Balázs R, Hajós F, Johnson A L, Reynierse G L, Tapia R, Wilkin G P
Brain Res. 1975 Mar 14;86(1):17-30. doi: 10.1016/0006-8993(75)90634-4.
By a combination of differential and sucrose density gradient (both discontinuous and linear) centrifugation, large fragments of the cerebellar glomeruli were isolated in high purity from hand homogenised tissue. The final preparation contained only about 1% of the tissue protein, but over 90% of its volume was accounted for by the glomerulus particles. The ultrastructure of the glomerulus particles was well preserved. The enzyme profile was characteristic: the glomerulus particles were enriched in glutamate decarboxylase (GAD) activity (relative specific activity (RSA), 2.54), but the RSA of choline acetyltransferase (ChAc) was only 1.05. These findings are consistent with the view that GAD activity is very high in the inhibitory Golgi terminals, which occupy only a small fraction of the total volume of the particles, and acetylcholine may be a transmitter only in a relatively small fraction of the mossy fibre terminals. The glomerulus particles also contained a high concentration of succinate dehydrogenase (SDH) activity (RSA, 1.91), whereas the RSA of glutamate dehydrogenase (GDH) was only 1.15. The great asset of this preparation for future investigations is that it is composed almost exclusively from pre- and postsynaptic neuronal structures. Fractions containing neuropil fragments of non-glomerular origin were also obtained, but the profile of the estimated enzymes did not indicate unique characteristics.
通过差速离心和蔗糖密度梯度离心（不连续和线性）相结合的方法，从小脑手工匀浆组织中高纯度分离出小脑小球的大片段。最终制备物仅包含约1%的组织蛋白，但其体积的90%以上由小球颗粒占据。小球颗粒的超微结构保存良好。酶谱具有特征性：小球颗粒富含谷氨酸脱羧酶（GAD）活性（相对比活性（RSA），2.54），但胆碱乙酰转移酶（ChAc）的RSA仅为1.05。这些发现与以下观点一致，即抑制性高尔基终末中的GAD活性非常高，而其仅占颗粒总体积的一小部分，并且乙酰胆碱可能仅在相对较小部分的苔藓纤维终末中作为神经递质。小球颗粒还含有高浓度的琥珀酸脱氢酶（SDH）活性（RSA，1.91），而谷氨酸脱氢酶（GDH）的RSA仅为1.15。该制备物对未来研究的最大优点是它几乎完全由突触前和突触后神经元结构组成。还获得了含有非小球来源神经毡片段的组分，但估计酶的谱图未显示独特特征。
