ISOLATION OF SYNAPTIC PLASMA-MEMBRANE FROM BRAIN BY COMBINED FLOTATION-SEDIMENTATION DENSITY GRADIENT CENTRIFUGATION
ISOLATION OF SYNAPTIC PLASMA-MEMBRANE FROM BRAIN BY COMBINED FLOTATION-SEDIMENTATION DENSITY GRADIENT CENTRIFUGATION
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DOI：
10.1016/0005-2736(74)90268-5
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发表时间：
1974-01-01
期刊：
影响因子：
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通讯作者：
MATUS, AI
中科院分区：
文献类型：
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作者：
JONES, DH;MATUS, AI
描述了一种用于脑的亚细胞分级分离以获得高度富集突触质膜的制剂的方法。从两步蔗糖密度梯度的界面回收富集的级分，在所述界面上通过同时沉降和浮选离心分离来自脑的低渗裂解的粗线粒体级分。与神经元质膜相关的酶标记物活性在含有突触质膜的部分中富集，而与主要可能的污染物髓鞘和线粒体相关的酶标记物的不到10%在同一部分中被发现。富集部分的形态学检查表明，约80%的配置文件是可识别的突触起源。与先前描述的用于获得等效纯度的突触血浆富集级分的方法相比，这里报道的程序更简单、更短并且容量更大。
A method is described for the subcellular fractionation of brain to obtain a preparation highly enriched in synaptic plasma membranes. The enriched fraction is recovered from the interface of a two-step sucrose density gradient on which a hypotonically lysed crude mitochondrial fraction from brain has been separated by simultaneous sedimentation and flotation centrifugation. Enzyme marker activities associated with the neuronal plasma membrane are enriched in the synaptic plasma membrane-containing fraction while less than 10% of enzyme markers associated with the major probable contaminants, myelin and mitochondria, are found in the same fraction. Morphological examination of the enriched fraction suggests that about 80% of the profiles are recognisably synaptic in origin. Compared to previously described methods for obtaining synaptic plasma-enriched fractions of equivalent purity, the procedure reported here is simpler, shorter, and of greater capacity.