Development of spinal cord projections from neocortical transplants heterotopically placed in the neocortex of newborn hosts is highly dependent on the embryonic locus of origin of the graft

Previous experiments based on heterotopic transplantation paradigms have indicated that the distribution of efferents developed by layer V pyramidal cells seems to be related to where in the neocortex the cells develop and not to where they were generated. The present study was undertaken in an attempt to obtain a quantitative estimation of the weight of extrinsic factors in the development of neocortical efferents. Fragments of embryonic (E15–E19) frontal or occipital cortex were grafted homotopically or heterotopically into the frontal or occipital cortex of newborn rats. As adults, the hosts received an injection of a retrograde tracer into the pyramidal tract decussation, and the distribution of the subsequent cell labeling was examined in each category of transplant. The mean numbers of labeled cells were 725 in frontal-to-frontal transplants and 250 in frontal-to-occipital transplants. In occipital-to-frontal transplants, the numbers of labeled cells were extremely low, ranging from 0 to 14. Finally, as expected, practically no cell labeling was found in occipital-to-occipital transplants. Thus, transplants of presumptive frontal origin systematically develop and maintain in adulthood a spinal cord projection even though they are placed in the host occipital cortex. Conversely, transplants of presumptive occipital origin are practically incapable of maintaining a spinal cord projection in adulthood even though they are placed in the host frontal cortex. It seems, therefore, that the generation of regional differences in efferent connectivity found in the mature cortex depends on early regional specification within the neocortical neuroepithelium. © 1996 Wiley-Liss, Inc.     

Timing and plasticity of specification of CaM-Kinase II alpha expression by neocortical neurons

In this work, the differential expression of a chemical marker, the alpha-isoform of the calcium/calmodulin-dependent protein kinase II (CaM-Kinase II alpha) and the development of the spinal cord projection were used to determine in vivo the embryonic stages at which different aspects of the phenotype of neocortical cells are specified. We first performed a quantitative, immunocytochemical study on the levels of CaM-Kinase II alpha expression in the frontal, parietal and occipital cortical areas of control adult rats. We found that the levels of expression of CaM-Kinase II alpha were larger in the frontal and parietal areas than in the occipital areas. In addition, all layer V neurons identified as projecting to the spinal cord were CaM-Kinase II alpha immunopositive. We then grafted embryonic day (E) 12 or 14 cells from the presumptive frontal or occipital cortex of donor fetuses into the frontal or occipital cortex of newborn hosts. Cortical cells grafted at E12 differentiate neurons with molecular (CaM-Kinase II alpha) and connectivity (spinal cord projection) phenotypes appropriate to the cortical area where they complete their development whereas cells taken at E14 differentiate neurons with molecular and connectivity phenotypes appropriate to their cortical locus of origin. These findings suggest that E12 progenitors destined to generate layer V neurons are multipotent. The final phenotype of their progeny depends on regionalizing signals expressed in the environment. Later in corticogenesis, committed progenitors become unable to respond to regionalizing signals and generate neurons whose phenotype is appropriate to the initial cortical position of the precursor.     

Abnormalities in the Development of the Tectal Projection from Transplants of Embryonic Occipital Cortex Placed in the Damaged Occipital Cortex of Newborn Rats

We have examined the degree of precision in the topographic arrangement of the tectal projection developed by homotopic transplants of embryonic occipital cortex and tried to determine whether the development of the corticotectal projection is exclusively dependent on environmental cues or is also controlled by intrinsic factors. Transplants of embryonic (E16) occipital cortex were grafted into various areas of the occipital cortex (Oc1 or Oc2) of newborn rats and the organization of the tectal projection arising from the transplants was subsequently examined by injecting different neurotracers into the transplants. Our results indicate that in most cases the laminar and tangential distributions of the tectal projections from the transplants were abnormal. Indeed, whatever the location of the transplant in the host occipital cortex and whatever the placement of the injection into the transplant, a hybrid distribution of the tectal labeling was found, reminiscent of the pattern observed following tracer deposits in both Oc1 and Oc2 in intact animals. Since the grafts were composed of cells of both Oc1 and Oc2 embryonic origin, it is likely that the hybrid pattern of efferents reflects the heterogeneity of the embryonic origin of the cells composing the graft. These findings provide evidence that the development of the topographic distribution of neocortical efferents is not only dependent on factors extrinsic to the cortex and further indicate that even within one single cortical region, the occipital cortex, different areas (Oc1 vs Oc2) are not totally interchangeable. These findings might have important implications in transplantation experiments aiming at the reconstruction of damaged neocortical circuitry where a precise “point-to-point” reconstruction of the circuitry is expected.     

Regeneration of adult dorsal root axons into transplants of fetal spinal cord and brain: a comparison of growth and synapse formation in appropriate and inappropriate targets

Cut dorsal root axons regenerate into transplants of embryonic spinal cord and form synapses that resemble those found in the dorsal horn of normal spinal cord. One aim of the present study was to determine whether these axons also regenerate into and establish synapses within transplants of embryonic brain. A second aim was to compare the patterns of growth in embryonic brain and spinal cord transplants. Embryonic spinal cord or brain was transplanted into the lumbar enlargement of adult Sprague-Dawley rats, the L4 or L5 dorsal root was cut, and the cut root was juxtaposed to the transplant. The transplants included whole pieces or dissociated cell suspensions of embryonic day 14 (E14) spinal cord, or whole pieces of E14 neocortex, E18 occipital cortex, E15 cerebellum, or E18 hippocampus. One month later the regenerated dorsal root axons were labeled by immunocytochemical methods to demonstrate calcitonin gene-related peptide (CGRP). CGRP-immunoreactive axons regenerated into all the transplants examined and formed synapses in the neocortex and cerebellum transplants in which they were sought. Synapses were far rarer in neocortex and cerebellum than we had observed previously in transplanted spinal cord, and the patterns of growth differed in transplants of spinal cord and brain. In solid transplants of spinal cord, regenerated axons remained relatively close to the interface with the dorsal root, branched, and formed bundles. Areas of dense ingrowth were separated by regions with few labeled axons. In transplants of brain regions, the regenerated axons were few, unbranched, and appeared as individual fibers rather than in bundles, but they were distributed widely in neocortex transplants. The results of quantitative studies confirmed these observations. The area fraction occupied by regenerated axons in solid spinal cord transplants was significantly larger than in occipital cortex or cerebellum transplants. Distribution histograms of the area occupied in transplants demonstrated that regenerated axons were distributed sparsely but homogeneously in transplants of brain, whereas spinal cord transplants were heterogeneous for regenerated axons and contained areas in which growth was dense or sparse. In contrast, several measurements of axon distribution, including area, longest axis, and length of lateral extension, indicated that CGRP-labeled axons spread more widely in occipital cortex transplants than in solid transplants of spinal cord or cerebellum. The results indicate that embryonic CNS tissues that are not normal targets support or enhance the growth of severed dorsal roots and suggest that the conditions that constitute a permissive environment for regenerating axons are relatively nonspecific.(ABSTRACT TRUNCATED AT 400 WORDS)     

Efferent connections of homotopic and heterotopic transplants of embryonic neocortical tissue placed in the occipital neocortex of newborn rats

We examined (i) the capacity of transplants of embryonic neocortex to restore corticofugal systems disrupted following neonatal damage to the occipital cortex and (ii) the influence of the embryonic origin of the transplanted neurons on the reconstruction of the corticofugal circuitry. Transplants of embryonic occipital or frontal cortex were grafted homo- or heterotopically into the damaged occipital cortex of newborn rats. Several months after grafting, an anterograde tracer was injected into each category of transplants. Homotopic transplants developed a set of projections directed exclusively towards most of the cortical and subcortical visual targets normally contacted by occipital cortical neurons. Heterotopic transplants formed a hybrid system of efferent projections that reflected both their embryonic origin and their new location within the host cortex. These findings are consistent with previous results indicating that fetal frontal and occipital neurons are not interchangeable. Consequently, transplantations aiming at the reconstruction of neural circuits disrupted following neonatal damage affecting a given cortical area should only use fetal cortical cells taken from the same cortical locale.     

Specific neurotrophic interactions between cortical and subcortical visual structures in developing rat: in vitro studies

We investigated the influence of different subcortical structures on the survival of specific populations of occipital cortex neurons developing in vitro. Explants of embryonic day 14-15 (E14-15) rat cortex were cultured for 5 days with explants of either diencephalon or optic tectum or another occipital cortex explant. Stereological analysis of the explants revealed that after 5 days in vitro (5 DIV) all the cortical explants contained equal proportions of healthy neurons, glia, neuropil, and degenerating profiles, regardless of the culturing conditions. In order to determine if different neuronal populations survived preferentially in the cortical explants as a result of the presence of potential target or afferent structures, we used HRP filling and 3H-thymidine labeling techniques. Specific differences in the morphology of the cells and their time of origin are found in the cortical explants. In the cortical explants cocultured with diencephalon (Cx + D) the cortical cells that survive tend to be round with small cross-sectional areas and have few neurites. These cells are generated late in the culturing period. The surviving cortical neurons in the cortex plus tectum (Cx + T) cultures are larger--many with a pyramidal-shaped soma and several neurites. These cells are generated earlier in vitro. The cortex cultured with other cortex (Cx + Cx) gives values intermediate to the Cx + D and Cx + T cultures. The results of these experiments suggest that there are diffusible trophic factors that arise from subcortical structures that selectively support the survival of neuron populations in the developing neocortex.     

Maintenance of transient occipitospinal axons in the rat

A developmentally transient occipitospinal projection arising from the visual association cortex persists if a fetal tectum is transplanted to the spinal cord at birth and at the same time, the host tectum is removed. The pathway is not sustained if the transplant is placed in the cord without a tectal lesion or if the tectum alone is damaged. Thus the tectal transplant promotes the survival of normally transient occipitospinal axons as long as another regular target of these axons is removed.     

A transient pyramidal tract projection from the visual cortex in the hamster and its removal by selective collateral elimination

During the early postnatal development of the neocortex in rats there is an axonal projection from the occipital cortex (which includes the visual cortex) to the spinal cord which is subsequently completely removed through a process of selective collateral elimination. In order to determine whether a similar phenomenon occurs during the development of the hamster cortex, we have injected the retrogradely transported fluorescent dye Fast Blue (FB) into the pyramidal decussation of hamsters at various ages. In adult hamsters such an injection results in a band of labeled neurons confined to layer V and to about the rostral two-thirds of the neocortex; no labeled cells are seen in the occipital cortex. However, a similar FB injection made during the first postnatal week results after a 4-day survival in a continuous band of FB-labeled layer V neurons spread throughout the tangential extent of the neocortex, including the occipital cortex. A similar continuous band of FB labeled layer V neurons is seen throughout the tangential extent of the neocortex including the occipital region in hamsters injected during the first postnatal week but allowed to survive until the fourth week (i.e., after the restriction of the widespread neonatal pattern has occurred). Injections of the anterograde tracer wheat germ agglutinin conjugated to horseradish peroxidase made into the occipital cortex, or for comparison, into more rostral cortical regions in hamsters ranging in age from neonates to adults, reveal that the extension of pyramidal tract axons is staggered along the anterioposterior axis of the cortex such that axons originating from the posterior regions lag behind those arising from more rostral areas. The transient occipital projection appears to reach a maximum around the end of the first postnatal week: a large number of labeled occipital axons is seen in the medullary pyramidal tract, and some of these can be followed through the pyramidal decussation and into the dorsal funiculus of the spinal cord. Injections into the occipital cortex on P16 label only a few fibers in the medullary pyramidal tract, and none is labeled in hamsters injected as adults.(ABSTRACT TRUNCATED AT 400 WORDS)     

Connections made by transplants to the cerebral cortex of rat brains damaged in utero

Homografts consisting of pieces of neocortex or dissociated cortical cells were transplanted from fetal rats into the cortex of newborn, microcephalic hosts. The cortex of the hosts lacked cells of the superficial layers, as a result of prenatal administration of a cytotoxic drug methylazoxymethanol. Grafts exhibited an internal organization with a tendency to form a molecular layer, and alternating cell and fiber zones, although these were not consistently oriented with respect to the host cortex. Both pyramidal and nonpyramidal cells survived. Some grafts were shown to receive callosal connections. Axonal outgrowth from transplanted neurons to several host brain areas was demonstrated with retrograde tracers. Outgrowth occurred not only to the contralateral cortex, in which the host's own callosal projection was deficient, but also to the thalamus and spinal cord, in which the host projection was intact. Thus, grafted fetal cortex is capable of making connections in the methylazoxymethanol-damaged host, but the pattern of connections made is not influenced by host deficiencies.     

Attraction Exerted in Vivo by Grafts of Embryonic Neocortex on Developing Thalamic Axons

In a previous study we provided evidence that embryonic (E) day 16 frontal cortical cells grafted into the occipital cortex of newborn rats receive inputs from the ventrolateral (VL) and ventromedial (VM) thalamic nuclei which, normally, project to the frontal cortex (25). The present study was designed to examine further the conditions of development of the thalamic innervation of heterotopic neocortical grafts. We demonstrate that VL/VM axons do not provide transitory aberrant input to the occipital cortex either in intact newborn animals or in rats having received neonatal occipital lesion and subsequent graft of E16 occipital cells. These findings indicate, therefore, that the VL/VM projection to the graft does not result from the stabilization of an initial widespread cortical projection from these thalamic nuclei occurring either spontaneously or in response to the lesion and homotopic transplantation procedures. We also show that the VL/VM projection to frontal-to-occipital grafts develops within a few days posttransplantation and is maintained in adulthood. Finally, this study establishes that most VL/VM axons which enter the grafts are not collaterals of thalamofrontal axons. After having reached the cortex, they proceed caudally primarily within the infragranular layers. The findings of this and previous (25) in vivo studies for the first time provide evidence that developing thalamic axons have the capacity to respond to signals from grafts of E16 cortical cells and are capable of deviating their trajectory to establish contact with the grafts. Only those axons arising from thalamic nuclei appropriate for the cortical locus of origin of the grafted cells respond to the guidance signals. The mechanisms by which the thalamic axons find their way to the graft probably rely on cell-contact signaling and/or long-range attraction exerted by diffusible molecules.     

Organization of raphe-cortical projections in rat: A quantitative retrograde study

Retrograde transport of a fluorescent dye was employed to study the projections from raphe nuclei to neocortex in the rat. The spatial distributions of labeled raphe cells were analyzed quantitatively to determine whether the nuclei are topographically organized with respect to different cortical targets. The dorsal raphe nucleus (DRN), exclusive of the lateral wing regions, has a predominantly (3:1) ipsilateral projection with decreasing numbers of cells projecting to frontal, parietal, and occipital cortex. Overlapping cell groups within the DRN project differentially to these three cortical areas: DRN cells innervating frontal cortex extend more rostrally and laterally than those to either parietal or occipital cortex. The medium raphe and B9 projections are bilaterally symmetric, with equal cell numbers projecting to frontal, parietal, and occipital cortex. The rostro-caudal distributions of cells that project to disparate cortical areas differ in B9 but not in MR. The percentage of cortically projecting cells that are serotonergic is 80% for the DRN, 60% in the MR and 33% in the B9 cell group. The dorsal raphe nucleus and the B9 cell group are organized heterogeneously, and overlapping sets of neurons project differentially upon particular areas of neocortex. In contrast, the median raphe nucleus projects uniformly upon the neocortex and does not exhibit topographic organization. The three rostral raphe nuclei (DR, MR and B9) are each organized according to different rules with regard to their efferent projections to cortex. The differential organization of the raphe nuclei suggests that groups of cells within these three raphe nuclei are likely to innervate different combinations of cortical targets and thus to have different functional effects.     

Attraction exerted in vivo by grafts of embryonic neocortex on developing thalamic axons

In a previous study we provided evidence that embryonic (E) day 16 frontal cortical cells grafted into the occipital cortex of newborn rats receive inputs from the ventrolateral (VL) and ventromedial (VM) thalamic nuclei which, normally, project to the frontal cortex (25). The present study was designed to examine further the conditions of development of the thalamic innervation of heterotopic neocortical grafts. We demonstrate that VL/VM axons do not provide transitory aberrant input to the occipital cortex either in intact newborn animals or in rats having received neonatal occipital lesion and subsequent graft of E16 occipital cells. These findings indicate, therefore, that the VL/VM projection to the graft does not result from the stabilization of an initial widespread cortical projection from these thalamic nuclei occurring either spontaneously or in response to the lesion and homotopic transplantation procedures. We also show that the VL/VM projection to frontal-to-occipital grafts develops within a few days posttransplantation and is maintained in adulthood. Finally, this study establishes that most VL/VM axons which enter the grafts are not collaterals of thalamofrontal axons. After having reached the cortex, they proceed caudally primarily within the infragranular layers. The findings of this and previous (25) in vivo studies for the first time provide evidence that developing thalamic axons have the capacity to respond to signals from grafts of E16 cortical cells and are capable of deviating their trajectory to establish contact with the grafts. Only those axons arising from thalamic nuclei appropriate for the cortical locus of origin of the grafted cells respond to the guidance signals. The mechanisms by which the thalamic axons find their way to the graft probably rely on cell-contact signaling and/or long-range attraction exerted by diffusible molecules.     

Changes in the topographically organized connections between the nucleus isthmi and the optic tectum after partial tectal ablation in adult goldfish

The projection of the nucleus isthmi to the ipsilateral optic tectum was examined in normal goldfish. This was compared to the projection in animals in which the entire visual field had been induced to compress onto a rostral half tectum by caudal tectal ablation. The isthmo-tectal projection was examined by making localized injections of horseradish peroxidase into the optic tecta and observing the patterns of labeled cells within the nucleus isthmi. The teleost nucleus isthmi consists of a cell sparse medulla covered by a cellular cortex, which is thick on the rostral, medial, and dorsal surfaces of the nucleus. Almost all isthmic cells projecting to the tectum were located in the area of thick cortex. In normal fish, rostral tectal injections labeled cells in the rostroventral portion of the thick cortex; injections midway in the rostrocaudal tectal axis labeled more caudodorsally located cells, and caudal tectal injections labeled cells a little further caudally in extreme dorsal cortex. The rostroventral to caudodorsal isthmic axis was therefore seen to project rostrocaudally along the tectum. This topography contrasts somewhat with the situation seen in amphibia where the rostrocaudal tectal axis receives projections from the rostrocaudal isthmic axis. In fish with half-tectal ablations, injections near the caudal edge of the half tectum (at a site that had originally been midtectal) labeled cells that had previously projected to caudal tectum. Rostral tectal injections in fish with compression of the visual field gave a normal pattern of labeled isthmic cells. The results indicate that a topographically ordered isthmo-tectal projection exists in goldfish that may be induced to compress onto a half tectum.     

Early (E12) cortical progenitors can change their fate upon heterotopic transplantation

To help understand how the cortical map is set up during the early stages of corticogenesis, we have examined the developmental fate of embryonic day (E) 12 cortical progenitors in the rat. We have analysed the pattern of thalamic connections and cytoarchitectonic organization developed by progenitor cells removed at E12 from the presumptive parietal or occipital cortex and grafted into the parietal cortex of newborn hosts. Occipital progenitors grafted into the parietal cortex differentiated into neurons that developed reciprocal connections with the ventrobasal complex of the host thalamus. They could also form barrel-like structures, within which axons of the ventrobasal complex were distributed in dense patches. Some of these barrel-like structures were arranged in rows. Moreover, these progenitors failed to develop characteristic traits of occipital cortex cells as they did not establish connections with the dorsal lateral geniculate nucleus. We propose that cortical progenitors are not committed at E12 and, upon heterotopic transplantation, have the capacity to respond to local cues and to subsequently differentiate and maintain major phenotypic characteristics of neurons in their new environment. Only early progenitors are multipotent. By E13/E14, indeed, most cortical cells become irreversibly committed and upon heterotopic transplantation differentiate neurons with phenotypic characteristics of their cortical site of origin (Pinaudeau et al., 2000, Eur. J. Neurosci., 12, 2486-2496).     

Selective elimination of axons extended by developing cortical neurons is dependent on regional locale: experiments utilizing fetal cortical transplants

In adult rats, cortical neurons that extend an exon through the pyramidal tract (a major subcortical efferent projection of the neocortex) are limited to layer V of about the rostral two-thirds of the neocortex. In neonates, however, pyramidal tract neurons are distributed throughout the neocortex, but all of those found in certain areas, such as the posterior occipital region (including primary visual cortex) selectively lose their pyramidal tract axon (Stanfield et al., 1982) yet maintain axon collaterals to other subcortical targets (O'Leary and Stanfield, 1985). To determine if the regional location of a developing pyramidal tract neuron critically influences the maintenance or elimination of the axon collaterals it initially extends, pieces of cortex from embryonic day 17 (E17) rat fetuses (exposed to 3H-thymidine on E15) were transplanted heterotopically into the cortex of newborn (PO) rats; rostral cortex was placed into the posterior occipital region (R----O), or posterior occipital cortex into a rostral cortical locale (O----R). The retrograde tracers Fast blue (FB) and Diamidino yellow (DY) were used to assay for the presence of specific populations of cortical projection neurons within the autoradiographically identified transplants. In terms of the extension and maintenance of pyramidal tract axons, the transplanted neurons behave like the host neurons of the recipient cortical region rather than like those of their site of origin. At P40, following FB injections into the pyramidal decussation on P34, pyramidal tract neurons are labeled within the O----R transplants, but none can be labeled within R----O transplants, although in the same R----O cases transplanted neurons are labeled by an injection of DY in the superior colliculus. However, at P13 pyramidal tract neurons can be identified within the R----O transplants, as well as in the host occipital cortex, following injections made on P9, a period when the distribution of pyramidal tract neurons in normal rats is widespread (Stanfield and O'Leary, 1985b). In a second series of host rats, on P34 FB was injected in the pyramidal decussation of the O----R cases, or in the superior colliculus of the R----O cases, and in both groups DY was injected into the region of contralateral cortex homotopic for the new location of the transplant. On P40, in both the O----R and R----O transplants, many neurons singly labeled with FB or DY are found, but no double dye-labeled cells are seen.(ABSTRACT TRUNCATED AT 400 WORDS)     

Timing and origin of the first cortical axons to project through the corpus callosum and the subsequent emergence of callosal projection cells in mouse

A precise knowledge of the timing and origin of the first cortical axons to project through the corpus callosum (CC) and of the subsequent emergence of callosal projection cells is essential for understanding the early ontogeny of this commissure. By using a series of mouse embryos and fetuses of the hybrid cross B6D2F₂/J weighing from 0.36 g to 1.0 g (embryonic day E15.75–E17.25), we examined the spatial and temporal distribution of callosal projection cells by inserting crystals of the lipophilic dye (DiI: 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate) into the contralateral white matter just lateral to the midsagittal plane. Around 0.4 g or E15.8, retrogradely labeled cells were found restricted to a discrete cluster continuously distributed from the most ventral part of presumptive cingulate cortex to the hippocampus. During subsequent development, however, the tangential distribution of these labeled cells in ventromedial cortex did not extend further dorsally, and in fetuses where the CC became distinct from the hippocampal commissure (HC), labeled axons of cells in the ventral cingulate cortex were observed to intersect the callosal pathway and merge with labeled axons of the HC derived from cells in the hippocampus. The first cortical axons through the CC crossed the midline at about 0.64 g or E16.4, and these axons originated from a scattered neuronal population in the dorsal to lateral part of the presumptive frontal cortex. The earliest callosal cells were consistently located in the cortical plate and showed an immature bipolar appearance, displaying an ovoid- or pearl-shaped perikaryon with an apical dendrite coursing in a zig-zagging manner toward the pial surface and a slender axon directed toward the underlying white matter. Callosal projection cells spread progressively with development across the tangential extent of the cerebral cortex in both lateral-to-medial and rostral-to-caudal directions. In any cortical region, the first labeled cells appeared in the cortical plate and their number in the subplate was insignificant compared to that in the cortical plate. Thus, these results clarify that the CC is pioneered by frontal cortical plate cells, and the subsequent ontogeny of callosal projection cells proceeds according to the gradient of cortical maturation. J. Comp. Neurol. 400:197–206, 1998. © 1998 Wiley-Liss, Inc.     

Specific neurotrophic interactions between cortical and subcortical visual structures in developing rat: in vivo studies

The specificity of trophic interactions in the rat visual system is investigated in vivo by using a combination of tissue culture and CNS transplantation methods. In a companion paper (Repka and Cunningham: '87) we showed that explants of embryonic day 14 (E14) occipital cortex are biased to contain different cortical cell populations depending on whether the explants develop in culture with diencephalon or with optic tectum. In this study we transplanted these precultured cortical explants into the cavity created by a lesion of the occipital cortex in newborn rats and then measured the neuron-occupied volume and the numbers of thymidine-labeled cells in the surviving ipsilateral dorsal lateral geniculate nucleus (dLGN) of the host rats. The results were compared to animals with lesions but no transplants, animals with transplants of E14 cortical tissue that had not been precultured, and animals with cerebellar transplants that had been similarly precultured either with other cerebellar tissue or with diencephalon. At 5 days postlesion, both the largest dLGN volume and the greatest number of labeled dLGN neurons survive in animals with cortical transplants precultured with diencephalon or other cortex. The surviving dLGN neurons that are rescued by these transplants are generated on E15 or E16, a period that corresponds to the latter part of geniculate neurogenesis. Relatively few cells generated on E14 survive in any group of animals. Furthermore, animals with all types of cortical transplants have significantly larger volumes of surviving dLGN than animals with either lesions only or cerebellar transplants.(ABSTRACT TRUNCATED AT 250 WORDS)     

Tectal transplants into the occipital cortex of the newborn rat

Embryonic day 15 rostral tectum (presumptive superior colliculus) was transplanted into the occipital cortex of newborn rats. One to two months later, the transplants were visualized and injected with either horseradish peroxidase (HRP) or wheat germ agglutinin conjugated with HRP (WGA/HRP). After the appropriate survival time and processing with either diaminobenzidine (DAB) or tetramethylbenzidine tetrahydrochloride (TMB), HRP-labelled pyramidal cells were found in layer V of the host ipsilateral occipital cortex. Thus, the occipitotectal connections are formed between host and graft despite the fact that the fibers must grow in a direction opposite to their normal course to reach the aberrantly positioned tectal graft.     

The transient corticospinal projection from the occipital cortex during the postnatal development of the rat

The transient occipital cortical component of the pyramidal tract which we previously had identified during the postnatal development of the rat (Stanfield et al., '82) has been studied with anterograde as well as retrograde techniques. A continuous band of retrogradely labeled layer V neurons which spans the entire cortex including the occipital cortex is seen following injections of the fluorescent marker Fast Blue into the pyramidal decussation during the first postnatal week. No labeled cells are found in the occipital cortex following similar injections made on postnatal day 20 (P20), although such injections label many neurons in the more rostral cortical fields. However, if the Fast Blue injection is made on P2 and the animal is allowed to survive until P25 a large number of Fast Blue-labeled layer V neurons is found in the occipital cortex, even though an acute, second injection of the retrograde tracer Nuclear Yellow made into the pyramidal decussation shortly before the animal is killed results in no occipital cortical labeling. When Fast Blue injections confined to the mid- or upper-cervical spinal cord are made on P4 and the animals are killed on P9, again many retrogradely labeled neurons are found in the occipital cortex. Further, when injections of 3H-proline or wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) confined to the occipital cortex are made during the first 2 postnatal weeks, anterogradely transported label is seen within the pyramidal tract. At all stages examined the amount of such label and its caudal extent are less than that seen following similar injections into the parietal or frontal cortex. The greatest extent of the labeled occipital cortical fibers is reached at about the end of the first postnatal week and the number of these fibers seems to peak at about this same time. At this stage many of these labeled axons extend for a considerable distance down the spinal cord with some reaching as far caudal as lower lumbar levels, and at this stage some of these labeled occipital corticospinal fibers enter into the spinal gray. Over the next week the number of occipital cortical fibers in the pyramidal tract rapidly decreases and by P17 occipital cortical injections of 3H-proline or WGA-HRP result in virtually no transported label caudal to the pons.(ABSTRACT TRUNCATED AT 400 WORDS)     

Laminar and areal differences in the origin of the subcortical projection neurons of the rat somatosensory cortex

Fluorescent retrograde tracing techniques were employed in a double-labelling paradigm to determine the distribution of corticospinal, corticotectal, and corticotrigeminal projection neurons in layer Vb of the adult and neonatal rat somatosensory cortex. The double-labelling paradigm allowed a direct comparison of the cortical distribution of neurons projecting to each target and identification of neurons projecting to more than one target. In the adult rat, each population of projection neurons was found to have a unique laminar and/or areal distribution. Corticospinal projection neurons were located throughout the width of layer Vb in the medial granular portion of somatosensory cortex, while corticotrigeminal projection neurons were distributed throughout the width of layer Vb in the more laterally located dysgranular portion of somatosensory cortex. Corticotectal projection neurons were located more superficially in layer Vb than either corticospinal or corticotrigeminal projection neurons and found scattered throughout both dysgranular and granular somatosensory cortex. Each combination of subcortical injections also resulted in double labelling a small percentage of uniquely distributed neurons. These distribution differences coupled with measurements of cell size allowed us to identify the parent population of the dual projection neurons. Subpopulations of corticotectal neurons also project to the brainstem trigeminal complex and to the spinal cord. Subpopulations of corticotrigeminal neurons also project to the spinal cord, and a proportion of corticotrigeminal neurons projects to at least two targets within the brainstem trigeminal complex (nucleus principalis and subnucleus interpolaris). In the adult rat, corticospinal neurons (as defined by either laminar position or somal size) did not appear to give off collaterals to either the superior colliculus or brainstem trigeminal complex. In the neonatal rat, double-labelled neurons which project to both the spinal cord and the tectum are distributed throughout the full width of layer Vb, rather than restricted to the superficial portion of the layer as in the adult rat. Further, it appears as if the ontogenetic change in the laminar distribution of corticospinal and tectal projection neurons is achieved by mechanisms of selective process elimination rather than cell death. These results are discussed in terms of both the developmental factor which may contribute to the discrete distribution of cortical projection neurons found in the adult and the functional significance of bifurcating projection neurons.