Alteration in glutathione content and associated enzyme activities in the synaptic terminals but not in the non-synaptic mitochondria from the frontal cortex of Parkinson's disease brains.

Altered redox dynamics contribute to physiological aging and Parkinson's disease (PD). This is reflected in the substantia nigra (SN) of PD patients as lowered antioxidant levels and elevated oxidative damage. Contrary to this observation, we previously reported that non-SN regions such as caudate nucleus and frontal cortex (FC) exhibited elevated antioxidants and lowered mitochondrial and oxidative damage indicating constitutive protective mechanisms in PD brains. To investigate whether the sub-cellular distribution of antioxidants could contribute to these protective effects, we examined the distribution of antioxidant/oxidant markers in the neuropil fractions [synaptosomes, non-synaptic mitochondria and cytosol] of FC from PD (n = 9) and controls (n = 8). In the control FC, all the antioxidant activities [Superoxide dismutase (SOD), glutathione (GSH), GSH peroxidase (GPx), GSH-S-transferase (GST)] except glutathione reductase (GR) were the highest in cytosol, but several fold lower in mitochondria and much lower in synaptosomes. However, FC synaptosomes from PD brains had significantly higher levels of GSH (p = 0.01) and related enzymes [GPx (p = 0.02), GR (p = 0.06), GST (p = 0.0001)] compared to controls. Conversely, mitochondria from the FC of PD cases displayed elevated SOD activity (p = 0.02) while the GSH and related enzymes were relatively unaltered. These changes in the neuropil fractions were associated with unchanged or lowered oxidative damage. Further, the mitochondrial content in the synaptosomes of both PD and control brains was ≥five-fold lower compared to the non-synaptic mitochondrial fraction. Altered distribution of oxidant/antioxidant markers in the neuropil fractions of the human brain during aging and PD has implications for (1) degenerative and protective mechanisms (2) distinct antioxidant mechanisms in synaptic terminals compared to other compartments.

Differences between unipolar and bipolar I depression in the quantitative analysis of glutamic acid decarboxylase-immunoreactive neuropil.

Alterations in GABAergic neurotransmission are assumed to play a crucial role in the pathophysiology of mood disorders. Glutamic acid decarboxylase (GAD) is the key enzyme in GABA synthesis. This study aimed to differentiate between unipolar and bipolar I depression using quantitative evaluation of GAD-immunoreactive (GAD-ir) neuropil in several brain regions known to be involved in the pathophysiology of mood disorders. Immunohistochemical staining of GAD 65/67 was performed in the orbitofrontal, anterior cingulate and dorsolateral prefrontal cortex (DLPFC), the entorhinal cortex, the hippocampal formation and the medial dorsal and lateral dorsal (LD) thalamic nuclei, with a quantitative densitometric analysis of GAD-ir neuropil. The study was performed on paraffin-embedded brains from 9 unipolar and 12 bipolar I depressed patients (8 and 6 suicidal patients, respectively) and 18 matched controls. In unipolar patients, compared with controls, only the increased relative density of GAD-ir neuropil in the right LD was different from the previous results in depressed suicides from the same cohort (Gos et al. in J Affect Disord 113:45-55, 2009). On the other hand, the left DLPFC was the only area where a significant decrease was observed, specific for bipolar I depression. Significant differences between both diagnostic groups were found in these regions. By revealing abnormalities in the relative density of GAD-ir neuropil in brain structures, our study suggests a diathesis of the GABAergic system in mood disorders, which may differentiate the pathophysiology of unipolar from that of bipolar I depression.

Metabolic enzymes in glial cells of the honeybee brain and their associations with aging, starvation and food response.

The honey bee has been extensively studied as a model for neuronal circuit and memory function and more recently has emerged as an unconventional model in biogerontology. Yet, the detailed knowledge of neuronal processing in the honey bee brain contrasts with the very sparse information available on glial cells. In other systems glial cells are involved in nutritional homeostasis, detoxification, and aging. These glial functions have been linked to metabolic enzymes, such as glutamine synthetase and glycogen phosphorylase. As a step in identifying functional roles and potential differences among honey bee glial types, we examined the spatial distribution of these enzymes and asked if enzyme abundance is associated with aging and other processes essential for survival. Using immunohistochemistry and confocal laser microscopy we demonstrate that glutamine synthetase and glycogen phosphorylase are abundant in glia but appear to co-localize with different glial sub-types. The overall spatial distribution of both enzymes was not homogenous and differed markedly between different neuropiles and also within each neuropil. Using semi-quantitative Western blotting we found that rapid aging, typically observed in shortest-lived worker bees (foragers), was associated with declining enzyme levels. Further, we found enzyme abundance changes after severe starvation stress, and that glutamine synthetase is associated with food response. Together, our data indicate that aging and nutritional physiology in bees are linked to glial specific metabolic enzymes. Enzyme specific localization patterns suggest a functional differentiation among identified glial types.

Differential effects of methylmercury intoxication in the rat's barrel field as evidenced by NADPH diaphorase histochemistry.

In the present study, we investigated the effects of mercury intoxication on the structure of the posteromedial barrel subfield (PMBSF) in the primary somatosensory cortex (SI) of adult rats, as revealed by histochemical reactivity to the enzyme NADPH diaphorase (NADPH-d). Enzymatic reactivity in the neuropil inside barrels was drastically reduced in intoxicated animals, suggesting that the synthesis and/or transport of the nitric oxide synthase enzyme can be altered in acute mercury intoxication. However, the cell bodies and dendrites of barrel neurons, also strongly reactive to the enzyme, were spared from the mercury's deleterious effects.

Identification of subdivisions in the medial geniculate body of the guinea pig.

The accurate and reliable identification of subdivisions within the auditory thalamus is important for future studies of this nucleus. However, in the guinea pig, there has been no agreement on the number or nomenclature of subdivisions within the main nucleus of the auditory thalamus, the medial geniculate body (MGB). Thus, we assessed three staining methods in the guinea pig MGB and concluded that cytochrome oxidase (CYO) histochemistry provides a clear and reliable method for defining MGB subdivisions. By combining CYO with acetylcholinesterase staining and extensive physiological mapping we defined five separate divisions, all of which respond to auditory stimuli. Coronal sections stained for CYO revealed a moderate to darkly-stained oval core. This area (the ventral MGB) contained a high proportion (61%) of V-shaped tuning curves and a tonotopic organisation of characteristic frequencies. It was surrounded by four smaller areas that contained darkly stained somata but had a paler neuropil. These areas, the dorsolateral and suprageniculate (which together form the dorsal MGB), the medial MGB and the shell MGB, did not have any discernable tonotopic frequency gradient and contained a smaller proportion of V-shaped tuning curves. This suggests that CYO permits the identification of core and belt areas within the guinea pig MGB.

Localization of nitric oxide synthase in the central complex and surrounding midbrain neuropils of the locust Schistocerca gregaria.

Nitric oxide (NO), generated enzymatically by NO synthase (NOS), acts as an important signaling molecule in the nervous systems of vertebrates and invertebrates. In insects, NO has been implicated in development and in various aspects of sensory processing. To understand better the contribution of NO signaling to higher level brain functions, we analyzed the distribution of NOS in the midbrain of a model insect species, the locust Schistocerca gregaria, by using NADPH diaphorase (NADPHd) histochemistry after methanol/formalin fixation; results were validated by NOS immunohistochemistry. NADPHd yielded much higher sensitivity and resolution, but otherwise the two techniques resulted in corresponding labeling patterns throughout the brain, except for intense immunostaining but only weak NADPHd staining in median neurosecretory cells. About 470 neuronal cell bodies in the locust midbrain were NADPHd-positive positive, and nearly all major neuropil centers contained dense, sharply stained arborizations. We report several novel types of NOS-expressing neurons, including small ocellar interneurons and antennal sensory neurons that bypass the antennal lobe. Highly prominent labeling occurred in the central complex, a brain area involved in sky-compass orientation, and was analyzed in detail. Innervation by NOS-expressing fibers was most notable in the central body upper and lower divisions, the lateral accessory lobes, and the noduli. About 170 NADPHd-positive neurons contributed to this innervation, including five classes of tangential neuron, two systems of pontine neuron, and a system of columnar neurons. The results provide new insights into the neurochemical architecture of the central complex and suggest a prominent role for NO signaling in this brain area.

Sexual dimorphism of vocal control nuclei in budgerigars (Melopsittacus undulatus) revealed with Nissl and NADPH-d staining.

Nissl staining and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry were used to explore the existence of sexual dimorphism in vocal control nuclei of adult budgerigars (Melopsittacus undulatus), a parrot species capable of lifelong vocal learning. Behavioral studies indicate that adult males possess larger vocal repertoires than adult females and learn new calls more quickly. The results of the present study show that the volumes of all vocal nuclei, as measured using both Nissl-stained and NADPH-d-stained material, as well as the total numbers of NADPH-d neurons, were 35-110% greater in males. Furthermore, all vocal nuclei exhibit conspicuous NADPH-d staining compared to surrounding fields in both adult males and females. Nevertheless, there were no significant gender differences in either the intensity of neuropil staining or the densities of NADPH-d neurons in vocal nuclei. Moreover NADPH-d neuron somal shapes were similar in males and females. Diameters of NADPH-d neurons in vocal nuclei were 8.5-32% larger in males than in females. Greater size of NADPH-d neuronal somata in males may be a general property of this cell type in budgerigars because a similar gender difference was found in a visual nucleus, the entopallium, which is not directly associated with the vocal control system and does not exhibit sexual dimorphism in total volume or total NADPH-d neuron numbers. Taken together, the results of the present study favor the hypothesis that superior lifelong vocal learning ability in male budgerigars rests largely on larger volumes of vocal control nuclei in males rather than on sexual dimorphism in the internal composition of vocal nuclei.

Postnatal development of alkaline phosphatase activity correlates with the maturation of neurotransmission in the cerebral cortex.

We have shown previously that the tissue nonspecific alkaline phosphatase (TNAP) is selectively expressed in the synaptic cleft of sensory cortical areas in adult mammals and, by using sensory deprivation, that TNAP activity depends on thalamocortical activity. We further analyzed this structural functional relationship by comparing the developmental pattern of TNAP activity to the maturation of the thalamocortical afferents in the primate brain (Callithrix jacchus). Cortical expression of alkaline phosphatase (AP) activity reflects the sequential maturation of the modality-specific sensory areas. Within the visual cortex, the regional and laminar distribution of AP correlates with the differential maturation of the magno- and parvocellular streams. AP activity, which is transiently expressed in the white matter, exhibits a complementary distributional pattern with myelin staining. Ultrastructural analysis revealed that AP activity is localized exclusively to the myelin-free axonal segments, including the node of Ranvier. It was also found that AP activity is gradually expressed in parallel with the maturation of synaptic contacts in the neuropile. These data suggest the involvement of AP, in addition to neurotransmitter synthesis previously suggested in the adult, in synaptic stabilization and in myelin pattern formation and put forward a role of AP in cortical plasticity and brain disorders.

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.

[Localization of NADPH-diaphorase in the brain of the shore crab Hemigrapsus sanguineus].

The presence and localization of NADPH-diaphorase in the cerebral ganglion of the shore crab Hemigrapsus sanguineus was investigated with histochemical and electron histochemical methods. The reactivity of this enzyme was found in the deutrocerebrum, mainly in neuropils of olfactory lobes, the lateral antennular neuropil, a laterodorsal group of cells, and in the oculomotor nerve nucleus. Ultrastructural localization of the enzyme was detected in neurons on the perinuclear membrane, and in membranes of endoplasmic reticulum, in mitochondria and cytosol. The enzyme was found in axons of the antennular nerve, and in terminals of receptor axons in the glomerulus. The obtained data testify to participation of NO in perception and processing of the olfactory information.

Ultrastructural localization of NADPH diaphorase and nitric oxide synthase in the neuropils of the snail CNS.

Comparative studies on the nervous system revealed that nitric oxide (NO) retains its function through the evolution. In vertebrates NO can act in different ways: it is released solely or as a co-transmitter, released from presynaptic or postsynaptic site, spreads as a volumetric signal or targets synaptic proteins. In invertebrates, however, the possible sites of NO release have not yet been identified. Therefore, in the present study, the subcellular distribution of the NO synthase (NOS) was examined in the central nervous system (CNS) of two gastropod species, the terrestrial snail, Helix pomatia and the pond snail, Lymnaea stagnalis, which are model species in comparative neurobiology. For the visualization of NOS NADPH-diaphorase histochemistry and an immunohistochemical procedure using a universal anti-NOS antibody were applied. At light microscopic level both techniques labeled identical structures in sensory tracts ramifying in the neuropils of central ganglia and cell bodies of the Lymnaea and Helix CNS. At ultrastructural level NADPH-d reactive/NOS-immunoreactive materials were localized on the nuclear envelope and membrane segments of the rough and smooth endoplasmic reticulum, as well as the cell membrane and axolemma of positive perikarya. NADPH-d reactive and NOS-immunoreactive varicosities connected to neighboring neurons with both unspecialized and specialized synaptic contacts. In the varicosities, the majority of the NADPH-d reactive/NOS-immunoreactive membrane segments were detected in round and pleomorph agranular vesicles of small size (50-200 nm). However, only a small portion (16%) of the vesicles displayed the NADPH-d reactivity/NOS-immunoreactivity. No evidence for the postsynaptic location of NOS was found. Our results suggest that the localization of NADPH-diaphorase and NOS is identical in the snail nervous system. In contrast to vertebrates, however, NO seems to act exclusively in an anterograde way possibly released from membrane segments of the presynaptic transmitter vesicle surface. Based on the subcellular distribution of NOS, NO could be both a volume and a synaptic mediator, in addition NO may function as a co-transmitter.

Concentrated expression of Ca2+/ calmodulin-dependent protein kinase II and protein kinase C in the mushroom bodies of the brain of the honeybee Apis mellifera L.

We have previously used the differential display method to identify a gene that is expressed preferentially in the mushroom bodies of worker honeybees and to show that it encodes a putative inositol 1,4,5-trisphosphate receptor (IP3R) homologue (Kamikouchi et al. [1998] Biochem. Biophys. Res. Commun. 242:181-186). In the present study, we examined whether the expression of some of the genes for proteins involved in the intracellular Ca2+ signal transduction is also concentrated in the mushroom bodies of the honeybee by isolating cDNA fragments that encode the Ca2+/calmodulin-dependent protein kinase II (CaMKII) and protein kinase C (PKC) homologues of the honeybee. In situ hybridization analysis revealed that the expression of these genes was also concentrated in the mushroom bodies of the honeybee brain: The CaMKII gene was expressed preferentially in the large-type Kenyon cells of the mushroom bodies, whereas that for PKC was expressed in both the large and small types of Kenyon cells. The expression of the genes for IP3R and CaMKII was concentrated in the mushroom bodies of the queen and drone as well as in those of the worker bee. Furthermore, the enzymatic activities of CaMKII and PKC were found to be higher in the mushroom bodies/central bodies than in the optic and antennal lobes of the worker bee brain. These results suggest that the function of the intracellular Ca2+ signal transduction is enhanced in Kenyon cells in comparison to other neuronal cell types in the honeybee brain.

The normal distribution and projections of constitutive NADPH-d/NOS neurons in the brainstem vestibular complex of the rat.

The vestibular system is a highly conserved sensory system in vertebrates that is largely responsible for maintenance of one's orientation in space, posture, and balance and for visual fixation of objects during motion. In light of the considerable literature indicating an involvement of nitric oxide (NO) in sensory systems, it is important to determine whether NO is associated with vestibular pathways. To study the relationship of NO to vestibular pathways, we first examined the normal distribution of constitutive NADPH-diaphorase (NADPH-d), a marker for nitric oxide synthase (NOS), in the vestibular complex (VC) and then examined its association with selected vestibular projection neurons. Survey of the four major vestibular nuclei revealed that only the medial vestibular nucleus contained significant numbers of perikarya stained for NADPH-d/NOS. By contrast, all the vestibular nuclei contained a network of fine processes that stained positive for NADPH-d, although the density of this network varied among the individual nuclei. To determine whether NADPH-d/NOS neurons project to vestibular efferent targets, injections of the retrograde tracer Fluoro-Gold were made into known targets of second-order vestibular neurons. Vestibular neurons containing constitutive NADPH-d/NOS were found to project predominantly to the oculomotor nucleus. A small number of neurons also participate in vestibulothalamic and intrinsic vestibular connections. These results indicate that NADPH-d/NOS neurons are prevalent in the MVN and that a subpopulation of these neurons project to the oculomotor complex. Nitric oxide is probably released locally from axons located throughout the vestibular complex but may play a particularly important role in vestibulo-ocular pathways.

Conductive hearing loss results in changes in cytochrome oxidase activity in gerbil central auditory system.

Conductive hearing loss (CHL) restricts auditory input to an intact peripheral auditory system. Effects of deprivation on the central auditory system (CAS) have been debated, although a number of studies support the hypothesis that CHL can cause modification of CAS structure and function. The present study was designed to test the hypothesis that unilateral CHL results in a decrease in cytochrome oxidase (CO) activity in CAS nuclei that receive major afferent input from the affected ear. Gerbils at postnatal day 12 (P21) or 6-8 weeks underwent left unilateral CHL (malleus removal), cochlear ablation, or a sham surgical procedure. After a survival time of 48 hours or 3 weeks, animals were sacrificed and tissue was processed for cytochrome oxidase histochemistry. Optical density (OD) measurements were made from individual neurons in the anteroventral cochlear nucleus (AVCN) and from medial and lateral dendritic fields in the medial superior olivary nucleus (MSO), the lateral superior olivary nucleus, and the inferior colliculus. The width of the CO-stained neuropil in MSO was also measured as an estimate of dendritic length. OD measures were corrected to neutral areas of the brain. Cochlear ablation caused significant decreases in CO activity in left lower brainstem nuclei, particularly in adult animals. Following CHL, a significant decrease in CO activity was observed in the ipsilateral AVCN and a significant increase was observed in the contralateral AVCN. Cochlear ablation resulted in decreased width of MSO neuropil containing dendrites that receive primary input from the ablated ear. CHL resulted in a significant increase in the width of MSO neuropil on both sides of the brain in the P21 animals that survived 3 weeks but not in P21 animals that survived only 48 hours or in the adult animals. Unilateral CHL is associated with changes in CO activity in the AVCN and may affect MSO dendritic length in younger animals.

Cerebral expression of the alpha2-subunit of soluble guanylyl cyclase is linked to cerebral maturation and sensory pathway refinement during postnatal development.

Soluble guanylyl cylase (sGC) has been identified for being a receptor for the gaseous transmitters nitric oxide and carbon monoxide. Currently four subunits alpha1, alpha2, beta1, and beta2 have been characterized. Heterodimers of alpha and beta-subunits as well as homodimers of the beta2-subunit are known to constitute functional sGC which use GTP to form cGMP a potent signal molecule in a multitude of second messenger cascades. Since NO-cGMP signaling plays a pivotal role in neuronal development we analyzed the maturational expression pattern of the newly characterized alpha2-subunit of sGC within the brain of Wistar rats by means of RNase protection assay and immunohistochemistry. alpha2-subunit mRNA as well as immunoreactive alpha2-protein increased during postnatal cerebral development. Topographical analysis revealed a selective high expression of the alpha2-subunit in the choroid plexus and within developing sensory systems involving the olfactory and somatosensory system of the forebrain as well as parts of the auditory and visual system within the hindbrain. In cultured cortical neurons the alpha2-subunit was localized to the cell membrane, especially along neuronal processes. During the first 11 days of postnatal development several cerebral regions showed a distinct expression of the alpha2-subunit which was not paralleled by the alpha1/beta1-subunits especially within the developing thalamo-cortical circuitries of the somatosensory system. However, at later developmental stages all three subunits became more homogenously distributed among most cerebral regions, indicating that functional alpha1/beta1 and alpha2/beta1 heterodimers of sGC could be formed. Our findings indicate that the alpha2-subunit is an essential developmentally regulated constituent of cerebral sensory systems during maturation. In addition the alpha2-subunit may serve other functions than forming a functional heterodimer of sGC during the early phases of sensory pathway refinement.

Effect of auditory cortex lesions on NADPH-diaphorase staining in the inferior colliculus of rat.

Projections from the auditory cortex (AC) in the rat terminate in the dorsal cortex (DC) and in the external cortex (EC) of the inferior colliculus (IC), areas which exhibit a moderate number of nicotinamide-adenine dinucleotide phosphate-diaphorase (NADPH-d) positive neurons. NADPH-d co-localizes with nitric oxide synthase, which is responsible for the production of the transcellular messenger, nitric oxide. Changes in NADPH-d staining in the IC were found after unilateral lesions of the AC. Lesions resulted in a reduction in NADPH-d staining in neurons and neuropil within the ipsilateral DC and EC with the maximum reduction occurring 3-4 days after lesion. The reduction in NADPH-d staining in the contralateral IC was less pronounced. Lesions affecting auditory areas Te 1 and Te 3 produced the largest decrease in NADPH-d staining in neurons and neuropil. This finding may be related to the abolition of the influence of glutamatergic corticocollicular and commissural pathways.

Alterations of GABA(A)beta2/3 immunoreactivity in the dentate gyrus after perforant pathway lesion.

Immunocytochemical techniques were employed to examine the changes in the GABA receptor subunits beta2/3 within the dentate gyrus of the rat brain 1, 3, 7, 14, 30 and 90 days after a unilateral perforant pathway lesion. Three days post-lesion we observed a decrease in beta2/3 immunolabeling in the inner molecular layer of the dentate gyrus followed by a comparable decrease in the outer molecular layer 7 days post-lesion. These decreases were transient; 30 and 90 days post-lesion, beta2/3 immunolabeling appeared similar to controls in the inner portion of the molecular layer, while in the outer region the labeling was increased. In this latter region we also observed a dense band of AChE fibers. Following survival times of 3 days we observed a diffuse staining of the neuropil in the hilar region, and a dense amorphous accumulation of peroxidase reaction product in the polymorphic region. These responses were transient and by 14 days the hilar/polymorphic region appeared indistinguishable from controls. These data suggest a unique pattern of immunoabeling in the molecular and polymorphic region in response to perforant pathway lesion. A putative explanation for this response is discussed.

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.

Effect of age and neuroleptics on tyrosine hydroxylase-IR in sector CA2 of schizophrenic brain.

The density of TH-IR varicosities was analyzed in the hippocampus of 15 normal controls and 11 schizophrenics. The average density of varicosities in apposition with pyramidal cells and in the neuropil was 30-35% lower in CA2, but not other sectors of schizophrenics. Age was correlated with varicosity density in all sectors, particularly in CA2 where young patients showed a 50% reduction on non-pyramidal cells. Neuroleptic dose showed a negative correlation with the density of varicosities, and notably the dose of young schizophrenics was four times higher than that of older subjects. Thus, antipsychotic dose appears to be associated with a suppression of a normal age-related increase of dopamine projections to CA2 during the early phases of schizophrenia.

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.