Columnar specificity of intrinsic horizontal and corticocortical connections in cat visual cortex

A prominent and stereotypical feature of cortical circuitry in the striate cortex is a plexus of long-range horizontal connections, running for 6-8 mm parallel to the cortical surface, which has a clustered distribution. This is seen for both intrinsic cortical connections within a particular cortical area and the convergent and divergent connections running between area 17 and other cortical areas. To determine if these connections are related to the columnar functional architecture of cortex, we combined labeling of the horizontal connections by retrograde transport of rhodamine-filled latex microspheres (beads) and labeling of the orientation columns by 2-deoxyglucose autoradiography. We first mapped the distribution of orientation columns in a small region of area 17 or 18, then made a small injection of beads into the center of an orientation column of defined specificity, and after allowing for retrograde transport, labeled vertical orientation columns with the 2-deoxyglucose technique. The retrogradely labeled cells were confined to regions of orientation specificity similar to that of the injection site, indicating that the horizontal connections run between columns of similar orientation specificity. This relationship was demonstrated for both the intrinsic horizontal and corticocortical connections. The extent of the horizontal connections, which allows single cells to integrate information over larger parts of the visual field than that covered by their receptive fields, and the functional specificity of the connections, suggests possible roles for these connections in visual processing.     

Development of intrinsic connections in cat somatosensory cortex

We explored the development of clustered connections in cat somatosensory cortex by using intracortical injections of biocytin and carbocyanine dye (DiI). Biocytin injections in adults revealed clusters of retrogradely labeled cells, affirming earlier reports of the patchy nature of corticocortical connectivity in the adult cat somatosensory cortex. On postnatal days (PNDs) 1 and 3, a diffuse distribution of axons and labeled cells were found after DiI injections. A dramatic rearrangement of connections had taken place by PND 6. Well-defined clusters of labeled cells surrounded the injection site. Intercluster zones were relatively free of labeled cells and contained few labeled axons. At later ages (PNDs 12 and 15), a clear patchy distribution of intrinsic connections was seen. We analyzed neuronal clustering by using spatial point process analysis, which corroborated our qualitative observations. The density of labeled neurons was significantly higher in clusters than in the intercluster zones on PND 6. Autocorrelations run on profile plots of optical density values, along paths parallel to the edge of the injection sites (reflecting axonal distributions), revealed significant periodicity (P < .05; center-to-center ∼ 750 μm) by PND 6. These data demonstrate that corticocortical connections in cat somatosensory cortex develop from a diffuse network at birth, which transforms into a patchy network by the end of the 1st postnatal week. This is substantially earlier than the development of local circuits in the cat visual cortex and may reflect fundamental differences in the organization of the two cortices. J. Comp. Neurol. 384:501–516, 1997. Published 1997 Wiley-Liss, Inc.     

Specificity of intrinsic connections in primate primary visual cortex

Several recent studies have suggested a patchy system of intrinsic lateral connections in area 17 of the macaque monkey. To see whether this pattern bore any relationship to the cytochrome oxidase blobs we made multiple tiny injections of horseradish peroxidase into layers 2 and 3 of area 17, small enough so that some of the injections (or their cores) were entirely inside a single blob, or entirely outside. When the injection centers were entirely in blobs, the label in layers 2 and 3 was transported preferentially to nearby blobs, avoiding nonblob areas. When the injections were in nonblob areas, the label was found predominantly in surrounding nonblob areas, avoiding the blobs. Besides this lateral transport, label was also present in the layers below 2 and 3: the label in layers 4 and 6 was very restricted, occupying roughly the diameter of the injection core and presumably representing axons of cell bodies at the injection site; in layer 5 diffusely labeled patches observed the same blob/nonblob segregation seen above layer 4.     

Neuronal connections underlying orientation selectivity in cat visual cortex

How do neurons of the visual cortex acquire their acute sensitivity to the orientation of a visual stimulus? The question has preoccupied those who study the cortex since Hubel and Wiesel¹ first described orientation selectivity over twenty-five years ago. At the time, they proposed an elegant and enduring model for the origin of orientation selectivity. Fig. 1A, which is adapted from their original paper and which contains the essence of their model, is by now familiar to most students of the visual system and to many others besides. Yet the model, and the central question that it addresses, is still the subject of intense debate. Competing models have arisen in the intervening years, along with diverse experiments that bear on them.     

Development of the extrinsic connections of the visual cortex in the cat

Lesions were made in the visual system in a series of kittens, and fiber degeneration stained by the Eager or Fink-Heimer methods. Stereotaxic lesions made in the lateral geniculate nucleus (LGN) in one day old kittens led to the appearance of finely dotted fiber degenerationn in area 17 (concentrated in layer IV) two days later. Thalamic lesions involving the LGN in older kittens caused fiber degeneration in the other visual areas. This was first seen in the lateral suprasylvian gyrus (SSG) at six days, in area 19 at 14 days, and in area 18 at 21 days. Lesions of visual cortex affecting areas 17 and 18 caused degeneration of descending connections to the LGN, lateralis posterior and superior colliculus when made in kittens one day old. However, ipsilateral cortical connections to area 19 were not seen until the kittens were 19 days old at operation, and connections to the suprasylvian gyrus appeared at 21 days. Contralateral cortical connections carried by the corpus callosum were not seen until the kittens were 26 days old at operation, and a distribution of degeneration comparable with that seen in adults was present at 40 days. This time sequence of development is discussed in relation to what is known about the time of development of functioning in the cat's visual system.     

Cortical hierarchy reflected in the organization of intrinsic connections in macaque monkey visual cortex

Neuronal response properties vary markedly at increasing levels of the cortical hierarchy. At present it is unclear how these variations are reflected in the organization of the intrinsic cortical circuitry. Here we analyze patterns of intrinsic horizontal connections at different hierarchical levels in the visual cortex of the macaque monkey. The connections were studied in tangential sections of flattened cortices, which were injected with the anterograde tracer biocytin. We directly compared the organization of connections in four cortical areas representing four different levels in the cortical hierarchy. The areas were visual areas 1, 2, 4 and Brodman's area 7a (V1, V2, V4 and 7a, respectively). In all areas studied, injections labeled numerous horizontally coursing axons that formed dense halos around the injection sites. Further away, the fibers tended to form separate clusters. Many fibers could be traced along the way from the injection sites to the target clusters. At progressively higher order areas, there was a striking increase in the spread of intrinsic connections: from a measured distance of 2.1 mm in area V1 to 9.0 mm in area 7a. Average interpatch distance also increased from 0.61 mm in area V1 to 1.56 mm in area 7a. In contrast, patch size changed far less at higher order areas, from an average width of 230 m?M in area V1 to 310 m?m in area 7a. Analysis of synaptic bouton distribution along axons revealed that average interbouton distance remained constant at 6.4 m?m (median) in and out of the clusters and in the different cortical areas. Larger injections resulted in a marked increase in the number of labeled patches but only a minor increase in the spread of connections or in patch size. Thus, in line with the more global computational roles proposed for the higher order visual areas, the spread of intrinsic connections is increased with the hierarchy level. On the other hand, the clustered organization of the connections is preserved at higher order areas. These clusters may reflect the existence of cortical modules having blob-like dimensions throughout macaque monkey visual cortex. © 1993 Wiley-Liss, Inc.     

Intrinsic connections in cat visual cortex: a combined anterograde and retrograde tracing study

Area 18 of cat visual cortex was examined for intrinsic axons following small, columnar injections of an anterograde tracer,Phaseolus vulgaris leucoagglutinin (PHA-L). Locally projecting axons radiated from the injection site and branched to form 10–15 discrete, approximately circular patches 500–750 μm in diameter consisting of many bouton-studded terminal arborizations. Labeled fibers and boutons ramified densely in layers I, II/II, V, and VI, and were noticeably less dense in layer IV. Afferent and efferent pathways originating from the same cortical columns were studied by injecting a mixture of PHA-L and wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). Between 10 and 15 patches of cells retrogradely labeled by WGA-HRP surrounded each injection site. Within a patch, labeled cells were found in all layers and included both pyramidal and non-pyramidal cells. The distribution of PHA-L labeling was similar to that obtained when PHA-L was injected alone. Most often, the labeled patches resulting from injections of such mixtures contained both anterograde and retrograde labeling. However, patches consisting of retrograde labeling alone and of anterograde labeling alone were also observed, indicating that the local connections linking neighboring cortical columns were not always reciprocal.     

Topography of orientation centre connections in the primary visual cortex of the cat.

The functional topography of lateral connections to orientation-centre zones was studied by optical imaging of intrinsic signals in combination with tracer injections (fluorescent beads and biocytin) and electrophysiological recordings. Three-dimensional reconstruction of anterogradely labelled axon terminals and retrogradely labelled somata revealed a uniform distribution across all orientations in a non-patchy manner. The overall lateral extent of the labelling was 3-4 mm in layer 3, that is about half of the extent observed for orientation domain connections in the same layer. These bulk injection data are in contrast with the reportedly sharp orientation tuning of neurons of centre zones and suggest that orientation specificity here does not require highly specific connections. Nonetheless, another plausible scenario is that orientation centre connections are orientation specific but their specificity present at the single cell level cannot be revealed by bulk labelling due to their large spatial overlap.     

The postnatal development of clustered intrinsic connections in area 18 of the visual cortex in kittens

Retrogradely transported neuronal markers were injected into area 18 of the visual cortex in normal kittens of various ages and in animals that had been binocularly deprived of patterned visual stimulation by eyelid suture. In normal kittens aged 20 days or more, distinct clusters of retrogradely labelled cells were identified in area 18 surrounding the injection sites; these cells lay mainly in cortical layers II, III, and the upper part of IV, but with some in layers V and VI. In kittens younger than 10 days, labelled cells were observed around injection sites, although they were not organized into clusters. The results suggest that the typical clustered pattern of neurones forming intrinsic connections in area 18 emerges during the second week postnatally from an immature non-patchy distribution. Binocular deprivation did not prevent the appearance of these patches of cells.     

Patterns of connections in rat visual cortex

The definition of visual areas is one of the central problems in visual cortex research. Rodent extrastriate cortex offers a striking example of the complexity of this issue, in that different parcelation schemes identify within it from 2 to as many as 13 separate visual areas. In the experiments reported here, patterns of connections within rat visual cortex were studied in an effort to better define its organizational layout. The experimental paradigm used consisted of the following steps: first, the pattern of callosal connections was revealed in vivo with the fluorescent tracer bisbenzimide. Then, using the callosal pattern as a landmark, single injections of WGA-HRP were placed at various sites in striate and extrastriate cortex. Subsequently, the relation between the tangential distribution of ipsilateral corticocortical connections, the callosal connections, and the borders of striate cortex were examined in the flattened cortex preparation. The experiments revealed widespread, patchy connections within rat visual cortex. These connections appeared to reflect 3 organizational trends. First, neighboring sites were more extensively connected than distant ones. Second, extrastriate sites receiving common striate cortex inputs tended to be interconnected. Finally, projections from opposite poles in striate cortex tended to form interdigitating patterns of connections in regions of overlap. Altogether these trends suggest that the extrastriate band adjoining striate cortex has a single, global map organization. However, within the global map, a clear modular organization was evident, which appeared to correspond to the multiple visuotopic representations reported for this region. Based on its location, and some organizational similarities. it is suggested that the global map may constitute the rat homolog of area V2 in cat and monkey.     

Corticocortical connections among visual areas in the cat

The cortical interconnections of 17 visual areas in the cat were studied by making single injections through recording micropipettes of the neuroanatomical tracers 3H-leucine and horseradish peroxidase (HRP) into the visual cortex of 40 adult animals. Coronal sections from each of the brains were analyzed for location of silver grains and HRP-filled neurons. There are five main results: (1) all corticocortical connections among visual areas are reciprocal. (2) Each cortical visual area has a unique set of cortical connections; the cortical targets of no two cortical visual areas are identical. (3) There is a vast and complicated pattern of connections among the visual areas which implies that there are numerous parallel circuits which run through any one visual area. (4) The connections among the cortical visual areas link retinotopically similar loci and are consistent with the visuotopic maps which microelectrode recording experiments have provided. (5) The connections among visual cortical areas often originate from, or terminate in, discontinuous patches within each area; this result obtains not only for areas 17, 18, 19, and posteromedial lateral suprasylvian area (PMLS), but for at least 13 other areas as well. The data reveal many parallel pathways and suggest multiple functional circuits interconnecting visual cortical areas. Since each visual area has multiple inputs and outputs it may have multiple functions, a different one for each of the circuits of which it is a part.     

Postnatal development of neuronal connections in cat visual cortex studied by intracellular recording in slice preparation

Postnatal development of neuronal connections in cat visual cortex (area 17) was studied in slice preparations obtained from kittens aged 1–18 weeks after birth and adult cats by recording intracellularly excitatory (EPSP) and inhibitory postsynaptic potentials (IPSP) evoked in cortical cells by stimulation of white matter. The EPSPs were already present in all cells at 1 week of age. Their efficiency assessed by their maximum rate of rise was low initially and increased progressively with age. In contrast, the IPSPs were absent in half of the cells at 1 week and almost all of the cells came to demonstrate inhibition by 9 weeks except for a few layer II-III cells. At all ages about three-quarters of the IPSPs had GABA_A-mediated early and GABA_B-mediated late components with different time course, reversal potential and sensitivity to GABA antagonists, while the remaining IPSPs had only the early component. The efficiency of both IPSPs assessed by the associated conductance increase showed an increase of more than twice from 1 to 5 weeks, reaching the same level as adults. The time course of the development of inhibition demonstrated in this study paralleled the time course of the development of selective visual responsiveness in cortical cells, suggesting that the postnatal maturation of inhibitory connections is a basis of maturation of visual responsiveness.     

Plasticity of cat visual cortex

Monocular eyelid closure in kittens mimics certain visual deficits in humans that result in amblyopia ex anopsia. I have now studied the effects of monocular eyelid closure in cat upon the slow-wave response recorded from visul cortex. The pattern of changes in the response closely paralleled the changes in visual function of the amblyopic eye, in particular the suppression during binocular vision.     

Synaptic organization of cortico-cortical connections from the primary visual cortex to the posteromedial lateral suprasylvian visual area in the cat

The synaptic organization of the projection from the cat striate visual cortex to the posteromedial lateral suprasylvian cortical area (PMLS) was examined. The anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) was iontophorectically delivered into area 17, and anterogradely labeled fibers were revealed in PMLS by means of an immunocytochemical detection method. Most axons and presumptive terminal swellings were found in layers III and IV. The neuronal elements (n = 190) that were postsynaptic to anterogradely labeled boutons were quantitatively analyzed. All anterogradely labeled cortico-cortical boutons (n = 182) established type 1 synapses. The results show that 83% of the postsynaptic targets were dendritic spines, probably belonging to pyramidal cells. Dendritic shafts constituted 17% of the targets. The dendritic shafts postsynaptic to cortico-cortical boutons were studied for the presence of gamma-aminobutyric acid (GABA) with a postembedding immunogold method. Most dendritic shafts (85%) that were tested were found to be GABA-positive, demonstrating that they originate from local inhibitory neurons. Taking into account that most postsynaptic targets were spines and extending the results of the immunocytochemical testing to the total population of postsynaptic dendrites, it was calculated that at least 14% of targets originated from GABA-positive cells. Thus cortico-cortical axons establish direct monosynpatic connections mainly with pyramidal and to a lesser extent with GABAergic nonpyramidal neurons in area PMLS, providing both feedforward excitation and feedforward inhibition to a visual associational area known to be involved in the processing of motion information. The results are consistent with previously demonstrated deficits in physiological properties of neurons in PMLS following removal of cortico-cortical afferents.     

Intrinsic laminar lattice connections in primate visual cortex

Intracortical injections of horseradish peroxidase (HRP) reveal a system of periodically organized intrinsic connections in primate striate cortex. In layers 2 and 3 these connections form a reticular or latticelike pattern, extending for about 1.5-2.0 mm around an injection. This connectional lattice is composed of HRP-labeled walls (350-450 microns apart Saimiri and about 500-600 microns in macaque) surrounding unlabeled central lacunae. Within the lattice walls there are regularly arranged punctate loci of particularly dense HRP label, appearing as isolated patches as the lattice wall labeling thins further from the injection site. A periodic organization has also been demonstrated for the intrinsic connections in layer 4B, which are apparently in register with the supragranular periodicities, although separated from these by a thin unlabeled region. The 4B lattice is particularly prominent in squirrel monkey, extending for 2-3 mm from an injection. In both layers, these intrinsic connections are demonstrated by orthogradely and retrogradely transported HRP and seem to reflect a system of neurons with long horizontal axon collaterals, presumably with arborizations at regularly spaced intervals. The intrinsic connectional lattice in layers 2 and 3 resembles the repetitive array of cytochrome oxidase activity in these layers; but despite similarities of dimension and pattern, the two systems do not appear identical. In primate, as previously described in tree shrews (Rockland et al., '82), the HRP-labeled anatomical connections resemble the pattern of 2-deoxyglucose accumulation resulting from stimulation with oriented lines, although the functional importance of these connections remains obscure.     

Hypothalamic connections of the insular cortex in the cat

We have investigated the anatomical organization of the connections between the hypothalamus and the insular cortex of the cat by using retrograde and anterograde transport of horseradish peroxidase-wheat germ agglutinin. The rostroventral sectors of the insular cortex are strongly interconnected with the supramammillary and caudal lateral hypothalamic areas. The possible relationship of these connections with the well-known participation of the insular cortex in visceral and autonomic integration is discussed.     

Postnatal growth of intrinsic connections in mouse barrel cortex.

Surprisingly little is known about the development of connections within a functional area of the cerebral cortex. We examined the postnatal growth of connections in mouse barrel cortex during the second and third weeks after birth, coinciding with the period of rapid synaptogenesis that occurs just after the barrels first form. A barrel is a group of neurons in layer 4 of somatosensory cortex that is part of a cortical column. Each whisker/barrel column is linked anatomically and functionally to a homotopic whisker on the contralateral face. Radial groups of cortical neurons were labeled with the neuronal tracer biotinylated dextran amine in mice ranging in age from postnatal day 8 (P8; P0 is the date of birth) to adulthood. The spatial distributions of retrogradely labeled neurons in different laminae were analyzed. The barrel map in layer 4 was used as a template to compare quantitative data from different animals and to account for substantial changes in barrel and barrel field size during development. Intrinsic projections 1) innervate increasingly more distant targets within barrel cortex up to 3 weeks of age; 2) continue to form in targets after 3 weeks, effectively strengthening existing connections; 3) follow a timetable for growth that is layer-specific; 4) link more distant barrel columns in layer 4 from neurons that are found preferentially in the barrel side and the septa between barrels; and 5) form over the shortest distances between the barrel columns. These data indicate that intrinsic connections in mouse barrel cortex develop by the progressive addition of neuronal connections rather than by sculpting preliminary connections. We describe statistically significant changes in connectivity during development that may be applied to model and assess the development of connections after a variety of experimental perturbations, such as to the environment and/or the genome.     

Stimulus-analyzing mechanisms in the cat visual cortex

Units in the cat's visual cortex were observed while the orientation and intensity of illumination of a “preferred” visual stimulus was varied. In addition to showing the typical sensitivity to stimulus orientation the units were also sensitive to intensity. Almost all (52 of 56) increased firing rate with increased intensity. Thus, even at higher levels of the visual pathway where neurons are noted for their ability to abstract elements of form from the total visual input, intensity continues to exert a major influence. For a given neuron various combinations of intensity and orientation produced the same response. The results do not support a “place” theory of pattern recognition.     

Laminar origins of visual corticocortical connections in the cat

The interconnections among visual areas in cat cortex were studied with respect to the specific laminae in which the cortically projecting neurons are located. Single injections of HRP were made through recording micropipettes into nine different visual areas. In 15 cortical areas the laminar distribution of neurons which were retrogradely filled with HRP was plotted. In this way we determined the laminar origins of the cortical projections to the nine separate cortical visual areas which were injected. There are three major observations. First, areas 17 and 18 are the only two visual areas in which layers II and III are the primary site of cortically projecting cells; in the other 13 areas the deeper layers of cortex provide a large percentage of such neurons. Second, within any one cortical area, cortically projecting neurons may be distributed among different layers; the specific layer depends upon the cortical target of those neurons. Third, any one cortical area receives projections from several different cortical layers, the specific layers being dependent upon the area from which the projection originates. An individual cortical area, therefore, contributes to several different cortical visual circuits, with each of these circuits defined by the laminar connections of its neurons.     

Synaptic plasticity of feedback connections in rat visual cortex

The issue we want to address in the present paper is to establish whether electrical stimulation of latero medial (LM) area, a secondary visual area in the rat, is able to induce Long Term Potentiation (LTP) and Long Term Depression (LTD) in primary visual cortex (V1). To this aim rat slices containing area V1 and LM were prepared at P23 and P40 and field potentials in layers 2/3 of area V1 were recorded stimulating LM. We showed that it was never possible to induce LTP in area V1, unless bicuculline, a gamma-aminobutyric acid (GABA) receptors blocker, was applied to the slice. In contrast, LTD was normally inducible. Thus, cortical gabaergic circuitry in area V1 controls LTP but not LTD elicited by stimulation of feedback connections from LM.