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首页> 外文期刊>Molecular biology of the cell >Visualizing lipid raft dynamics and early signaling events during antigen receptor-mediated B-lymphocyte activation
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Visualizing lipid raft dynamics and early signaling events during antigen receptor-mediated B-lymphocyte activation

机译：在抗原受体介导的B淋巴细胞激活过程中可视化脂质筏动力学和早期信号事件

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Recent biochemical evidence indicates that an early event in signal transduction by the B-cell antigen receptor (BCR) is its translocation to specialized membrane subdomains known as lipid rafts. We have taken a microscopic approach to image lipid rafts and early events associated with BCR signal transduction. Lipid rafts were visualized on primary splenic B lymphocytes from wild-type or anti-hen egg lysozyme BCR transgenic mice, and on a mature mouse B-cell line Bal 17 by using fluorescent conjugates of cholera toxin B subunit or a Lyn-based chimeric protein, which targets green fluorescent protein to the lipid raft compartment. Time-lapse imaging of B cells stimulated via the BCR with the antigen hen egg lysozyme, or surrogate for antigen anti-IgM, demonstrated that lipid rafts are highly dynamic entities, which move laterally on the surface of these cells and coalesce into large regions. These regions of aggregated lipid rafts colocalized with the BCR and tyrosine-phosphorylated proteins. Microscopic imaging of live B cells also revealed an inducible colocalization of lipid rafts with the tyrosine kinase Syk and the receptor tyrosine phosphatase CD45. These two proteins play indispensable roles in BCR-mediated signaling but are not detectable in biochemically purified lipid raft fractions. Strikingly, BCR stimulation also induced the formation of long, thread-like filopodial projections, similar to previously described structures called cytonemes. These B-cell cytonemes are rich in lipid rafts and actin filaments, suggesting that they might play a role in long-range communication and/or transportation of signaling molecules during an immune response. These results provide a window into the morphological and molecular organization of the B-cell membrane during the early phase of BCR signaling. [References: 45]
机译：最近的生化证据表明，B细胞抗原受体（BCR）进行信号转导的早期事件是其易位到称为脂筏的特定膜亚结构域。我们采用了显微镜方法来对脂筏和与BCR信号转导相关的早期事件进行成像。通过使用霍乱毒素B亚基或基于Lyn的嵌合蛋白的荧光偶联物，可以在野生型或抗鸡蛋溶菌酶BCR转基因小鼠的原代脾脏B淋巴细胞和成熟的小鼠B细胞系Bal 17上看到脂质筏。 ，将绿色荧光蛋白靶向脂质筏区室。通过BCR刺激的B细胞的时移成像用鸡蛋鸡蛋溶菌酶或抗原抗IgM的替代物，证明脂质筏是高度动态的实体，它们在这些细胞的表面横向移动并聚结成大区域。聚集的脂质筏的这些区域与BCR和酪氨酸磷酸化蛋白共定位。活B细胞的显微成像还显示脂质筏与酪氨酸激酶Syk和受体酪氨酸磷酸酶CD45的可诱导共定位。这两种蛋白在BCR介导的信号传导中起着不可或缺的作用，但在生物化学纯化的脂质筏部分中却无法检测到。引人注目的是，BCR刺激还诱导了长的线状丝状突起的形成，类似于先前描述的称为细胞因子的结构。这些B细胞细胞因子富含脂质筏和肌动蛋白丝，表明它们可能在免疫应答过程中在信号分子的长距离通讯和/或转运中发挥作用。这些结果为了解BCR信号传导早期B细胞膜的形态和分子组织提供了一个窗口。 [参考：45]

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