Morphometric variability of nicotinamide adenine dinucleotide phosphate diaphorase neurons in the primary sensory areas of the rat.

Even though there is great regional variation in the distribution of inhibitory neurons in the mammalian isocortex, relatively little is known about their morphological differences across areal borders. To obtain a better understanding of particularities of inhibitory circuits in cortical areas that correspond to different sensory modalities, we investigated the morphometric differences of a subset of inhibitory neurons reactive to the enzyme nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) within the primary auditory (A1), somatosensory (S1), and visual (V1) areas of the rat. One hundred and twenty NADPH-d-reactive neurons from cortical layer IV (40 cells in each cortical area) were reconstructed using the Neurolucida system. We collected morphometric data on cell body area, dendritic field area, number of dendrites per branching order, total dendritic length, dendritic complexity (Sholl analysis), and fractal dimension. To characterize different cell groups based on morphology, we performed a cluster analysis based on the previously mentioned parameters and searched for correlations among these variables. Morphometric analysis of NADPH-d neurons allowed us to distinguish three groups of cells, corresponding to the three analyzed areas. S1 neurons have a higher morphological complexity than those found in both A1 and V1. The difference among these groups, based on cluster analysis, was mainly related to the size and complexity of dendritic branching. A principal component analysis (PCA) applied to the data showed that area of dendritic field and fractal dimension are the parameters mostly responsible for dataset variance among the three areas. Our results suggest that the nitrergic cortical circuitry of primary sensory areas of the rat is differentially specialized, probably reflecting peculiarities of both habit and behavior of the species.

Neuropil reactivity, distribution and morphology of NADPH diaphorase type I neurons in the barrel cortex of the adult mouse.

The mouse, like a few other rodent and marsupial species, displays a striking modular architecture in its primary somatosensory cortex (SI). These modules, known as barrels, are mostly defined by the peculiar arrangement of granule cells and thalamic axons in layer IV. In the present work, we studied both the distribution and morphology of neurons stained for NADPH diaphorase (NADPH-d) and neuropil reactivity in the posteromedial barrel subfield (PMBSF), which represents the mystacial whiskers. We then compared our results with previous descriptions of NADPH-d distribution in both neonatal and young mice. We found two types of neurons in the PMBSF: type I neurons, which have large cell bodies and are heavily stained by the NADPH-d reaction; and type II neurons, characterized by relatively small and poorly stained cell bodies. The distribution of type I cells in the PMBSF was not homogenous, with cells tending to concentrate in septa between barrels. Moreover, the cells found in septal region possess both a larger and more complex dendritic arborization than cells located inside barrels. Our findings are at variance with results from other groups that reported both an absence of type II cells and a homogeneous distribution of type I cells in the PMBSF of young animals. In addition, our results show a distribution of type I cells which is very similar to that previously described for the rat's barrel field.

A morphometric study of the progressive changes on NADPH diaphorase activity in the developing rat's barrel field.

The distribution of NADPH diaphorase (NADPH-d)/nitric oxide synthase (NOS) neurons was evaluated during the postnatal development of the primary somatosensory cortex (SI) of the rat. Both cell counts and area measurements of barrel fields were carried out throughout cortical maturation. In addition, NADPH-d and cytochrome oxidase (CO) activities were also compared in both coronal and tangential sections of rat SI between postnatal days (P) 10 and 90. Throughout this period, the neuropil distributions of both enzymes presented a remarkable similarity and have not changed noticeably. Their distribution pattern show the PMBSF as a two-compartmented structure, displaying a highly reactive region (barrel hollows) flanked by less reactive regions (barrel septa). The number of NADPH-d neurons increased significantly in the barrel fields between P10 and P23, with peak at P23. The dendritic arborization of NADPH-d neurons became more elaborated during barrel development. In all ages evaluated, the number of NADPH-d cells was always higher in septa than in the barrel hollows. Both high neuropil reactivity and differential distribution of NADPH-d neurons during SI development suggest a role for nitric oxide throughout barrel field maturation.

The barrel field of the adult mouse SmI cortex as revealed by NADPH-diaphorase histochemistry.

The main goal of the present work was to investigate the pattern of NADPH-diaphorase activity in the somatosensory cortex of the adult mouse. Our results show that this enzyme, which is responsible for the production of the neuronal messenger nitric oxide, is abundant within the neuropil of SmI cortex, revealing the complete pattern of barrel fields. A previous study, however, had reported that NADPH-diaphorase reactivity within the barrels was transient, disappearing after the second postnatal week. We hypothesize that the massive occurrence of NADPH-diaphorase in the barrel fields of the adult mouse brain is related to the potential for plastic changes in the somatosensory cortex that is maintained throughout maturity.

NADPH-d positive neurons in the developing somatosensory cortex of the rat: effects of early and late environmental enrichment.

The effects of environmental enrichment upon the topographic arrangement of NADPH diaphorase-positive neurons (NADPH-d+ neurons) was studied in the somatosensory cortex of 56 Sprague-Dawley albino rats during early stages of development (18th, 24th, 30th and 60th postnatal days). This diaphorase is easily demonstrable, providing a convenient marker for quantitative studies. Environmental enrichment diminished the number of NADPH-d+ neurons and exerted its maximal influence during lactation, a time of exceptional cortical susceptibility. This implies that the magnitude of such effects on the density of NADPH-d+ neurons is age-dependent. Furthermore, it was found that the experience-dependent cortical changes persisted after a subsequent period without environmental stimulation. The effects of early environmental enrichment did not occur uniformly throughout the cerebral hemispheres but, instead, such effects were maximal in the latero-ventral sector of the cerebral cortex where a dramatic reduction in the number of NADPH-d+ neurons was observed. Particularly striking was the existence of a latero-medial sequence of NADPH-d+ neurons in the infragranular layer and a reversed distribution of labeled cells, in the supragranular layer. Both ontogenetic sequences of NADPH-d+ neurons remained unchanged during postnatal development in controls and enriched subjects (18th-60th postnatal days).

Distribution of NADPH-d positive neurons during postnatal development of the rat somatosensory cortex correlates with gradients of neurogenesis and development.

The expression of nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) was studied in the rat somatosensory cortex during postnatal development from day 6 to 120. Distribution of labeled neurons was quantified in dorso-medial and ventro-lateral aspects of the cortex, and correlated with known tridimensional gradients of histogenetic development and maturation of cortical neurons. NADPH positive neurons were non-pyramidal cells that in all developmental periods were more numerous in infragranular than in supragranular layers of the cortex. Additionally, more labeled cells were found in ventro-lateral than dorso-medial infragranular layers and in anterior than posterior aspects of the cerebral cortex. These patterns of distribution correlate well with the gradients of histogenetic development and with the pattern of maturation of cortical neurons.