Sarkar P K, Ray A K
Department of Animal Physiology, Bose Institute, Calcutta, India.
Methods Find Exp Clin Pharmacol. 1992 Sep;14(7):493-7.
A simple and rapid low speed density gradient centrifugation method has been described to isolate relatively pure synaptosomes and non-synaptic mitochondria from rat brain cerebral cortex. The purity of the fractions isolated were tested biochemically by considering some marker enzymes, i.e., lactate dehydrogenase, plasma membrane Na+K+ATPase and acetylcholinesterase for the synaptosomes and succinate dehydrogenase for the mitochondrial fraction. The adrenergic receptor properties in the synaptosomal membrane were evaluated by observing the effect of different beta- and alpha 2-adrenergic receptor agonists and antagonists as function of synaptic plasma membrane Na+K+ATPase activity. Isoproterenol (beta-agonist) and yohimbine (alpha 2-antagonist)-induced changes in the synaptosomal Na+K+ATPase activity were counteracted by beta-antagonist propranolol and alpha 2-agonist clonidine, respectively. In the non-synaptic mitochondria the corresponding effects were insignificant. The study illustrates an easy, rapid and low-speed preparatory method to obtain synaptosomal and non-synaptic mitochondrial fractions of high purity.
已描述了一种简单快速的低速密度梯度离心法，用于从大鼠脑皮层中分离出相对纯净的突触体和非突触线粒体。通过检测一些标记酶，即乳酸脱氢酶、质膜钠钾ATP酶和乙酰胆碱酯酶（用于突触体）以及琥珀酸脱氢酶（用于线粒体部分），对分离出的各部分纯度进行了生化检测。通过观察不同的β-和α2-肾上腺素能受体激动剂和拮抗剂对突触体膜钠钾ATP酶活性的影响，评估了突触体膜中的肾上腺素能受体特性。异丙肾上腺素（β-激动剂）和育亨宾（α2-拮抗剂）诱导的突触体钠钾ATP酶活性变化分别被β-拮抗剂普萘洛尔和α2-激动剂可乐定抵消。在非突触线粒体中，相应的影响不显著。该研究说明了一种简便、快速且低速的制备方法，可获得高纯度的突触体和非突触线粒体部分。
