Ultrastructural Research of Platelet Raft

血小板筏的超微结构研究

• 批准号: 17591022
• 负责人: SUZUKI Hidenori
• 金额: $ 2.24万
• 依托单位: Tokyo Metropolitan Organization for Medical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591022/
• 关键词: Platelet; Raft; Glycoprotein; GPVI; Cytoskeleton; Electron microscopy; シグナル分子

Previous studies have shown critical and functional role of membrane rafts in initiating signaling through platelet activating collagen receptor, the GPVI/FcRγ complex upon collagen stimulation. However, little is known about how GPVI-anchoring membrane rafts are regulated upon collagen stimulation. Since platelet cytoskeleton has been known to be involved in localization and accumulation of signaling molecules to critical locations within activated platelets, we examined effects of cytochalasin D (CytD) on morphological changes of membrane rafts in collagen-stimulated platelets. Washed platelets were pretreated with or without 100μM CytD for 1 hr at room temperature followed by addition of 1 mM RGDS and 1 mM CaCl_2 before stimulation and then stimulated with 2 mg/ml collagen for 30 sec at 37℃ under stirring conditions. After the reaction was stopped, membrane rafts were isolated as Brij98 detergent-insoluble membranes by a modification of previously described method to ensure more acc … More urate isolation of physiological raft structure. Immunogold Electron microscopic analyses of Brij98-insoluble rafts were performed and revealed vesicle structure of individual raft using negative-staining.When the diameters of approximately 200 vesicles of rafts in each sample were measured, those after collagen stimulation were significantly (p<0.01) enlarged than those before stimulation (160.97±65.38 vs 105.91±34.94 nm). Immunogold-stained particles of GPVI were also significantly (p<0.01) increased in rafts isolated after collagen stimulation than in those before stimulation (4.0±3.8 vs 1.6±2.1). Pretreatment of platelets with CytD before collagen stimulation almost completely inhibited all those changes observed in GPVI-anchoring membrane rafts after stimulation. Thus, present study indicates that GPVI-anchoring membrane rafts are enlarged possibly by fusing each other in cytoskeleton-dependent manner after collagen stimulation, thereby resulting in accumulation of GPVI in individual membrane raft. Less

先前的研究已经显示膜筏在通过血小板活化胶原蛋白受体（胶原蛋白刺激后的GPVI/FcRγ复合物）启动信号传导中的关键和功能性作用。然而，鲜为人知的是如何GPVI锚定膜筏调节胶原蛋白刺激。由于血小板细胞骨架已被称为参与本地化和积累的信号分子的关键位置内活化的血小板，我们研究了细胞松弛素D（CytD）对胶原蛋白刺激的血小板膜筏的形态变化的影响。洗涤后的血小板在室温下用或不用100μM CytD预处理1小时，然后在刺激前加入1 mM RGDS和1 mM CaCl_2，然后在37℃搅拌条件下用2 mg/ml胶原刺激30秒。在反应停止后，通过对先前描述的方法进行修改以确保更高的准确性，将膜筏分离为Brij 98洗涤剂不溶性膜。 ...更多信息 对生理筏结构进行合理隔离。对Brij 98不溶性筏进行免疫金电子显微镜分析，采用负染法揭示了单个筏的囊泡结构，测量每个样本中约200个筏囊泡的直径时，胶原刺激后的直径比刺激前明显增大（p<0.01）（160.97±65.38 vs 105.91±34.94 nm）。在胶原刺激后分离的筏中，免疫金染色的GPVI颗粒也显著增加（p<0.01）（4.0±3.8 vs 1.6±2.1）。在胶原刺激前用CytD预处理血小板几乎完全抑制了刺激后在GPVI锚定膜筏中观察到的所有这些变化。因此，目前的研究表明，GPVI锚定膜筏扩大可能是通过融合后，在胶原蛋白刺激后，在细胞因子依赖的方式，从而导致GPVI在单个膜筏的积累。少

项目成果

Separation of a cholesterol-enriched microdomain involved in T-cell signal transduction

• DOI: 10.1111/j.1742-4658.2005.04938.x
• 发表时间: 2005-11-01
• 期刊: FEBS JOURNAL
• 影响因子: 5.4
• 作者: Shimada, Y;Inomata, M;Ohno-Iwashita, Y
• 通讯作者: Ohno-Iwashita, Y

図説 血栓・止血・血管学 血栓症制圧のために(担当:血小板の形態)

血栓、止血、血管学图解：控制血栓（负责：血小板形态）

• 发表时间: 2005
• 作者: Shimada Y;Inomata M;Suzuki H;Hayashi M;Waheed AA;Ohno-Iwashita Y;鈴木英紀;鈴木 英紀(分担執筆);鈴木英紀(分担執筆)
• 通讯作者: 鈴木英紀(分担執筆)

別冊・医学のあゆみ 血液疾患-state of arts Ver.3(担当:先天性血小板機能異常症/storage pool病)

单册/病史血液疾病-state of arts Ver.3（负责人：先天性血小板功能障碍/储存池疾病）

• 发表时间: 2005
• 作者: Shimada Y;Inomata M;Suzuki H;Hayashi M;Waheed AA;Ohno-Iwashita Y;鈴木英紀;鈴木 英紀(分担執筆);鈴木英紀(分担執筆);鈴木英紀(分担執筆)
• 通讯作者: 鈴木英紀(分担執筆)

New strategy of platelet substitutes for enhancing platelet aggregation at high shear rates : cooperative effects of a mixed system of fibrinogen γ-chain dodecapeptide-or glycoprotein Ibα-conjugated latex beads under flow conditions.

血小板替代品在高剪切速率下增强血小板聚集的新策略：纤维蛋白原γ链十二肽或糖蛋白Ibα缀合乳胶珠混合系统在流动条件下的协同作用。

• 发表时间: 2006
• 期刊: J Artif Organs 9
• 作者: Okamura Y;Handa M;Suzuki H;Ikeda Y;Takeoka S
• 通讯作者: Takeoka S

血小板活性化とシグナル伝達

血小板激活和信号转导

• 发表时间: 2005
• 期刊: 顕微鏡 40
• 作者: Higashi T;Tsukada J;Yoshida Y;Mizobe T;Mouri F;Minami Y;Tanaka Y;鈴木 英紀
• 通讯作者: 鈴木 英紀