Results from several electrophysiological studies predict that the neuronal NO-synthesizing enzyme, nNOS, resides within spines formed by pyramid-to-pyramid axo-spinous synaptic junctions of the cortex. On the other hand, light microscopic neuroanatomical detection of nNOS within pyramidal neurons has been difficult, suggesting that these neurons contain nNOS at levels below threshold for detection. Our results obtained by electron microscopic immunocytochemistry indicate that nNOS occurs within spiny neurons, such as those of pyramidal neurons, albeit discretely within their spines. Dual electron microscopic immunocytochemistry, whereby antigenic sites to the NR1 subunit of NMDA receptors are probed simultaneously with sites immunoreactive for nNOS, reveals that some, although not all, nNOS within spines co-exist with NR1 subunits. Additionally, immunoreactivity for the NR1 subunit is detectable within nNOS-axons, indicating that NO may be generated in response to axo-axonic interactions with glutamatergic axons in the vicinity and independently of action potential propagation. Immunoreactivity for NR1 subunits within axons (with or without nNOS- immunoreactivity) may additionally serve to confer receptivity of these axons to NO generated coincidentally with activity. Analysis of the visual cortex of monocular adult animals indicates that the level of nNOS within neurites is dependent on chronic activity levels of the surrounding neuropil and independent of somatic input level. Together, these findings point to plasticity of nNOS neurons within adult brain tissue, involving regulation of subcellular nNOS distribution.
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