Light and electron microscopic study of neuronal nitric oxide synthase-immunoreactive neurons in the rat subiculum

Abstract

Neurons in the rat subiculum that are capable of producing nitric oxide were studied by using an antibody to the neuronal isoform of nitric oxide synthase (nNOS). In the light microscope, the staining pattern with the nNOS antibody closely resembled that seen following histochemical processing with nicotinamide adenine dinucleotide phosphate diaphorase. Immunostained neurons were found in all layers, and, in addition, large dendrites in the apical dendrite layer were also immunopositive. Although a few immunolabelled cells had the typical morphology of interneurons, most were found to have the characteristics of pyramidal neurons. In the subiculum, these immunoreactive pyramidal neurons were concentrated mainly in the most superficial cell layers and closest to the CA1 region, but pyramidal neurons in the CA1 layer of the hippocampus were consistently immunonegative. Immunopositive profiles in the subiculum were studied in the electron microscope and compared with unlabelled structures. Ultrastructural criteria suggest that both pyramidal and nonpyramidal subicular neurons are immunopositive for nNOS. Large, spiny dendrites and smaller, varicose dendrites were found to be immunoreactive for nNOS. Vesicle-containing profiles were probably presynaptic axons, and immunopositive boutons were seen to make symmetrical and asymmetrical synaptic contacts.