THE PREPARATION OF CHOLINERGIC SYNAPTOSOMES FROM BOVINE SUPERIOR CERVICAL GANGLIA
THE PREPARATION OF CHOLINERGIC SYNAPTOSOMES FROM BOVINE SUPERIOR CERVICAL GANGLIA
复制标题
牛颈上神经节胆碱能突触体的制备
作者：
W. S. Wilson;J. Cooper
本文介绍了从牛颈交感神经节制备较纯的胆碱能突触体的方法。通过与胶原酶孵育分散组织后，差速离心产生粗突触体部分（P2），通过在蔗糖或Ficoll-蔗糖密度梯度上离心进一步纯化。乙酰胆碱和胆碱乙酰转移酶的测定，和电子显微镜证实，突触前神经末梢存活的孵育过程中，出现在P2馏分，此后沉降到约密度。1.13在蔗糖中为1.06，在聚蔗糖中为1.06。洗涤的P2级分具有2.3nmol/mg蛋白质的乙酰胆碱含量，其比来自脑的相应级分高至少三倍。
—A method is described for the preparation of relatively pure cholinergic synaptosomes from bovine cervical sympathetic ganglia. After dispersion of the tissue by incubation with collagenase, differential centrifugation yielded a crude synaptosomal fraction (P2), which was further purified by centrifugation on sucrose or Ficoll‐sucrose density gradients. Assay of acetylcholine and choline acetyltransferase, and electron microscopy confirmed that presynaptic nerve endings survived the incubation procedure, appeared in the P2 fraction and thereafter sedimented to a density of approx. 1.13 in sucrose and 1.06 in Ficoll‐sucrose. The washed P2 fraction had an acetylcholine content of 2.3 nmol/mg of protein, which is at least three times higher than the corresponding fraction from brain.