepsy.

关键词：脑化学/遗传学脑化学/生理学钙信号/生理学螯合剂/药理学克隆分子依他酸/药理学电生理学胞吞作用/生理学胞吐作用/生理学外显子海马/细胞学海马/生理学内含子膜糖蛋白类/遗传学膜糖蛋白类/生理学小鼠基因敲除分子序列数据突变神经组织蛋白质类/遗传学神经组织蛋白质类/生理学神经递质药/遗传学神经递质药/代谢神经递质药/生理学发作/病理生理学突触/生理学体重减轻/生理学动物小鼠

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Integrators of the cytoskeleton that stabilize microtubules

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CELL 1999年第2期98卷 229-238页

作者： Yang, YM Bauer, C Strasser, G Wollman, R Julien, JP Fuchs, E Univ Chicago Pritzker Sch Med Howard Hughes Med Inst Dept Mol Genet & Cell Biol Chicago IL 60637 USA Univ Chicago Pritzker Sch Med Howard Hughes Med Inst Dept Neurol Chicago IL 60637 USA McGill Univ Neurosci Res Ctr Montreal PQ H3G 1A4 Canada

Sensory neurodegeneration occurs in mice defective in BPAG1, a gene encoding cytoskeletal linker proteins capable of anchoring neuronal intermediate filaments to actin cytoskeleton. While BPAG1 null mice fail to anchor neurofilaments (NFs), BPAG1/NF null mice still degenerate in the absence of NFs. We report a novel neural splice form that lacks the actin-binding domain and instead binds and stabilizes microtubules. This interaction is functionally important;in mice and in vitro, neurons lacking BPAG1 display short, disorganized, and unstable microtubules defective in axonal transport. Ironically, BPAG1 neural isoforms represent microtubule-associated proteins that when absent lead to devastating consequences. Moreover, BPAG1 can functionally account for the extraordinary stability of axonal microtubules necessary for transport over long distances. Its isoforms interconnect all three cytoskeletal networks, a feature apparently central to neuronal survival.

关键词：轴突运输/生理学轴突/化学轴突/代谢轴突/超微结构载体蛋白质类细胞培养的细胞支架蛋白质类/化学细胞支架蛋白质类/遗传学细胞支架蛋白质类/代谢桥粒斑蛋白质类小鼠基因敲除微丝蛋白质类/代谢微丝/代谢微丝/超微结构显微镜检查免疫电子微管/代谢微管/超微结构分子序列数据神经组织蛋白质类/化学神经组织蛋白质类/遗传学神经组织蛋白质类/代谢神经微丝蛋白质类/代谢神经元传入/细胞学神经元传入/代谢神经元传入/超微结构蛋白质结构三级动物小鼠

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The ins and outs of a polyglutamine neurodegenerative disease: Spinocerebellar ataxia type 1 (SCA1)

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NEUROBIOLOGY OF DISEASE 2000年第3期7卷 129-134页

作者： Orr, HT Univ Minnesota Inst Human Genet Minneapolis MN 55455 USA Univ Minnesota Dept Genet Minneapolis MN 55455 USA Univ Minnesota Dept Cell Biol & Dev Lab Med & Pathol Minneapolis MN 55455 USA

Polyglutamine neurodegenerative disorders are characterized by the expansion of a glutamine tract within the mutant disease-causing protein. Expression of the mutant protein induces a progressive loss of neuronal function and the subsequent neurodegeneration of a set of neurons characteristic to each disease. Spinocerebellar ataxia type 1 (SCA1) is one polyglutamine disease where various experimental model systems, in particular transgenic mice, have been utilized to dissect the molecular and cellular events important for disease. This review summarizes these findings and places them in a context of potential future research directions, (C) 2000 Academic Press.

关键词：疾病模型动物小鼠转基因/遗传学神经组织蛋白质类/遗传学神经变性疾病/遗传学核蛋白质类/遗传学肽类/遗传学脊髓小脑共济失调/遗传学脊髓小脑共济失调/病理学脊髓小脑共济失调/病理生理学动物人类小鼠

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Cloning and functional expression of a novel degenerin-like Na⁺ channel gene in mammals

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JOURNAL OF PHYSIOLOGY-LONDON 1999年第2期519卷 323-333页

作者： Sakai, H Lingueglia, E Champigny, G Mattei, MG Lazdunski, M CNRS Inst Pharmacol Mol & Cellulaire UPR 411 F-06560 Valbonne France INSERM U491 F-13385 Marseille France

1. A degenerate polymerase chain reaction (PCR) homology screening procedure was applied to rat brain cDNA in order to identify novel genes belonging to the amiloride-sensitive Na+ channel and degenerin (NC/DEG) family of ion channels. A single gene was identified that encodes a protein related to but clearly different from the already cloned members of the family (18-30% amino acid sequence identity). Phylogenetic analysis linked this protein to the group of ligand-gated channels that includes the mammalian acid-sensing ion channels and the Phe-Met-Arg-Phe-amide (FMRFamide)-activated Na+ channel. 2. Expression of gain-of-function mutants after cRNA injection into Xenopus laevis oocytes or transient transfection of COX cells induced large constitutive currents. The activated channel was amiloride sensitive (IC50, 1.31 mu M) and displayed a low conductance (9-10 pS) and a high selectivity for Na+ over K+ (ratio of the respective permeabilities, PNa+ / Pk+ greater than or equal to 10), all of which are characteristic of NaC/DEG channel behaviour. 3. Northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed a predominant expression of its mRNA in the small intestine, the liver (including hepatocytes) and the brain. This channel has been called the brain-liver-intestine amiloride-sensitive Na+ channel (BLINaC). 4. Corresponding gain-of-function mutations in Caenorhabditis elegans degenerins are responsible for inherited neurodegeneration in the nematode. Besides the BLINaC physiological function that remains to be established, mutations in this novel mammalian degenerin-like channel might be of pathophysiological importance in inherited nenrodegeneration and liver or intestinal pathologies.

关键词：氨基酸序列生物物理学印迹法 RNA COS细胞染色体图克隆分子原位杂交荧光离子通道/生物合成离子通道/遗传学肝/细胞学肝/药物作用肝/代谢小鼠近交BALBC 分子序列数据神经组织蛋白质类/生物合成神经组织蛋白质类/遗传学卵母细胞/代谢 RNA 信使/生物合成 RNA 信使/分离和提纯大鼠 Wistar 逆转录聚合酶链反应钠通道/生物合成钠通道/遗传学组织分布爪蟾属动物小鼠大鼠

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Otx genes in corticogenesis and brain development

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CEREBRAL CORTEX 1999年第6期9卷 533-542页

作者： Acampora, D Barone, P Simeone, A CNR Int Inst Genet & Biophys I-80125 Naples Italy Univ Naples Federico II Dept Neurol Sci I-80131 Naples Italy

A large number of genes have been isolated in the last few years that seem to be involved in the processes of induction, specification and regionalization of the brain. The generation of mouse mutant models for such genes has provided a powerful tool of analysis into their in-vivo properties. Among these genes, Otx1 and Otx2 play a crucial role in several processes of brain morphogenesis. Otx2 is required in the early specification of the neuroectoderm destined to become the fore-midbrain, Otx1 is required mainly for correct corticogenesis, and both genes co-operate in patterning the developing brain through a gene-dosage dependent mechanism. Furthermore, an extensive functional equivalence occurs between the mammalian Otx genes and their Drosophila homolog, orthodenticle. This appears particularly fascinating when considering the huge differences between the brains of mice and flies. This review deals with the major aspects related to the involvement of Otx1 and Otx2 in the development of the mammalian brain and, in particular, of the cerebral cortex.

关键词：脑化学/遗传学大脑皮质/胚胎学基因表达调控发育期同源盒结构域蛋白质类小鼠基因敲除神经组织蛋白质类/遗传学 Otx转录因子类反式激活因子类/遗传学转录因子动物小鼠

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Expression of a novel RNA-splicing factor, RA301/Tra2β, in vascular lesions and its role in smooth muscle cell proliferation

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AMERICAN JOURNAL OF PATHOLOGY 2001年第5期158卷 1685-1694页

作者： Tsukamoto, Y Matsuo, N Ozawa, K Hori, O Higashi, T Nishizaki, J Tohnai, N Nagata, I Kawano, K Yutani, C Hirota, S Kitamura, Y Stern, DM Ogawa, S Natl Cardiovasc Ctr Dept Pathol Suita Osaka 5658565 Japan Natl Cardiovasc Ctr Dept Neurosurg Suita Osaka 5658565 Japan Dainippon Pharmaceut Co Ltd R&D Operat Suita Osaka Japan Kanazawa Univ Sch Med Dept Neuroanat Kanazawa Ishikawa 920 Japan Osaka Rosai Hosp Dept Pathol Sakai Osaka Japan Osaka Univ Grad Sch Med Dept Pathol Suita Osaka Japan Columbia Univ Coll Phys & Surg Dept Physiol & Cellular Biophys New York NY 10032 USA

RA301/Tra2 beta, a sequence-specific RNA-binding protein, was first cloned as a stress molecule in re-oxygenated astrocytes. In human vascular tissues, we have found enhanced RA301/Tra2 beta expression in coronary artery with intimal thickening, and atherosclerotic aorta. Balloon injury to the rat carotid artery induced RA301/Tra2 beta transcripts followed by expression of the antigen, which was detected in medial and neointimal vascular smooth muscle cells (VSMCs). In cultured VSMCs, hypoxia/re-oxygenation caused induction of RA301/Tra2 beta and was accompanied by cell proliferation, both of which were blocked by the addition of either diphenyl iodonium, a NADPH oxidase inhibitor, PD98059, a mitogen-activated protein kinase kinase inhibitor, or antisense oligonucleotide for RA301/Tra2 beta, Consistent with a link between RA301/Tra2 beta and cell proliferation, platelet-derived growth factor also induced expression of RA301/Tra2 beta in cultured VSMCs. These data suggest a possible role for RA301/Tra2 beta in the regulation of VSMC proliferation, especially in the setting of hypoxia/reoxygenation-induced cell stress.

关键词：联苯化合物/药理学颈动脉/代谢颈动脉/病理学细胞分裂/药物作用细胞分裂/生理学 DNA 反义/药理学剂量效应关系药物类黄酮物质/药理学基因表达肌平滑血管/细胞学肌平滑血管/药物作用肌平滑血管/代谢神经组织蛋白质类/遗传学神经组织蛋白质类/生理学鎓类化合物/药理学氧/药理学 RNA剪接 RNA 信使/遗传学 RNA 信使/代谢 RNA结合蛋白质类/遗传学 RNA结合蛋白质类/生理学大鼠 Sprague-Dawley 超氧化物类/代谢青少年成年人老年人动物女(雌)性人类婴儿男(雄)性中年人大鼠

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Interaction of S-SCAM with neural plakophilin-related armadillo-repeat protein/δ-catenin

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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 1999年第3期256卷 456-461页

作者： Ide, N Hata, Y Deguchi, M Hirao, K Yao, I Takai, Y Osaka Univ Sch Med Dept Mol Biol & Biochem Suita Osaka 5650871 Japan JCR Pharmacuet Co Ltd Takai Biotimer Project ERATO Japan Sci & Technol Corp Kobe Hyogo 6512241 Japan

Synaptic scaffolding molecule (S-SCAM) is a multiple PDZ domain-containing protein, which interacts with neuroligin, a cell adhesion molecule, and the NMDA receptor. In this study, we searched for S-SCAM-interacting proteins and obtained a neural plakophilin-related armadillo-repeat protein (NPRAP)/delta-catenin, NPRAP/delta-catenin bound to the last PDZ domain of S-SCAM via its carboxyl-terminus in three different cell-free assay systems, was coimmunoprecipitated with S-SCAM from rat crude synaptosomes, and was localized at the excitatory synapses in rat hippocampal neurons. NPRAP/delta-catenin may be implicated in the molecular organization of synaptic junctions through the interaction with S-SCAM, (C) 1999 Academic Press.

关键词： Armadillo域蛋白质类结合部位生物传感技术 COS细胞载体蛋白质类/化学载体蛋白质类/遗传学载体蛋白质类/代谢基因文库海马/细胞学海马/代谢动力学分子量神经组织蛋白质类/化学神经组织蛋白质类/遗传学神经组织蛋白质类/代谢神经元/代谢肽碎片/化学合成肽碎片/代谢沉淀素试验蛋白质结合受体 N-甲基-D-天冬氨酸/代谢重组融合蛋白质类/生物合成重组融合蛋白质类/遗传学重组融合蛋白质类/代谢突触体/代谢转染酵母菌/遗传学动物大鼠

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Otx2, Gbx2 and Fgf8 interact to position and maintain a mid-hindbrain organizer

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CURRENT OPINION IN CELL BIOLOGY 2000年第6期12卷 736-741页

作者： Joyner, AL Liu, AM Millet, S NYU Sch Med Dept Cell Biol New York NY 10016 USA NYU Sch Med Howard Hughes Med Inst New York NY 10016 USA NYU Sch Med Skirball Inst Biomol Med Dev Genet Program New York NY 10016 USA

A decade ago, chick-quail transplantation studies demonstrated that the junction between the midbrain and hindbrain has the properties of an organizing center capable of patterning the midbrain and cerebellum. Many of the genes that function to pattern these tissues have been identified and extensively studied. Recent experiments have shown that Otx2, Gbx2 and Fgf8 genes play a major role in the positioning and functioning of this organizing center.

关键词：躯体发育模式/遗传学脑/胚胎学脑/代谢小脑/胚胎学小脑/代谢胚胎诱导/遗传学反馈/生理学成纤维细胞生长因子8 成纤维细胞生长因子/遗传学成纤维细胞生长因子/代谢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/代谢中脑/胚胎学中脑/代谢神经组织蛋白质类/遗传学神经组织蛋白质类/代谢组织调控区胚胎/代谢 Otx转录因子类前脑/胚胎学前脑/代谢原癌基因蛋白质类/代谢菱脑/胚胎学菱脑/代谢反式激活因子类/遗传学反式激活因子类/代谢 Wnt蛋白质类斑马鱼蛋白质类动物小鼠

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Mapping and developmental expression analysis of the WD-repeat gene Preb

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GENOMICS 2000年第3期63卷 391-399页

作者： Clelland, CLT Craciun, L Bancroft, C Lufkin, T CUNY Mt Sinai Sch Med Dept Physiol & Biophys New York NY 10029 USA CUNY Mt Sinai Sch Med Brookdale Ctr Dev & Mol Biol New York NY 10029 USA

We have isolated from mouse a novel WD-motif-containing gene designated Preb. This gene encodes a predicted protein of 416 amino acids and has significant homology with other members of the WD-motif gene superfamily that play a role in cell fate determination. Preb maps to the proximal end of chromosome 5 in mouse, near the Hmx1 homeobox gene. Preb is detectable in early stage embryos in the peripheral nervous system, developing liver, and surface ectoderm. Later, Preb is expressed in the anterior portion of Rathke's pouch, which gives rise to the anterior pituitary, the organ responsible for the production of prolactin and other hormones. In midgestation embryos, the most extensive expression of Preb is observed in the perichondrium of the craniofacial, axial, and appendicular skeleton. The expression pattern of Preb in murine embryos suggests a potential role in the specification of multiple cell types, in particular, the fetal skeleton. (C) 2000 Academic Press.

关键词：氨基酸序列碱基序列骨发育/遗传学染色体图交叉遗传 DNA 互补/遗传学 DNA结合蛋白质类/生物合成 DNA结合蛋白质类/遗传学外胚层/代谢胚胎和胎儿发育/遗传学胎儿蛋白质类/生物合成胎儿蛋白质类/遗传学基因表达调控发育期基因鸟嘌呤核苷酸交换因子类原位杂交肝/胚胎学肝/代谢小鼠近交C57BL 分子序列数据鼠科神经组织蛋白质类/生物合成神经组织蛋白质类/遗传学垂体/胚胎学垂体/代谢 RNA 信使/生物合成 RNA 信使/遗传学转录因子/生物合成转录因子/遗传学动物女(雌)性人类男(雄)性小鼠大鼠

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Highwire regulates synaptic growth in Drosophila

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NEURON 2000年第2期26卷 313-329页

作者： Wan, HI DiAntonio, A Fetter, RD Bergstrom, K Strauss, R Goodman, CS Univ Calif Berkeley Dept Mol & Cell Biol Howard Hughes Med Inst Berkeley CA 94720 USA Theodor Boveri Inst Biowissensch LS Genet D-97074 Wurzburg Germany

The formation, stabilization, and growth of synaptic connections are dynamic and highly regulated processes. The glutamatergic neuromuscular junction (NMJ) in Drosophila grows new boutons and branches throughout larval development. A primary walking behavior screen followed by a secondary anatomical screen led to the identification of the highwire (hiw) gene. In hiw mutants, the specificity of motor axon pathfinding and synapse formation appears normal. However, NMJ synapses grow exuberantly and are greatly expanded in both the number of boutons and the extent and length of branches. These synapses appear normal ultrastructurally but have reduced quantal content physiologically. hiw encodes a large protein found at presynaptic terminals. Within presynaptic terminals, HIW is localized to the periactive zone surrounding active zones;Fasciclin II (Fas II), which also controls synaptic growth, is found at the same location.

关键词：秀丽新小杆线虫/遗传学染色体图果蝇属/遗传学果蝇属/生长和发育果蝇属/生理学果蝇蛋白质类生长锥/代谢分子序列数据突变/生理学神经组织蛋白质类/化学神经组织蛋白质类/遗传学神经组织蛋白质类/代谢神经组织蛋白质类/生理学神经肌肉接头/超微结构肽碎片/遗传学肽碎片/生理学突触前末梢/代谢突触前末梢/超微结构序列同源性氨基酸突触/生理学突触/超微结构组织分布转录遗传动物人类

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