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Journal of Histochemistry and Cytochemistry, Vol. 45, 1611-1622, Copyright © 1997 by The Histochemical Society, Inc.

ARTICLE

Regional and Ultrastructural Immunolocalization of Copper-Zinc Superoxide Dismutase in Rat Central Nervous System

Sandra Morenoa, Roberta Nardaccib, and Maria Paola Cerùa
a Dipartimento di Biologia di Base ed Applicata, Università dell'Aquila, Coppito, L'Aquila, Italy
b Dipartimento di Biologia Cellulare e dello Sviluppo, Università di Roma "La Sapienza," Rome, Italy

Correspondence to: Maria Paola Cerù, Dip. di Biologia di Base ed Applicata, Università dell’Aquila, Via Vetoio, 10 - Coppito, 67010 L’Aquila, Italy.

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. (J Histochem Cytochem 45:1611-1622, 1997)

Key Words: free radicals, antioxidants, catalase, peroxisomes, immunohistochemistry, biotin amplification, tyramine conjugates, neurons, rat brain


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