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首页> 美国卫生研究院文献>other >Real-time imaging of plasma membrane deformations reveals pre-fusion membrane curvature changes and a role for dynamin in the regulation of fusion pore expansion
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Real-time imaging of plasma membrane deformations reveals pre-fusion membrane curvature changes and a role for dynamin in the regulation of fusion pore expansion

机译：血浆膜变形的实时成像显示出融合膜曲率变化和动力学在融合孔膨胀调节中的作用

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摘要

Assays for real-time investigation of exocytosis typically measure what is released from the granule. From this inferences are made about the dynamics of membrane remodeling as fusion progresses from start to finish. We have recently undertaken a different approach to investigate the fusion process, by focusing not primarily on the granule, but rather its partner in exocytosis – the plasma membrane. We have been guided by the idea that biochemical interactions between the granule and plasma membranes before and during fusion, cause changes in membrane conformation. To enable study of membrane conformation, a novel imaging technique was developed combining polarized excitation of an oriented membrane probe (diI) with Total Internal Reflection Fluorescence Microscopy (pTIRFM). Because this technique measures changes in membrane conformation (or deformations) directly, its usefulness persists even after granule cargo reporter (catecholamine, or protein), is no longer present. In this mini-review, we first summarize the workings of pTIRFM. We then discuss the application of the technique to investigate deformations in the membrane preceding fusion, and later, during fusion pore expansion. Finally, we discuss how expansion of the fusion pore may be regulated by the GTPase activity of dynamin.
机译：用于实时调查的胞尿量的测定通常测量颗粒中释放的内容。从这种推断下，由于融合从开始完成而进行了膜重塑的动态。我们最近通过主要注重颗粒，而是其在卵尿量的伴侣 - 血浆膜上，我们最近进行了一种不同的方法来调查融合过程。我们一直在指导的想法，即融合前和期间颗粒和血浆膜之间的生化相互作用，导致膜构象的变化。为了能够实现膜构象的研究，开发了一种新的成像技术，与总内反射荧光显微镜（PTRIRFM）相结合的偏振激励。因为这种技术措施直接测量膜构象（或变形）的变化，即使在颗粒货物报告机（儿茶酚胺或蛋白质）之后，其有用性仍不再存在。在这个迷你审查中，我们首先总结了PTIRFM的工作。然后，我们讨论该技术的应用来研究熔融孔隙前后膜的膜中的变形。最后，我们讨论如何通过动力学的GTP酶活性来调节融合孔的膨胀。

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