Live-cell imaging of receptors around postsynaptic membranes
Live-cell imaging of receptors around postsynaptic membranes
复制标题
突触后膜周围受体的活细胞成像
作者：
Hiromitsu Tanaka;Shumpei Fujii;T. Hirano
该方案描述了如何成像谷氨酸受体围绕在黏附蛋白涂层玻璃表面上形成的兴奋性突触后膜的运输。该方案是为了阐明受体在突触可塑性诱导过程中的运动方式。将分离的神经元培养在包被神经瑞新的玻片上，诱导玻璃表面形成突触后膜样结构。然后将标记有荧光蛋白的谷氨酸受体导入神经元，并在全内反射荧光显微镜下观察。整个过程需要大约3周的时间。神经元活动引起的细胞表面受体数量的变化可以被量化，并且可以清楚地观察到假突触后膜周围受体的个别胞吐事件。该方法可用于研究膜蛋白在细胞膜的其他特殊区域的运动，如抑制性突触后膜、突触前膜或免疫突触。
This protocol describes how to image the trafficking of glutamate receptors around excitatory postsynaptic membrane formed on an adhesion protein–coated glass surface. The protocol was developed to clarify how receptors move during the induction of synaptic plasticity. Dissociated neurons are cultured on a coverslip coated with neurexin, which induces the formation of postsynaptic membrane-like structures on the glass surface. A glutamate receptor tagged with a fluorescent protein is then transfected into neurons, and it is observed with total internal reflection fluorescence microscopy. The whole process takes about 3 weeks. Changes in the amount of cell-surface receptors caused by neuronal activities can be quantified, and individual exocytosis events of receptors can be clearly observed around the pseudo-postsynaptic membrane. This protocol has potential applications for studies of movements of membrane proteins around other specialized regions of the cell membrane, such as the inhibitory postsynaptic membrane, the presynaptic membrane or the immunological synapses.
影响因子：
56.9
作者：
Shi, SH;Hayashi, Y;Malinow, R
通讯作者：
Malinow, R