Impact of Monocular Retinal Lesions on Blob Size in Adult Human V1.

We studied the attributes of cytochrome c oxidase (CytOx)-rich blobs and ocular dominance columns (OD) in human V1 associated with monocular retinal lesions. Interblob distance, blob cross-sectional area, OD width, and OD arrangement pattern were analyzed in CytOx-reacted tangential sections of flat-mounted V1 preparations. Monocular deprivation induces differential expression of CytOx in the corresponding ODs in V1. We were thereby able to identify the V1 regions associated with the lesioned area in the retina, assign which OD was associated with each eye, and assign the corresponding blob in Layer III as deprived or nondeprived of visual input. We found that nondeprived blobs are more conspicuously stained than blobs outside the lesioned area. Notably, we found a selective expansion of blobs associated with the nonlesioned eye, whereas blobs associated with the deprived eye showed no significant change in size. Blob size in the latter condition was similar to the one observed in normal participants. These effects were present throughout the representation of the lesion in V1, suggesting that the underlying plasticity mechanisms do not depend on eccentricity. Retinal lesion caused no change in interblob distance, which was comparable to the normal brain (i.e., participants with no retinal lesion). This indicates that blob center is a stable hallmark of cortical organization. Finally, the width of ODs associated with the nonlesioned eye tended to be larger compared with ODs of the lesioned eye. However, this effect did not reach statistical significance. The stability of ODs thereby contrasts with blob plasticity, suggesting that the retinal lesion-triggered imbalance in the thalamocortical projection to Layer IVc has a limited impact on OD CytOx reactivity. On the other hand, we argue that ocular imbalance supports intracortical lateral competition that increases CytOx reactivity in the periblob region associated with the nonlesioned eye, accounting for the blob expansion we observe.

Cortical maps as a fundamental neural substrate for visual representation.

Visual perception is the product of serial hierarchical processing, parallel processing, and remapping on a dynamic network involving several topographically organized cortical visual areas. Here, we will focus on the topographical organization of cortical areas and the different kinds of visual maps found in the primate brain. We will interpret our findings in light of a broader representational framework for perception. Based on neurophysiological data, our results do not support the notion that vision can be explained by a strict representational model, where the objective visual world is faithfully represented in our brain. On the contrary, we find strong evidence that vision is an active and constructive process from the very initial stages taking place in the eye and from the very initial stages of our development. A constructive interplay between perceptual and motor systems (e.g., during saccadic eye movements) is actively learnt from early infancy and ultimately provides our fluid stable visual perception of the world.

E.L., a modern-day Phineas Gage: Revisiting frontal lobe injury.

Background: How the prefrontal cortex (PFC) recovers its functionality following lesions remains a conundrum. Recent work has uncovered the importance of transient low-frequency oscillatory activity (LFO; < 4 Hz) for the recovery of an injured brain. We aimed to determine whether persistent cortical oscillatory dynamics contribute to brain capability to support 'normal life' following injury. Methods: In this 9-year prospective longitudinal study (08/2012-2021), we collected data from the patient E.L., a modern-day Phineas Gage, who suffered from lesions, impacting 11% of his total brain mass, to his right PFC and supplementary motor area after his skull was transfixed by an iron rod. A systematic evaluation of clinical, electrophysiologic, brain imaging, neuropsychological and behavioural testing were used to clarify the clinical significance of relationship between LFO discharge and executive dysfunctions and compare E.L.´s disorders to that attributed to Gage (1848), a landmark in the history of neurology and neuroscience. Findings: Selective recruitment of the non-injured left hemisphere during execution of unimanual right-hand movements resulted in the emergence of robust LFO, an EEG-detected marker for disconnection of brain areas, in the damaged right hemisphere. In contrast, recruitment of the damaged right hemisphere during contralateral hand movement, resulted in the co-activation of the left hemisphere and decreased right hemisphere LFO to levels of controls enabling performance, suggesting a target for neuromodulation. Similarly, transcranial magnetic stimulation (TMS), used to create a temporary virtual-lesion over E.L.'s healthy hemisphere, disrupted the modulation of contralateral LFO, disturbing behaviour and impairing executive function tasks. In contrast to Gage, reasoning, planning, working memory, social, sexual and family behaviours eluded clinical inspection by decreasing LFO in the delta frequency range during motor and executive functioning. Interpretation: Our study suggests that modulation of LFO dynamics is an important mechanism by which PFC accommodates neurological injuries, supporting the reports of Gage´s recovery, and represents an attractive target for therapeutic interventions. Funding: Fundação de Amparo Pesquisa Rio de Janeiro (FAPERJ), Universidade Federal do Rio de Janeiro (intramural), and Fiocruz/Ministery of Health (INOVA Fiocruz).

Genetic variability of the sws1 cone opsin gene among New World monkeys.

Vision is a major sense for Primates and the ability to perceive colors has great importance for the species ecology and behavior. Visual processing begins with the activation of the visual opsins in the retina, and the spectral absorption peaks are highly variable among species. In most Primates, LWS/MWS opsins are responsible for sensitivity to long/middle wavelengths within the visible light spectrum, and SWS1 opsins provide sensitivity to short wavelengths, in the violet region of the spectrum. In this study, we aimed to investigate the genetic variation on the sws1 opsin gene of New World monkeys (NWM) and search for amino acid substitutions that might be associated with the different color vision phenotypes described for a few species. We sequenced the exon 1 of the sws1 opsin gene of seven species from the families Callitrichidae, Cebidae, and Atelidae, and searched for variation at the spectral tuning sites 46, 49, 52, 86, 90, 93, 114, 116, and 118. Among the known spectral tuning sites, only residue 114 was variable. To investigate whether other residues have a functional role in the SWS1 absorption peak, we performed computational modeling of wild-type SWS1 and mutants A50I and A50V, found naturally among the species investigated. Although in silico analysis did not show any visible effect caused by these substitutions, it is possible that interactions of residue 50 with other sites might have some effect in the spectral shifts in the order of ~14 nm, found among the NWM. We also performed phylogenetic reconstruction of the sws1 gene, which partially recovered the species phylogeny. Further studies will be important to uncover the mutations responsible for the phenotypic variability of the SWS1 of NWM, and how spectral tuning may be associated with specific ecological features such as preferred food items and habitat use.

A subacute model of glaucoma based on limbal plexus cautery in pigmented rats.

Glaucoma is a neurodegenerative disorder characterized by the progressive functional impairment and degeneration of the retinal ganglion cells (RGCs) and their axons, and is the leading cause of irreversible blindness worldwide. Current management of glaucoma is based on reduction of high intraocular pressure (IOP), one of its most consistent risk factors, but the disease proceeds in almost half of the patients despite such treatments. Several experimental models of glaucoma have been developed in rodents, most of which present shortcomings such as high surgical invasiveness, slow learning curves, damage to the transparency of the optic media which prevents adequate functional assessment, and variable results. Here we describe a novel and simple method to induce ocular hypertension in pigmented rats, based on low-temperature cauterization of the whole circumference of the limbal vascular plexus, a major component of aqueous humor drainage and easily accessible for surgical procedures. This simple, low-cost and efficient method produced a reproducible subacute ocular hypertension with full clinical recovery, followed by a steady loss of retinal ganglion cells and optic axons, accompanied by functional changes detected both by electrophysiological and behavioral methods.

Distribution of cytochrome oxidase-rich patches in human primary visual cortex.

We studied the tangential distribution of cytochrome oxidase (CytOx)-rich patches (blobs) in the striate cortex (V1) of normally sighted Homo sapiens. We analyzed the spatial density and cross-sectional area of patches in CytOx-reacted tangential sections of flat-mounted preparations of V1 and surrounding areas. CytOx-rich patches were most clearly defined in the supragranular cortical layers of V1, particularly at middle levels of layer III. Variations in patch spatial density were subtle across different visual eccentricity representations. Within the binocular representation of V1, the average patch spatial density decreased slightly with increasing cortical eccentricity, from around 1.0 patch/mm2 in the foveal representation to 0.6 patch/mm2 at the representation of ∼60° eccentricity, but seemed to increase again at the representation of the monocular crescent. Across the entire sample, the cross-sectional area of patches (i.e., patch size) varied from approximately 0.2-0.8 mm2 , with a mean value of 0.32 mm2 . Notably, there was no significant variation in the mean patch size across eccentricity representations. Human patches are on average larger than those reported for nonhuman primate brains, and analysis of species with different brain sizes suggests an approximately linear relationship between V1 area and patch size. The relative constancy of patch metrics across eccentricities is in stark contrast with the exponential variation in V1 cortical magnification, suggesting a nearly invariant modular organization throughout human V1.

Neuronal response properties across cytochrome oxidase stripes in primate V2.

Cytochrome oxidase histochemistry reveals large-scale cortical modules in area V2 of primates known as thick, thin, and interstripes. Anatomical, electrophysiological, and tracing studies suggest that V2 cytochrome oxidase stripes participate in functionally distinct streams of visual information processing. However, there is controversy whether the different V2 compartments indeed correlate with specialized neuronal response properties. We used multiple-electrode arrays (16 × 2, 8 × 4 and 4 × 4 matrices) to simultaneously record the spiking activity (N = 190 single units) across distinct V2 stripes in anesthetized and paralyzed capuchin monkeys (N = 3 animals, 6 hemispheres). Visual stimulation consisted of moving bars and full-field gratings with different contrasts, orientations, directions of motion, spatial frequencies, velocities, and color contrasts. Interstripe neurons exhibited the strongest orientation and direction selectivities compared to the thick and thin stripes, with relatively stronger coding for orientation. Additionally, they responded best to higher spatial frequencies and to lower stimulus velocities. Thin stripes showed the highest proportion (80%) of neurons selective to color contrast (compared to 47% and 21% for thick and interstripes, respectively). The great majority of the color selective cells (86%) were also orientation selective. Additionally, thin stripe neurons continued to increase their firing rate for stimulus contrasts above 50%, while thick and interstripe neurons already exhibited some degree of response saturation at this point. Thick stripes best coded for lower spatial frequencies and higher stimulus velocities. In conclusion, V2 CytOx stripes exhibit a mixed degree of segregation and integration of information processing, shedding light into the early mechanisms of vision.

Frontal Alpha Asymmetry and Theta Oscillations Associated With Information Sharing Intention.

Social media has gained increasing importance in many aspects of everyday life, from building relationships to establishing collaborative networks between individuals worldwide. Sharing behavior is an essential part of maintaining these dynamic networks. However, the precise neural factors that could be related to sharing behavior in online communities remain unclear. In this study, we recorded electroencephalographic (EEG) oscillations of human subjects while they were watching short videos. The subjects were later asked to evaluate the videos based on how much they liked them and whether they would share them. We found that, at the population level, subjects watching videos that would not be shared had higher power spectral density (PSD) amplitudes in the theta band (4-8 Hz), primarily over the frontal and parietal sites of the right hemisphere, than subjects watching videos that would be shared. Previous studies have associated task disengagement with an increase in scalp-wide theta activation, which can be interpreted as a mind-wandering effect. This might suggest that the decision to not share the video may lead to a more automatic/effortless neural pattern. We also found that watching videos that would be shared was associated with lower PSD amplitudes in the alpha band (8-12 Hz) over the central and right frontal sites, and with more negative scores of frontal alpha asymmetry (FAA) index scores. These results may be related to previous work linking right-sided frontal EEG asymmetry to the pursuit of social conformity and avoidance of negative outcomes, such as social isolation. Finally, using support vector machine (SVM) algorithms, we show that these EEG parameters and preference rating scores can be used to improve the predictability of sharing information behavior. The information sharing-related EEG pattern described here could therefore improve our understanding of the neural markers associated with sharing behavior and contribute to studies about stimuli propagation.

Precise visuotopic organization of the blind spot representation in primate V1.

The optic disk is a region of the retina consisting mainly of ganglion cell axons and blood vessels, which generates a visual scotoma known as the blind spot (BS). Information present in the surroundings of the BS can be used to complete the missing information. However, the neuronal mechanisms underlying these perceptual phenomena are poorly understood. We investigate the topography of the BS representation (BSR) in cortical area V1 of the capuchin monkey, using single and multiple electrodes. Receptive fields (RFs) of neurons inside the BSR were investigated using two distinct automatic bias-free mapping methods. The first method (local mapping) consisted of randomly flashing small white squares. For the second mapping method (global mapping), we used a single long bar that moved in one of eight directions. The latter stimulus was capable of eliciting neuronal activity inside the BSR, possibly attributable to long-range surround activity taking place outside the borders of the BSR. Importantly, we found that the neuronal activity inside the BSR is organized topographically in a manner similar to that found in other portions of V1. On average, the RFs inside the BS were larger than those outside. However, no differences in orientation or direction tuning were found between the two regions. We propose that area V1 exhibits a continuous functional topographic map, which is not interrupted in the BSR, as expected by the distribution of photoreceptors in the retina. Thus V1 topography is better described as "visuotopic" rather than as a discontinuous "retinotopic" map.

Automatic mapping of visual cortex receptive fields: a fast and precise algorithm.

An important issue for neurophysiological studies of the visual system is the definition of the region of the visual field that can modify a neuron's activity (i.e., the neuron's receptive field - RF). Usually a trade-off exists between precision and the time required to map a RF. Manual methods (qualitative) are fast but impose a variable degree of imprecision, while quantitative methods are more precise but usually require more time. We describe a rapid quantitative method for mapping visual RFs that is derived from computerized tomography and named back-projection. This method finds the intersection of responsive regions of the visual field based on spike density functions that are generated over time in response to long bars moving in different directions. An algorithm corrects the response profiles for latencies and allows for the conversion of the time domain into a 2D-space domain. The final product is an RF map that shows the distribution of the neuronal activity in visual-spatial coordinates. In addition to mapping the RF, this method also provides functional properties, such as latency, orientation and direction preference indexes. This method exhibits the following beneficial properties: (a) speed; (b) ease of implementation; (c) precise RF localization; (d) sensitivity (this method can map RFs based on few responses); (e) reliability (this method provides consistent information about RF shapes and sizes, which will allow for comparative studies); (f) comprehensiveness (this method can scan for RFs over an extensive area of the visual field); (g) informativeness (it provides functional quantitative data about the RF); and (h) usefulness (this method can map RFs in regions without direct retinal inputs, such as the cortical representations of the optic disc and of retinal lesions, which should allow for studies of functional connectivity, reorganization and neural plasticity). Furthermore, our method allows for precise mapping of RFs in a 30° by 30° area of the visual field for an array of microelectrodes of any size in less than 6 min.

Quantification of early stages of cortical reorganization of the topographic map of V1 following retinal lesions in monkeys.

We quantified the capacity for reorganization of the topographic representation of area V1 in adult monkeys. Bias-free automated mapping methods were used to delineate receptive fields (RFs) of an array of neuronal clusters prior to, and up to 6 h following retinal lesions. Monocular lesions caused a significant reorganization of the topographic map in this area, both inside and outside the cortical lesion projection zone (LPZ). Small flashed stimuli revealed responses up to 0.85 mm inside the boundaries of the LPZ, with RFs representing regions of undamaged retina immediately surrounding the lesion. In contrast, long moving bars that spanned the scotoma resulting from the lesion revealed responsive units up to 1.87 mm inside the LPZ, with RFs representing interpolated responses in this region. This reorganization is present immediately after monocular retinal lesioning. Both stimuli showed a similar and significant (5-fold) increase of the RF scatter in the LPZ, 0.56 mm (median), compared with the undamaged retina, 0.12 mm. Our results reveal an array of preexisting subthreshold functional connections of up to 2 mm in V1, which can be rapidly mobilized independently from the differential qualitative reorganization elicited by each stimulus.

GABA inactivation of area V4 changes receptive-field properties of V2 neurons in Cebus monkeys.

To investigate the contribution of feedback circuits from area V4 to the receptive-field properties of V2 neurons, we used tungsten microelectrodes to record extracellular single units in these visual areas, before and after pressure injections of a solution of 0.25 mol/L of GABA in two anesthetized and paralyzed Cebus apella monkeys. The visual stimulus consisted of a single bar moving in one of eight directions. Using a device made of four stainless steel pipettes and one central tungsten electrode, we inactivated, with different amounts of GABA, topographically corresponding areas of V4, while studying V2 neurons. We studied a total of 36 V2 neurons during six sessions of GABA injections into area V4. GABA inactivation of visual area V4 produced a general decrease in the excitability of the neurons, which included a decrease in spontaneous and driven activities, followed by changes in direction selectivity. The changes in selectivity were toward an increase in directional selectivity and decrease in orientation selectivity. Thus, feedback connections arising from V4, an area of the ventral steams of visual information processing, are capable of not only modulating the spontaneous and driven activity of V2 neurons, but also of modifying V2 receptive field properties, such as its direction and/or orientation selectivity.

GABA inactivation of visual area MT modifies the responsiveness and direction selectivity of V2 neurons in Cebus monkeys.

We investigated the contribution of the projections from area MT to the receptive field properties of cells in visual area V2 in anesthetized and paralyzed Cebus apella monkeys. We recorded extracellular single-unit activity using tungsten microelectrodes in three monkeys before and after pressure injection of a 0.25-mol/l GABA solution. The visual stimulus consisted of a single bar moving in one of eight directions. In total, 72 V2 neurons were studied in 18 sessions of GABA injection into area MT. A group of 22 neurons was investigated over a shorter period of time ranging from 15 to 60 min, during which the activity did not return to baseline levels. The remaining 50 neurons were studied over a period of at least 2 h, and no statistical difference was observed in the neuronal response before and long after GABA inactivation. The effects on these 50 neurons consisted of an early (1-20 min) significant general decrease in excitability with changes in either orientation or direction selectivity. The differential decrease in excitability resulted in an intermediate improvement (20-40 min) of the signal-to-noise ratio for the stimulus-driven activity. The inactivation depended on the quantity of GABA injected into area MT and persisted for a period of 2 h. The GABA inactivation in area MT produced inhibition of most cells (72%) and a significant change of direction tuning in the majority (56%) of V2 neurons. Both increases and also decreases in the direction tuning of V2 neurons were observed. These feedback projections are capable of modulating not only the levels of spontaneous and driven activity of V2 neurons but also the V2 receptive field properties, such as direction selectivity.

Cone photopigment variations in Cebus apella monkeys evidenced by electroretinogram measurements and genetic analysis.

We investigated the color vision pattern in Cebus apella monkeys by means of electroretinogram measurements (ERG) and genetic analysis. Based on ERG we could discriminate among three types of dichromatic males. Among females, this classification is more complex and requires additional genetic analysis. We found five among 10 possible different phenotypes, two trichromats and three dichromats. We also found that Cebus present a new allele with spectral peak near 552nm, with the amino acid combination SFT at positions 180, 277 and 285 of the opsin gene, in addition to the previously described SYT, AFT and AFA alleles.

Distribution of neurofilament proteins in the lateral geniculate nucleus, primary visual cortex, and area MT of adult Cebus monkeys.

We investigated the distribution pattern of SMI-32-immunopositive cells in the lateral geniculate nucleus (LGN) and in the primary (V1) and middle temporal (MT) cortical visual areas of the adult New World monkey Cebus apella. In the LGN, the reaction for SMI-32 labeled cells in both the magnocellular (M) and parvocellular (P) layers. However, the cellular label was heavier in M layers, which also showed a more intense labeling in the neuropil. In V1, the reaction showed a lamination pattern, with the heaviest labeling occurring in layer 4B and upper layer 6 (layers that project to area MT). Area MT shows a dense band of labeled neuropil and large pyramidal neurons in layer 3, large darkly labeled but less densely packed neurons in layer 5, and a population of small, lightly labeled cells in layer 6. These results resemble those found in other New and Old World monkeys, which suggest that the preferential labeling of projection neurons associated with fast-conducting pathways to the extrastriate dorsal stream is a common characteristic of simian primates. In the superficial layers of V1 in Cebus monkeys, however, SMI-32-labeled neurons are found in both cytochrome oxidase blobs and interblob regions. In this aspect, our results in Cebus are similar to those found in the Old World monkey Macaca and different from those described for squirrel monkey, a smaller New World Monkey. In Cebus, as well as in Macaca, there is no correlation between SMI-32 distribution and the blob pattern.

Parallel evolution of cortical areas involved in skilled hand use.

Dexterous hands, used to manipulate food, tools, and other objects, are one of the hallmarks of primate evolution. However, the neural substrate of fine manual control necessary for these behaviors remains unclear. Here, we describe the functional organization of parietal cortical areas 2 and 5 in the cebus monkey. Whereas other New World monkeys can be quite dexterous, and possess a poorly developed area 5, cebus monkeys are the only New World primate known to use a precision grip, and thus have an extended repertoire of manual behaviors. Unlike other New World Monkeys, but much like the macaque monkey, cebus monkeys possess a proprioceptive cortical area 2 and a well developed area 5, which is associated with motor planning and the generation of internal body coordinates necessary for visually guided reaching, grasping, and manipulation. The similarity of these fields in cebus monkeys and distantly related macaque monkeys with similar manual abilities indicates that the range of cortical organizations that can emerge in primates is constrained, and those that emerge are the result of highly conserved developmental mechanisms that shape the boundaries and topographic organizations of cortical areas.

Distribution of calbindin-28kD and parvalbumin in V1 in normal adult Cebus apella monkeys and in monkeys with retinal lesions.

Several proteins have their normal patterns of distributions altered by monocular visual deprivation. We studied the distribution of the calcium-binding proteins calbindin-28kD (Cb) and parvalbumin (Pv) in V1 in normal adult Cebus apella monkeys and in monkeys with monocular retinal lesions. In normal monkeys, the interblobs regions in layers 2/3 and the layer 4B are intensely labeled for Cb, while Pv reaction showed a complementary labeling pattern with a stronger staining in layers 4A, 4C and in the blob regions in layers 2/3. In monkeys with monocular retinal lesion, the laminar distribution of these proteins was differentially affected, although both reactions resulted in stronger labeling in non-deprived ocular dominance columns. While Cb reaction resulted in stronger labeling in layers 1 through 5, Pv labeling was heavier in layers 2/3, 4A and 4C. There was a clear reduction in the intensity of neuropil staining for both Pv and Cb in deprived ocular dominance columns with little or no reduction in number of labeled cells. This reduction could thus be attributed to activity-dependent changes at synapses level.

Cortical visual areas in monkeys: location, topography, connections, columns, plasticity and cortical dynamics.

The visual system is constantly challenged to organize the retinal pattern of stimulation into coherent percepts. This task is achieved by the cortical visual system, which is composed by topographically organized analytic areas and by synthetic areas of the temporal lobe that have more holistic processing. Additional visual areas of the parietal lobe are related to motion perception and visuomotor control. V1 and V2 represent the entire visual field. MT represents only the binocular field, and V4 only the central 30 degrees-40 degrees. The parietal areas represent more of the periphery. For any eccentricity, the receptive field grows at each step of processing, more at anterior areas in the temporal lobe. Minimal point image size increases towards the temporal lobe, but remains fairly constant toward the parietal lobe. Patterns of projection show asymmetries. Central V2 and V4 project mainly to the temporal lobe, while peripherals V2 (more than 30 degrees) and V4 (more than 10 degrees) also project to the parietal lobe. Visual information that arrives at V1 projects to V2, MT and PO, which then project to other areas. Local lateral propagation and recursive loops corroborate to perceptual completion and filling in. Priority connections to temporal, parietal and parieto-temporal cortices help construct crude early representations of objects, trajectories and movements.

Differential expression of Zif268 and c-Fos in the primary visual cortex and lateral geniculate nucleus of normal Cebus monkeys and after monocular lesions.

The transcription factors c-Fos and Zif268 have been used as markers of neuronal activity, and they also have been implicated in neuronal plasticity. In this study, we investigated the expression of c-Fos and Zif268 proteins in the lateral geniculate nucleus (LGN) and in the cortical primary visual area (V1) of normal adult Cebus apella monkeys and in animals with monocular lesions. In the LGN, the reaction for c-Fos showed immunopositive cells in both magnocellular (M) and parvocellular (P) layers; however, the label was heavier in P layers. In animals that suffered monocular lesions, the immunocytochemistry for c-Fos showed more labeling in layers related to the normal eye compared with those of the lesioned eye. No specific label was observed after the reaction for Zif268 in the LGN. In V1, the reaction for both Zif268 and c-Fos showed a pattern of lamination in which heavier labeling was found in layers 2/3, 4A, 4C, and 6. After monocular lesions, we observed a clear pattern of ocular dominance columns in V1 for both c-Fos and Zif268, in which the columns related to the normal eye are more heavily labeled than those related to the lesioned eye. This pattern is more evident in layer 4C after c-Fos reaction, whereas, after Zif268, it is more clearly observed in layers 2/3. These results suggest that, in addition to be regulated by functional activity, these transcription factors are involved in different processes during cortical reorganization.

Three streams of visual information processing in V2 of Cebus monkey.

Gattass and collaborators (Gattass R, Rosa MGP, Souza APB, Piñon MCG, Neuenschwander S [1990a] Braz J Med Biol Res 23:375-393) proposed that the dorsal stream of visual processing, as defined by Ungerleider and Mishkin (Ungerleider LG, Mishkin M [1982] In: Ingle DJ, Goodale MA, Mansfield RJW, editors. Analysis of visual behavior. Cambridge: Massachusetts Institute of Technology. p 549-586), can be subdivided into dorsolateral and dorsomedial streams, and suggested that they may be involved in different aspects of the processing of motion and spatial perception, respectively. The goal of the present study was to provide additional evidence for this hypothesis by using cytochrome oxidase immunohistochemistry combined with retrograde tracing techniques. In Old World monkeys, the locations of visual area 4 (V4; ventral stream) and middle temporal area (MT; dorsal stream) projecting neurons in V2 supports the hypothesis that the cytochrome oxidase (CytOx)-rich thin stripes and the CytOx-poor interstripes are associated with the ventral stream, and that the CytOx-rich thick stripes belong to the dorsal stream. In this study we describe, in the New World monkey Cebus, the distribution of retrogradely labeled cells in V2 relative to the CytOx compartments after fluorescent tracers were placed in areas V4, MT, and the parietooccipital area (PO). We found PO-projecting neurons in CytOx-rich thick stripes and CytOx-poor interstripes in V2, whereas MT-projecting neurons appeared almost exclusively in thick stripes. In contrast, V4-projecting neurons were located mostly in CytOx-poor interstripes and CytOx-rich thin stripes. In addition, V4- and MT-projecting neurons were located mainly in supragranular layers, whereas PO-projecting neurons were located in supragranular and infragranular layers. These results support the hypothesis for the existence of three distinct streams of visual processing: ventral (including V4), dorsolateral (including MT), and dorsomedial (including PO).