Connectional evidence for dorsal and ventral V3, and other extrastriate areas in the prosimian primate,Galago garnetti

Previously we described patterns of connections that support the concept of V3 in small New World marmoset monkeys, three species of larger New World monkeys, and two species of Old World macaque monkeys. Here we describe a pattern of V1 connections with extrastriate visual cortex in Galago garnetti (also known as Otolemur garnetti) that demonstrates the existence of a V3 in a strepsirhine (prosimian) primate. Injections of fluorochromes or cholera toxin subunit-B (CTB) in V1 labeled cells and terminals in retinotopically matched regions in V2, V3, DL (V4), and MT. Labeled axon terminations were more focused primarily in middle layers of cortex, likely representing 'feedforward' input from V1, whereas labeled cells were more widespread and found in both superficial and deeper cortical layers, indicative of feedback projections. Averaged across injections, V3 had the third largest percentage of labeled cells (11%), following only V2 (47%) and the middle temporal area (MT; 19%). The dorsolateral area (DL, or V4; 9%) also contained a relatively large number of retrogradely labeled cells. These results indicate that V2, V3, DL (V4), and MT are retinotopically connected with V1, and provide major sources of feedback. Other extrastriate areas were less densely connected to V1, and there was no clear indication of labeled terminals. Inferotemporal cortex (IT) provided nearly 7% of feedback connections, whereas the dorsomedial area (DM) contributed about 3%. The remaining areas that have been proposed for galago extrastriate cortex, MTc, MST, FST, LPP and VPP, each accounted for about 1% or less of the total number of labeled cells. Thus, six extrastriate areas, V2, MT, V3, DL (V4), IT, and DM provide over 96% of visual cortex projections to V1. These areas also provide most of the projections to V1 in New and Old World monkeys.     

Neuronal projections from V1 to V2 in amblyopia

The mechanism of amblyopia in children with congenital cataract is not understood fully, but studies in macaques have shown that geniculate synapses are lost in striate cortex (V1). To search for other projection abnormalities in amblyopia, the pathway from V1 to V2 was examined using a triple-label technique in three animals raised with monocular suture. [(3)H]proline was injected into one eye to label the ocular dominance columns. Cholera toxin B subunit conjugated to gold (CTB-Au) was injected into V2 to label V1 projection neurons. Alternate sections were processed for cytochrome oxidase (CO) and CTB-Au, or dipped for autoradiography. Eight fields of CTB-Au-labeled cells in V1 opposite injection sites were plotted in layers 2/3 or 4B. After thin stripe injection, labeled cells were concentrated in CO patches. Despite column shrinkage, cells in deprived and normal columns were equal in size and density in both layers 2/3 and 4B. After pale or thick stripe injection, labeled cells were concentrated in interpatches. Only 23% of projection neurons originated from deprived columns. This reduction exceeded the degree of column shrinkage, a result explained by the fact that column shrinkage causes disproportionate loss of interpatch territory. These data indicate that early monocular form deprivation does not alter the segregation of patch and interpatch pathways to V2 stripes or cause selective loss or atrophy of V1 projection neurons. The effect of shrinkage of geniculocortical afferents in layer 4C following visual deprivation is not amplified further by attenuation of the amblyopic eye's projections from V1 to V2.     

Distribution of neurons projecting to the superior colliculus correlates with thick cytochrome oxidase stripes in macaque visual area V2

In visual area V2 of macaque monkeys, cytochrome oxidase (CO) histochemistry reveals a pattern of alternating densely labeled thick and thin stripe compartments and lightly labeled interstripe compartments. This modular organization has been associated with functionally separate pathways in the visual system. We examined this idea further by comparing the pattern of CO stripes with the distribution of neurons in V2 that project to the superior colliculus. Visually evoked activity in the superior colliculus is known to be greatly reduced by blocking magnocellular but not parvocellular layers of the lateral geniculate nucleus (LGN). From previous evidence that V2 thick stripes are closely associated with the magnocellular LGN pathway, we predicted that a significant proportion of V2 neurons projecting to the superior colliculus would reside in the thick stripes. To test this prediction, the tangential distribution of retrogradely labeled corticotectal cells in V2 was compared with the pattern of CO stripes. We found that neurons projecting to the superior colliculus accumulated preferentially into band-like clusters that were in alignment with alternate CO dense stripes. These stripes were identified as thick stripes on the basis of their physical appearance and/or by their affinity to the monoclonal antibody Cat-301. A significantly smaller proportion of labeled cells was observed in thin and interstripe compartments. These data provide further evidence that the spatial distribution of subcortically projecting neurons can correlate with the internal modular organization of visual areas. Moreover, they support the notion that CO compartments in V2 are associated with functionally different pathways. J. Comp. Neurol. 377:313–323, 1997. © 1997 Wiley-Liss, Inc.     

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). J. Comp. Neurol. 466:104–118, 2003. © 2003 Wiley‐Liss, Inc.     

Pale cytochrome oxidase stripes in V2 receive the richest projection from macaque striate cortex

Once the visual pathway reaches striate cortex, it fans out to a number of extrastriate areas. The projections to the second visual area (V2) are known to terminate in a patchy manner. V2 contains a system of repeating pale-thin-pale- thick stripes of cytochrome oxidase (CO) activity. We examined whether the patchy terminal fields arising from primary visual cortex (V1) projections are systematically related to the CO stripes in V2. Large injections of an anterograde tracer, [(3)H]proline, were made into V1 of both hemispheres in 5 macaques. The resulting V2 label appeared in layers 2-6, with the densest concentration in layer 4. In 21/29 injections, comparison of adjacent flatmount sections processed either for autoradiography or CO activity showed that the heaviest [(3)H]proline labeling was located in pale CO stripes. In 7/29 injections, there was no clear enrichment of labeling in the CO pale stripes. In 1 injection, the proline label correlated with dark CO stripes. On a fine scale, CO levels vary within V2 stripes, giving them an irregular, mottled appearance. In all stripe types, the density of proline label would often wax and wane in opposing contrast to these local fluctuations in CO density. Our data showed that V1 input is generally anti-correlated with the intensity of CO staining in V2, with strongest input to pale stripes. It is known that the pulvinar projects preferentially to dark stripes. Therefore, V2 receives interleaved projections from V1 and the pulvinar. Because these projections favor different stripe types, they may target separate populations of neurons.     

Pulvinar and other subcortical connections of dorsolateral visual cortex in monkeys

The present study used injections of neuroanatomical tracers to determine the subcortical connections of the caudal and rostral subdivisions of the dorsolateral area (DL) and the middle temporal crescent area (MT(C)) in owl monkeys (Aotus trivirgatus), squirrel monkeys (Saimiri sciureus), and macaque monkeys (Macaca fascicularis and M. radiata). Emphasis was on connections with the pulvinar. Patterns of corticopulvinar connections were related to subdivisions of the inferior pulvinar (PI) defined by histochemical or immunocytochemical architecture. Connections of DL/MT(C) were with the PI subdivisions, PICM, PICL, and PIp; the lateral pulvinar (PL); and, more sparsely, the lateral portion of the medial pulvinar (PM). In squirrel monkeys, there was a tendency for caudal DL to have stronger connections with PICL than PICM and for rostral DL/MT(C) to have stronger connections with PICM than PICL. In all three primates, DL/MT(C) had reciprocal connections with the pulvinar and claustrum; received afferents from the locus coeruleus, dorsal raphe, nucleus annularis, central superior nucleus, pontine reticular formation, lateral geniculate nucleus, paracentral nucleus, central medial nucleus, lateral hypothalamus, basal nucleus of the amygdala, and basal nucleus of Meynert/substantia innominata; and sent efferents to the pons, superior colliculus, reticular nucleus, caudate, and putamen. Projections from DL/MT(C) to the nucleus of the optic tract were also observed in squirrel and owl monkeys. Similarities in the subcortical connections of the dorsolateral region, especially those with the pulvinar, provide further support for the conclusion that the DL regions are homologous in the three primate groups.     

A reticular pattern of intrinsic connections in primate area V2 (area 18)

A system of periodic intrinsic connections is demonstrated in area V2 (area 18) of squirrel and macaque monkeys by large injections of tritiated amino acids, horseradish peroxidase (HRP), and fluorescent latex beads. These connections originate from pyramidal neurons concentrated in layers 3 and 5. Terminations occur in all cortical layers, largely coextensive with labeled neurons but more restricted in layer 4. This multilaminar distribution contrasts with the mainly supragranular localization of periodic intrinsic connections in V1 (area 17), and may imply a close interaction, in V2, of periodic intrinsic connections with pulvinocortical, as well as with corticocortical terminations (concentrated, respectively, in layers 3 and 5, and in lower 3 and 4). As in V1, the tangential configuration of these connections in V2 is reticular or latticelike, and is detectable for 2.5-3.0 mm from an injection site of HRP, 3H amino acids, or latex beads. Cross-sectional widths of labeled regions vary from 250 to 800 micron in squirrel monkey and from 400 to 1,000 micron in macaque, depending on which portion of the lattice is measured. When periodic intrinsic connections are compared with stripes labeled histochemically by cytochrome oxidase (CO), no clear relationship is obvious between the two systems. This result contrasts with the orderly tangential alignment reported between CO-reactive zones in V2 and certain extrinsic connections; namely, pulvinocortical terminations (Livingstone and Hubel, '82) and clusters of neurons projecting to area V4 (DeYoe and Van Essen, '84). Other extrinsic connections, however, such as backgoing connections from V2 to V1, do not seem to have a periodic distribution. Thus, although some discontinuous cortical connections relate to each other in a precise mosaic fashion, intrinsic and some extrinsic connections may observe different modes of organization.     

Cortical projections to the two retinotopic maps of primate pulvinar are distinct

Comprised of at least five distinct nuclei, the pulvinar complex of primates includes two large visually driven nuclei; one in the dorsal (lateral) pulvinar and one in the ventral (inferior) pulvinar, that contain similar retinotopic representations of the contralateral visual hemifield. Both nuclei also appear to have similar connections with areas of visual cortex. Here we determined the cortical connections of these two nuclei in galagos, members of the stepsirrhine primate radiation, to see if the nuclei differed in ways that could support differences in function. Injections of different retrograde tracers in each nucleus produced similar patterns of labeled neurons, predominately in layer 6 of V1, V2, V3, MT, regions of temporal cortex, and other visual areas. More complete labeling of neurons with a modified rabies virus identified these neurons as pyramidal cells with apical dendrites extending into superficial cortical layers. Importantly, the distributions of cortical neurons projecting to each of the two nuclei were highly overlapping, but formed separate populations. Sparse populations of double-labeled neurons were found in both V1 and V2 but were very low in number (<0.1%). Finally, the labeled cortical neurons were predominately in layer 6, and layer 5 neurons were labeled only in extrastriate areas. Terminations of pulvinar projections to area 17 was largely in superficial cortical layers, especially layer 1.     

Patterns of inter- and intralaminar GABAergic connections distinguish striate (V1) and extrastriate (V2, V4) visual cortices and their functionally specialized subdivisions in the rhesus monkey.

Local GABAergic connections are undoubtedly important for the operation of cerebral cortex, including the tuning of receptive field properties of visual cortical neurons. In order to begin to correlate specific configurations of GABAergic networks with particular receptive field properties, we examined the arrangement of GABAergic neurons projecting to foci in compartments of known functional specialization in striate (area V1) and extrastriate (areas V2, V4) cortices of rhesus monkeys. GABAergic cells were detected autoradiographically following microinjections into supragranular, granular, or infragranular layers of 5, 10, or 50 nl of 3H-nipecotic acid, which selectively exploits the GABA reuptake mechanism. These injections produced complex inter- and intralaminar distributions of retrograde perikaryal labeling that was selective for GABA-immunopositive neurons and glia. The pattern of retrograde labeling depended on both the laminar and cytoarchitectonic location of injection sites. In all cases, a high density of labeled neurons was present in the immediate vicinity of injection sites, with the density of labeled neurons decreasing for the most part uniformly with horizontal distance. Injections in supragranular layers produced relatively widespread labeling (up to 1.5-1.7 mm from the center of injections) in upper layers, whereas in granular and infragranular layers, labeling was confined to a radius of 0.25-0.5 mm. Conversely, injections in infragranular layers produced labeling that was widest (up to 1 mm) in lower layers, but more laterally restricted in supragranular layers. Injections in granular layers, on the other hand, produced an even distribution of labeling, 0.6-1.0 mm in diameter, throughout all layers. Comparably placed injections in V1, V2, and V4 resulted in patterns of labeling that were distinguished by features including stepwise increases in the lateral extent of labeling from striate to extrastriate areas, and the circular versus markedly elongated intralaminar distribution of labeled neurons in V1 and V4 versus V2. Further, for superficial injections, labeling was present in all layers in V1 and V2, but did not extent below the top layer V in area V4. These findings offer clear examples of organizational differences in the intrinsic inhibitory connections of visual cortices. The results also demonstrate that the number of GABAergic neurons projecting to any spot in cortex decreases systematically with horizontal distance from the spot, and that radiolabeled cells do not coalesce to form slabs, columns, or clusters. This relatively even distribution of retrogradely labeled cells in the tangential plane is consistent with recent computer simulations (Worgotter and Koch, 1991) that suggest that inhibitory neurons broadly tuned as a population can produce the specific response properties of cortical neurons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Nasotemporal asymmetries in V1: Ocular dominance columns of infant,adult, and strabismic macaque monkeys

To quantify asymmetries of input from the two eyes into each cerebral hemisphere, we measured ocular dominance column (ODC) widths and areas in the striate visual cortex (area V1) of macaque monkeys. Ocular dominance stripes in layer 4C were labeled by using transneuronal transport of intraocularly injected wheat germ agglutinin-horseradish peroxidase (WGA-HRP) or cytochrome oxidase (CO) histochemistry, after deafferentation of one eye or even by leaving afferent input intact. In infant monkey aged 4 and 8 weeks, ocular dominance stripes labeled by WGA-HRP appeared adultlike with smooth, sharply defined borders. In normal infant and normal adult macaque, ocular dominance stripes driven by the nasal retina (i.e., contralateral eye) were consistently wider than stripes driven by the temporal retina (i.e., ipsilateral eye). Asymmetries in the percentage of area V1 driven by nasal vs. temporal ODCs showed a similar “nasal bias”: in infant macaque, approximately 58% of ODCs in V1 were driven by nasal retina, and in adult macaque approximately 57%. The asymmetries tended to be slightly smaller in opercular V1 and greater in calcarine V1. “Spontaneous” ocular dominance stripes were revealed by CO staining of V1 in a naturally strabismic monkey and in a monkey made strabismic by early postnatal alternating monocular occlusion. In these animals, ocular dominance stripes and CO blobs corresponding to the nasal retina stained more intensely for CO in both the right and left V1. ODC spacing and the nasotemporal asymmetry in ODC width and area were similar in strabismic and normal monkeys. Our results in normal monkeys extend the observations of previous investigators and verify that nasotemporal inputs to opercular and calcarine V1 are unequal, with a consistent bias favoring inputs from the nasal retina. The CO results in strabismic macaque suggest that the nasal ODC bias promotes interocular suppression when activity in neighboring ODCs is decorrelated by abnormal binocular experience in infancy. J. Comp. Neurol. 388:32–46, 1997. © 1997 Wiley-Liss, Inc.     

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.     

Topographic patterns of v2 cortical connections in a prosimian primate (Galago garnetti)

Topographic patterns of cortical connections of the second visual area (V2) were examined in a lorisiform prosimian primate (Galago garnetti). Up to five different tracers were injected into dorsal and ventral V2. Tracers included wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) and up to four fluorochromes. Tracer injections consistently labeled neurons and terminals in primary visual cortex (V1), V2, the middle temporal area (MT), and the dorsolateral visual area (DL). Labeled neurons were also found in other proposed extrastriate areas such as the dorsomedial visual area (DM), dorsointermediate area (DI), middle temporal crescent (MTc), medial superior temporal area (MST), ventral posterior parietal area (VPP), and caudal inferotemporal cortex (ITc), but these connections were more variable and less dependent on the retinotopic position of injection sites in V2. Areal boundaries were identified by differences in cytochrome oxidase (CO) and myelin staining. We conclude that V2 cortical connections in prosimian galagos are similar to those in simian primates, suggesting that prosimians and other lines of primate evolution have retained several visual areas from a common ancestor that relate to V2 in similar ways. Architectural features of striate and extrastriate areas in prosimian galagos are similar to simian primates, with notable exceptions such as stripes in V2, which appear to be less differentiated in galagos.     

Cortical integration of parallel pathways in the visual system of primates

Injections of wheatgerm agglutinin conjugated with horseradish peroxidase (WGA-HRP) were placed in the middle temporal visual area (MT) of squirrel monkeys to reveal the distributions of interconnections with functionally distinct modules in areas 17 and 18. In agreement with previous reports, brain sections cut parallel to the surface of manually flattened cortex and reacted for cytochrome oxidase (CO) revealed CO dense blobs in area 17 and alternating CO dense thick and thin bands separated by CO light interbands in area 18. Alternate sections stained for myelin indicated that the CO light interblobs and interbands are more densely myelinated than the CO dense blobs and bands. Our major finding is that projections from MT to areas 17 and 18 are both to modules projecting to MT and modules projecting to other targets. In area 17, the cells in the middle layers projecting to injection sites in MT typically were distributed in several short merging and diverging rows, suggesting the convergence of projections from several matched orientation columns in area 17 to the restricted injection site in MT. Backward projections from MT to more superficial layers in area 17 were distributed more evenly across cortex and over a wider area of cortex. These terminations were dense throughout the interblob cortex which includes all orientation columns and neurons projecting to area 18, but were light over the blobs. As previously reported, neurons in area 18 projecting to MT were located in one set of the CO dense bands. However, these bands appeared to be thin rather than thick.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lack of robust LGN label following transneuronal rabies virus injections into macaque area V4

In primates, retinal inputs are relayed through the magno- and parvocells of the lateral geniculate nucleus (LGN) indirectly to extrastriate visual cortex. The most direct pathway identified to the extrastriate cortex is a disynaptic one that provides robust magno- and parvocellular inputs to the middle temporal area (MT). The inclusion of parvocells in this projection is somewhat surprising because of their importance for color and form vision, whereas MT is more strictly tuned to velocity. This raises the question of whether areas more involved in color and form processing, such as V4, receive similar projections. We report here on experiments that use rabies virus injections into V4 to retrogradely label mono- and disynaptic inputs. We find only a small number of labeled neurons in the LGN in a pattern consistent with monosynaptic labeling of koniocells, rather than disynaptic labeling of magno- and parvocells. The lack of robust magno- and parvocellular label was not due to ineffective viral transport because in the same cases we find hundreds of neurons labeled in the thalamic reticular nucleus, a structure that can only be labeled disynaptically from the cortex. We also find a complete absence of neurons labeled in V1, but thousands in adjacent areas V2 and V3. This result helps explain the absence of labeled magno- and parvocells in the LGN because disynaptic transport from an extrastriate visual area should require a relay through V1. Taken together, these results suggest that ascending magno/parvocellular inputs to V4 are more hierarchically organized than the relatively direct inputs to MT.     

The origin of the spinomesencephalic tract in the rat: An anatomical study using the retrograde transport of horseradish peroxidase

Projections of the middle temporal visual area, MT, and of visual cortex adjoining MT were investigated with autoradiographic methods in the prosimian primate, Galago senegalensis. Ipsilateral cortical targets of MT included area 17, area 18, cortex caudal to MT, cortex ventral to MT, and parietal-occipital cortex dorsal to MT. This pattern of projections suggests that extrastriate cortex contains a number of visual subdivisions in addition to MT. Contralateral projections were to MT and parietal-occipital cortex. Projections from MT to areas 17 and 18 connected regions representing similar parts of the visual hemifield while the location of callosal projections in MT matched the location of the injection site in the other hemisphere. Label in area 17 wac concentrated in layers I, III, and VI whereas other cortical areas were most densely labeled in the granular and supragranular layers. Subcortical projections of MT included the reticular nucleus of the thalamus, the lateral posterior nucleus, the superior pulvinar, the inferior pulvinar, the superior colliculus, and the pontine nuclei. The projection pattern to the superior and inferior pulvinar nuclei suggests that MT projects in a topographic manner to two subdivisions within each of these structures. Injections in cortex just outside of MT labeled area 18, inferotemporal cortex, parietal-occipital cortex, and, to a lesser extent, MT. The projections to inferotemporal cortex clearly distinguish the bordering cortex from MT. Contralateral cortical terminations were in locations corresponding to the injection site. Subcortical targets were generally similar to those seen after MT injections, although additional projections were observed depending on the location of the injection. Comparison of these results from the prosimian galago with studies in New and Old World monkeys indicates there are substantial similarities in projections. Thus, some of the cortical and thalamic subdivisions described for monkeys appear to exist in prosimians.     

Feedforward and feedback connections and their relation to the cytox modules of V2 in Cebus monkeys

To study the circuitry related to the ventral stream of visual information processing and its relation to the cytochrome oxidase (CytOx) modules in visual area V2, we injected anterograde and retrograde cholera toxin subunit B (CTb) tracer into nine sites in area V4 in five Cebus apella monkeys. The injection site locations ranged from 2° to 10° eccentricity in the lower visual field representation of V4. Alternate cortical sections, cut tangentially to the pial surface or in the coronal plane, were stained for CTb immunocytochemistry or for CytOx histochemistry or for Nissl. Our results indicate that the V4‐projecting cells and terminal‐like labeling were located in interstripes and thin CytOx‐rich stripes and avoided the CytOx‐rich thick stripes in V2. The feedforward projecting cell bodies in V2 were primarily located in the supragranular layers and sparsely located in the infragranular layers, whereas the feedback projections (i.e., the terminal‐like labels) were located in the supra‐ and infragranular layers. V4 injections of CTb resulted in labeling of the thin stripes and interstripes of V2 and provided an efficient method of distinguishing the V2 modules that were related to the ventral stream from the CytOx‐rich thick stripes, related to the dorsal stream. In V2, there was a significant heterogeneity in the distribution of projections: feedforward projections were located in CytOx‐rich thin stripes and in the CytOx‐poor interstripes, whereas the feedback projections were more abundant in the thin stripes than in the interstripes. J. Comp. Neurol. 522:3091–3105, 2014. © 2014 Wiley Periodicals, Inc.     

A double-labeling investigation of the afferent connectivity to cortical areas V1 and V2 of the macaque monkey

The afferent connectivity of areas V1 and V2 was investigated using the fluorescent dyes fast blue and diamidino yellow. Simultaneous injection of each dye in retinotopically corresponding regions of these areas gave rise to two afferent populations of labeled neurons in subcortical and cortical structures which project to both areas. These two populations showed a variable degree of overlap in their spatial distribution. Neurons labeled by both dyes (double-labeled neurons) which, therefore, project to both areas, were found in substantial numbers in these overlap zones. When the injections were made in non-retinotopically corresponding regions in the two areas, both populations of labeled cells overlapped extensively in the cortex but not in subcortical structures, suggesting that the laws governing the topography of these two types of connections are different. In the cortex, the labeled neurons extended from the fundus of the lunate sulcus to the fundus of the superior temporal sulcus. A few labeled neurons were also found in the inferior temporal cortex and the parahippocampal gyrus. In all cortical regions, corticocortical neurons projecting to V1 and V2 were found in both supra- and infragranular layers, although double-labeled neurons were more numerous in infragranular layers. With increasing distance from V1 there was an increase in the proportion of neurons labeled in infragranular layers. The comparative strength of input to V1 and V2 was computed and was found to be higher to V2 in all cortical regions except the superior temporal sulcus which projected equally heavily to both areas. The superior temporal sulcus also stood out in that of all cortical regions it contained the highest proportion of double-labeled neurons. Single- and double-labeled neurons were found in a number of subcortical structures including the lateral geniculate nucleus, the inferior and lateral pulvinar, the intralaminar nuclei, the nucleus basalis of Meynert, and the amygdala. The pattern of labeling in the lateral pulvinar was in agreement with the suggestion that this structure has a complex topographical organization containing at least a dual representation of the visual field (Bender, D. B. (1981) J. Neurophysiol. 46: 672-693). In the pulvinar complex, densities of labeled neurons permitted evaluation of the strength of input to V1 and V2, the latter being the strongest. These results demonstrate that areas V1 and V2 share a vast amount of common input from the same cortical and subcortical structures and that a number of neurons project to both areas via branching axons.     

Visual cortical projections and chemoarchitecture of macaque monkey pulvinar

We investigated the patterns of projections from the pulvinar to visual areas V1, V2, V4, and MT, and their relationships to pulvinar subdivisions based on patterns of calbindin (CB) immunostaining and estimates of visual field maps (P(1), P(2) and P(3)). Multiple retrograde tracers were placed into V1, V2, V4, and/or MT in 11 adult macaque monkeys. The inferior pulvinar (PI) was subdivided into medial (PI(M)), posterior (PI(P)), central medial (PI(CM)), and central lateral (PI(CL)) regions, confirming earlier CB studies. The P(1) map includes PI(CL) and the ventromedial portion of the lateral pulvinar (PL), P(2) is found in ventrolateral PL, and P(3) includes PI(P), PI(M), and PI(CM). Projections to areas V1 and V2 were found to be overlapping in P(1) and P(2), but those from P(2) to V2 were denser than those to V1. V2 also received light projections from PI(CM) and, less reliably, from PI(M). Neurons projecting to V4 and MT were more abundant than those projecting to V1 and V2. Those projecting to V4 were observed in P(1), densely in P(2), and also in PI(CM) and PI(P) of P(3). Those projecting to MT were found in P(1)- P(3), with the heaviest projection from P(3). Projections from P(3) to MT and V4 were mainly interdigitated, with the densest to MT arising from PI(M) and the densest to V4 arising from PI(P) and PI(CM). Because the calbindin-rich and -poor regions of P(3) corresponded to differential patterns of cortical connectivity, the results suggest that CB may further delineate functional subdivisions in the pulvinar.     

Effects of experimental strabismus on the architecture of macaque monkey striate cortex

Strabismus, a misalignment of the eyes, results in a loss of binocular visual function in humans. The effects are similar in monkeys, where a loss of binocular convergence onto single cortical neurons is always found. Changes in the anatomical organization of primary visual cortex (V1) may be associated with these physiological deficits, yet few have been reported. We examined the distributions of several anatomical markers in V1 of two experimentally strabismic Macaca nemestrina monkeys. Staining patterns in tangential sections were related to the ocular dominance (OD) column structure as deduced from cytochrome oxidase (CO) staining. CO staining appears roughly normal in the superficial layers, but in layer 4C, one eye's columns were pale. Thin, dark stripes falling near OD column borders are evident in Nissl-stained sections in all layers and in immunoreactivity for calbindin, especially in layers 3 and 4B. The monoclonal antibody SMI32, which labels a neurofilament protein found in pyramidal cells, is reduced in one eye's columns and absent at OD column borders. The pale SMI32 columns are those that are dark with CO in layer 4. Gallyas staining for myelin reveals thin stripes through layers 2-5; the dark stripes fall at OD column centers. All these changes appear to be related to the loss of binocularity in cortical neurons, which has its most profound effects near OD column borders.