Docherty M, Bradford H F, Cash C D, Ehret M, Maitre M, Joh T H
Department of Biochemistry, Imperial College of Science, Technology and Medicine, London, England.
J Neurochem. 1991 May;56(5):1569-80. doi: 10.1111/j.1471-4159.1991.tb02053.x.
Monoaminergic synaptosomes have been isolated and purified from rat brain by immunomagnetophoresis. This novel technique uses magnetic beads to which Protein A is bound. Noradrenergic, dopaminergic, and serotonergic synaptosomes (previously cell-surface labelled with anti-dopamine-beta-hydroxylase, anti-tyrosine hydroxylase, and anti-tryptophan hydroxylase, respectively) may be isolated in a highly purified state. The synaptosomal subpopulations are recovered in a viable metabolic state and show glucose-stimulated respiration and Ca2(+)-dependent neurotransmitter release. A novel subtype of dopamine-beta-hydroxylase was found in dopaminergic terminals. No evidence for glutamate corelease from monoaminergic synaptosomes was obtained.
已通过免疫磁泳从大鼠脑中分离并纯化了单胺能突触体。这项新技术使用结合了蛋白A的磁珠。去甲肾上腺素能、多巴胺能和5-羟色胺能突触体(之前分别用抗多巴胺-β-羟化酶、抗酪氨酸羟化酶和抗色氨酸羟化酶进行细胞表面标记)可以高度纯化的状态分离出来。突触体亚群以存活的代谢状态回收,并显示出葡萄糖刺激的呼吸作用和Ca2(+)依赖性神经递质释放。在多巴胺能终末发现了一种新型的多巴胺-β-羟化酶亚型。未获得单胺能突触体共释放谷氨酸的证据。