Screening of drugs for neurodegenerative disorders using neurotransmitter release machinery expressed in Xenopus oocytes

利用非洲爪蟾卵母细胞表达的神经递质释放机制筛选治疗神经退行性疾病的药物

基本信息

批准号：
05557010
负责人：
SHUNTOH Hisato
金额：
$ 3.58万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

1. Complementary DNA clones encoding neural visinin-like calcium-binding proteins (NVP-1, NVP-2 and NVP-3) were isolated from rat brain cDNA library. NVP-1, NVP-2 and NVP-3 mRNAs are most highly expressed in brain and are differentially expressed in various regions of rat brain. Furthermore, alterations in NVP-1 may be involvedin the pathology of Phecyclidine-induced psychosis. 2. The effect of phorbol ester on the Na+-dependent uptake of GABA by rat cortical synaptosomes and Xenopus oocytes expressing GAT1 (neuronal GABA transporter) was examined. Protein kinase C may modulate the GABA uptake into presynaptic terminals through the inhibition of GAT1 activity. 3. The serotonin release from RBL-2H3 cells is subject to tonic, Ca2+-independent, inhibition by protein phosphatase 1.4. In the Schaffer collateral-CA1 pathway in rat hippocampal slices, calyculin A induced a long-lasting potentiation of the extracellularfield potentials with a transient increase in glutamate release. 5. Heat shock transiently suppresses the inositol trisphosphate-mediated intracellular calcium signal transduction system and that HSP72 is involved in the recovery of receptor-stimulated calcium rise in the neuronal cells. 6. By screening the mutagen-treated fission yeast cells, immunosuppressant- and temperature- sensitive (its1-6) mutant strains were isolated. Gene products of its1+ and its3+ were beta-adaptin homolog, which is a subunit of adapter protein AP-2, and Pl (4) P5-kinase homolog, which is required for exocytosis, respectively. These results suggest that calcineurin plays an important role in vesicular recycling. In conclusion, our several results provide important clues about the molecular basis of neurotransmitter release and secretory vesicle recycling or endocytosis.

1。从大鼠脑cDNA文库中分离出编码神经粘蛋白钙结合蛋白（NVP-1，NVP-2和NVP-3）的互补DNA克隆（NVP-1，NVP-2和NVP-3）。 NVP-1，NVP-2和NVP-3 mRNA在大脑中最高表达，并且在大鼠脑的各个区域差异表达。此外，NVP-1的变化可能涉及磷酸肽诱导的精神病的病理。 2。研究了大鼠皮质突触体和表达GAT1（神经元GABA转运蛋白）的大鼠皮质突触体和爪蟾卵母细胞对Na+依赖性摄取的影响。蛋白激酶C可以通过抑制GAT1活性来调节GABA摄取到突触前末端。 3。从RBL-2H3细胞中释放的5-羟色胺释放受蛋白质磷酸酶1.4的滋补，Ca2+非依赖性的抑制作用。在大鼠海马切片中的Schaffer侧支CA1途径中，钙化蛋白A诱导了细胞外场电位的持久增强，并且谷氨酸释放的短暂增加。 5。热休克瞬时抑制肌醇三磷酸介导的细胞内钙信号转导系统，并且HSP72参与了神经元细胞中受体刺激的钙升高的恢复。 6。通过筛选诱变的裂变酵母细胞，免疫抑制剂和温度敏感（ITS1-6）突变体菌株。 ITS1+和ITS3+的基因产物是β-ADAPTIN同源物，它是衔接蛋白AP-2的亚基和PL（4）P5-激酶同源物，分别是胞吐作用所必需的。这些结果表明钙调神经蛋白在囊泡回收中起重要作用。总之，我们的几个结果提供了有关神经递质释放和分泌囊泡回收或内吞作用的分子基础的重要线索。