The layout of functional maps in area 18 of strabismic cats

Strabismus (or squint) is both a well-established model for developmental plasticity and a frequent clinical symptom. To analyze experience-dependent plasticity of functional maps in the brain we used optical imaging of intrinsic signals to visualize both orientation and ocular dominance domains in cat area 18. In strabismic animals, iso-orientation domains exhibited a pinwheel-like organization, as previously described for area 18 of normally raised animals and for area 17 of both normally raised and strabismic cats. In area 18, mean pinwheel density was similar in the experimental (2.2 pinwheel centers per mm2 cortical surface) and control animals (2.3/mm2 in normally raised animals), but significantly lower than in area 17 of both normally raised and strabismic cats (2.7-3.4/mm2). A comparison of orientation and ocular dominance domains revealed that iso-orientation domains were continuous across the borders of ocular dominance domains and tended to cross these borders at steep angles. Thus, the orientation map does not seem to be modified by experience-dependent changes in afferent activity. Together with our recent observation that strabismus does not enhance the segregation of ocular dominance domains in cat area 18, the present data indicate that the layout of functional maps in area 18 is less susceptible to experience-dependent manipulations than in area 17.     

Two-dimensional analysis of the spacing of ocular dominance columns in normally raised and strabismic kittens

In the primary visual cortex of cats, ferrets and macaque monkeys, the thalamocortical afferents conveying signals from the two eyes terminate in alternating regions of layer IV known as ocular dominance columns. Previous experiments have indicated that the periodicity of these columns can be influenced by visual experience: compared to normally raised animals both strabismic cats and cats raised with alternating monocular exposure displayed an increased spacing of adjacent ocular dominance columns in the primary visual cortex (area 17). However, recently it was shown that the formation of ocular dominance columns begins much earlier than previously supposed, indicating that early visual experience might only have a limited influence on the development of the spatial pattern of ocular dominance columns. We therefore visualized the complete pattern of ocular dominance columns in area 17 of normally raised and strabismic kittens during early postnatal development (age 3-6 weeks), particularly focussing on littermates. In addition, we used a previously developed spatial analysis (period statistics) to quantify columnar spacing two-dimensionally. We observed a pronounced interindividual variability in both normally raised and strabismic animals, with column spacings ranging from 783 to 1362 microm. In contrast to previous reports, there were no significant differences in columnar periodicity between normally raised and strabismic cats. These data indicate that rearing has less influence on column spacing while the interindividual variability is much greater than previously supposed, suggesting that genetic differences have an influence on column spacing.     

Interhemispheric connections of areas 17 and 18 in the cortical columns of cats with unilateral strabismus

The interhemispheric connections of areas 17 and 18 of the cerebral cortex were investigated in cats with experimental unilateral strabismus. Single cortical columns were microiontophoretically injected with horseradish peroxidase, and retrogradelly labeled cells were demonstrated in the opposite brain hemisphere. After tracer injection in area 18 columns, the labeled callosal cells were located in area 17/18 transitional zone, similar to what was found in normal cats. However, in some cases the expansion of the region of callosally labeled cells distribution, was found. It is proposed that the extent of the region of callosally-connected cells may vary depending on whether the cells receive their input from intact or strabismic eye.     

Spatial reorganization of horizontal connections of cat cortical area 17 induced by eye rotation

In six cats with experimental unilateral or bilateral strabismus, surgically induced early in postnatal life, torsion eye rotation with a deviation angle of 10 to 20 degrees was also detected. Spatial distribution of retrogradely labeled neurons in area 17 was studied following microiontophoretic injection of horseradish peroxidase in area 17 or 18 cortical columns. Eye rotation was shown to cause the increase of the length of horizontal neuronal connections in area 17 along the projection of visual field vertical meridian. The reorganization of neuronal connections, detected in this work, may promote the functional changes, described in the literature, that compensate for the eye rotation.     

Neuronal connections of eye-dominance columns in the cat cerebral cortex after monocular deprivation

Plastic changes in intrahemisphere neuronal connections of the eye-dominance columns of cortical fields 17 and 18 were studied in monocularly deprived cats. The methodology consisted of microintophoretic administration of horseradish peroxidase into cortical columns and three-dimensional reconstruction of the areas of retrograde labeled cells. The eye dominance of columns was established, as were their coordinates in the projection of the visual field. In field 17, the horizontal connections of columns receiving inputs from the non-deprived eye via the crossed-over visual tracts were longer than the connections of the "non-crossed" columns of this eye and were longer than in normal conditions; the connections of the columns of the deprived eye were significantly reduced. Changes in the spatial organization of horizontal connections in field 17 were seen for the columns of the non-deprived eye (areas of labeled cells were rounder and the density of labeled cells in these areas were non-uniform). The longest horizontal connections in deprived cats were no longer than the lengths of these connections in cats with strabismus. It is suggested that the axon length of cells giving rise to the horizontal connections of cortical columns has a limit which is independent of visual stimulation during the critical period of development of the visual system.     

Exogenous supply of nerve growth factor prevents the effects of strabismus in the rat.

It has recently been reported that exogenous supply of nerve growth factor prevents the effects of monocular deprivation both in rats and in cats. Here we have extended these experiments to the case of strabismus. Repeated intraventricular injections of nerve growth factor were performed in rats made surgically strabismic early in the critical period. At the end of the critical period the ocular dominance distribution of visual cortical neurons was assessed in strabismic untreated, strabismic nerve growth factor-treated and strabismic Cytochrome C-treated (control) rats by means of extracellular recordings. We found that in rats surgical strabismus causes a consistent loss of binocular neurons. By contrast the treatment with nerve growth factor maintains the normal ocular dominance distribution of neurons in the primary visual cortex. We conclude that nerve growth factor exogenously supplied prevents the effects induced by surgical strabismus in rats and suggest that nerve growth factor has a role in visual cortical plasticity.     

The structure of internal intraneuronal connections in the area 17 of the cat cerebral cortex

Spatial distribution of horizontal intrinsic neuronal connections in area 17 of cat cerebral cortex was studied following HRP microion tophoretic injections into the single cortical columns. Cluster analysis of labelled cell distribution in superficial layers was performed in the tangential cortical plane. Area 17 was found to contain 7 +/- 1 clusters each consisting of 1-5 cells. Clusters were arranged in two rows, separated by a distance of 1.2 +/- 0.3 mm. The distance between the centers of the clusters that form the rows was equal to 0.8 +/- 0.3 mm. The spatial characteristics of cell clusters sending axons to cortical column, found in this study, were compared with the known data on the optical imaging of the activity of neurons of orientation and ocular dominance columns of visual cortex. It is suggested that area 17 cortical column receives inputs from cells of 6-8 hypercolumns that have similar ocular dominance and orientation preference.     

Topographic relations between ocular dominance and orientation columns in the cat striate cortex

Summary In the visual cortex of four adult cats ocular dominance and orientation columns were visualized with (³H)proline and (¹⁴C)deoxyglucose autoradiography. The two columnar systems were reconstructed from serial horizontal sections or from flat-mount preparations and graphically superimposed. They share a number of characteristic features: In both systems the columns have a tendency to form regularly spaced parallel bands whose main trajectory is perpendicular to the border between areas 17 and 18. These bands frequently bifurcate or terminate in blind endings. The resulting irregularities are much more pronounced in the ocular dominance than in the orientation system. The periodicity of the columnar patterns was assessed along trajectories perpendicular to the main orientation of the bands and differed in the two columnar systems. The spacing of the ocular dominance stripes was significantly narrower than the spacing of orientation bands. The mean periodicity of a particular columnar system was virtually identical in the two hemispheres of the same animal but it differed substantially in different animals. However, the spacing of orientation columns covaried with that of the ocular dominance columns, the ratios of the mean spacings of the two columnar systems being similar in the four cats. The superposition of the two columnar systems revealed no obvious topographic relation between any of the organizational details such as the location of bifurcations, blind endings and intersections. We suggest the following conclusions: 1. The developmental processes generating the two columnar systems seem to obey the same algorithms but they act independently of each other. 2. The space constants of the two systems are rigorously specified and appear to depend on a common variable. 3. The main orientation of the bands in both columnar systems is related to a) the representation of the vertical meridian, b) the anisotropy of the cortical magnification factor, and c) the tangential spread of intracortical connections.     

Effects of convergent strabismus on spatio-temporal response properties of neurons in cat area 18

Summary Single-cell recording experiments were carried out to determine whether rearing kittens with surgically induced convergent strabismus (esotropia) alters the development of receptive field (RF) properties of neurons in area 18. In agreement with previous work on kittens with divergent strabismus (exotropia), there was a marked loss of binocularly driven cells in area 18 of esotropic cats. In contrast to the striate cortex of strabismic cats, the spatial properties of area 18 neurons, including receptive-field size and spatial frequency tuning, did not differ from those in normal controls. On the other hand, we found that contrast thresholds, measured at an optimal spatial frequency, were significantly elevated, and that the contrast gain in many cells was reduced in strabismic cats. These deficits were observed in both eyes, though the cells dominated by the deviating eye had a lower response amplitude at all contrasts. Furthermore, temporal frequency tuning curves were abnormal in strabismic cats in that the optimal frequencies and temporal resolutions were shifted to lower values. These effects were also bilateral. Velocity tuning, measured with a high-contrast bar stimulus, revealed that area 18 neurons in strabismic cats were unable to respond to very high velocities compared to normals. This reduced response was more severe when measured with the deviating eye in spite of the bilateral nature of the deficit. Finally, latencies to electrical stimulation of the optic chiasm or the optic radiation were significantly longer in strabismic cats. The magnitude of these effects was virtually the same for both eyes. From these observations, we conclude that the temporal properties of area 18 neurons, particularly the cells abilities to follow fast temporal modulations, are affected by raising kittens with surgically induced convergent strabismus.     

Relation between intrinsic connections and isofrequency contours in the inferior colliculus of the big brown bat, Eptesicus fuscus

Information processing in the inferior colliculus depends on interactions between ascending pathways and intrinsic circuitry, both of which exist within a functional tonotopic organization. To determine how local projections of neurons in the inferior colliculus are related to tonotopy, we placed a small iontophoretic injection of biodextran amine at a physiologically characterized location in the inferior colliculus. We then used electrophysiological recording to place a grid of small deposits of Chicago Sky Blue throughout the same frequency range to specify an isofrequency contour. Using three-dimensional computer reconstructions, we analyzed patterns of transport relative to the physiologically determined isofrequency contour to quantify the extent of the intrinsic connection lamina in all three dimensions. We also performed a quantitative analysis of the numbers of cells in different regions relative to the biodextran amine injection. Biodextran amine-labeled fibers were mainly located dorsomedial to the injection site, confined within the isofrequency contour, but biodextran amine-labeled cells were mainly located ventrolateral to the injection site. When we counted numbers of labeled cells classified by morphological type, we found that both elongate and multipolar cells were labeled within the isofrequency contour. Because the dendrites of multipolar cells typically extend outside the isofrequency lamina, it is likely that they receive input from other isofrequency contours and relay it to more dorsomedial portions of their specific isofrequency contour, along with the frequency-specific projections of the elongate cells. Within a given isofrequency contour, there is a consistent organization in which intrinsic connections ascend from the ventrolateral portion to more dorsomedial points along the contour, forming a cascaded system of intrinsic feedforward connections that seem ideally suited to provide the delay lines necessary to produce several forms of selectivity for temporal patterns in inferior colliculus neurons.     

Intrinsic collaterals of layer 6 Meynert cells and functional columns in primate V1

Meynert cells are a distinct type of large neuron which project to area MT/V5 and to subcortical targets, including the superior colliculus. They have recently been shown to have extensive intrinsic collaterals spreading up to 8.0 mm within layer 6 of area V1 [J Comp Neurol 441 (2001) 134]. Using intrinsic signal imaging combined with tracer injections, this study investigates how Meynert cell collaterals are mapped in relation to the functional architecture of area V1 in macaque monkeys. In particular, we examined whether terminations of individual axon segments are selective for same-eye or opposite-eye domains. Analysis of 39 anterogradely labeled axon segments (from six injection sites in four hemispheres) showed that terminal segments cross over several pairs of ocular dominance columns (ODCs) and contact both left- and right-eye ODCs, with a slight bias for the contralateral eye. This contrasts with the same-eye bias previously reported for intrinsic collaterals of pyramidal neurons in layer 3. The suggestion is that the system of Meynert intrinsic collaterals is involved with binocular interactions over wide sectors of the visual field. This might be related to processes such as optic flow or, especially given the wide-field spread, even contour completion or interpolation.     

Thalamic connections of the second somatic sensory area in cats studied with anterograde and retrograde tract-tracing techniques.

The thalamic connections of the second somatosensory area in the anterior ectosylvian gyrus of cats have been investigated using the retrograde tracer horseradish peroxidase and the anterograde tracer Phaseolus vulgaris leucoagglutinin. Horseradish peroxidase was injected iontophoretically in several somatotopic zones of the second somatosensory area map of six cats. Sites of horseradish peroxidase delivery were identified preliminarily by recording with microelectrodes the responses of neurons to skin stimulation. Phaseolus vulgaris leucoagglutinin was iontophoretically injected within the ventrobasal complex (one cat) or in the posterior complex (one cat). Horseradish peroxidase injections into cytoarchitectonic area SII retrogradely labeled neurons in the ipsilateral ventrobasal complex and in the posterior complex. Counts of labeled neurons from the ipsilateral thalamus showed that the overwhelming majority of horseradish peroxidase-labeled neurons were in the ventrobasal complex (96.3-96.9%) and few were in the posterior complex (3.1-3.7%). Neurons labeled in the ventrobasal complex were observed throughout the anteroposterior extent of the nucleus, while their mediolateral distribution varied with the site of horseradish peroxidase delivery in the body map of the second somatosensory area, which indicates that the projections from the ventrobasal complex to the second somatosensory area are somatotopically organized. In the cat in which the horseradish peroxidase injection involved both the second somatosensory area proper and the second somatosensory area medial, which lies in the lower bank of suprasylvian sulcus, labeled neurons were almost as numerous in the ventrobasal complex as in the posterior complex. Phaseolus vulgaris leucoagglutinin injected in the ventrobasal complex anterogradely labeled thalamocortical fibers in the ipsilateral anterior ectosylvian gyrus. In this case, patches of labeled fibers and terminals were distributed exclusively within the cytoarchitectonic borders of the second somatosensory area proper. Labeled terminals were numerous in layer IV and lower layer III, but terminal boutons and fibers with axonal swellings, probably forming synapses en passant, were frequently observed also in layers VI and I. Injection of Phaseolus vulgaris leucoagglutinin in the posterior complex labeled thalamocortical fibers in two distinct regions in the ipsilateral anterior ectosylvian gyrus, one lying laterally and the other medially, which correspond, respectively, to the fourth somatosensory area and the second somatosensory area medial. In both areas the densest plexus of labeled fibers and axon terminals was in layer IV and lower layer III, but numerous labeled fibers and terminals were also observed in layer I. In this case, only rare fragments of labeled fibers were present in second somatosensory area proper, but no labeled terminals could be observed.     

A Method for the Detection and Quantitative Analysis of Two-Dimensional Distribution Patterns of Labeled Neurons in the Cerebral Cortex

The aim of the present work was to develop an algorithm for analyzing two-dimensional patterns of labeled neurons and their structure. Studies of corticocortical connections between fields 17 and 18 and the posteromedial area of the suprasylvian sulcus in cats as an example showed that the algorithm was suitable for the reconstruction and analysis of patterns of initial neurons and can be used as an algorithm for developing highly specialized computer programs.     

The postnatal development of visual callosal connections in the absence of visual experience or of the eyes

Summary Counts of callosal neurons retrogradely labeled by horseradish peroxidase (visualized using multiple substrates) were obtained in areas 17 and 18 of five kittens reared with their eyelids bilaterally sutured and of three kittens which had undergone bilateral enucleation on postnatal days 1–4. These counts were compared with those obtained in normal adult cats.The normal adult distribution of the callosal neurons results from the gradual postnatal reduction of a more widespread juvenile population. Binocular visual deprivation by lid suturing dramatically decreases the final number of callosal neurons and narrows their region of distribution (callosal zone) in areas 17 and 18. A less severe reduction in the final number of callosal neurons is caused by bilateral enucleation, which also increases the width of the callosal zone compared to that of normal cats. Thus, visual experience is necessary for the normal stabilization of juvenile callosal connections. However, since some callosal neurons form connections in the absence of vision, other influences capable of stabilizing juvenile callosal neurons also exist. These influences are probably antagonized by destabilizing influences or inhibited, when the eyes are intact.This work was supported by Swiss National Science Foundation grant 2.219.0.78 to Dr. G.M. Innocenti; Dr. D.O. Frost received a fellowship from the American-Swiss Foundation for Scientific Exchange     

The reciprocal connections of the suprageniculate nucleus and the superior colliculus in the rat

Reciprocally bilateral connections between the superior colliculus and the suprageniculate nucleus have been studied in the rat, using the anterograde and retrograde transport techniques of HRP. In those cases where the HRP deposit was restricted to the superficial layers of the colliculus, anterogradely labeled fibers and retrogradely labeled neurons were observed in the ipsilateral suprageniculate nucleus. However, upon HRP injection extended into the intermediate layers of the colliculus, the number of labeled fibers and neurons was not only increased ipsilaterally but were also observed in the contralateral suprageniculate nucleus. The density of the labeled fibers and the number of labeled neurons was always greater in the ipsilateral side. These results show that the suprageniculate nucleus and the superior colliculus are connected reciprocally and bilaterally, with an ipsilateral dominance.     

Interhemisphere Connections of Eye Dominance Columns in the Cat Visual Cortex in Conditions of Impaired Binocular Vision

Data from studies of interhemisphere connections in fields 17 and 18 of cats reared in conditions of impaired binocular vision (monocular deprivation, uni- and bilateral strabismus) are presented. Monosynaptic connections between neurons were studied by microiontophoretic application of horseradish peroxidase into cortical eye dominance columns and the distributions of retrograde labeled callosal cells were analyzed. Spatial asymmetry and eye-specific interhemisphere neuron connections persisted in conditions of monocular deprivation and strabismus. Quantitative changes in connections were less marked in monocular deprivation than strabismus. In cats with impaired binocular vision, as in intact animals, the widths of callosal-receiving zones were greater than the widths of the callosal cell zones, which is evidence for the non-reciprocity of interhemisphere connections in cortical areas distant from the projection of the vertical meridian. Morphofunctional differences between cells mediating connections in the opposite directions are proposed. Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 94, No. 6, pp. 627–636, June, 2008.     

Structure of Reciprocal Connections of Visual Cortical Fields 17 and 18 in the Cat

Microiontophoretic application of horseradish peroxidase to individual columns in fields 17 and 18 of the cat cortex was used to identify the distribution by area and layer of retrograde labeled cells in both fields. After application of marker to fields 17 or 18, the area of labeled cells in field 17 (in the tangential plane) was extended and orientated along the projection of the horizontal meridian of the field of vision. The area of labeled cells in field 18 in these cases was orientated along the projections of the vertical meridian. Similar differences in the organization of the connections of fields 17 and 18 were seen in the projection zone of the central 10° of the field of vision at different elevations. Thus, the spatial distributions of internal and external connections in each field coincide and their orientations in fields 17 and 18 are mutually perpendicular. It is suggested that field 17 performs the more detailed analysis of information on the horizontal components of an image and communicates this to field 18, while field 18 is responsible for the more detailed analysis of information about the vertical components of the same image, communicating this to field 17.     

The retinotopic match between area 17 and its targets in visual suprasylvian cortex

Summary It is widely believed that cells in area 17 send axons specifically to neurons in other cortical areas whose receptive fields coincide with their own. We asked whether this was true in cats for area 17's projection to a large suprasylvian visual area, the Clare-Bishop area. Receptive fields were plotted at multiple sites in the Clare-Bishop area. Then, in area 17, anterograde tracer was injected at a retinotopically-characterized site, giving rise to patches of labeled terminals in the Clare-Bishop area. Receptive field centers recorded within these patches were located close to the visual field location at the injection site in area 17. Receptive fields recorded outside of labeled patches, on the other hand, were never in register with that plotted in area 17. However, due to their large size, even fields located outside of labeled patches often encompassed the visual field point injected in area 17. In other experiments, receptive fields for both neurons and presumed cortical afferents were recorded at the same site in the Clare-Bishop area. The centers of such pairs of receptive fields were on average less than 1° apart. Finally, the gaps between widely separated patches of label were investigated. Both physiological and anatomical evidence indicated that a different part of the visual field was represented in gaps than in the adjacent patches.     

Intrinsic inter- and intralaminar connections and their relationship to the tonotopic map in cat primary auditory cortex

Summary Small iontophoretic injections of the lectin, Phaseolus vulgaris leucoagglutinin (PHA-L), were made into different layers of the primary auditory cortex (AI) of cats. Injections in layer I labeled two types of morphologically distinct fibers in layer I as well as a smaller number of axons in layers II and III. Layer II injections labeled descending axons that produced a dense plexus of terminal fibers in layers I–III of both AI and adjacent auditory fields. Injections in layer III also labeled a dense plexus of axon collaterals at the junction of layers V and VI and labeled patches of terminal fibers in both AI and adjacent auditory fields. These were densest in layers I–III but usually extended into layers IV and V as well. The patches were partly formed by axon collaterals of layer III pyramidal cells that traveled for over 4 mm in the gray matter. Injections confined to layer IV labeled axons in all layers of the cortex but none of these axons appeared to reach the white matter. The axons spread laterally in layer IV and up into the superficial layers and ramified especially layer I. Injections in layers V and VI labeled axons in all layers of the cortex but these were densest in the deep layers where labeling was fairly homogeneous. In the upper layers the labeling was arranged in semi-discrete patches. Large injections involving layers I–III were studied in tangential sections. Between 3 and 8 patches of terminal labeling were observed in AI and these were mainly arranged in a band with its long axis aligned approximately in the dorsoventral direction. However dense patches of terminal labeling also occurred both anterior and posterior to the injection site. In selected experiments portions of the tonotopic map in AI were mapped by single unit recording and subsequently the map was related to patches of anterogradely labeled fibers that surrounded injections of PHA-L. Rows of dorsoventrally oriented patches were among cells with a similar best frequency to those in the injection site. However patches located anterior or posterior to the injection site were among cells with higher or lower best frequencies. Two injections of PHA-L close together produce different patterns of labeling. One of the injections usually produces one or more patches that has no correlate among the patches of fibers labeled by the adjacent injection. This is clearest when one of the injections is made with biotinylated PHA-L that can be visualized directly without the use of primary antibodies. Thus the intrinsic connections of AI arising from nearby cylinders of neurons are not homogenous and clusters of cells can be identified by their unique pattern of connections within AI.     

Eye-rotation-induced spatial reorganization of horizontal connections in field 17 of the cat cortex

Six cats with rotation of one or both eyes (strabismus) produced surgically in the early postnatal period demonstrated torsional deviation of the eyes by 10–20 degrees in addition to the rotation. The spatial distribution of retrograde labeled neurons in field 17 was studied by microiontophoretic administration of horseradish peroxidase into individual cortical columns in fields 17 and 18. These studies showed that rotation of the eyes increased the extent of horizontal neuronal connections in field 17 along the projection of the vertical meridian of the field of vision. It is suggested that this reorganization of neuronal connections may support functional changes compensating for eye rotation, as described in the literature. __________ Translated from Morfologiya, Vol. 127, No. 2, pp. 69–71, March–April, 2005.