Projecting Ganglion Cells Research Articles

Architecture of the optic chiasm and the mechanisms that sculpt its development.

At the optic chiasm the two optic nerves fuse, and fibers from each eye cross the midline or turn back and remain uncrossed. Having adopted their pathways the fibers separate to form the two optic tracts. Research into the architecture and development of the chiasm has become an area of increasing interest. Many of its mature features are complex and vary between different animal types. It is probable that numerous factors sculpt its development. The separate ganglion cell classes cross the midline at different locations along the length of the chiasm, reflecting their distinct periods of production as the chiasm develops in a caudo-rostral direction. In some mammals, uncrossed axons are mixed with crossed axons in each hemi-chiasm, whereas in others they remain segregated. These configurations are the product of different developmental mechanisms. The morphology of the chiasm changes significantly during development. Neurons, glia, and the signals they produce play a role in pathway selection. In some animals fiber-fiber interactions are also critical, but only where crossed and uncrossed pathways are mixed in each hemi-chiasm. The importance of the temporal dimension in chiasm development is emphasized by the fact that in some animals uncrossed ganglion cells are generated abnormally early in relation to their retinal location. Furthermore, in albinos, where many cells do not exit the cell cycle at normal times, there are systematic chiasmatic abnormalities in ganglion cell projections.

Slit Inhibition of Retinal Axon Growth and Its Role in Retinal Axon Pathfinding and Innervation Patterns in the Diencephalon

We have analyzed the role of the Slit family of repellent axon guidance molecules in the patterning of the axonal projections of retinal ganglion cells (RGCs) within the embryonic rat diencephalon and whether the slits can account for a repellent activity for retinal axons released by hypothalamus and epithalamus. At the time RGC axons extend over the diencephalon, slit1 and slit2 are expressed in hypothalamus and epithalamus but not in the lateral part of dorsal thalamus, a retinal target. slit3 expression is low or undetectable. The Slit receptors robo2, and to a limited extent robo1, are expressed in the RGC layer, as are slit1 and slit2. In collagen gels, axon outgrowth from rat retinal explants is biased away from slit2-transfected 293T cells, and the number and length of axons are decreased on the explant side facing the cells. In addition, in the presence of Slit2, overall axon outgrowth is decreased, and bundles of retinal axons are more tightly fasciculated. This action of Slit2 as a growth inhibitor of retinal axons and the expression patterns of slit1 and slit2 correlate with the fasciculation and innervation patterns of RGC axons within the diencephalon and implicate the Slits as components of the axon repellent activity associated with the hypothalamus and epithalamus. Our findings suggest that in vivo the Slits control RGC axon pathfinding and targeting within the diencephalon by regulating their fasciculation, preventing them or their branches from invading nontarget tissues, and steering them toward their most distal target, the superior colliculus.

T Differential Effects of Acetylcholine and Glutamate Blockade on the Spatiotemporal Dynamics of Retinal Waves

In the immature vertebrate retina, neighboring ganglion cells express spontaneous bursting activity (SBA), resulting in propagating waves. Previous studies suggest that the spontaneous bursting activity, asynchronous between the two eyes, controls the refinement of retinal ganglion cell projections to central visual targets. To understand how the patterns encoded within the waves contribute to the refinement of connections in the visual system, it is necessary to understand how wave propagation is regulated. We have used video-rate calcium imaging of spontaneous bursting activity in chick embryonic retinal ganglion cells to show how glutamatergic and cholinergic connections, two major excitatory synaptic drives involved in spontaneous bursting activity, contribute differentially to the spatiotemporal patterning of the waves. During partial blockade of cholinergic connections, cellular recruitment declines, leading to spatially more restricted waves. The velocity of wave propagation decreases during partial blockade of glutamatergic connections, but cellular recruitment remains substantially higher than during cholinergic blockade, thereby altering correlations in the activity of neighboring and distant ganglion cells. These findings show that cholinergic and glutamatergic connections exert different influences on the spatial and temporal properties of the waves, raising the possibility that they may play distinct roles during visual development.

Extrinsic Modulation of Retinal Ganglion Cell Projections: Analysis of the Albino Mutation in Pigmentation Mosaic Mice

Tyrosinase is a key enzyme involved in the synthesis of melanin in the retinal pigment epithelium (RPE). Mice that are homozygous for the albino allele at the tyrosinase locus have fewer retinal ganglion cells with uncrossed projections at the optic chiasm. To determine the site of the albino gene action we studied the projections of retinal ganglion cells in two types of pigmentation mosaic mice. First, we generated mosaic mice that contain a translocated allele of the wild-type tyrosinase on one X chromosome but that also have the lacZ reporter transgene on the opposite X chromosome. In these lacZ/tyrosinase mice, which are homozygous for the albino allele on chromosome 7, X-inactivation ensures that tyrosinase cannot be functional within 50% of the retinal ganglion cells and that these individual cells can be identified by their expression of the lacZ reporter gene product, β-galactosidase. The proportion of uncrossed retinal ganglion cells expressing β-galactosidase was found to be identical to the proportion that did not express it, indicating that the albino mutation associated with axonal behavior at the optic chiasm must affect ganglion cells in a cell-extrinsic manner. Second, to determine whether the RPE is the source of the extrinsic signal, we generated aggregation chimeras between pigmented and albino mice. In these mosaic mice, the extent of the uncrossed projection corresponded with the amount of pigmented cells within the RPE, but did not correspond with the genotypes of neural retinal cells. These studies demonstrate that the albino mutation acts indirectly upon retinal ganglion cells, which in turn respond by making axonal guidance errors at the optic chiasm.

Asymmetric growth and development of the Xenopus laevis retina during metamorphosis is controlled by type III deiodinase.

During the metamorphosis of the Xenopus laevis retina, thyroid hormone (TH) preferentially induces ventral ciliary marginal zone (CMZ) cells to both increase their proliferation and give rise to ipsilaterally projecting ganglion cells. Here we show that dorsal CMZ cells express type III deiodinase (D3), an enzyme that inactivates TH. The dorsal CMZ cells can be induced to proliferate if deiodinase activity is inhibited. D3 or dominant-negative thyroid hormone receptor transgenes inhibit both TH-induced proliferation of the ventral CMZ cells and the formation of the ipsilateral projection. Thus, the localized expression of D3 in the dorsal CMZ cells accounts for the asymmetric growth of the frog retina.

Dynamics of primate P retinal ganglion cells: responses to chromatic and achromatic stimuli.

1. The majority of primate retinal ganglion cells (RGCs) project to the parvocellular layers of the lateral geniculate nucleus (LGN). These P cells play a central role in early visual processing. 2. An improved method of systems analysis has allowed us to explore the dynamics of the colour-opponent subregions of P-cell receptive fields with a single chromatic stimulus. The data show that the centre and surround subregions of the P-cell receptive field have similar temporal responses, but the surround is slightly delayed. The centre and surround demonstrate a large degree of chromatic selectivity. 3. The responses of the centre and surround subregions were fitted with a linear model and the model was used to predict the responses of P cells to new chromatic and achromatic stimuli. Although linear models predict the chromatic responses well, simple linear combinations of centre and surround responses fail to predict P-cell responses to achromatic stimuli. 4. The temporal responses of the different subpopulations of P cells, such as ON/OFF or L-centre/M-centre were not significantly different.

Development and regulation of substance P expression in neurons of the tadpole optic tectum.

Activity-dependent synaptic plasticity is characteristic of developing visual systems. Using the frog retinotectal system, we investigated the extent to which afferent input affects neurotransmitter expression in a target structure. We have concentrated on a particular subpopulation of tectal cells that is immunoreactive to substance P (SP). Early in development, SP expression in tectal neurons was restricted to the anterior lateral region of the tectum. As tadpoles developed, this expression expanded into progressively more posterior and medial regions in a manner that closely followed the gradient of tectal maturation. At all times, however, anterior and lateral tectal regions had a greater percent of SP-like immunoreactive (SP-ir) cells than posterior and medial ones. Horseradish peroxidase (HRP) labeling of the retinal ganglion cell projection in conjunction with SP immunocytochemistry demonstrated that innervation by retinal ganglion cell terminals preceded the expression of SP by tectal cells. This suggested that the optic nerve may influence SP differentiation and/or expression. In support of this idea, transection of the optic nerve resulted in a decrease in SP expression in the deafferented tectal lobe of tadpoles. This result, opposite to that seen previously in the adult, also indicates that optic nerve-dependent regulation of SP expression in the developing and mature systems occurs through different pathways.

Projections of single retinal ganglion cells to the visual centers: an intracellular staining study in a plethodontid salamander.

The projection specificity of retinal ganglion cells and the morphology of their terminals were studied in the plethodontid salamander Plethodon jordani. In an in vitro approach, ganglion cells were stained with biocytin and reconstructed by means of light microscopy. Single retinal ganglion cells often have multiple terminal structures in the thalamus, pretectum, and tectum. The projection pattern in the diencephalic neuropils is related to the depth of the terminal arbor within the tectal fiber layer. Terminal arbors in the tectum differ in location, size, and branching pattern. The following types could be distinguished: The most superficial of the optic terminals in layer 1 are relatively small with a diameter of about 100 microm. With the exception of a few varicosities (beads) in the pretectal neuropils, their stem axons have no further collaterals or terminal arbors in the diencephalic neuropils. Intermediate terminals in layer 2 fan out to form a dense plexus with a medio-lateral extent of 180 microm on average. Some terminals in this layer show obvious antenna-like fibers reaching toward the surface of the tectum. The axons of layer 2 projecting neurons have additional collaterals and terminal arbors in the thalamus and pretectum. The deep layer 3 terminals spread out over a diameter of 400 microm on average and their degree of branching is moderate. The axons of layer 3 projecting ganglion cells have dense additional terminal arbors in the thalamus and pretectum. The deepest retinal terminals in the tectum are found within the predominantly efferent fiber layers. This type consists of an unbranched, but beaded axon which runs rostro-caudally with several bends and loops. The stem axon has an additional very dense terminal arborization in the neuropil of the nucleus Bellonci pars medialis and additional sparse collaterals in the pretectal area.

Peripherin-like immunoreactivity in type II spiral ganglion cell body and projections

Peripherin, an intermediate filament protein, is present in neuronal subpopulations of both peripheral and central nervous systems. The distribution of peripherin was studied in the adult rat cochlea using immunohistochemistry on whole mount material, in cryostat sections and sections of plastic embedded tissue. In the spiral ganglion, peripherin labeling was restricted to the perikarya of a subpopulation of neurons and their peripheral and central processes. Peripherin positive neurons had the following features: (i) they have a large eccentric nucleus, they were often found in a cluster of 2 or 3 cells, (ii) they were often located near the intraganglionic spiral bundle fibers, (iii) they represented roughly 8% of the whole ganglion population and (iv) on the average they had smaller perikarya than non-immunoreactive cells. Immunostaining on semithin plastic sections revealed positive reactivity on Type II ganglion cells, while Type I neurons were negative. Double labeling using peripherin and three neurofilament (NF) subunit antibodies confirmed the presence of both markers within the same spiral ganglion cell type. Type II neurons have been previously documented as the only subpopulation of the spiral ganglion that presents a strong positive NF immunoreactivity within their perikarya. In the organ of Corti, peripherin-positive fibers formed bundles that course beneath the outer hair cells and send branches that end as boutons contacting the outer hair cells. All these characteristics suggest that peripherin-positive cells are Type II neurons, and that peripherin constitutes a reliable marker for this spiral ganglion subpopulation, as well as their peripheral and central processes.

γ-Aminobutyric acid enhances the light response of ganglion cells in the trout pineal organ

Electrical recordings from achromatic second order neurons of intact superfused pineal organs of the rainbow trout were used to investigate the role of γ-aminobutyric acid (GABA) in the transmission of photoreceptor signals onto centrally projecting ganglion cells. Bath-applied GABA decreased the spike discharge rate of 98% of achromatic ganglion cells in a dose-dependent manner. GABA was also active if applied during synaptic blockade, demonstrating the presence of GABAergic receptors at the ganglion cell level. Responsiveness of ganglion cells to light was reversibly enhanced by GABA. The light response curve of ganglion cells, which was obtained by plotting spike rate versus light intensity, was significantly shifted to lower frequencies by GABA, indicating that GABA is an important inhibitory modulator of ganglion cell activity in the trout pineal organ.

Nasotemporal overlap investigated in a case of agenesis of the corpus callosum

In primates fibres from nasal retinae project contralaterally and those from temporal retinae ipsilaterally. At the border between these two regions an area of overlap where ipsi- and contralaterally projecting ganglion cells intermingle has been demonstrated in monkeys and cats. However, behavioural studies have failed to provide confirmatory evidence in humans. In this experiment simple reaction times to lateralized light flashes at four points of eccentricity (1/2, 1, 2 and 4 degrees) were recorded in an acallosal female. Responses made by the directly stimulated hemisphere were subtracted from those made by the indirectly stimulated hemisphere to arrive at estimates of interhemispheric transmission time. If present, a region of overlap should result in information being available to both hemispheres and consequently alleviate the need for any interhemispheric crossing. However, a large transmission time was found, even with stimuli presented close to fixation, thereby providing no evidence for the existence of such a region in humans.

Morphological diversity in terminals of W-type retinal ganglion cells at projection sites in cat brain.

The morphological features of retinal terminals in cat brain were examined at sites where projections of W-type ganglion cells predominate. These included the parvicellular C laminae of the dorsal lateral geniculate nucleus, the ventral lateral geniculate nucleus, stratum griseum superficiale of the superior colliculus, and the suprachiasmatic nucleus. Positive identification of retinal terminals was achieved following anterograde transport of intravitreally injected native or wheat germ agglutinin-conjugated horseradish peroxidase. In contrast to the classic features of retinal terminals as defined from sites where X- and Y-type ganglion cells predominate, i.e. round synaptic vesicles, large profiles, and pale mitochondria, substantial numbers of terminals in W-cell rich areas were found to contain dark mitochondria. Synaptic vesicles, although consistently round, were typically smaller in terminals with dark mitochondria than in those with pale mitochondria. These findings indicate a diversity among terminals of W-cells and suggest that such terminals cannot be distinguished on the basis of limited morphological criteria.

Increased serotonin in the developing superior colliculus does not alter the number or distribution of retinotectal ganglion cells.

Administration of a single subcutaneous dose of 5,7-dihydroxytryptamine (5,7-DHT) to newborn hamsters results in a significant increase in the density of serotoninergic (5-HT) fibers in the superficial layers of the superior colliculus (SC) and marked abnormalities in both the crossed and uncrossed retinotectal projections when these animals reach adulthood (R. Rhoades, C. Bennett-Clarke, R. Lane, M. Leslie, and R. Mooney, 1993, J. Comp. Neurol. 334:397-409). The present study was undertaken to determine whether changes in the retinotectal projection of 5,7-DHT-treated animals were associated with alterations in the number or distribution of retinal ganglion cells in these animals. Nissl staining of retinae from normal adult and 5,7-DHT-treated hamsters revealed no differences between them in the number or average diameter of cells in the retinal ganglion cell layer. Retrograde labeling with horseradish peroxidase (HRP) demonstrated no effect of 5,7-DHT treatment on the number or distribution of ipsilaterally or contralaterally projecting ganglion cells. Neonatal 5,7-DHT administration also had no effect on the distribution of soma diameters for HRP-labeled retinal ganglion cells. Electron microscopic analysis demonstrated no significant difference between the number of optic nerve fibers in the normal and 5,7-DHT-treated hamsters. The results are consistent with the conclusion that the effect of 5,7-DHT on the retinotectal projection may primarily be a function of this toxin, or the increase in 5-HT it induces, on the terminal arbors of retinotectal axons rather than on their parent cells.

Bifurcated projections of retinal ganglion cells bilaterally innervate the lateral geniculate nuclei in the cat

Cats were injected with the fluorescent retrograde tracers, Fluoro-Gold (FG) and Evans Blue (EB), into the left and right lateral geniculate nuclei (LGN), respectively. About 4.56% of the ganglion cells in the temporal retina were double-labeled by these dyes. 4.7% of these cells were of the large type, 30.3% were of the medium type, and 65% were classified as cells of the small type. These results indicate that members of all three ganglion cell size classes, mainly those of small type, bilaterally innervate the LGN via axonal bifurcation.

Morphological diversity and glutamate immunoreactivity of retinal terminals in the suprachiasmatic nucleus of the cat.

Although the cat visual system has been the subject of intensive investigation, little attention has been given to the morphological features of ganglion cell projections to the suprachiasmatic nucleus. The present study has utilized anterograde transport of horseradish peroxidase and wheat germ agglutinin-conjugated horseradish peroxidase to label ganglion cell terminals in the cat suprachiasmatic nucleus. Visualization of the reaction product was facilitated through the use of gold-substituted silver intensification. Ganglion cell terminals were found to be morphologically diverse, making both asymmetric and symmetric contacts with postsynaptic processes. Synaptic vesicles were either scattered or densely packed, sometimes forming paracrystalline arrays. In contrast to other retinorecipient areas in which ganglion cell terminals have been characterized by the presence of lightly staining mitochondria, many of the retinal terminals in the suprachiasmatic nucleus were seen to contain darkly stained mitochondria. Postembedding antiglutamate immunocytochemistry was used to evaluate the level of endogenous glutamate in these ganglion cell terminals. Although morphologically diverse, all of the retinal terminals in the suprachiasmatic nucleus were glutamate positive, consistent with the postulated role of glutamate as the neurotransmitter of retinal ganglion cells.

Genetic control of retinal projections in inbred strains of albino mice.

Mutations in the tyrosinase gene are often associated with a misrouting of retinal ganglion cell axons at the optic chiasm. In albinos, tyrosinase activity is lost and some ganglion cell axons that would normally project into the ipsilateral optic tract instead cross midline and project into the contralateral tract. The developmental mechanisms that cause this modification in neuronal connectivity are unknown. In this study, we screened six diverse strains of albino mice (strains 129, A, AKR, BALB/c, C57BL/6-c/c, and CD-1) to discover genetically determined variations and possible gene loci that might affect the severity of the albino decussation abnormality. Ganglion cells were retrogradely labeled with horseradish peroxidase, and the ipsilaterally and contralaterally projecting cells were counted. The average number of ipsilaterally projecting ganglion cells in the six albino strains varies from 1,000 to 1,300. Despite this variation, 1.8-1.9% of the total population projects ipsilaterally in each strain. In comparison, 2.8% project ipsilaterally in the pigmented strain, C57BL/6(-)+/+. However, the percentage of displaced, ipsilaterally projecting cells varies substantially among albino strains--from a low of 4% in strain CD-1 to a high of nearly 10% in C57BL/6-c/c. We conclude that even with large differences in genetic background and in absolute numbers of ganglion cells, there is no appreciable variation in the magnitude of decussation error among albino mice. The consistent effect of null alleles at tyrosinase suggests a comparably tight linkage between the biochemical activity of this enzyme and the mechanisms that control decussation phenotype.

Do retinal ganglion cells project bilaterally in ground squirrels?

Bilaterally projecting retinal ganglion cells have recently been reported in the Japanese monkey Macaca fuscata and albino rat. In the present experiments, we sought to determine whether ganglion cells project bilaterally in the 13-lined ground squirrel Spermophilus tridecemlineatus. We injected rhodamine-conjugated latex microspheres (red beads) into the right optic tract, superior colliculus and the dorsal lateral geniculate nucleus and FITC-conjugated latex microspheres (green beads) into these structures on the left side. In retinal wholemounts of all subjects, we found cells filled with red beads throughout the left retina and cells with green beads throughout the right retina. Ipsilaterally projecting ganglion cells were confined to the temporal retina where they were mixed with cells projecting contralaterally. In no case did we observe a doubly labeled cell. We interpret these observations as indicating that ganglion cells in this species do not project bilaterally.

The Visual System of the Florida Garfish, <i>Lepisosteus platyrhincus</i>(Ginglymodi) (Part 2 of 2)

Infusion of cobaltous-lysine into the optic nerve of juvenile Florida garfish reveals that the preoptic area, pretectum, thalamus and the mediorostral and ventrolateral poles of the optic tectum each receive bilateral, monosynaptic input. The bilateral input to the mediorostral pole of the tectum projects via the dorsal optic tract and terminates in the rostral half of the tectum. The bilateral input to the ventrolateral tectum projects via the ventral optic tract and extends the whole length of the tectum. The incidence of direct ipsilateral input to the tectum suggests binocular vision may play a functional role in the survival of this species. In order to test this hypothesis, we examined the retinal distribution of ipsilateral retinal ganglion cells that project to the mediorostral tectum and the size and location of the binocular visual field. Cell distribution was examined by retrograde labelling with rhodamine-conjugated dextran amine delivered by microinjections into the mediorostral pole of the right optic tectum. In the ipsilateral retina, ganglion cells are distributed in a narrow temporoventral area with a mean of 0.44±0.14 x 10³ cells per mm² apposing the retinal margin. In the contralateral retina, ganglion cells are also distributed within this temporoventral region with a mean peak density of 2.33 ± 0.47 x 10³ cells per mm². Three broad classes of ipsilaterally projecting retinal ganglion cells (orthotopic cells within the ganglion cell layer, displaced cells within the inner nuclear layer, and giant cells within the ganglion and inner nuclear layers) are identified, intermingled in both the ipsilateral and contralateral retinae after retrograde labelling from the mediorostral pole of the tectum. Ophthalmoscopic mapping of the monocular and binocular visual fields reveals two small frontal wedges of binocular overlap. A dorsal wedge (12° wide) extends from approximately 7° above the horizontal axis to 10° beyond the vertical axis, and a ventral wedge (20° wide) extends approximately 10° below the horizontal axis to 15° beyond the vertical axis. The dorsal binocular visual field is subtended by the temporoventral region of the retina possessing both ipsilaterally and contralaterally projecting ganglion cells which terminate in the mediorostral pole of the optic tectum. Therefore, the partial decussation of retinal ganglion cell axons at the optic chiasm brings information from corresponding regions of the binocular visual field into register in the mediorostral pole of the optic tectum. In addition, the size of the binocular visual field matches the relative proportion of the retina containing ipsilaterally projecting ganglion cells. The ontogeny and phylogeny of ipsilateral retinofugal projections in actinopterygians is discussed in relation to the putative secondary pathways mediating binocular vision.

The visual system of the Florida garfish, Lepisosteus platyrhincus (Ginglymodi). IV. Bilateral projections and the binocular visual field.

Infusion of cobaltous-lysine into the optic nerve of juvenile Florida garfish reveals that the preoptic area, pretectum, thalamus and the mediorostral and ventrolateral poles of the optic tectum each receive bilateral, monosynaptic input. The bilateral input to the mediorostral pole of the tectum projects via the dorsal optic tract and terminates in the rostral half of the tectum. The bilateral input to the ventrolateral tectum projects via the ventral optic tract and extends the whole length of the tectum. The incidence of direct ipsilateral input to the tectum suggests binocular vision may play a functional role in the survival of this species. In order to test this hypothesis, we examined the retinal distribution of ipsilateral retinal ganglion cells that project to the mediorostral tectum and the size and location of the binocular visual field. Cell distribution was examined by retrograde labelling with rhodamine-conjugated dextran amine delivered by microinjections into the mediorostral pole of the right optic tectum. In the ipsilateral retina, ganglion cells are distributed in a narrow temporoventral area with a mean of 0.44 +/- 0.14 x 10(3) cells per mm2 apposing the retinal margin. In the contralateral retina, ganglion cells are also distributed within this temporoventral region with a mean peak density of 2.33 +/- 0.47 x 10(3) cells per mm2. Three broad classes of ipsilaterally projecting retinal ganglion cells (orthotopic cells within the ganglion cell layer, displaced cells within the inner nuclear layer, and giant cells within the ganglion and inner nuclear layers) are identified, intermingled in both the ipsilateral and contralateral retinae after retrograde labelling from the mediorostral pole of the tectum. Ophthalmoscopic mapping of the monocular and binocular visual fields reveals two small frontal wedges of binocular overlap. A dorsal wedge (12 degrees wide) extends from approximately 7 degrees above the horizontal axis to 10 degrees beyond the vertical axis, and a ventral wedge (20 degrees wide) extends approximately 10 degrees below the horizontal axis to 15 degrees beyond the vertical axis. The dorsal binocular visual field is subtended by the temporoventral region of the retina possessing both ipsilaterally and contralaterally projecting ganglion cells which terminate in the mediorostral pole of the optic tectum. Therefore, the partial decussation of retinal ganglion cell axons at the optic chiasm brings information from corresponding regions of the binocular visual field into register in the mediorostral pole of the optic tectum.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of cycloheximide and ganglioside GM1 on the viability of retinotectally projecting ganglion cells following ablation of the superior colliculus in neonatal rats.

The time-course and extent of death of retinal ganglion cells (RGCs) following ablation of the superior colliculus (SC) in neonatal Wistar rats has recently been described [Harvey, A. R. and Robertson, D. (1992) J. Comp. Neurol., 325, 83-94]. Normal and pyknotic nuclei of retinotectally projecting ganglion cells were visualized using the fluorescent retrograde tracer diamidino yellow (DY), which had been injected into the SC at P2 (day of birth = P0), 2 days prior to tectal removal. The present report sets out to determine whether cycloheximide, an inhibitor of protein synthesis, or ganglioside GM1 reduced this lesion-induced RGC death. All surgery was carried out under ether anaesthesia; DY was injected into the left SC at P2 and the injected area was removed at P4. Cycloheximide (20-500 ng) was injected into the vitreous chamber of the right eye immediately after the lesion and again 11-12 h later. In some rats, cycloheximide administration was delayed until 12 h after the SC ablation. Control rats received SC lesions alone or lesions plus sham eye injections of saline. Different doses of GM1 were applied i.p. or intraocularly. Rats were perfused 24 h after the SC lesion, at the time of peak RGC death. Retinae of lesion only or sham eye injected rats contained approximately 11% pyknotic RGCs and the density of normal RGCs was approximately 3400/mm2. The rate of pyknosis in cycloheximide treated retinae was reduced to approximately 3%. Normal RGC density in these retinae was approximately 5500/mm2, similar to that found in retinae of unlesioned animals.(ABSTRACT TRUNCATED AT 250 WORDS)