Moreno S, Nardacci R, Cerù M P
Dipartimento di Biologia di Base ed Applicata, Università dell'Aquila, Coppito, L'Aquila, Italy.
J Histochem Cytochem. 1997 Dec;45(12):1611-22. doi: 10.1177/002215549704501204.
We examined the distribution of copper-zinc superoxide dismutase (CuZnSOD) in adult rat central nervous system by light and electron microscopic immunocytochemistry, using an affinity-purified polyclonal antibody. The enzyme appeared to be exclusively localized in neurons. No immunoreactivity was seen in non-neuronal cells. The staining intensity was variable, depending on the brain region and, within the same region, on the neuron type. Highly immunoreactive elements included cortical neurons evenly distributed in the different layers, hippocampal interneurons, neurons of the reticular thalamic nucleus, and Golgi, stellate, and basket cells of the cerebellar cortex. Other neurons, i.e., pyramidal cells of the neocortex and hippocampus, Purkinje and granule cells of the cerebellar cortex, and the majority of thalamic neurons, showed much weaker staining. In the spinal cord, intense CuZnSOD immunoreactivity was present in many neurons, including motor neurons. Pre-embedding immunoelectron microscopy of the neocortex, hippocampus, reticular thalamic nucleus, and cerebellar cortex showed cytosolic and nucleoplasmic labeling. Moreover, single membrane-limited immunoreactive organelles identified as peroxisomes were often found, even in neurons that appeared weakly stained at the light microscopic level. In double immunogold labeling experiments, particulate CuZnSOD immunoreactivity co-localized with catalase, a marker enzyme for peroxisomes, thus demonstrating that in neural tissue CuZnSOD is also present in peroxisomes.
我们使用亲和纯化的多克隆抗体,通过光镜和电镜免疫细胞化学方法研究了成年大鼠中枢神经系统中铜锌超氧化物歧化酶(CuZnSOD)的分布。该酶似乎仅定位于神经元中。在非神经元细胞中未观察到免疫反应性。染色强度各不相同,这取决于脑区,并且在同一区域内还取决于神经元类型。高免疫反应性的细胞包括均匀分布在不同层的皮质神经元、海马中间神经元、丘脑网状核神经元以及小脑皮质的高尔基细胞、星状细胞和篮状细胞。其他神经元,即新皮质和海马的锥体细胞、小脑皮质的浦肯野细胞和颗粒细胞,以及大多数丘脑神经元,染色则弱得多。在脊髓中,许多神经元,包括运动神经元,都存在强烈的CuZnSOD免疫反应性。对新皮质、海马、丘脑网状核和小脑皮质进行包埋前免疫电镜检查显示,胞质和核质均有标记。此外,即使在光镜下染色较弱的神经元中,也经常发现被鉴定为过氧化物酶体的单膜限定免疫反应性细胞器。在双重免疫金标记实验中,颗粒状CuZnSOD免疫反应性与过氧化物酶体的标记酶过氧化氢酶共定位,从而证明在神经组织中CuZnSOD也存在于过氧化物酶体中。