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R D Lund

Publications and source records attributed to R D Lund.

At least 145 records · Page 8Linked to original sources

Prenatal development of the optic projection in albino and hooded rats.

The development of retinofugal projections has been examined in albino and hooded rat embryos from embryonic day 16 to birth (E21.5). Horseradish peroxidase (HRP) was injected intraocularly through the uterine wall and its anterograde transport revealed with TMB and DAB. The retrograde transport of HRP or the fluorescent dyes Nuclear yellow, Fast blue and propidium iodide from optic tract, superior colliculus (SC) or lateral geniculate body (LG) injections was used to demonstrate the origin of the projections. Superficial projections to the contralateral SC were first identified at E16. A light projection to the entire medio-lateral extent of the ipsilateral SC could be detected a day later. The optic axons grow over the surface of the diencephalon at E16 and it was only at later stages that the fibers were observed to invade successively deeper parts of the LG. A superficial projection to the ipsilateral LG could first be detected at E17. Both the ipsilateral and contralateral projections grew through the entire dorso-ventral extent of the lateral geniculate body: some restriction of the axons to their normal adult termination zones could be detected by E21. No difference in the distribution of projections could be detected between the albino and pigmented rats although the projections were lighter, and possibly because of this were detected later, in the albino rats. At all the ages examined in this study labeled retinal ganglion cells were observed in the non-injected eyes after injection of label into the contralateral eye. The use of persistent fluorescent dyes showed that these retinal ganglion cells did not survive for more than 5 days postnatally. The projection to the uninjected eye came preferentially from ganglion cells in the lower nasal retina while the ipsilateral central projections came predominantly but not exclusively from the lower temporal retina of the injected eye. It appears, therefore, that the initial projections of optic axons in the rat are not limited to their normal termination zones and that the choice of pathway at the chiasm appears to be only loosely controlled.

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Modification of visual callosal projections in rats.

The effects of a variety of developmental manipulations on the distribution of the callosal pathway to visual cortex were examined by using the Fink Heimer technique in adult rats. First, the callosal projections in albino and pigmented rats were compared and found to be similar. The callosal pathway was limited in area 17 to a region adjoining its lateral border with area 18a. Second, dark-reared rats were found to have normal callosal projections. Third, rats bilaterally enucleated at birth and expanded callosal inputs within area 17. Fourth, monocular enucleation at birth produced an expanded callosal pathway to area 17 contralateral to the enucleation and normal callosal projections to the opposite hemisphere. The expanded callosal inputs after enucleation showed a patchy distribution and usually avoided the most medial part of area 17. Fifth, a reduction in the callosal projections to the area 17/18a border was found after neonatal unilateral optic tract lesions. Sixth, expanded callosal inputs to area 17 were found following unilateral thalamic lesions at birth. The abnormal projection occupied mainly layers IV and III. The results of the different experiments indicate that the detailed distribution of the visual callosal projection within area 17 depends heavily on the organization of the retinogeniculocortical pathways to each hemisphere.

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Alterations of the crossed parabigeminotectal projection induced by neonatal eye removal in rats.

Projections of the parabigeminal nucleus to the contralateral superior colliculus and dorsal lateral geniculate nucleus were examined in normal adult pigmented rats and in adult rats from which one or both eyes had been removed at birth. In normal rats the crossed parabigeminotectal projection is restricted to the superficial layers in anterior and medial areas of colliculus, regions innervated also by the lower temporal portion of the ipsilateral retina. In unilaterally enucleated animals the crossed parabigeminotectal projection to the "denervated" colliculus is expanded, as is the retinal projection from the ipsilateral eye. In addition, there is a crossed parabigeminal projection to the "denervated" dorsal lateral geniculate nucleus in these rats. In bilaterally enucleated animals the parabigeminotectal projection is expanded, but not as greatly as in unilateral enucleation cases; there is a crossed parabigeminothalamic projection in these animals as well. The corresponding termination patterns of the contralateral parabigeminal nucleus and the ipsilateral retina in the normal superior colliculus may indicate a functional and/or developmental interdependence between the projections from these two regions. The existence of an expanded parabigeminotectal projection in bilaterally enucleated rats shows that a sustained ipsilateral retinotectal projection is not necessary for the establishment of a crossed parabigeminotectal projection, but points to the possibility that ipsilateral retinal input may constrain the parabigeminal projection to terminate within certain boundaries. The even greater expansion of the projection from the parabigeminal nucleus to the colliculus which receives an expanded projection from the ipsilateral retina of unilaterally enucleated rats suggests that the functional organization of the ipsilateral retinotectal projection may be capable of restricting the size of the terminal field of the crossed parabigeminotectal projection.

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Transplantation of reaggregates of embryonic neural retinae to neonatal rat brain: differentiation and formation of connections.

Embryonic day 14 neural retinae were dissociated into single cells and reaggregated prior to transplantation over the superior colliculi of newborn rats. One month after transplantation, the brains of the host rats were examined for transplant differentiation and the formation of projections from the transplant to the host brain. All reaggregated retinal transplants differentiated in the host brain, showing normal lamination and cellular morphology. Electron microscopic examination demonstrated normal synaptology with the transplanted retinal neuropil. Horseradish peroxidase injections into the host superior colliculus retrogradely filled cells within the transplant. These cells were found in the lamina corresponding to the ganglion cell layer and displayed a morphology characteristic of normal ganglion cells. Lesions of the transplants confirmed the projection of the reaggregated retinal transplants to superior colliculus. Degeneration was also traced into a number of ther nuclei that are normally retinorecipient. The presence of degenerating synaptic terminals resembling those seen after eye removal in control rats was confirmed by electron microscopic examination. It appears that despite disruption of initial cell-cell associations early in retinal development and prior to transplantation, the reaggregated transplants retain the ability to differentiate and form appropriate connection with the host brain.

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A crossed parabigemino-lateral geniculate projection in rats blinded at birth.

Anatomical techniques are used to demonstrate a projection from the parabigeminal nucleus (PBG) to the contralateral dorsal lateral geniculate nucleus (dLGN) in adult pigmented rats which had been bilaterally enucleated at birth. No similar pathway is detected in normal animals. Ultrastructural examination of degeneration in dLGN of blinded rats after contralateral PBG lesion reveals a similarity of the PBG terminals to "replacement terminals" seen in dLGN of anophthalmic or neonatally enucleated mice (Cullen and Kaiserman-Abramof 1976).

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Transient retinofugal pathways in the developing chick.

The central retinal projections have been examined in normal chick embryos early in development to determine if a transient ipsilateral projection is present. Anterograde transport of HRP identified using tetramethyl benzidine as a chromogen and degeneration techniques were used on embryos of successive ages between 6 and 16 days of incubation to show the central distribution of the retinal axons from one eye. Besides the anticipated contralateral projection, a small projection was identified to primary visual nuclei on the side of the brain ipsilateral to the injected or lesioned eye in embryos between days 6 and 12 of incubation. A projection from the injected eye into the contralateral optic nerve was also identified in a number of embryos. By embryonic day 15 the retinal projection to the ipsilateral side of the brain and into the contralateral optic nerve had disappeared. This loss of the anomalous projections coincides with a period of substantial cell death in the retinal ganglion cell layer. It appears, therefore, that in the chick, like the rat, the ipsilateral retinofugal projection resulting from an embryonic enucleation may in part be due to retention of a normal ipsilateral projection.

Afferent Pathways↗

Transplantation of tectal tissue in rats. III. Functional innervation of transplants by host afferents.

Fetal tectal tissue was transplanted adjacent to the superior colliculus of neonatal rats. In mature recipient animals, ultrastructural studies provided evidence of host cortical synapses on cells within the transplants. To determine whether these synapses were functionally effective in driving transplant cells, tungsten in glass microelectrodes were used to record from single units in the transplants and the host cerebral cortex was electrically stimulated. Of the 214 tested units which were shown histologically to be within the transplants, 25 (11.7%) were orthodromically excited from host cortex at latencies of 15 ms or less (mean 7.3 ms). A further four transplant units were activated at relatively long latencies (40-300 ms). The presence of a functional host retinal input was also tested by electrically stimulating the host optic nerves. No transplant units were activated from these sites, apparently because very few electrode penetrations passed through the localized zones of optic axon distribution. The possible functional interrelationships between transplant and host are discussed.

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Influence of grafted glia cells and host mossy fibers on anomalously migrated host granule cells surviving in cortical transplants.

Embryonic cerebral cortex was transplanted over the cerebellum of one- to two-day-old rats. In mature rats, clusters of granule cells that failed to migrate into the internal granule cell layer were now found within the graft tissue. Immunocytochemical staining of astrocytic glial cells in the cortical transplants revealed that glial processes were distributed in an unusual polarized orientation in those regions that contained host granule cells. Other areas of the graft exhibited glia cells with processes that projected radially from their cell body, thus resembling fibrous astrocytes. Fibrous astrocytes in transplants, however, were more heavily stained than similar glial cells in the intact cerebral cortex of the host, indicating a quantitative difference in the glia fibrillary acidic protein. Acetylcholinesterase-positive fibers were observed between the clusters of granule cells in the cortical grafts. Such fibers traversed the molecular layer and, since they could be traced from the white matter of the host cerebellum, they were presumed to be mossy fibers. It is concluded that migration of external granule cells in the cerebellum can altered by placing embryonic cerebral cortex next to the developing cerebellum. Granule cells that have migrated into the grafted cerebral cortical tissue nevertheless receive afferent fibres.

Acetylcholinesterase↗

Cultured embryonic retinae transplanted to rat brain: differentiation and formation of projections to host superior colliculus.

Retinae of rats on embryonic day 14 were placed in explant culture for 2-14 days prior to transplantation adjacent to the superior colliculus of newborn rats. In explant culture cell division and neuronal differentiation continued unabated. One month after transplantation host brains were examined for transplant survival, differentiation and formation of projections to the host brain. The cultured retinal transplants survived and developed a morphology typical of mature retina, with normal cell and fiber laminae present. HRP injections into the host superior colliculus labeled neurons in the ganglion cell layer of the transplant which closely resembled ganglion cells in vivo. A small number of transplants received lesions. Degeneration material was traced into the superior colliculus and pretectal nuclei confirming that the cultured transplants had projections appropriate for retina entering the host brain. These results correlate closely with those seen after transplantation of embryonic rat retinae that had not been cultured. Thus, a period of explant culture of up to two weeks does not affect the ability of embryonic retinal transplants to differentiate and form projections into the host brain.

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Transplantation of tectal tissue in rats. II. Distribution of host neurons which project to transplants.

Tectal tissue was dissected from fetal rats and transplanted adjacent to the superior colliculus of newborn rats. The recipient animals were then allowed to survive for 6 or more weeks. Subsequent examination revealed that the transplants generally lay over the host inferior colliculus and rostral part of the cerebellum and had substantial fiber connections with the host superior colliculus. To determine which host areas projected to the transplants, horseradish peroxidase (HRP) was injected into the transplants, and the host brain was examined for the presence of retrogradely filled neurons. Labeled cells were found in nearly 50 host areas. Most of these areas are known to project to normal superior colliculus. There was a consistency between one animal and another in the frequency and density of cell label in the various areas. The projection from host cortex (particularly from visual cortical areas) was the densest and most consistent projection. Other areas which commonly projected into the transplants included pretectum, parabigeminal nucleus, superior colliculus, and the brachial region of the inferior colliculus. Sparse and infrequent projections were found from ventral lateral geniculate nucleus, substantia nigra, zona incerta, and catecholaminergic nuclei. No unequivocally labeled retinal ganglion cells were found. The results indicate that the host projection into the transplants is limited to those areas with axons in the vicinity of the host/transplant interconnection. However, the data also suggest that (1) the relative maturity of particular host pathways at the time of transplantation and (2) some form of preferential or absolute affinity expressed between host axons and transplant cells are also factors which influence the pattern of connections formed between host and transplant.

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Transplantation of tectal tissue in rats. I. Organization of transplants and pattern of distribution of host afferents within them.

We have examined the maturation of tectal tissue transplanted from fetal rats to the midbrain of newborns and have characterized the distribution of host retinal and cortical afferents within the transplants. The transplants develop characteristic internal order and connections which distinguish them from either embryonic cortex or retina placed in the same region. Host retinal afferents project to clearly circumscribed regions, where they synapse mainly on small dendrites or dendritic spines, and only rarely on vesicle-containing profiles. The retinorecipient areas contain few stained axons in neurofibrillar preparations and are almost always located at the surface of the transplant. There is very little overlap in the input from the two eyes into a single transplant even though the projections from each eye may lie adjacent to one another. Cortical afferents spread more broadly in the transplants, but are largely absent from areas of optic termination and from other more deeply located regions with sparse fiber staining properties. The observations suggest that when placed close to its normal location, tectal tissue can develop a number of features characteristic of normal superior colliculus. Appreciation of the internal order of the transplants makes it possible to investigate the cortical and retinal afferent pathways using physiological techniques.

Afferent Pathways↗

Transplantation of embryonic occipital cortex to the brain of newborn rats: a Golgi study of mature and developing transplants.

Segments of the occipital cortex were taken from rat embryos (E16-E19) and transplanted to the cerebral cortex or the tectal region of a newborn rat host. With the aid of Golgi impregnation techniques, neuron morphology was studied in cortical transplants which had survived for 1 week or more in the host brain. In mature transplants (greater than 4 weeks) three main groups of neurons, termed groups I-III, were identified. Group I neurons resembled pyramidal neurons of the intact cerebral cortex. No preferential orientation of either soma or dendrites of group I neurons was observed in the transplants, and some group I neurons had curved apical dendrites. Group II neurons had predominantly stellate form and their dendrites were densely covered with spines. Paucity or absence of dendritic spines characterized group III neurons which exhibited various dendritic topologies. Different neuron types were also recognized in immature transplants growing for 1 and 2 weeks in the host brain. The sequence of dendritic maturation of transplanted cortical neurons is similar to that seen in intact cortex, although the stage reached related more to the actual age of the transplant than to that of the host. Thus, group I neurons in the 1-week-old transplants taken from E16 embryos had not attained the same complexity of branching as pyramidal neurons in the surrounding host cortex, but rather resembled slightly younger cells more like those found in the cerebral cortex of the newborn rat. These results show, therefore, that at least the basic cell classes identified in intact visual cortex can also be recognized in the cortical transplants. This will provide a foundation for studies defining which cells project to the host brain and which are involved in particular intrinsic connections.

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The distribution of the callosal projection to the occipital visual cortex in rats and mice.

The principal finding in this study is that the callosal projection to the occipital cortex in rats and mice follows a complex and highly reproducible pattern which has not previously been described in detail. In some regions, the callosal projection is associated with well defined cytoarchitectonic boundaries such as the border between areas 17 and 18a. However, extrastriate cortex lateral to area 17 receives callosal inputs which are not related to previously defined cytoarchitectonic boundaries. Following intraocular injections of [3H]fucose, transneuronal label occupies area 17 and mainly the posterior part of area 18a. A region in posterolateral area 18a which is 'subdivided' into callosal and sparsely callosal regions appears to receive an input from the lateral geniculate nucleus, based on transneuronal autoradiography. Comparison of the distribution of callosal axons and transneuronal label suggests that regions of murid cortex similar to areas 18, 19 and lateral suprasylvian cortex in cats may be located posteriorly in area 18a.

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Development of a transient retino-retinal pathway in hooded and albino rats.

Intraocular HRP injections in E16-21 embryos show that during the normal development of the central optic projections in hooded and albino rats many optic axons grow through the chiasm into the contralateral eye. This retino-retinal projection disappears shortly after birth. This suggests that an initial, imprecise guidance of growing axons is followed by a selective elimination of axons taking aberrant pathways and failing to make appropriate synapses.

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Experimentally induced enlargement of the uncrossed retinotectal pathway in rats.

The present experiments investigate the effects of neonatal lesions upon projection patterns of the uncrossed retinotectal pathway in albino rats. The results indicate that enlargement of the terminal field from one eye can be induced either by a contralateral optic tract lesion or by removal of the opposite eye at birth. The extent of the enlargement is more prominent in the latter case. If an optic tract lesion is accompanied by eye enucleation on the side ipsilateral to the tract lesion, the uncrossed retinotectal projection from the remaining eye will undergo further enlargement. However, optic fiber counts show that such an enlargement of the terminal field is not due to a significant increase in the number of uncrossed optic axons which contribute to the enlarged projection, but rather to an increased terminal arbor of individual axons (as shown by results from fiber counts). While a severe ganglion cell loss was observed in the retina contralateral to a tract lesion, a substantial population of cells persists in the ganglion cell layer and the number of cells appear much higher than the number of uncrossed optic axons arising from the same eye. The implications of these findings are discussed in relation to results reported in previous studies.

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Transplantation of embryonic occipital cortex to the tectal region of newborn rats: a light microscopic study of organization and connectivity of the transplants.

Occipital cortex was taken from fetal rats and transplanted to the tectal region of newborn rats, where it developed a specific structural identity reflecting in part its cortical origin. The implants showed locally distributing intratransplant connections, and the majority formed connections with defined regions of te host cerebral cortex and the brainstem. A sparse afferent projection from the host had its origin in visual, somatosensory, and cingulate areas of te cortex, pretectum, superior colliculus, central gray, hypothalamus, pontine reticular formation, raphe nuclei, and the locus coeruleus. No input was identified from either the retina or the dorsal thalamus. Efferent fibers were observed in normal fiber preparations as compact bundles running through the host brainstem along two main routes, one group of bundles in a dorsal position and a second group more ventral. Efferent fibers traveling rostrally along the first pathway distributed in the lower part of the stratum griseum superficiale and in the intermediate laminae of the superior colliculus, and in some cases they reached the pretectum and the lateral posterior thalamic nucleus. Deep efferent fibers ran rostrally and caudally in the central gray, and in some cases laterally directed fibers were seen to distribute in the midbrain tegmentum and reticular formation, in one case reaching the pontine gray. The finding that most afferent and many efferent connections of cortical transplants are uncharacteristic of normal cortex stands in marked contrast to retina and tectum, which, when transplanted to the same region, make relatively normal connections.

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