Webster J C, Klingman J D
Neurochem Res. 1979 Apr;4(2):137-53. doi: 10.1007/BF00964140.
Synaptosomal plasma membrane (SPM) and other subcellular fractions were isolated from the forebrain of 1-day-old chickens by a procedure based on that of Davis and Bloom (16) and Cotman and Taylor (13). The procedure involves the centrifugation through a discontinuous sucrose gradient of a crude synaptosomal-mitochondrial fraction which has been lysed and weighed with iodonitrotetrazolium. SPM isolated by this method contains only small amounts of lysosomal or mitochondrial membranes and is practically devoid of contaminating microsomal membranes, as estimated by enzyme marker assays. The purity of chick-brain SPM prepared by this method is compared to the purity of chick-brain fractions obtained by two other laboratories, using different methods (4, 59). The SPM were extracted with Triton X-100 and all fractions solubilized in sodium dodecyl sulfate (sds). The delipidated proteins of all fractions were subjected to SDS-polyacrylamide electrophoresis on slab gels and stained for protein. A distinct difference was observed between the patterns given by the Triton-soluble and -insoluble fractions. Electron microscopy of the synaptic junction fraction showed numerous junctional complexes.
通过基于戴维斯和布卢姆（16）以及科特曼和泰勒（13）的方法，从1日龄鸡的前脑中分离出突触体细胞膜（SPM）和其他亚细胞组分。该方法包括将已裂解并用碘硝基四氮唑称重的粗突触体 - 线粒体组分通过不连续蔗糖梯度离心。通过这种方法分离的SPM仅含有少量溶酶体或线粒体膜，并且根据酶标记测定估计几乎没有污染的微粒体膜。使用不同方法（4,59）将通过该方法制备的鸡脑SPM的纯度与另外两个实验室获得的鸡脑组分的纯度进行比较。用Triton X - 100提取SPM，并将所有组分溶解在十二烷基硫酸钠（SDS）中。所有组分的脱脂蛋白在平板凝胶上进行SDS - 聚丙烯酰胺电泳并进行蛋白质染色。在Triton可溶性和不溶性组分给出的模式之间观察到明显差异。突触连接组分的电子显微镜显示出许多连接复合体。
