Cellular dynamics of functional molecules and second messengers during synaptic transmission

突触传递过程中功能分子和第二信使的细胞动力学

• 批准号: 07508004
• 负责人: MIKOSHIBA Katsuhiko
• 金额: $ 13.82万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07508004/
• 关键词: neurotransmitter; neurotransmission; synaptotagmin; inositol polyphosphate; synapse; sinaptic vesicle; IP_3; IP_3レセプター; カルシウムイオン; 情報伝達; カルシウムチャネル; 小胞体; セカンドメッセンジャー

The inositol high-polyphodphate series (IP4, IP5 and IP6 ; IHPS) inhibit the neurotransmission through binding to the 2nd C2 domain of synaptotagmin (Syt), synaptic vesicle membrane proteins using superior cervical ganglion and squid giant synapse. Now we have revealed several proteins, including alpha adaptins which are specific subunits of clathrin AP2, from mouse brain were eluted by affinity elution chromatography from the C2 domain of Syt II-immobilized Sepharose using 50muM of inositol hexakisphosphate (IP6). The interaction between Syt II and AP2 was inhibited more strongly by IHPS than by the same concentration of IP3. Limited digestion of mouse crude synaptosomal fractions with trypsin revealed different cleavage pattern between in the presenc and the absence of 50muM IP6. These results suggests that IHPS-binding to C2B domain of synaptotagmin alter the state of protein-protein interaction such as synaptotagmin-AP2 interaction, which may induce the inhibition of some events involved in the neurotransmission, e.g. endocytosis.

肌醇高聚磷酸盐系列（IP 4、IP 5和IP 6; IHPS）通过与突触结合蛋白（Syt）的第二个C2结构域、突触囊泡膜蛋白结合抑制上级颈神经节和鱿鱼巨突触的神经传递。现在，我们已经揭示了几种蛋白质，包括网格蛋白AP 2的特异性亚基α衔接素，从小鼠脑中通过亲和洗脱色谱从Syt II固定的Sepharose的C2结构域使用50 μ M的肌醇六磷酸（IP 6）洗脱。IHPS对Syt II和AP 2之间的相互作用的抑制作用强于相同浓度的IP 3。用胰蛋白酶对小鼠粗突触体组分进行有限消化，发现在存在和不存在50 μ M IP 6的情况下存在不同的切割模式。这些结果表明，IHPS与突触结合蛋白C2B结构域的结合改变了蛋白质-蛋白质相互作用的状态，如突触结合蛋白-AP 2相互作用，这可能导致抑制参与神经传递的一些事件，如内吞作用。

项目成果

Inoue, Yoshiro: "Cell death of oligodendrocytes or demyelination induced by overexpression of proteolipid protein depending on expressed gene dosage." Neuroscience Research. 25. 161-172 (1996)

Inoue, Yoshiro：“蛋白脂质蛋白过度表达诱导少突胶质细胞死亡或脱髓鞘，具体取决于表达基因的剂量。”

Matsumoto, Mineo: "Ataxia and epileptic seizures in mice lacking type 1 inositol 1,4,5-trisphoshate receptor." Nature. 379. 168-171 (1996)

Matsumoto, Mineo：“缺乏 1 型肌醇 1,4,5-三磷酸受体的小鼠会出现共济失调和癫痫发作。”

Monkawa,T.,et al.: "Heterotetrameric complex formation of inositol 1,4,5-trisphosphate receptor subunits." J.Biol.Chem.270. 14700-14704 (1995)

Monkawa,T.,et al.：“肌醇 1,4,5-三磷酸受体亚基的异四聚体复合物的形成。”

Fukuda,M.et al.: "Functional diversity of C2 domains of synaptotagmin family." J.Biol.Chem.270(44). 26523-26527 (1995)

Fukuda,M.et al.：“突触结合蛋白家族 C2 结构域的功能多样性。”

Matsumoto, Mineo et al.: "Ataxia and epileptic seizures in mice lacking type 1 inositol 1,4,5-triphosphate receptor." Nature. 379. 168-171 (1996)

Matsumoto、Mineo 等人：“缺乏 1 型肌醇 1,4,5-三磷酸受体的小鼠出现共济失调和癫痫发作。”