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We used biotinylated dextran amine (BDA) to anterogradely label individual axons projecting from primary somatosensory cortex (S1) to four different cortical areas in rats. A major goal was to determine whether axon terminals in these target areas shared morphometric similarities based on the shape of individual terminal arbors and the density of two bouton types: en passant (Bp) and terminaux (Bt). Evidence from tridimensional reconstructions of isolated axon terminal fragments (n=111) did support a degree of morphological heterogeneity establishing two broad groups of axon terminals. Morphological parameters associated with the complexity of terminal arbors and the proportion of beaded Bp vs stalked Bt were found to differ significantly in these two groups following a discriminant function statistical analysis across axon fragments. Interestingly, both groups occurred in all four target areas, possibly consistent with a commonality of presynaptic processing of tactile information. These findings lay the ground for additional work aiming to investigate synaptic function at the single bouton level and see how this might be associated with emerging properties in postsynaptic targets.

It has been demonstrated that rat litter size affects the immune cell response, but it is not known whether the long-term effects aggravate age-related memory impairments or microglial-associated changes. To that end, we raised sedentary Wistar rats that were first suckled in small or large litters (6 or 12pups/dam, respectively), then separated into groups of 2-3 rats from the 21st post-natal day to study end. At 4months (young adult) or 23months (aged), all individual rats were submitted to spatial memory and object identity recognition tests, and then sacrificed. Brain sections were immunolabeled with anti-IBA-1 antibodies to selectively identify microglia/macrophages. Microglial morphological changes in the molecular layer of the dentate gyrus were estimated based on three-dimensional reconstructions. The cell number and laminar distribution in the dentate gyrus was estimated with the stereological optical fractionator method. We found that, compared to young rat groups, aged rats from large litters showed significant increases in the number of microglia in all layers of the dentate gyrus. Compared to the microglia in all other groups, microglia in aged individuals from large litters showed a significantly higher degree of tree volume expansion, branch base diameter thickening, and cell soma enlargement. These morphological changes were correlated with an increase in the number of microglia in the molecular layer. Young adult individuals from small litters exhibited preserved intact object identity recognition memory and all other groups showed reduced performance in both spatial and object identity recognition tasks. We found that, in large litters, brain development was, on average, associated with permanent changes in the innate immune system in the brain, with a significant impact on the microglial homeostasis of aged rats., (Copyright © 2013 IBRO. Published by Elsevier Ltd. All rights reserved.)

The cerebral cortex is often described as a composite of repeated units or columns, integrating the same basic circuit. The 'ice-cube' model of cortical organization, and 'canonical' circuit, born from insights into functional architecture, still require systematic comparative data. Here we probed the anatomy of an individual neuronal type within V1 to determine whether or not its dendritic trees are consistent with the 'ice-cube' model and theories of canonical circuits. In a previous report we studied the morphometric variability of NADPH-diaphorase (NADPH-d) neurons in the rat auditory, visual and somatosensory primary cortical areas. Our results suggested that the nitrergic cortical circuitry of primary sensory areas are differentially specialized, probably reflecting peculiarities of both habit and behavior of the species. In the present report we specifically quantified the dendritic trees of NADPH-d type I neurons as a function of eccentricity within V1. Individual neurons were reconstructed in 3D, and the size, branching and space-filling of their dendritic trees were correlated with their location within the visuotopic map. We found that NADPH-d neurons became progressively smaller and less branched with progression from the central visual representation to the intermediate and peripheral visual representation. This finding suggests that aspects of cortical circuitry may vary across the cortical mantle to a greater extent that envisaged as natural variation among columns in the 'ice-cube' model. The systematic variation in neuronal structure as a function of eccentricity warrants further investigation to probe the general applicability of columnar models of cortical organization and canonical circuits., (Copyright © 2012 IBRO. Published by Elsevier Ltd. All rights reserved.)

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., (Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.)

The integration of cutaneous, proprioceptive, and motor information in area S2 seems to be essential for manual object recognition and motor control. Part of the inputs to S2 comes from area S1. However no detailed investigations of the morphology of this projection are available. In the present study we describe and quantify the morphology of axon fragments of S1 to S2 ipsilateral projections in the agouti somatosensory cortex. Two groups of projecting axon arbors in S2 were individually reconstructed in three dimensions using Neurolucida, after a single electrophysiological guided BDA injection in either the forelimb (n=4) or the hindlimb (n=4). Electrophysiological mapping was performed 15 days after injections, allowing the localization of S2. Cluster analysis of 40 fragments after hindlimb and 40 after forelimb distinguished two clusters of terminals designated as type I and type II. On average, Type I fragments had greater surface areas and segment lengths than type II fragments, whereas type II fragments had higher number of terminal boutons, number of segments and branching points/mm than type I fragments. Type I corresponded to 58% of the axons projecting from the hindlimb representation in S1 whereas 63% of the sample originating from the forelimb representation in S1 corresponded to type II axons. The results suggest possible parallel processing by two stereotyped classes of axon terminals in the S1 to S2 projections that may represent at least part of the circuitry groundwork associated with distinct somatomotor skills of these limbs in agoutis., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Introduction: Acute neurodegenerative diseases, including stroke and traumatic brain and spinal cord injury, possess an elevated worldwide incidence. Two distinct lesive patterns can be identified after these destructive events: primary damage, an early consequence of the primary pathological event, and secondary neural degeneration (SND), a group of pathological events inducing late degeneration in cells not or even only partially affected by the primary damage. This pathological mechanism is an important contributing factor for functional deficits and target for therapeutic approaches. Several factors are involved on the SND etiology, including excitotoxicity, inflammation, and oxidative stress., Aim: To review the main mechanisms underlying the SND occurring after acute neural disorders., Development: The more recent findings about the eliciting processes of SND degeneration are discussed, as well as their significance to degeneration of white matter tracts., Conclusions: The characterization of the events underlying SND is of fundamental importance for the development of new therapeutic approaches effective enough to decrease the functional deficits, contributing to the improvement of the quality of life of people suffering neurological diseases. These therapeutic approaches must be validated in experimental models of both brain and spinal cord diseases, which effectively simulate human neural disorders protecting both gray and white matters for a better neuroprotective efficacy.

Following acute and chronic neurodegenerative disorders, a cascade of pathological events including inflammatory response, excitotoxicity and oxidative stress induces secondary tissue loss in both gray and white matter. Axonal damage and demyelination are important components of the white matter demise during these diseases. In spite of this, a few studies have addressed the patterns of inflammatory response, axonal damage and demyelination following focal ischemic damage to the central nervous system (CNS). In the present study, we describe the patterns of inflammatory response, axonal damage and myelin impairment following microinjections of 10 pmol of endothelin-1 into the rat striatum. Animals were perfused at 1 day, 3 days and 7 days after injection. 20 mum sections were stained by hematoxylin and immunolabeled for neutrophils (anti-MBS-1), activated macrophages/microglia (anti-ED1), damaged axons (anti-betaAPP) and myelin (anti-MBP). The evolution of acute inflammation was quantitatively assessed by cell counts in different survival times. There was recruitment of both neutrophils and macrophages to the damaged striatal parenchyma with maximum recruitment at 1 day and 7 days, respectively. Progressive myelin impairment in the striatal white matter tracts has been observed mainly at later survival times. beta-APP+ endbulbs were not present in all evaluated time points. These results suggest that progress myelin impairment in the absence of damage to axonal cylinder is a feature of white matter pathology following endothelin-1-induced focal striatal ischemia.

Inflammatory response, axonal damage and demyelination are important components of the pathophysiology of acute neurodegenerative diseases. We have investigated the outcome of these pathological events following an excitotoxic or an ischemic damage to the spinal nucleus of adult rats at 1 and 7 days postinjury. Microinjections of 80 nmol of NMDA or 40 pmol of endothelin-1 into the rat spinal nucleus induced differential histopathological events. NMDA injection induced intense tissue loss in the gray matter (GM) without significant tissue loss in the white matter (WM). There was a mild inflammatory response, with recruitment of a few neutrophils and macrophages. Axonal damage was present in the GM following NMDA injection, with negligible axonal damage in the WM. Myelin impairment was apparent at 7 days. Microinjections of endothelin-1 into the same region induced lesser tissue loss than NMDA injections, concomitant with an intense inflammatory response characterized by recruitment of macrophages, but not of neutrophils. There were more axonal damage and early myelin impairment after endothelin-1 injection. These results were confirmed by quantitative analysis. Microcysts were present in the WM of the trigeminothalamic tract at 7 days following injection of endothelin-1. These results show that an ischemic damage to the spinal nucleus affects both GM and WM with more bystander inflammation, axonal damage and myelin impairment, while excitotoxic damage induces effects more restricted to the GM. These pathological events may occur following acute damage to the human brain stem and can be an important contributing factor to the underlying functional deficits.

Rodentia is the largest mammalian order, with more than 2,000 species displaying a great diversity of morphological characteristics and living in different ecological niches (terrestrial, semi-aquatic, arboreal and fossorial). Analysis of the organization of the somatosensory areas in six species of rodents allowed us to demonstrate that although these species share a similar neocortical blueprint with other eutherian mammals, important differences exist between homologous areas across different species, probably as a function of both lifestyle and peripheral sensory specializations typical of each species. We based this generalization on a phylogenetic comparison of the intrinsic organization of the primary somatosensory area (SI) across representatives of different rodent suborders. This analysis revealed considerable structural variability, including the differential expansion of cortical representation of specific body parts (cortical amplification) as well as the parcellation of areas into processing modules.

Viral neurotropism is the ability of viruses to infect neuronal cells. This is well studied for herpesviruses, rabies-related viruses, and a few others, but it is poorly investigated among almost all arboviruses. In this study, we describe both the neurotropism and the neuropathological effects of Amazonian rhabdoviruses on the brains of experimentally infected-newborn mice. Suckling mice were intranasally infected with 10(-4) to 10(-8) LD50 of viruses. Animals were anaesthetized and perfused after they had become sick. Immunohistochemistry using specific anti-virus and anti-active caspase three antibodies was performed. All infected animals developed fatal encephalitis. Survival time ranged from 18 h to 15 days. Viruses presented distinct species-dependent neurotropism for CNS regions. Histopathological analysis revealed variable degrees of necrosis and apoptosis in different brain regions. These results showed that viruses belonging to the Rhabdoviridae family possess distinct tropism for CNS structures and induce different pattern of cell death depending on the CNS region.

The mechanisms of white matter (WM) damage during secondary degeneration are a fundamental issue in the pathophysiology of central nervous system (CNS) diseases. Our main goal was to describe the pattern of an acute inflammatory response and secondary damage to axons in different WM tracts of acutely injured rat spinal cord. Adult rats were deeply anesthetized and injected with 20 nmol of NMDA into the spinal cord ventral horn on T7. Animals were perfused after survival times of 1 day, 3 days and 7 days. Ten micrometer sections were submitted to immunocytochemical analysis for activated macrophages/microglia, neutrophils and damaged axons. There were inflammatory response and progressive tissue destruction of ventral WM (VWM) with formation of microcysts in both VWM and lateral WM (LWM). In the VWM, the number of beta-amyloid precursor protein (beta-APP) end-bulbs increased from 1 day with a peak at 3 days, decreasing by 7 days following the injection. APP end-bulbs were present in the dorsal WM (DWM) at 3 days survival time but were not in the LWM. Electron microscopic analysis revealed different degrees of myelin disruption and axonal pathology in the vacuolated WM up to 14 mm along the rostrocaudal axis. Quantitative analysis revealed a significant loss of medium and large axons (P < 0.05), but not of small axons (P > 0.05). Our results suggest that bystander axonal damage and myelin vacuolation are important secondary component of the pathology of WM tracts following rat SCI. Further studies are needed to understand the mechanisms of these pathological events.

We investigated the histochemical positivity to NADPH-diaphorase, which reveals nitric oxide synthase activity, in area 17 of rats malnourished early in life, both in the post-weaning period (group M1), and in adulthood after nutritional recovering (group M2). Control pups (C1 and C2 groups) received ad libitum after weaning the same diets as their mothers. Rats of group M2 were nutritionally recovered by receiving the control diet from post-natal day 42 until adulthood. Aldehyde-fixed sections (200-microm thick) through area 17 were processed for NADPH-diaphorase histochemistry following the malic enzyme indirect method. The features of NADPH-diaphorase-containing neurons of area 17 of malnourished young (M1) and adult (M2) rats were analyzed quantitatively in comparison to the matched groups C1 and C2. Permanent changes, represented by increase in the density and dendritic field areas of NADPH-diaphorase-positive cells, and transitory ones, represented by decreased values of soma areas, were observed in area 17 of the M1 and M2 cases. However, some other features, such as dendritic branch angle and number of dendrites per cell in the gray matter, remained unchanged after malnutrition. Thus, the findings indicate a possible relationship between early malnutrition and alterations in nitric oxide synthase-containing cells in the visual cortex. Physiological implications of these data may be related to synaptic plasticity and refinement of developmental brain circuits.

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.

The effects of methylmercury (MeHg) on histochemical demonstration of the NADPH-diaphorase (NADPH-d) activity in the striate cortex were studied in 4 adult cats. Two animals were used as control. The contaminated animals received 50 ml milk containing 0.42 microgram MeHg and 100 g fish containing 0.03 microgram MeHg daily for 2 months. The level of MeHg in area 17 of intoxicated animals was 3.2 micrograms/g wet weight brain tissue. Two cats were perfused 24 h after the last dose (group 1) and the other animals were perfused 6 months later (group 2). After microtomy, sections were processed for NADPHd histochemistry procedures using the malic enzyme method. Dendritic branch counts were performed from camera lucida drawings for control and intoxicated animals (N = 80). Average, standard deviation and Student t-test were calculated for each data group. The concentrations of mercury (Hg) in milk, fish and brain tissue were measured by acid digestion of samples, followed by reduction of total Hg in the digested sample to metallic Hg using stannous chloride followed by atomic fluorescence analysis. Only group 2 revealed a reduction of the neuropil enzyme activity and morphometric analysis showed a reduction in dendritic field area and in the number of distal dendrite branches of the NADPHd neurons in the white matter (P < 0.05). These results suggest that NADPHd neurons in the white matter are more vulnerable to the long-term effects of MeHg than NADPHd neurons in the gray matter.

This is the first report of the effects of malnutrition during brain development on biocytin-labelled axon terminals and histochemical pattern of NADPH-diaphorase (NADPH-d)-containing neurons in area 17 of the adult rat. Wistar rats (n = 6) were submitted early in life (from gestation up to 42 days of age) to a multideficient diet (the 'regional basic diet' (RBD) of low-income human populations of north-east Brazil, containing only 8% protein). From day 43 up to adulthood (135-212 days), they were switched to a commercial laboratory chow diet (Purina do Brasil Ltda), with 22% protein. These animals were compared to control rats (n = 11), fed the laboratory chow diet until adulthood. The brains of four adult malnourished and five controls were processed according to the indirect method of the malic enzyme to reveal NADPH-d-containing neurons. Five other adult subjects (three controls and two malnourished) received iontophoretic injections of the tracer biocytin in area 17and were processed according to the glucose oxidase-DAB-nickel protocol in order to visualize axon terminals filled with biocytin. Three other control rats were processed for both techniques. In these last brains, no double-labelled cells could be found, suggesting that the NADPH-d-containing-neurons and the biocytin-labelled ones belong to different groups of cells, in area 17. The appendages of the NADPH-d-positive cells showed minor qualitative and quantitative differences between undernourished and control rats. The soma area of these cells was reduced in the white matter of malnourished rats, as compared to the controls (468.6 ± 54.3 μm(2); n = 4 and 515.4 ± 30.5 μm(2);n = 8, respectively; p < 0.05). The cell density (cells/mm(2)) was greater in the malnourished group than in the control, both at the grey matter (16.6 ± 4.4; n = 4 and 11.3 ± 4.3; n = 8, respectively; p < 0.05) and at the white matter (55.9 ± 15.7; n = 4 and 24.4 ± 8.5; n = 8; p < 0.005). The number of potential synaptic sites in the biocytin-labelled axon terminals was reduced as compared to the control (126 ± 33 boutons/mm, n = 32 and 160 ± 37; n = 30, respectively; p < 0.01). The results indicate that the rat area 17 is affected differently by early malnutrition, regarding biocytin-labelled axon terminals, on the one hand, and NADPH-d-containing neurons, on the other. Concerning these last cells, the data also suggest that they are less sensitive to the injury represented by early malnutrition.

Metric features and modular and laminar distributions of intrinsic projections of area 17 were studied in Cebus apella. Anterogradely and retrogradely labeled cell appendages were obtained using both saturated pellets and iontophoretic injections of biocytin into the operculum. Laminar and modular distributions of the labeled processes were analyzed using Nissl counterstaining, and/or cytochrome oxidase and/or NADPH-diaphorase histochemistry. We distinguished three labeled cell types: pyramidal, star pyramidal and stellate cells located in supragranular cortical layers (principally in layers IIIa, IIIb alpha, IIIb beta and IIIc). Three distinct axon terminal morphologies were found, i.e., Ia, Ib and II located in granular and supragranular layers. Both complete and partial segregation of group I axon terminals relative to the limits of the blobs of V1 were found. The results are compatible with recent evidence of incomplete segregation of visual information flow in V1 of Old and New World primates.

Mercury is a neurotoxic agent that produces different effects on the brain. In the present work, the effects of chronic doses of methyl mercury (MeHg) were studied on the dopaminergic system of the rat striatum, using microdialysis coupled to high-performance liquid chromatography in order to quantify the in vivo release of dopamine (DA) and its acidic metabolites DOPAC and HVA. The administration of an equivalent total dose of 6 mg/kg of MeHg induced significant increases in the striatal release of DA and/or its acidic metabolites, independently of the pattern of administration. These effects are discussed on the base of the release and the metabolization of DA. In conclusion, the effect of MeHg administered under these experimental conditions on the in vivo release of DA and its metabolites seems to have a dose-dependent component and seems to be an accumulative process., (Copyright 1997 Academic Press.)

We studied the distribution of NADPH-diaphorase activity in the visual cortex of normal adult New World monkeys (Saimiri sciureus) using the malic enzyme "indirect" method. NADPH-diaphorase neuropil activity had a heterogeneous distribution. In coronal sections, it had a clear laminar pattern that was coincident with Nissl-stained layers. In tangential sections, we observed blobs in supragranular layers of V1 and stripes throughout the entire V2. We quantified and compared the tangential distribution of NADPH-diaphorase and cytochrome oxidase blobs in adjacent sections of the supragranular layers of V1. Although their spatial distributions were rather similar, the two enzymes did not always overlap. The histochemical reaction also revealed two different types of stained cells: a slightly stained subpopulation and a subgroup of deeply stained neurons resembling a Golgi impregnation. These neurons were sparsely spined non-pyramidal cells. Their dendritic arbors were very well stained but their axons were not always evident. In the gray matter, heavily stained neurons showed different dendritic arbor morphologies. However, most of the strongly reactive cells lay in the subjacent white matter, where they presented a more homogenous morphology. Our results demonstrate that the pattern of NADPH-diaphorase activity is similar to that previously described in Old World monkeys.

Metric features of the axon terminals and cell morphology of intrinsic projections of area 17 were studied in the Cebus apella. Anterograde and retrograde labeled cell appendages were obtained using saturated pellets and iontophoretic injections of biocytin in the operculum. Details of the histological and histochemical procedures have been described elsewhere (Amorim and Picanço-Diniz, 1996). We distinguished three labeled cell types: pyramidal, star pyramidal and stellate cells and three distinct morphologies of axon terminals were found: Ia, Ib and II, located at supragranular layers. Axon terminals of the group I innervate larger extent of striate cortex through longer intermediate segments, and acute branching angles compared to group II. Group II present on average similar characteristics of the smooth neurons axon terminals. The results taken together with the occurrence of only two types of synapses (I and II) from Gray's ultrastructural studies, seem to give an additional support to extend to the Cebus apella the major subdivision of neocortical neuronal morphology that classified them as smooth and spine neurons.

NADPH-diaphorase (NADPH-d) activity was studied comparatively in area 17 of four mammalian species, two primates and two rodents. Three brain hemispheres each from adult capuchin-monkeys, owl-monkeys, agoutis and guinea pigs were fixed with aldehyde fixatives by perfusion and 200 microns sections were submitted to NADPH-d histochemistry, using the indirect malic enzyme method. In all species studied the neuropil pattern of enzymes activity presented a clear layered appearance. In primates, histochemical staining was most intense in layer IVc, while in rodents the highest intensity of the neuropil reaction was in supragranular layers (II and III). Comparison of cell density in grey and white matter showed that the majority of NADPH-d-positive neurones were located in the white matter of primates but not of rodents. Since NADPH-d is a nitric oxide synthase the results are very important for comparative functional studies of neuromediators and their correlations with laminar and modular organization of area 17 of the mammalian brain.

A morphological study of intrinsic projection in area 17 of Cebus monkey was conducted after iontophoretic injection of biocytin. Thirty axon terminals located in supragranular layers were qualitatively and quantitatively analyzed using 3-d automatic microscopy. Three types of axon terminals could be identified: Ia, Ib and II. Group I was characterized by a sparse and/or long-distance branch pattern, while type II presented compact and localized arborization. Ia axon terminals formed "clusters" and "terminaux" boutons while Ib did not. On average, group II axon terminals tended to present straight or obtuse branching angles and a much more ramified pattern, and occupy a smaller cortical territory with shorter intermediate segments and higher density of synaptic potential sites than group I. The common characteristics of group I included innervation of larger cortical territories, longer intermediate segments, acute branching angles and lower synaptic density compared to group II. The results are compatible with the major subdivisions of neocortical neuronal morphology that classifies them as smooth and spine neurons. Smooth neurons may be related to axon terminals of group II while spine neurons may be related to group I.

Zif268 transcription factor is expressed throughout Cebus apella visual cortex at high basal levels. Monocular eyelid suture alters the levels of Zif268 on neurons connected to the deprived eye, revealing ocular dominance columns in the striate cortex of Cebus as previously demonstrated in Old World monkeys (Chaudhuri and Cynader, Brain Res., 605 (1993) 349-353). Zif268 ocular dominance columns are revealed in adult Cebus monkey after 24-48 h of monocular deprivation, but not in infant monkeys up to 3 months of age. In 6-month-old Cebus monkeys, Zif268 ocular dominance columns are still poorly defined. These results suggest that Zif268 ocular dominance columns establish late during normal primate visual system development, and that some degree of visual plasticity is still present at this age in the Cebus monkey.

The present report describes the activity of NADPH-diaphorase (NADPHd) in area 17 of autopsied normal human visual cortex. Four human brains from autopsy tissue (4-8 h postmortem) were fixed by immersion in 4% paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.2-7.4, or in 10% formalin for 24 h. NADPHd histochemistry was done using the malic enzyme indirect method. The neuropile pattern of enzyme activity presented a clear six-layer appearance. Cell morphology and the laminar distribution of 73 NADPHd-positive neurons are described. All neurons found in area 17 of human cortex were sparsely spiny or smooth cells, located in all cortical layers except layer 4c. Quantitative analysis of the branching pattern of the dendritic tree was carried out. A symmetrical pattern was observed with no particular dendritic bias except for a few white matter and layer 1 cells. Larger dendritic fields were found in white matter cells when compared to the other cortical layers. Comparison of cell densities for gray and white matters showed that 85% of the NADPHd-positive neurons were located in the white matter. NADPHd was colocalized with nitric oxide synthase which produces nitric oxide, a short-life neuromediator implicated in synaptic plasticity, neuroprotection, and neurotoxicity. Thus, the spatial distribution of the NADPHd cells is important for posterior functional studies of the neuromediators in the brain.

M and P retinal ganglion cell morphology revealed by biocytin retrograde labelling was compared in two closely related New-World monkeys, Cebus and Aotus, to investigate whether nocturnal and diurnal species of primates have similar cell classes. Monkey and cat ganglion cells from regions of matching cell class densities were also compared. Cat alpha, cat beta, Aotus M, and Cebus M cells were similar in many aspects, but Cebus M cells had higher branching density. Cebus and Aotus P cells formed a distinct group and represent a primate specialization common to diurnal and nocturnal simians.

We have examined the morphology of neurons in layer 1 by injecting them intracellularly with lucifer yellow in lightly fixed brain slices (250 microns thick) taken from the medial bank of area 17 in adult cats. Of 22 neurons with well-filled dendrites, 16 had smooth dendrites, two had sparsely spiny dendrites (less than 200 spines) and, unexpectedly, four had spiny dendrites typical of pyramidal cells. The axon was generally not well filled. Computer reconstructions showed that parts of the dendritic tree had been lost in the sectioning. Nevertheless, measurements of the length of intact dendrites suggested an average diameter of the dendritic tree of 220 microns. The density of the neurons was such that the dendritic trees of about six neurons cover each point in layer 1. Thus, despite the very low density of neurons that characterizes layer 1, there are more than sufficient neurons to sample from the entire representation of the visual field in area 17.

We have successfully used biocytin as a retrograde tracer in the mammalian visual system. Retinal ganglion cells, pyramidal and stellate cortical neurons were labelled. Both pressure injections and gel implants were used successfully for retrograde labelling. Biocytin was detected using avidin conjugates and horseradish peroxidase histochemistry. Retrograde filling with biocytin proved to be more reliable and to allow better morphological resolution than other commonly used neurotracers such as horseradish peroxidase. The fine details of cell morphology observable by this method are comparable in many cases to the results obtained with intracellular tracer injections. The morphological resolution obtained with this method allows the study of brain microcircuits using extracellular deposits of biocytin.

The cortical representation of the contralateral visual field in area 17 of the agouti's brain was studied by multiunit recording. The borders of area 17 were determined by electrophysiological, cytoarchitectonic and myeloarchitectonic criteria. The results were plotted in flat, bidimensional representations of the cerebral cortex to minimize perspective distortions. The V1 map, a first order topological transformation of the visual field, shows a magnified representation of the horizontal meridian that corresponds to a retinal specialization, the visual streak. The visual field representation has asymmetries that are not directly related to the topography of the retinal ganglion cell density. Whereas the ganglion cell density shows a plateau along the visual streak, the areal cortical magnification factor is higher in the region that corresponds to the intersection of the horizontal and vertical meridians. This suggests functional specializations that are not obvious when one considers the distribution of the whole ganglion cell population but which might be related to the distribution of specific ganglion cell classes.

The present report describes a head holder designed to be used for lateral-eyed species in vision research. The head-holder employs a prosthesis implanted on the skull and provides for adjustable movements in different planes, thus allowing precise positioning of the eye with respect to the visual space.

The Modulation Transfer Function (MTF) of the visual system of the opossum, D. marsupialis aurita, was determined using the amplitude of Visually Evoked Cortical Potentials (VECP) as response indicator. Stimuli consisted of a 180 degrees phase reversal of sinusoidally modulated gratings with an average luminance of 2.4 cd/m2. Contrast sensitivity was determined for various spatial frequencies and the MTF was calculated by the least square fit of an exponential function. The average acuity value obtained was 1.25 c/deg. The Fourier transform of the MTF was considered an approximation of the Line Spread Function of the visual system. The lowest value observed was 14 min of arc. The visual acuity observed in the mesopic range was not altered when stimulus intensity was raised to photopic levels.

The distribution of ganglion cells was determined in whole-mounted Cebus monkey retinae. Ganglion cell density along the horizontal meridian was asymmetric, being 1.2-4.3 higher in the nasal retinal region when compared to temporal retina at the same eccentricities. The total number of ganglion cells varied from 1.34 to 1.4 million. Ganglion cell density peaked at 49,000/mm2 about 0.5 mm nasal to the fovea. Comparison between ganglion cell density and areal cortical magnification factors for V1 and V2 reveals that the relative representation of the fovea increases in the visual cortex. This effect seems to be a general feature of the visual system of primates.

1. We have studied the cytoarchitecture and myeloarchitecture of the agouti's cortical surface that can be activated by visual stimulation. Five architectonic subdivisions that correspond to distinctive visuotopic representations were characterized. 2. The largest portion of the visual cortex is occupied by area 17 which is situated lateral to the cingulate cortex, medial to area 18, posterior to the parietal cortex, and anterior to the agranular retrosplenial cortex. Additionally, four architectonic subdivisions in the extrastriate visual cortex were distinguished, i.e., from medial to lateral: area 18, area 19, anterior lateral area, and temporal posterior area. 3. Along the border of the extrastriate cortex a ring of nonvisual cortical fields was encountered encompassing parietal (somatic sensorial) cortex, temporal anterior and temporal intermediate (auditory) areas, a band of pre-rhinal cortex, and agranular retrosplenial cortex.

The refractive state of the eye of the South American opossum Didelphis marsupialis aurita was investigated with electrophysiological techniques. Using adult specimens, trapped from the wild, averaged cortical evoked responses were recorded from the region of projection of the central visual field. Stimuli consisted of a phase reversal of a square wave grating generated on a CRO screen, with luminance of 2.4 cd/m2 and contrast 0.84. The refractive state of the eye was altered by means of trial lenses and the amplitude of the cortical responses thus obtained compared to those obtained with no lens (control values). Refraction "tuning curves" were determined for each animal. The average refractive state was found to be -2.27 D indicating that this species when raised in its habitat shows, at low ambient luminosity, some degree of myopia. Determination of the Contrast Sensitivity Function indicate that induced ametropias lead to a reduction of the cut-off value of the spatial frequency and a loss of contrast sensitivity.