Poduslo S E, Chechik T, Filbin M T, Rosenfeld J
J Neurosci Res. 1986;15(4):553-67. doi: 10.1002/jnr.490150412.
The properties of neuronal plasma membranes are highly specialized, since they include the receptors by which many hormones, neurotransmitters, drugs, and toxins initiate their actions, as well as the components necessary for receiving synaptic input from a variety of sources and for generating the appropriate pattern of action potentials. To be able to study plasma membranes of neurons, it is necessary to separate sufficient quantities of neurons from the other cell types, processes, and myelin found in brain. The methodology for obtaining neurons by bulk-isolation from 10-day-old rat brain has been established, and techniques for purifying the plasma membranes are presented here. The procedure involves swelling the cells in a buffered solution, homogenizing the cells vigorously, and separating the membrane fragments on discontinuous sucrose gradients. The plasma membrane fraction is enriched in plasma membrane marker enzymes and has low activity for contaminating enzymes for subcellular organelles. Electron micrographs show that the fraction consists of membrane vesicles and profiles of various sizes. The protein composition reveals over 60 proteins with molecular weights as high as 100,000 to a low of 14,000. This plasma membrane fraction now can be the focus for studying receptors, glycoproteins, and cell-specific markers of neurons in health and disease.
神经元质膜的特性高度专业化，因为它们包含许多激素、神经递质、药物和毒素借以启动其作用的受体，以及接收来自各种来源的突触输入并产生适当动作电位模式所需的成分。为了能够研究神经元的质膜，有必要从大脑中发现的其他细胞类型、突起和髓磷脂中分离出足够数量的神经元。从10日龄大鼠大脑中批量分离获得神经元的方法已经建立，本文介绍了纯化质膜的技术。该过程包括在缓冲溶液中使细胞膨胀，剧烈匀浆细胞，并在不连续蔗糖梯度上分离膜片段。质膜部分富含质膜标记酶，而污染亚细胞器的酶活性较低。电子显微镜照片显示，该部分由各种大小的膜泡和膜轮廓组成。蛋白质组成显示有60多种蛋白质，分子量高达100,000至低至14,000。现在，这种质膜部分可以成为研究健康和疾病状态下神经元的受体、糖蛋白和细胞特异性标志物的重点。
