Ariano M A
Neuroscience. 1983 Nov;10(3):707-23. doi: 10.1016/0306-4522(83)90212-9.
The cellular distribution of guanylate cyclase (EC 4.6.1.2), guanosine 3',5'-phosphate (cyclic GMP), cyclic GMP-dependent protein kinase (EC 2.7.1.38), and cyclic GMP phosphodiesterase (EC 3.1.4.17) have been examined in the rostral rat caudate-putamen complex. Immunofluorescent staining for guanylate cyclase, cyclic GMP, and cyclic GMP-dependent protein kinase in fresh frozen caudate-putamen tissues is analogous to the immunoperoxidase localization in perfusion-fixed striatal slices. Homologous immunoreactivity in the cytoplasm and processes of ovoid and rounded neurons, 15-20 microns in diameter can be seen for these three components of the cyclic GMP system. Immunoreactive neurons are uniformly distributed throughout the caudate-putamen complex of all experimental tissue examined. Occasional large neurons, greater than 25 microns in diameter, in the ventral region of the striatum show immunoreactivity. Enzyme histochemical determination of the activities of guanylate cyclase and cyclic GMP phosphodiesterase show the medium-sized neuronal population (15-20 microns) contain hydrolytic activity for these proteins. Large- to medium-sized capillaries demonstrate guanylate cyclase synthetic activity, but the endothelial cells do not exhibit immunohistochemical staining. This suggests that physiological activity of an enzyme cannot be completely discerned through application of immunohistochemical procedures. Additionally, enzymatically detected guanylate cyclase histochemical activity was not uniformly distributed throughout the striatal neuropil. Enzyme histochemical detection of cyclic GMP phosphodiesterase demonstrates homologous cellular staining to guanylate cyclase enzymatic reactivity. The activity of the phosphodiesterase hydrolytic enzyme could be detected evenly distributed throughout the neuropil within cells 15-20 microns in diameter, analogous in cytoarchitecture to immunohistochemically visualized guanylate cyclase, cyclic GMP, and protein kinase elements. Ultrastructural examination of rat caudate-putamen demonstrates that the immunoreactivity for the components of the cyclic GMP system is predominantly distributed within the medium-spiny neuron subtype of this structure. Occasional aspiny neurons demonstrate peroxidase immunoreactivity for the cyclase, cyclic GMP, and the protein kinase, as does the luminal surface of capillary endothelial cells. The subcellular distribution of the antigenic determinants for these three elements and the hydrolytic activity of the phosphodiesterase enzyme show proximity to one another and are confined to the postsynaptic region of asymmetrical, but not symmetrical, terminal boutons. The asymmetrical terminal population of the caudate-putamen is derived from striatal afferents from the neocortex, intralaminar thalamus, and substantia nigra, and to a lesser extent the intrinsic striatal circuitry.(ABSTRACT TRUNCATED AT 400 WORDS)
已对大鼠尾状核 - 壳核复合体前部中鸟苷酸环化酶(EC 4.6.1.2)、3',5'-环磷酸鸟苷(环磷酸鸟苷)、环磷酸鸟苷依赖性蛋白激酶(EC 2.7.1.38)和环磷酸鸟苷磷酸二酯酶(EC 3.1.4.17)的细胞分布进行了研究。新鲜冷冻的尾状核 - 壳核组织中鸟苷酸环化酶、环磷酸鸟苷和环磷酸鸟苷依赖性蛋白激酶的免疫荧光染色类似于灌注固定纹状体切片中的免疫过氧化物酶定位。在直径为15 - 20微米的卵圆形和圆形神经元的细胞质和突起中,可以看到环磷酸鸟苷系统的这三个成分具有同源免疫反应性。免疫反应性神经元均匀分布在所有检查的实验组织的尾状核 - 壳核复合体中。在纹状体腹侧区域偶尔可见直径大于25微米的大型神经元呈现免疫反应性。鸟苷酸环化酶和环磷酸鸟苷磷酸二酯酶活性的酶组织化学测定表明,中等大小的神经元群体(15 - 20微米)含有这些蛋白质的水解活性。大到中等大小的毛细血管显示出鸟苷酸环化酶合成活性,但内皮细胞未表现出免疫组织化学染色。这表明通过免疫组织化学方法不能完全识别酶的生理活性。此外,酶法检测到的鸟苷酸环化酶组织化学活性在整个纹状体神经毡中分布并不均匀。环磷酸鸟苷磷酸二酯酶的酶组织化学检测显示出与鸟苷酸环化酶酶反应性同源的细胞染色。磷酸二酯酶水解酶的活性可以在直径为15 - 20微米的细胞内整个神经毡中均匀分布,其细胞结构类似于免疫组织化学可视化的鸟苷酸环化酶、环磷酸鸟苷和蛋白激酶成分。大鼠尾状核 - 壳核的超微结构检查表明,环磷酸鸟苷系统成分的免疫反应性主要分布在该结构的中等棘状神经元亚型内。偶尔的无棘神经元以及毛细血管内皮细胞的腔表面对环化酶、环磷酸鸟苷和蛋白激酶表现出过氧化物酶免疫反应性。这三种成分的抗原决定簇的亚细胞分布以及磷酸二酯酶的水解活性彼此接近,并局限于不对称而非对称终末小体的突触后区域。尾状核 - 壳核的不对称终末群体来自新皮层、丘脑板内核和黑质的纹状体传入纤维,在较小程度上也来自纹状体内在回路。(摘要截断于400字)