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以胶体金为探针,对大脑皮质树突棘中肌动蛋白进行免疫细胞化学定位。

Immunocytochemical localization of actin in dendritic spines of the cerebral cortex using colloidal gold as a probe.

作者信息

Cohen R S, Chung S K, Pfaff D W

出版信息

Cell Mol Neurobiol. 1985 Sep;5(3):271-84. doi: 10.1007/BF00711012.

Abstract

Immunocytochemical localization of actin in rat cerebral cortex embedded in the resin LR White was performed using 5 nm colloidal gold as a probe. Antigenicity is maintained throughout the embedding procedure and the low electron opacity of LR White permits fine filamentous structures to be visualized. Control experiments included incubating the sections with normal goat serum or mouse IgG instead of the primary antibody, preadsorbing the antibody with actin from bovine muscle or liver acetone powder, and heat treating the primary antibody. Immunoreactive actin was identified primarily in dendritic spines, particularly in the postsynaptic density (PSD), the subsynaptic web, and the spine apparatus and endothelial and smooth muscle cells of blood vessels. Within dendritic spines, actin which is labeled in the PSD is in continuity with the filaments of the subsynaptic web. These filaments, in turn, are in continuity with the spine apparatus and/or the spine membranes adjacent to the PSD. The PSD may therefore function like other submembranous filamentous arrays which communicate events occurring at the membrane, in this case, the postsynaptic membrane, to the underlying cytoskeletal network, i.e., the subsynaptic web of the spine. It is also suggested that the actin present in the spine may play a role in changes in spine shape and synaptic curvature. Some actin was also seen in the presynaptic process in association with synaptic vesicles, the filamentous network that is contiguous with the synaptic vesicle membrane, and the presynaptic dense projections. Actin may be involved in dynamic processes in the presynaptic ending which include vesicle translocation.

摘要

利用5纳米胶体金作为探针,对包埋于LR White树脂中的大鼠大脑皮层肌动蛋白进行免疫细胞化学定位。在整个包埋过程中抗原性得以保持,且LR White的低电子不透明度使细丝状结构得以可视化。对照实验包括用正常山羊血清或小鼠IgG代替一抗孵育切片、用来自牛肌肉或肝脏丙酮粉的肌动蛋白预吸附抗体以及对一抗进行热处理。免疫反应性肌动蛋白主要在树突棘中被识别,特别是在突触后致密部(PSD)、突触下网、棘器以及血管的内皮细胞和平滑肌细胞中。在树突棘内,PSD中被标记的肌动蛋白与突触下网的细丝相连。这些细丝又与棘器和/或与PSD相邻的棘膜相连。因此,PSD可能像其他膜下丝状阵列一样发挥作用,将膜上发生的事件,在这种情况下是突触后膜上的事件,传递给潜在的细胞骨架网络,即棘的突触下网。还表明,棘中存在的肌动蛋白可能在棘形状和突触曲率的变化中起作用。在突触前过程中也观察到一些肌动蛋白与突触小泡、与突触小泡膜相邻的丝状网络以及突触前致密突起有关。肌动蛋白可能参与突触前末梢的动态过程,包括小泡转运。

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