Babitch J A, Breithaupt T B, Chiu T C, Garadi R, Helseth D L
Biochim Biophys Acta. 1976 Apr 16;433(1):75-89. doi: 10.1016/0005-2736(76)90179-6.
A method is described for the preparation of synaptosomes and synaptosomal membranes from chicken brain. Procedures for isolating rat synaptosomal membranes could not be used directly; several modifications of existing procedures are reported. Purity of the subcellular and subsynaptosomal fractions was monitored by electron microscopy and measurements of ferrocytochrome c: oxygen oxidoreductase (EC 1.9.3.)), monoamine: oxygen oxidoreductase (deaminating) EC 1.4.3.4), rotenone-insensitive NADH: cytochrome c oxidoreductase (EC 1.6.99.3), NADPH: cytochrome c oxidoreductase (EC 1.6.99.1), orthophosphoric monoester phosphohydrolase (EC 3.1.3.2), ATP phosphohydrolase (EC 3.6.1.4), and levels of RNA. Microsomes are the main contaminant of the synaptosomal membrane fraction. Mitochondrial and lysosomal enzymes occur in lesser amounts. No myelin contamination was observed. Marker enzymes for contaminants suggest that these synaptosomal membranes are as pure as membranes described by others, and the specific activity of a neuronal membrane marker, (Na+ -K+)-activated ATPase, is as high as other preparations. Levels of this enzyme in the membrane fraction are enriched 13-fold over homogenate ATPase levels.
本文描述了一种从鸡脑中制备突触体和突触体膜的方法。不能直接采用分离大鼠突触体膜的方法；本文报道了对现有方法的若干改进。通过电子显微镜以及对亚铁细胞色素c：氧氧化还原酶（EC 1.9.3.）、单胺：氧氧化还原酶（脱氨基）EC 1.4.3.4）、鱼藤酮不敏感的NADH：细胞色素c氧化还原酶（EC 1.6.99.3）、NADPH：细胞色素c氧化还原酶（EC 1.6.99.1）、正磷酸单酯磷酸水解酶（EC 3.1.3.2）、ATP磷酸水解酶（EC 3.6.1.4）的测量以及RNA水平监测亚细胞和亚突触体组分的纯度。微粒体是突触体膜组分的主要污染物。线粒体和溶酶体酶的含量较少。未观察到髓磷脂污染。污染物的标记酶表明，这些突触体膜与其他人描述的膜一样纯净，并且神经元膜标记物（Na + -K +）激活的ATPase的比活性与其他制剂一样高。膜组分中该酶的水平比匀浆ATPase水平富集了13倍。
