Callosal Pattern Research Articles

Selective plasticity of callosal neurons in the adult contralesional cortex following murine traumatic brain injury

Traumatic brain injury (TBI) results in deficits that are often followed by recovery. The contralesional cortex can contribute to this process but how distinct contralesional neurons and circuits respond to injury remains to be determined. To unravel adaptations in the contralesional cortex, we used chronic in vivo two-photon imaging. We observed a general decrease in spine density with concomitant changes in spine dynamics over time. With retrograde co-labeling techniques, we showed that callosal neurons are uniquely affected by and responsive to TBI. To elucidate circuit connectivity, we used monosynaptic rabies tracing, clearing techniques and histology. We demonstrate that contralesional callosal neurons adapt their input circuitry by strengthening ipsilateral connections from pre-connected areas. Finally, functional in vivo two-photon imaging demonstrates that the restoration of pre-synaptic circuitry parallels the restoration of callosal activity patterns. Taken together our study thus delineates how callosal neurons structurally and functionally adapt following a contralateral murine TBI.

Visual Interhemispheric and Striate-Extrastriate Cortical Connections in the Rabbit: A Multiple Tracer Study.

Previous studies in rabbits identified an array of extrastriate cortical areas anatomically connected with V1 but did not describe their internal topography. To address this issue, we injected multiple anatomical tracers into different regions in V1 of the same animal and analyzed the topography of resulting extrastriate labeled fields with reference to the patterns of callosal connections and myeloarchitecture revealed in tangential sections of the flattened cortex. Our results extend previous studies and provide further evidence that rabbit extrastriate areas resemble the visual areas in rats and mice not only in their general location with respect to V1 but also in their internal topography. Moreover, extrastriate areas in the rabbit maintain a constant relationship with myeloarchitectonic borders and features of the callosal pattern. These findings highlight the rabbit as an alternative model to rats and mice for advancing our understanding of cortical visual processing in mammals, especially for projects benefiting from a larger brain.

Retinal input influences the size and corticocortical connectivity of visual cortex during postnatal development in the ferret

Retinal input plays an important role in the specification of topographically organized circuits and neuronal response properties, but the mechanism and timing of this effect is not known in most species. A system that shows dramatic dependence on retinal influences is the interhemispheric connection through the corpus callosum. Using ferrets, we analyzed the extent to which development of the visual callosal pattern depends on retinal influences, and explored the period during which these influences are required for normal pattern formation. We studied the mature callosal patterns in normal ferrets and in ferrets bilaterally enucleated (BE) at postnatal day 7 (P7) or P20. Callosal patterns were revealed in tangential sections from unfolded and flattened brains following multiple injections of horseradish peroxidase in the opposite hemisphere. We also estimated the effect of enucleation on the surface areas of striate and extrastriate visual cortex by using magnetic resonance imaging (MRI) data from intact brains. In BEP7 ferrets we found that the pattern of callosal connections was highly anomalous and the sizes of both striate and extrastriate visual cortex were significantly reduced. In contrast, enucleation at P20 had no significant effect on the callosal pattern, but it still caused a reduction in the size of striate and extrastriate visual cortex. Finally, retinal deafferentation had no significant effect on the number of visual callosal neurons. These results indicate that the critical period during which the eyes influence the development of callosal patterns, but not the size of visual cortex, ends by P20 in the ferret.

Diffusion Tensor Imaging Detects Early Cerebral Cortex Abnormalities in Neuronal Architecture Induced by Bilateral Neonatal Enucleation: An Experimental Model in the Ferret

Diffusion tensor imaging (DTI) is a technique that non-invasively provides quantitative measures of water translational diffusion, including fractional anisotropy (FA), that are sensitive to the shape and orientation of cellular elements, such as axons, dendrites and cell somas. For several neurodevelopmental disorders, histopathological investigations have identified abnormalities in the architecture of pyramidal neurons at early stages of cerebral cortex development. To assess the potential capability of DTI to detect neuromorphological abnormalities within the developing cerebral cortex, we compare changes in cortical FA with changes in neuronal architecture and connectivity induced by bilateral enucleation at postnatal day 7 (BEP7) in ferrets. We show here that the visual callosal pattern in BEP7 ferrets is more irregular and occupies a significantly greater cortical area compared to controls at adulthood. To determine whether development of the cerebral cortex is altered in BEP7 ferrets in a manner detectable by DTI, cortical FA was compared in control and BEP7 animals on postnatal day 31. Visual cortex, but not rostrally adjacent non-visual cortex, exhibits higher FA than control animals, consistent with BEP7 animals possessing axonal and dendritic arbors of reduced complexity than age-matched controls. Subsequent to DTI, Golgi-staining and analysis methods were used to identify regions, restricted to visual areas, in which the orientation distribution of neuronal processes is significantly more concentrated than in control ferrets. Together, these findings suggest that DTI can be of utility for detecting abnormalities associated with neurodevelopmental disorders at early stages of cerebral cortical development, and that the neonatally enucleated ferret is a useful animal model system for systematically assessing the potential of this new diagnostic strategy.

Development of the visual callosal cell distribution in the rat: mature features are present at birth.

In the present study, the early postnatal distribution and subsequent fate of visual callosal neurons were studied in neonatal rat pups. Previous studies had indicated that the adult pattern of visual callosal neurons was sculpted from an initially uniform distribution in the neonatal cortex. To reexamine this issue, we used a sensitive tracer, latex microspheres conjugated either to rhodamine or fluorescein, that was injected into the occipital cortex of one hemisphere in pups on the day of birth (PND 1), PND 6, or PND 12. Examination of the resulting retrograde labeling of cortical neurons in the opposite hemisphere indicates that features of the mature visual callosal pattern are present as early as PND 1. At all stages of postnatal development, the relative density of callosal projection cells varies consistently across the mediolateral extent of primary visual cortex-it is always highest in the region of the 17/18a border and lowest in the body of area 17. These data strongly suggest that, from the outset, visual cortical neurons in the region of the 17/18a border preferentially make connections with the opposite hemisphere. The results of experiments in which callosal neurons were labeled on the day of birth indicate that only those neurons that have migrated to their final cortical destinations have extended callosal axons into the vicinity of the visual cortex in the opposite hemisphere. The initial pattern of callosal neurons resembles a dense, compact version of the mature one, and the present study suggests that much of the remaining change in the appearance of this pathway may be accounted for by the decrease in the overall density of neurons that is due to expansion of the cortical gray matter during postnatal life. Taken together, these results suggest that the development of the visual callosal pathway in the rat may be more similar to that in the monkey than has been reported previously.

The callosal pattern in striate cortex is more patchy in monocularly enucleated albino than pigmented rats

We investigated the effect of neonatal monocular enucleation on the pattern of interhemispheric connections through the corpus callosum in occipital cortex of pigmented and albino rats. Callosal connections were revealed in tangential sections through the flattened cortex following multiple injections of horseradish peroxidase into the opposite hemisphere. In pigmented rats, we found that monocular enucleation induces the development of an anomalous band-like accumulation of callosal connections in middle portions of striate cortex in the hemisphere ipsilateral to the remaining eye, as reported previously. In one-eyed albino rats, we also found callosal connections anomalously placed in middle portions of striate cortex, but they tended to form several patches of labeling rather than a single continuous band as in pigmented rats. Densitometric analysis of the callosal patterns revealed that this difference between rat strains was statistically significant. The increased patchiness in the callosal pattern of one-eyed albino rats may reflect differences in the ipsilateral retinal projections in albino versus pigmented rats.

Overall pattern of callosal connections in visual cortex of normal and enucleated cats.

The effect of neonatal bilateral enucleation on the overall distribution of callosal connections in striate and extrastriate visual cortex of the cat was studied using tangential sections from the physically unfolded and flattened cortex. Callosal neurons were labeled by administering the anatomical tracer horseradish peroxidase directly to the transected corpus callosum. The pattern of callosal connections in binocularly enucleated cats showed both consistent differences and consistent similarities with the pattern in normal cats. In agreement with previous studies, it was found that callosal labeling at the 17/18 border of enucleated cats was considerably sparser than in normal cats. Moreover, we found that the strip containing the majority of labeled cells at the 17/18 border was narrower than in normal cats. In both normal and enucleated cats, scattered cells were distributed on either side of the 17/18 callosal strip, well into areas 17 and 18. In much of extrastriate cortex, the pattern of callosal connectivity in enucleated cats looked surprisingly normal. Details of the callosal pattern that were consistently found in normal cats could also be recognized in binocularly enucleated cats, such as two to four bridges of labeling spanning areas 18 and 19. Also, four zones that were free of callosal connectivity in area 7, on the banks of the suprasylvian sulcus, and in the posterior suprasylvian sulcus were found in both normal and enucleated cats. Finally, as in normal cats, dense cell labeling occurred on the crown of the suprasylvian gyrus at its posterior end, from which it extended laterally across both banks of the suprasylvian sulcus and into the fundus of this sulcus. The results of this study suggest that, although the stabilization of callosal connections at the 17/18 border region appears to depend on visual input, this input plays a less prominent role in the stabilization of callosal connections in extrastriate visual cortex.

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.

Organization of visual cortex in the mouse revealed by correlating callosal and striate-extrastriate connections

In this study, we have investigated the organization of mouse visual cortex by correlating in detail the distribution of striate-extrastriate projections with the pattern of callosal connections revealed by the transport of horseradish peroxidase from the contralateral hemisphere. Single injections of 3H-proline into striate cortex produce 8-9 discrete projection fields in the belt of cortex surrounding area 17. The number and arrangement of these fields closely resemble the pattern of extrastriate visual areas in the rat. The callosal pattern is also very similar to that in the rat, and provides a set of landmarks for the location of the striate-recipient zones. Thus, cortical regions containing dense aggregations of callosal cells and terminations surround totally or partially the sparsely callosal striate-recipient zones. By comparing our results with previous accounts of the rat visual plan, we were able to identify in lateral extrastriate cortex of the mouse areas anterolateral (AL), lateromedial (LM), laterointermediate (LI), laterolateral (LL), posterolateral (PL), and posterior (P). We also observed 1-2 projections fields into anteromedial (AM) extrastriate cortex, and one field (S) into the posteromedial border of the head representation in primary somatosensory cortex. Our results support the notions that the visual cortex in the mouse is subdivided into multiple visual areas, and that these areas are arranged according to a plan that is common in rodents.

In vivo visualization of callosal pathways: a novel approach to the study of cortical organization

I describe here the successful visualization of interhemispheric callosal connections in the live mammalian cortex. The development of this method was prompted by the finding that fluorescent tracer labeling of groups of cortical neurons, when done under optimal conditions, is sufficiently intense to be visible even in the whole brain preparation. The new approach could provide a useful tool for enhanced precision in localizing cortical modules in vivo. In a typical experiment, rats had their left cortical hemisphere extensively injected with the fluorescent tract-tracer bis-Benzimide (BB). After appropriate survival, the right cortical hemisphere was illuminated with UV light and the fluorescing callosal pattern could be discerned under the network of blood vessels even with the unaided eye. The pattern, although diffuse, was grossly similar to the pattern of callosal connections as seen in flattened, sectioned cortex. Features that could be discerned were: the main callosal band straddling the lateral border of area 17, several rings and bands in extrastriate areas 18a and 18b, and a major band straddling the lateral border of area 3. The vitally visualized callosal pattern was used to guide injections of either wheat germ agglutinin conjugated to HRP (WGA-HRP) or rhodamine-labeled microspheres (RLM) into precisely localized sites in occipital cortex. There were numerous instances of doubly labeled neurons stained both with BB and WGA-HRP or RLM, suggesting that uptake of BB combined with UV exposure did not hinder the ability of stained neurons to take up and transport a second tracer. It is suggested that vital tract tracing be used as a tool for enhanced precision in studies of cortical connectivity.

The pattern of callosal connections in posterior neocortex of congenitally anophthalmic rats.

In an effort to assess the innate capacity of the central visual system to specify corticocortical connectivity in the absence of retinal afferents, we examined the tangential distribution of callosal cells and terminations in posterior neocortex of congenitally anophthalmic rats. Although our results indicate that the callosal pattern is clearly anomalous in these rats, all features of the normal visual callosal pattern are recognizable in mutant rats, indicating that central visual pathways can generate many aspects of normal interhemispheric connectivity in the absence of input from the periphery. On the other hand, the presence of anomalies in the pattern indicates that the eyes are necessary to fine-tune the distribution of callosal connections at some developmental stage. Moreover, the fact that abnormalities in the callosal pattern of mutant rats are the same as those previously described in rats enucleated at birth suggests that the eyes begin to exert their influence on callosal development after birth.

Topography of interhemispheric connections in neocortex of mice with congenital deficiencies of the callosal commissure.

Normally, axons within the corpus callosum are ordered according to the cortical regions from which they originate, and callosal cells and terminations form elaborate cortical patterns related to the underlying topographic representations of the sensory periphery. About 30% of mice of the BALB/c strain show congenital deficiencies of the callosal commissure which range from total absence of the corpus callosum to a moderate reduction in the size of this commissure. In the light of current theories about the origin of these callosal deficiencies, it seems likely that fibers crossing the midplane in mutant mice have to circumvent local disturbances along their migration path. Since these disturbances in fiber trajectory may, in turn, alter the overall pattern of callosal projections, we set out to investigate whether the distribution of callosal connections in mice with marked deficiencies of the corpus callosum is as ordered as in normal mice. In groups of normal and mutant mice, we used multiple injections of horseradish peroxidase to reveal the overall distribution of callosal connections and restricted injections of horseradish peroxidase conjugated with wheat germ agglutinin to reveal finer aspects of the organization of the callosal pathway in these animals. Our results show that the number of labeled cells is reduced in mice with a small corpus callosum and that no labeled cells are present in the neocortex of acallosal mice. Furthermore, the topographic distribution of fibers within the corpus callosum of mutant mice can be significantly less ordered than in normal mice. However, even in mice with extreme deficiencies of the corpus callosum, callosal fibers originate from and terminate in all major areas of the cortex, and, within these areas, callosal cells and terminations are distributed according to the normal plan. The laminar distribution of callosal cells also appears normal in these mice. These findings indicate that gross developmental anomalies of the corpus callosum do not prevent normal specification of the callosal pattern during development. Within the context of current theories about the origin of congenital callosal deficiencies, our findings suggest that callosal fibers are able to establish appropriate contralateral connections in spite of alterations of their migration route. They also suggest that fiber topography within the corpus callosum does not play an important role in guiding migrating axons to their correct contralateral targets. Finally, our failure to find labeled fibers within the anterior commissure indicates that this commissure does not serve as an alternative route for deviated callosal axons.

Interhemispheric connections in the visual cortex of the squirrel monkey (Saimiri sciureus).

The callosal connections within the posterior parietal and occipital cortices were studied in the squirrel monkey with horseradish peroxidase tracing techniques. The data were evaluated with particular emphasis on the relationship of major callosal connections along the 17-18 border. The overall pattern of callosal connections in the squirrel monkey also was compared with callosal patterns in other New World simians. Our results show that the dense band of callosal connections along the 17-18 border in the squirrel monkey differs from the connections observed in other New World monkeys in that it is virtually confined to area 18 and avoids area 17. In addition to a continuous band of callosal connections in area 18 that parallels the 17-18 border, rostral extensions of the band are oriented perpendicular to the 17-18 border and present an obvious periodicity. The remaining parieto-occipital cortex contains a complex pattern of callosal connections that is strikingly similar to patterns reported for other New World monkeys. Thus, it is likely that the dorsolateral extrastriate visual cortex in the squirrel monkey is organized in a manner similar to that found within other New World monkeys.