Postnatal development of NADPH-diaphorase/nitric oxide synthase positive nerve cells in the visual cortex of the rat

Abstract

The postnatal development of NADPH-diaphorase (NADPH-d)/nitric oxide synthase (NOS) positive nerve cells was studied in the visual cortex of rats on postnatal day 1, 5, 10, 15, 20, 30 and at the age of 1 year. NADPH-d was demonstrated enzymhistochemically and NOS immunohistochemically using a polyclonal antibody. NADPH-d is localized in nerve cell somata, dendrites, axons and blood vessels, whereas NOS immunoreactivity is only detectable in nerve cells. The identity of NADPH-d cells with those which contain NOS was proved in double labelling experiments in the cortex of rats on postnatal day 5, 15 and at the age of 1 year. The results of these experiments have shown that in the cortex of rats NADPH-d positive cells are identical with NOS-positive cells in the different stages. Therefore we have used NADPH-d histochemistry in all other postnatal stages as a marker for neurons which contain NOS. NOS positive nerve cells appear very early on postnatal day 1 in the intermediate (white matter) and subplate (layers V and VI) region as small undifferentiated neurons. During the following postnatal differentiation these neurons reached their typical morphology in the second week and appeared in all layers. Neurons in layers V and VI preceded those in the superficial layers. Nerve cells in the white matter seem to have their own differentiation pattern because they showed characteristic features of immaturated varicose dendrites for a longer time. The investigation of soma size with the computerized "Kontron Videoplan" system (Zeiss, Germany) showed the largest cell bodies on postnatal day 20 which then decreased towards adulthood. Between postnatal day 10 and 20 some NOS-positive neurons especially in the deep layers displayed symptoms of degeneration, like shrunken cell bodies, corkscrew and twisted dendrites. Furthermore, NOS-positive neurons in layer I are not detectable in adult neocortex. These observations could suggest that some NOS-positive cells in the cerebral cortex of rats may occur only transiently. Also in the neuropil some alterations in the localization of NOS positive axonal boutons were observed. On postnatal day 10 NOS negative cell somata were shadowy surrounded by boutons. During the further development from postnatal day 20 until adulthood this particular position was no longer visible. Beside the NOS cells which played a transient role, the majority of these cells survived to adulthood and are a morphological (Martinotti-cells with ascending axons) and chemical (GABAergic, NADPH-d/NOS positive, peptide containing cells) defined cell type in the neuronal network of the cortex of the rat.