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F Scalia

Publications and source records attributed to F Scalia.

At least 19 recordsLinked to original sources

Unequal representation of the temporal and nasal retina in an anomalous projection to the lateral thalamus.

Study of an anomalously regenerated, nontopographically organized retinal projection in the frog olfactory cortex revealed that the temporal retina is the main source of this projection, suggesting the existence of specific temporal fiber-directed attractant or trophic influences. In the present study, we examined the organization of an anomalous retinal projection that forms in the frog thalamus after ablation of the optic tectum. The projections from different sectors of the retina were studied by means of the anterograde transport of biotinylated dextran-amine (BDA) delivered to incisions made across the nerve fiber layer in frogs surviving ablation of the contralateral tectal hemisphere for 13-46 weeks. The projections from nasal retinal sectors were always lightly constructed in the aberrant terminal field, whereas their projections to the lateral geniculate complex remained reasonably strong. In contrast, the projections from temporal retinal sectors, though also weak initially, in time became robust and filled the aberrant field over most of its extent. The specific amplification of the temporal fiber projection now observed in two foreign targets provides further evidence for the existence of target-based, attractant/trophic molecules with functional specificity for temporal retinal fibers. That such agents can exist or be inducible in a foreign area would suggest that they belong to a family of molecules having natural biological activity in normal development or regeneration. However, the possibility that the augmented role of the temporal retina in these projections is a result of experience-based plasticity is also discussed.

Animals↗

Delivering drugs, via microdialysis, into the environment of extracellularly recorded hippocampal neurons in behaving primates.

Hippocampal neurons in primates have been extensively studied with electrophysiological and neuroanatomical methods. Much less effort has been devoted to examining these cells with contemporary pharmacological techniques. Therefore, we modified a recently developed integrative technique (N. Ludvig, P.E. Potter, S.E. Fox, Simultaneous single-cell recording and microdialysis within the same brain site in freely behaving rats: a novel neurobiological method, J. Neurosci. Methods 55 (1994) 31-40 [9] ) for cellular neuropharmacological studies in behaving monkeys. A driveable microelectrode-microdialysis probe guide assembly was implanted stereotaxically into the left hippocampus of squirrel monkeys (Saimiri sciureus) under isoflurane anesthesia. The assembly was covered with a protective cap. After 3 weeks of postsurgical recovery and behavioral training, the experimental subject was seated in a primate chair. For 4-5 h, single-cell recording and microdialysis were simultaneously performed in the hippocampal implantation site. The technique allowed the recording of both complex-spike cells and fast-firing neurons without the use of head restraint. The control microdialysis solution, artificial cerebrospinal fluid (ACSF), was replaced with either 1 M ethanol or 500 microM N-methyl-D-aspartate (NMDA) for 10-30 min intervals. The ethanol perfusions principally suppressed the firing of the neurons in the dialysis area. The NMDA perfusions initially increased the firing of local neurons, then caused electrical silence. These drug delivery/cell recording sessions were performed with 1-4 day intersession intervals over a 1-month period. The described method provides a tool to elaborate the pharmacology of primate hippocampal neurons during behavior and without the confounding effects of systemic drug administrations.

Animals↗

Biotinylated dextran amine and biocytin hydrochloride are useful tracers for the study of retinal projections in the frog.

Anatomical study of the topographic organization of retinal projections requires a tracer capable of resolving fine morphological detail and permitting analysis of the projection from either the whole retina or selected areas. To obtain a permanent record of the experiments and to have access to ultrastructural data, it is preferable for the tracer to be compatible with both brightfield microscopy and electron microscopy. Biotinylated dextran amine and biocytin hydrochloride, as employed in the present experiments, meet these needs exceptionally well for anterograde tracing studies on the frog visual system. Both tracers labeled axons and terminal arbors more prominently than comparable material studied by the widely used methods of anterograde fiber-filling with horseradish peroxidase or cobalt. When used to trace the projections from small sectors of retina, the finest unmyelinated fibers in layers A, C and E of the frog optic tectum and their synaptic boutons were made readily visible by the new tracers.

Animals↗

A compartment-based, asymmetric representation of the retina in an induced projection to the olfactory cortex.

Displacing the optic nerve into the telencephalon in adult Rana pipiens induces a projection to olfactory cortex. We have examined the topographic organization of this projection anatomically by injecting a mixture of biotin dextran (BDA) with 3H-amino acids into the affected eye immediately after making cuts across defined sectors of the nerve fiber layer to trace the complementary patterns of anterograde migration of BDA and 3H label in the cut and intact retinal axons, respectively. Fibers from the temporal side of the optic disc terminated in an oblique band along the posterior two-thirds or more of the ectopic projection field. In contrast, fibers arising in the nasal retina terminated in a parallel strip occupying the anterior one-third or less of the field. Varying the location of the cuts within each hemiretina did not reveal any further organization along the nasotemporal or dorsoventral axes of the retina. The retinal location of the cells involved in this projection was further studied with injections of wheat germ agglutinin conjugated to horseradish peroxidase into the olfactory cortex. Ganglion cells labeled by retrograde transport were found throughout the retina, but they were much more numerous on the temporal side, having a mean spatial density 3.7-7.4 times greater in the temporal hemiretina, whereas the overall ganglion cell density (labeled plus unlabeled) was roughly the same in the two halves of the retina. These data provide an example of a permanent projection in which the overall representation of the retina, though nontopological, is polarized in one axis (nasotemporal) and, therefore, compartmentally organized.

Animals↗

Concentration and storage of biotin in the amphibian brain.

Prominent displays of endogenous biotin reactivity can be observed at specific locations in histochemical preparations of the forebrain and midbrain in the northern leopard frog (Rana pipiens) and common American toad (Bufo americanus). At the light microscopic level, the biotin reactivity appears in clusters of darkly stained puncta of either spherical or rodlike shape in the olfactory cortex, nucleus isthmi, and hypothalamus. With the electron microscope, the biotin reactive spheres are identified as neuronal varicosities and synaptic boutons and the rods as short segments of axons. Appropriate controls demonstrate that the punctate biotin-reactive structures are sites of concentration of biotin or a biotin analog in the processes of certain neurons. These data represent the first observation on the selective concentration of a vitamin in vertebrate neurons and suggest that biotin may have specialized functions in anatomically delimited areas of the central nervous system. Localization of the densest concentration of the biotin-reactive puncta in the dorsolateral prominence of the olfactory cortex may have relevance to the functional organization of the olfactory system. The distributions of biotin-reactive puncta were observed in laboratory-housed frogs and in wild toads captured in the summer months but were sparse or absent in batches of commercially obtained frogs examined immediately upon arrival in the laboratory. Systemic administration of biotin or biocytin hydrochloride did not alter the appearance or numbers of the biotin-reactive structures either in newly received or laboratory-housed frogs. These findings suggest that the capacity of the biotin-storage mechanism in the amphibian brain may be set by environmental factors and may be readily saturable from natural dietary or enteric sources.

Animals↗

Functional properties of regenerated optic axons terminating in the primary olfactory cortex.

When the optic nerve of Rana pipiens is cut and deflected into the telencephalon, the regenerating fibers terminate selectively in the superficial neuropil of the primary olfactory cortex. These redirected fibers and their terminals on the dendrites of the cortical cells appear normal by LM and EM criteria. Electrical recording, done 2-16 months after surgery, shows visually evoked activity in the superficial neuropil (Layer I) of the olfactory cortex, and visually excited responses in the deep cortical cell layer (Layer II). In the normal frog, the electrical activity seen in the neuropil of the olfactory cortex consists of small transients about 2-3 x the noise level of the electrode contact. These occur spontaneously and are also excited by puffs of air to the nose. There is no such excitation by visual stimuli. Larger initially negative spikes cell above noise level are recorded in the cell layer next to the ependymal surface, and these are also spontaneous, or excited by puffs of air to the nose, but not by visual stimuli. In the operated frog, the small transients in the neuropil appear and are excited by the puffs of air and by visual stimuli. Similarly the responses in the cell layer are excited by both sorts of stimuli. But new types of electrical signals appear in the neuropil; they are driven only by visual stimuli presented to the affected eye. These are very large transients of the kind found in the tectal neuropil and have the two characteristic shapes which were classified as B and C types in the tectum. Such large transients are never seen in the neuropil of the olfactory cortex in normal frogs. The receptive fields of the small visually driven transients in the neuropil are not easy to make out because the signal levels are so close to the noise level that different units cannot be reliably distinguished from each other. But the receptive fields of the much larger B and C type unit responses are as easy to classify and plot as they are in tectum, even though on the average they are only about 2/3 as large as in tectum. The single-unit receptive fields belong to one or another of the several types of retinal ganglion cell classes distinguished in optic-nerve recordings. Four of the major classes normally project to the tectum and a fifth projects to the lateral geniculate complex. But all five are present in the ectopic projection to the olfactory cortex.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Potentially functional regions of nucleic acids recognized by a Kohonen's self-organizing map.

Computer recognition of short functional sites on DNA, such as promoter regions or intron-exon boundaries, has recently attracted much interest. In this paper we have focused our attention on the automatic recognition of relevant features of human nucleic acid sequences by means of an unsupervised artificial neural network model. Sixty messenger RNA and 31 genomic DNA sequences were analysed. The results showed that in mRNA, the minimal similarity 60 base pattern was guanine- and cytosine-rich and located in most sequences in a range of 250 bases from either the middle point of the signal peptide coding region or from the start of the coding region. On DNA sequences a region defined by a cluster of minimal similarity patterns was present in many of the analysed genes. This zone may be related to alternative splicing and DNA methylation.

Algorithms↗

Further study of the aberrant optic nerve projection to olfactory cortex.

When implanted into the cerebral hemisphere, the regenerating optic nerve of the adult frog (Rana pipiens) forms a well-defined terminal field in the pars ventralis of the lateral (olfactory) cortex, and sometimes expands medially into the postolfactory eminence. These adjacent areas receive their normal input from the main olfactory bulb. The aberrant projection extends caudally toward the core neuropil of the medial amygdaloid nucleus, which receives its normal input from the accessory olfactory bulb, but does not enter this vomeronasal sector of the amygdala. The present study tests whether: 1) optic fibers would innervate the vomeronasal amygdala after surgical ablation of the accessory olfactory bulb, 2) the projection would transpose into adjacent cortex after olfactory cortex lesions, and 3) the projection would overflow into adjacent areas after being amplified by hemisection at the di-telencephalic junction (to minimize escape of fibers into the diencephalon). The retinal projection always terminated in the olfactory cortex when this area was intact, or in spared fragments of it after radical cortical lesions, but never entered the vomeronasal amygdala in any specimen, as studied by autoradiographic and horseradish peroxidase tracing techniques. With forebrain hemisection, the cortical terminal field increased in thickness but remained confined to the olfactory area. However, the interruption of the lateral forebrain bundle induced a new projection to the striatum in a region neighboring but separate from the olfactory cortical field. These findings support the hypothesis that retinal fibers have a specific affinity for primary olfactory cortex that is not normally allowed expression in development. Retinal fibers may also have a latent affinity for the striatum that is unmasked after deafferentation.

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Optic nerve regeneration with return of vision through an autologous peripheral nerve graft.

The optic fiber termination layer in the contralateral optic tectum was reinnervated and useful vision was recovered in the adult frog, after successful optic nerve regeneration through an autologous peripheral nerve-bridge used to replace the optic nerve and optic chiasma. During their course through the nerve-bridge, the optic fibers were associated with Schwann cells in the usual relationship observed in peripheral nerve.

Animals↗

Quantitative study of the tectally projecting retinal ganglion cells in the adult frog. II. Cell survival and functional recovery after optic nerve transection.

It is known from previous work that ganglion cells disappear from the retina in significant numbers during optic nerve regeneration in the adult frog. In the present study, the population size of surviving ganglion cells that have returned axon terminals to the correct tectal loci was estimated by counts of retrogradely labeled cells in retina-flat-mounts after tectal injections of HRP. Bilaterally symmetric injections were delivered to allow comparison of the normal and affected retinas. The frogs studied had regenerated the left optic nerve and had visually guided behaviors initiated by the recovered eye (see below). The proportion of tectally projecting ganglion cells in the normal retinas and in retinas of normal frogs studied in parallel ranged from 83-86% (Singman and Scalia: J. Comp. Neurol. 302:792-809, 1991). In the affected retinas, the subpopulation of tectally projecting cells was reduced by 40-90% after regeneration, and the relative size of this subpopulation ranged from 67-86%. The optic tectum was injected unilaterally in one specimen, on the side ipsilateral to the regenerated (left) optic nerve. The HRP-labeled ganglion cells in the ipsilateral (left) retina accounted for only 0.8% of the surviving ganglion cells in this animal, whereas it was previously found that the ipsilateral tectally projecting ganglion cells normally amount to 0.9-2.3% (Singman and Scalia, op. cit.) In frogs recovering from transection of the left optic nerve, the frequency, latency, and accuracy of the prey-acquisition responses initiated by the recovering eye were compared with those initiated by the normal eye. Mealworms or lure dummies were used to stimulate prey acquisition. In one experiment, the stimuli were presented unilaterally in the monocular fields of frogs permitted to use both eyes. Prior to the fourteenth postoperative week, the affected eye initiated responses of abnormally long latency and low frequency. In contrast, responses initiated by the affected eye after 14 weeks appeared to be normal in all respects. In another experiment, the normal eye was sutured shut in some frogs recovering for at least 24 weeks and then the affected eye was retested. The affected eye was capable of consistently initiating brisk and accurate prey acquisition. In a final experiment, two stimuli were presented simultaneously in bilaterally symmetric regions of the monocular fields of frogs surviving at least 42 weeks. These fully recovered frogs showed no preference for using either the normal or the recovered eye. Despite severe loss of tectally projecting ganglion cells during optic nerve regeneration, frogs are capable of apparently normal visual responses in prey acquisition tests.

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A note on the organization of the amphibian olfactory bulb.

After horseradish peroxidase (HRP) injections were made in limited sectors of the main olfactory bulb in the adult frog Rana pipiens, the cellular morphology of mitral cells and granule cells impregnated with HRP were examined in uninjected regions of the bulb. Mitral cells were observed to possess glomerular dendrites and prominent secondary dendrites, both of which have smooth shafts. The glomerular dendrites may be multiple, are often branched, and may arise from secondary dendrites, as well as from the cell body. The axon may also arise from a secondary dendrite. Granule cells have simple or branched peripheral dendrites, and these are spiny, where they intermingle with the mitral cell secondary dendrites. The prominence of the secondary dendrites of frog mitral cells contrasts sharply with their reported insignificance in urodeles, as studied in earlier literature. The layers of the main olfactory bulb are not as fully concentric in the frog, as they are in mammals. The implantation cone and glomerular layer occupy a small part of the surface area of the olfactory bulb on its anteroventral aspect, while the perimeters of the subjacent layers extend farther posteriorly and dorsally in successive steps. The granule cell core extends well beyond the perimeter of the mitral cell layer in a posterior direction. Long secondary dendrites of mitral cells also extend posteriorly beyond the perimeter of the mitral cell-external plexiform layer and interlace with granule cell peripheral dendrites in a plexiform layer external to the posterior region of the granule cell core. This layer, the superficial plexiform layer, forms an apron around the posterior segment of the olfactory bulb and contributes to the interbulbar adhesion.(ABSTRACT TRUNCATED AT 250 WORDS)

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Differential projections of the main and accessory olfactory bulb in the frog.

The central projections of the main olfactory bulb and the accessory olfactory bulb of the adult leopard frog (Rana pipiens) were reexamined, by using a horseradish peroxidase anterograde tracing method that fills axons with a continuous deposit of reaction product. The fine morphology preserved by this method allowed the terminal fields of the projection tracts to be delineated reliably, and for the first time. Herrick's amygdala has been newly subdivided into cortical and medial nuclei on the basis of cytoarchitecture, dendritic morphology, and the differential projections of the main and accessory olfactory tracts. The main olfactory bulb projects through the medial and lateral olfactory tracts to the postolfactory eminence, the rostral end of the medial cortex, the rostral end of the medial septal nucleus, the cortical amygdaloid nucleus, the nucleus of the hemispheric sulcus, and both the dorsal and ventral divisions of the lateral cortex, including its retrobulbar fringe. The lateral olfactory tract overlaps the dorsal edge of the striatal plate along the ventral border of the lateral cortex, but it is not certain whether any striatal cells are postsynaptic to the tract fibers. The lateral cortex is the largest of these territories, and receives the terminals of the main olfactory projection throughout its extent. It extends from the olfactory bulb to the posterior pole, and from the striatum to the summit of the hemisphere, where it borders the dorsal cortex. The medial and lateral olfactory tracts combine in the region of the amygdala to form a part of the stria medullaris thalami. These fibers cross in the habenular commissure and terminate in the contralateral cortical amygdaloid nucleus and periamygdaloid part of the lateral cortex. Cells projecting to the main olfactory bulb are found in the diagonal band and adjacent cell groups, but there is no evidence of an interbulbar projection arising from either the olfactory bulb proper or a putative anterior olfactory nucleus. The accessory olfactory bulb projects through the accessory olfactory tract to the medial and cortical amygdaloid nuclei. A fascicle of the tract crosses in the anterior commissure to terminate in the contralateral amygdala. While the main and accessory olfactory projections may converge in the cortical amygdaloid nucleus, the medial amygdaloid nucleus is connected exclusively with the accessory olfactory bulb.

Afferent Pathways↗

Identification of a new motif on nucleic acid sequence data using Kohonen's self-organizing map.

Here we present a performance test of a Kohonen features map applied to the fast extraction of uncommon sequences from the coding region of the human insulin receptor gene. We used a network with 30 neurons and with a variable input window. The program was aimed at detecting unique or uncommon DNA regions present in crude sequence data and was able to automatically detect the signal peptide coding regions of a set of human insulin receptor gene data. The testing of this program with HSIRPR cDNA release (EMBL data bank) indicated the presence of unique features in the signal peptide coding region. On the basis of our results this program can automatically detect 'singularity' from crude sequencing data and it does not require knowledge of the features to be found.

Algorithms↗

Quantitative study of the tectally projecting retinal ganglion cells in the adult frog: I. The size of the contralateral and ipsilateral projections.

The proportion of ganglion cells connected to the several central targets of the retinal projection varies in different species. In the frog, the retinotectal projection is clearly the largest branch of the optic pathway and the relative size of the tectally projecting population can be expected to be correspondingly great. However, there have been no studies aimed at quantifying the size of this population and at partitioning its contralateral and ipsilateral components. We injected the tectum with horseradish peroxidase (HRP) dried onto fine needles to count the numbers of retinal ganglion cells labeled by retrograde transport. The retinas were prepared as flat-mounts to facilitate the cell counting. The tecta were injected either unilaterally or bilaterally in mirror-symmetric loci. Specimens included completely normal frogs and frogs which had undergone unilateral optic nerve regeneration, although only normal retinas are presented in the current study. The retrograde transport interval was varied progressively (from 3 to 5 days), and single or multiple injections of HRP were placed singly or as clusters, in order to increment the cell counts toward a level of saturation. Approximately 70.9% of the neurons in the ganglion cell layer could be labeled by this method. Correcting for the presence of displaced amacrine cells, estimated to comprise approximately 16% of the neurons in the ganglion cell layer (Scalia et al., '85, Brain Res. 344:267-280), we calculate that approximately 84.4% of the retinal ganglion cells project contralaterally to the optic tectum. Flat-mounted retinas ipsilateral to unilaterally injected tecta of completely normal frogs were also examined for labeled cells. The results of injections in the rostrolateral, caudomedial, and caudolateral tectum were studied. We found that ipsilaterally labeled cells comprised no more than 2.3% of the overall population of ganglion cells in the ganglion cell layer. The ipsilaterally projecting cells were found in loci which were approximately mirror-symmetric to the regions of maximal cell labeling in the contralateral retinas from the same animals. The ipsilateral population was always displaced toward the periphery of the retina with respect to the contralateral population, regardless of whether the contralateral locus was centered in the temporal, ventronasal, or dorsonasal sector of the retina. Because the ipsilaterally projecting ganglion cells form such a minor population, and because they exist in the monocular as well as the binocular parts of the retina, it seems likely that they may not play a significant role in visual function in the frog.

Afferent Pathways↗

Further study of the outward displacement of retinal ganglion cells during optic nerve regeneration, with a note on the normal cells of Dogiel in the adult frog.

In a previous study we observed massive retinal ganglion cell death in adult Rana pipiens after periods of optic nerve regeneration, and reported that large numbers of the surviving cells had become displaced bodily into the inner plexiform layer of the affected eye (Scalia et al.: Brain Research 344:267-280, 1985). The outwardly displaced cells could be identified as retinal ganglion cells because they could be back-filled with horseradish peroxidase (HRP) injected into the regenerated optic nerve. Quantitative observations on the abnormal outward displacement of ganglion cells are reported here. Parallel observations on normally displaced ganglion cells (cells of Dogiel) are also reported to clarify the distinctions between these two classes of cells. For the present work, injections of HRP of varying size were placed in the optic tectum bilaterally in 3 normal frogs and 9 frogs sustaining unilateral optic nerve regeneration. Most injections were centered at loci mapping the middle region of the nasal retina. The retinas were examined as flat-mounts and in-section. In 8 other frogs sustaining optic nerve regeneration, the HRP was administered bilaterally directly to the optic nerves in the orbit. Ganglion cells were labeled by retrograde transport of the HRP in the retinal ganglion cell layer in both the normal and affected eyes in areas topographically isomorphic with the tectal areas subtended by the injections. In the normal eyes, the orthotopic ganglion cells formed a strict monolayer, and virtually no cells existed in the inner plexiform layer. In the retinas sustaining optic nerve regeneration, the retinal ganglion cells abnormally displaced into the inner plexiform layer were also labeled topographically in correspondence with the injection sites. The abnormally displaced cells comprised 5.5% of the total population of surviving neurons in the retinal ganglion cell and inner plexiform layers. The mean outward dislocation of the displaced cells, as measured in one frog surviving optic nerve crush for 8 weeks, was 69.9 +/- 2.4% of the distance across the inner plexiform layer, which itself was uniformly 14.3 +/- 0.39 microns thick. Cells of Dogiel, which were embedded within the inner nuclear layer, were also labeled when the injections of HRP spread to include the area of representation of the optic disc. The labeled cells were restricted to a dorsal, peripapillary locus capping the optic disc. Therefore, some cells of Dogiel project to the tectum normally, but only from the central retina.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Ocular↗

Synapse formation in the olfactory cortex by regenerating optic axons: ultrastructural evidence for polyspecific chemoaffinity.

The optic nerve was severed at its entry into the optic chiasma and implanted into the striatal region of the ipsilateral cerebral hemisphere in adult frogs (Rana pipiens). Regenerating retinal ganglion cell axons, observed by the autoradiographic tracing method and by horseradish peroxidase (HRP) fiber filling, grew anteriorly along the olfactory tracts and posteriorly along the ipsilateral lateral forebrain bundle and stria medullaris. Many of the regenerating axons ultimately joined the ipsilateral optic tract. The optic axons formed terminal plexuses in the olfactory cortex, lateral geniculate complex, pretectum, tectum, and basal optical nucleus but not in the amygdala or other cerebral territories not postsynaptic to the olfactory bulb, nor in the cell groups associated with the lateral forebrain bundle or stria medullaris. Optic axon terminals labeled with HRP were observed by electron microscopy in the ipsilateral olfactory cortex and in the normal projection areas of the optic nerve, although they were misplaced to the ipsilateral side. They contained clear, spherical synaptic vesicles and pale mitochondria and made Gray type I, asymmetric contacts on dendrites. The retinal projection to the olfactory cortex was formed early in regeneration and was maintained to some degree for periods up to 39 weeks. It was absent in a specimen surviving 50 weeks. Retinal innervation appeared earlier in the lateral geniculate complex and pretectum than in the tectum. These observations suggest that regenerating retinal ganglion cell axons have an affinity for neurons in the olfactory cortex, as well as for the neurons in the optic pathway to which they are normally postsynaptic. Unless the apparent selectivity of the aberrant projection is regulated by principles other than those that bring about the reinnervation of the normal optic centers, the data further suggest that the nature of the molecular mechanisms conveying synaptic specificity must be broad enough to permit the formation of limited sets of alternative synaptic connections. The ability to innervate selectively targets other than those normally specified is termed, here, polyspecificity. Since polyspecificity refers to the the affinity of retinal ganglion axons, as a class, for target structures considered as unit aggregates, it is conceptually different from the graded affinity of ganglion cells in different regions of the retina for target neurons in different regions of the tectum.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Ultrastructural evidence of the formation of synapses by retinal ganglion cell axons in two nonstandard targets.

After unilateral ablation of the optic tectum in the frog (Rana pipiens), retinal ganglion cell axons enter the lateral thalamic neuropil in large numbers. This area is normally a target of the tectal efferent projection but is not innervated directly from the retina in normal frogs nor in frogs undergoing optic nerve regeneration in the presence of an intact tectum. The ability of retinal axons to form synaptic contacts in this nonstandard target, previously suspected only from light microscope studies, has been ultrastructurally verified in the present investigation. Retinal axon terminals were selectively labeled for light and electon microscope study by introducing horseradish peroxidase (HRP) into the optic nerve 73-413 days after unilateral ablation of the contralateral optic tectum. In some of the frogs, the optic nerve had also been crushed to test the ability of retinal axons regenerating over a long distance to form this connection. The HRP-labeled retinal axon terminals had the same untrastructural morphology whether located in the lateral thalamic neuropil or in the correct regions of projection, e.g., the lateral geniculate complex. They contained clear, spherical synaptic vesicles and made Gray type I synapses on the unlabeled postsynaptic dendrites. The magnitude of the projection was disproportionately greater in animals having complete or nearly complete tectal ablation than in a specimen in which the lesion was significantly incomplete. An aberrant projection was also observed in the nucleus isthmi in some of the specimens. These findings have significance for chemoaffinity theories of the specification of synaptic connections in that the ability of retinal axons to synapse in nonstandard targets in this experimental context may be considered evidence for the expression of appropriate cell-surface recognition-molecules by the abnormally targeted postsynaptic neurons. The likelihood that the expression of these postsynaptic labels is normally repressed transynaptically by molecular signals from the intact tectal input is discussed.

Animals↗

Loss and displacement of ganglion cells after optic nerve regeneration in adult Rana pipiens.

After studying pathway selection in the brain of Rana pipiens during unilateral optic nerve regeneration, several frogs were allowed to survive for lengthy periods for use in the present investigation. Retina flat-mounts were prepared from both eyes at 42-50 weeks postoperation. In some cases, HRP was infiltrated into both optic nerves prior to sacrifice to assist in identifying retinal ganglion cells. All specimens showed reduced cell-densities in the ganglion cell layer of the eye that had sustained the nerve regeneration. In addition, many ganglion cells were displaced, abnormally, into the inner plexiform layer, and the normally-situated cells formed irregular bands and islands in some parts of the retina. Cell-counts showed an apparently time-related change in neuron number ranging from a loss of 41% compared with the unaffected eye at 42 weeks, to losses as great as 71% at 50 weeks. The probable number of displaced amacrine cells in the ganglion cell layer, assumed to be unaffected by the experiment, was estimated at a maximum of 16%. Possible factors underlying the loss and displacement of ganglion cells are discussed.

Animals↗