The retinohypothalamic tract in the cat: retinal ganglion cell morphology and pattern of projection

The pattern of retinal projection to the hypothalamus and the morphological properties of the retinal ganglion cells that comprise the retinohypothalamic tract have been examined in the cat. Intraocular injections of horseradish peroxidase revealed a dense retinal projection to the ventral suprachiasmatic nucleus; however, lighter projections were seen in the dorsal suprachiasmatic nucleus, and in hypothalamic regions both dorsal and lateral to the suprachiasmatic nucleus. Intrasuprachiasmatic nucleus injections of horseradish peroxidase retrogradely labelled retinal ganglion cells that were small to medium in soma size. The labelled ganglion cells exhibited long thin dendrites that were sparsely branched. The labelled retinal ganglion cells exhibited a significant change in soma size associated with retinal eccentricity. The morphological characteristics of the ganglion cells that project to the suprachiasmatic nucleus are similar to those of gamma cells.     

Glutamate-like Immunoreactivity in Retinal Terminals of the Mouse Suprachiasmatic Nucleus

With a view to identifying the neurotransmitter content of retinal terminals within the mouse suprachiasmatic nucleus, a highly specific antiserum to glutaraldehyde-coupled glutamate was used in a postembedding immunogold procedure at the ultrastructural level. Retinal terminals were identified by cholera toxin–horseradish peroxidase transported anterogradely from the retina and reacted with tetramethyl benzidine/tungstate/H₂O₂, or by their characteristically pale and distended mitochondria with irregular cristae. Controls included model ultrathin sections containing high concentrations of various amino acids. Alternate serial sections were labelled with anti-glutamate and anti-γ-aminobutyric acid (GABA). Data were analysed by computer-assisted image analysis. Density of glutamate labelling (gold particles per μm²) on whole retinal terminals was > 3 times higher than that on postsynaptic dendrites, and > 5 times higher than that on miscellaneous non-retinal non-glutamatergic terminals in the suprachiasmatic nucleus. The overall density of gold particles over retinal terminals was ～ 3 times higher than that over GABAergic terminals, in which glutamate-like immunoreactivity was mainly mitochondrial. Labelling of vesicles in retinal terminals was almost 5 times greater than the apparent labelling of vesicles in GABAergic terminals, underscoring the location of transmitter glutamate within synaptic vesicles in retinal terminals. In the retino-recipient region of the suprachiasmatic nucleus there was also a small population of non-retinal glutamatergic terminals. Their overall immunoreactivity was similar to or exceeded that of retinal terminals, but morphological features clearly distinguished between these two types of glutamate-containing terminals. The present results indicate that the vast majority of retinal terminals may use glutamate as a transmitter, in keeping with electrophysiological and neuropharmacological data from other sources. The possibility of cotransmitters within retinal terminals, suggested by the presence of dense-core vesicles among the glutamate-containing synaptic vesicles, has still to be addressed.     

Response of retinal terminals to loss of postsynaptic target neurons in the dorsal lateral geniculate nucleus of the adult cat.

We have used the neurotoxin kainic acid to produce rapid degeneration of neurons in the dorsal lateral geniculate nucleus (dLGN) of the adult cat. This degeneration mimics the rapid loss of geniculate neurons seen after visual cortex ablation in the neonate. Subsequent anterograde transport of horseradish peroxidase injected into the eye was used to reveal the projection patterns of retinal ganglion cell axons at different survival periods after the kainic acid injection. The density of retinal projections to the degenerated regions of the geniculate was reduced considerably at 4 and 6 months survival, but at 2 months was not significantly different from normal. The laminar pattern of projections to degenerated regions of the geniculate did not change in any animals studied, even when an adjacent lamina contained surviving cells. Electron microscopic examination of degenerated dLGN revealed intact retinal (RLP) and RSD terminals at all survival times, although the density of terminals appeared much reduced when compared to controls. Some RLP terminals exhibited the "dark reaction" of degeneration and these degenerating terminals were most numerous at 2 months survival. These findings demonstrate that, in response to degeneration of their usual target cells, mature retinal ganglion cells with withdraw their axon terminals from these regions of degeneration. We conclude that mature retinal ganglion cells continue to be dependent on target integrity for the maintenance of a normal axonal arborization.     

The projections of different morphological types of ganglion cells in the cat retina.

The central projections of the retinal ganglion cells of the cat were examined using the method of retrograde transport of horseradish peroxidase. Peroxidase was injected into the lateral geniculate nucleus and into the superior colliculus by means of a recording micropipette. After injections at retinotopically homologous points in these two structures in separate animals, tha patterns of retinal ganglion cell labeling were compared. We found that there were three populations of ganglion cells: small cells, that projected predominantly to the superior colliculus; medium-sized cells, that projected predominantly to the lateral geniculate nucleus; and large cells, some of which projected to both structures, and some of which projected to the lateral geniculate nucleus alons. Quantitative studies showed that the average size of the cells in each population was smaller at the area centralis than in the periphery. These results could be directly related to physiological classifications of retinal ganglion cells proposed by other authors.     

Retinal ganglion cell degeneration following loss of postsynaptic target neurons in the dorsal lateral geniculate nucleus of the adult cat.

Kainic acid was used to produce selective degeneration of neurons in the dorsal lateral geniculate nucleus of the adult cat. This degeneration mimics the rapid loss of geniculate neurons seen after visual cortex ablation in the neonate. Following survivals of 2, 4, or 6 months, the geniculate was injected with horseradish peroxidase and the retinae were examined for the presence of retrogradely labeled cells. Analysis of ganglion cell density in peripheral nasal retina revealed a 58% loss of cells overall at 6 months. The proportion of cells labeled with horseradish peroxidase decreased more rapidly, until none were labeled at 6 months. Separate analysis of small, medium, and large ganglion cell populations revealed that only medium-sized cells were lost at 2 months whereas both medium and large cells were lost at 4 and 6 months. By 6 months, 92% of medium cells and 65% of large cells had degenerated. These results show that mature retinal ganglion cells in the cat maintain a dependence on target integrity for their continued survival. When the appropriate target is lost, the ganglion cells respond first by axon terminal retraction and then by cell death.     

Anterograde axoplasmic transport of peroxidase in the rat visual system after intraocular neurotoxin injection

The effect of intraocular injection of the neurotoxin, kainic acid, on the retinofugal pathway has been examined by anterograde transport of horseradish peroxidase. Although less peroxidase was present in all the primary optic centers after the intraocular injection of kainic acid that in the controls, the area occupied by the retinal terminals remained fairly constant except for the pretectal region where there is a smaller area of peroxidase precipitate. These results suggest that certain ganglion cells are killed by kainic acid but some others survive and their terminals are present in the majority of the terminal fields.     

Development and characterization of glucocorticoid receptors in rat superior cervical ganglion

The effect of intraocular injection of the neurotoxin, kainic acid, on the retinofugal pathway has been examined by anterograde transport of horseradish peroxidase. Although less peroxidase was present in all the primary optic centers after the intraocular injection of kainic acid that in the controls, the area occupied by the retinal terminals remained fairly constant except for the pretectal region where there is a smaller area of peroxidase precipitate. These results suggest that certain ganglion cells are killed by kainic acid but some others survive and their terminals are present in the majority of the terminal fields.     

The retinal projection to the thalamus in the cat: a quantitative investigation and a comparison with the retinotectal pathway

The projection of cat retinal ganglion cells to the thalamus was examined using the method of retrograde axonal transport of horseradish peroxidase (HRP). After the injection site was determined physiologically, HRP was applied by one of three methods: iontophoretic injection of minimal amounts, single pressure injections and multiple pressure injections. Iontophoretic injections into single laminae of the dorsal part of the lateral geniculate nucleus (LGNd) revealed that laminae A and A1 receive almost exclusively axon terminals from alpha and beta cells. Single pressure injections elucidated the retinotopic organization of the LGNd. Multiple injections lead to HRP uptake in the whole LGNd including parts of adjacent thalamic nuclei and revealed that at least 77% of all retinal ganglion cells project to the thalamus. This pathway is made up of all alpha cells, all beta cells and almost half of the gamma cells. The thalamus receives its visual input predominantly from the ipsilateral temporal and the contralateral nasal retina; some alpha cells were also labeled in the contralateral temporal retina. The shape of the decussation line was analyzed and its width was found to be proportional to the average ganglion cell spacing along the dorsoventral axis of the retina. From a comparison of the retinothalamic and retinotectal pathways, an estimate of the number of cells with bifurcating axons could be given. The axons of all alpha cells, 10% of the beta cells, and every second gamma cell bifurcate; this amounts to 30% of the retinal ganglion cells.     

An anterograde-retrograde transneuronal transport of conjugates of wheat germ agglutinin with horseradish peroxidase (WGA-HRP): labeling of neurons in the reticular nucleus of the thalamus with WGA-HRP injected into the posterior column nuclei in the cat

Neuronal cell bodies in the reticular thalamic nucleus (R) were labeled with wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) which was injected contralaterally into the posterior column nuclei (PCN) in the cat. The tracer was assumed to be transported to the posterolateral ventral thalamic nucleus (VPL), where it could escape from axon terminals of the PCN neurons and then be taken up by axon terminals of R neurons to label retrogradely the cell bodies of the R neurons.     

Ultrastructural and immunocytochemical characterization of terminals of postsynaptic ascending dorsal column fibers in the rat cuneate nucleus

The morphology, synaptic contacts, and neurotransmitter enrichment of postsynaptic dorsal column terminals in the cuneate nucleus of rats were investigated and compared with those of identified primary afferents. For this purpose, anterograde transport of horseradish peroxidase–based tracers injected in the spinal cord was combined with postembedding immunogold labeling for glutamate and gamma-aminobutyric acid (GABA). Anterogradely labeled postsynaptic dorsal column terminals were morphologically homogeneous: they were small (mean area = 1. 37 μM2) and dome-shaped, contacted single dendritic shafts or cell bodies, and were not involved in axoaxonic synapses. The majority of them were not enriched in glutamate or GABA immunoreactivity compared with other tissue components. Their morphology, size, and neurotransmitter content thus differed from that of primary afferents. These differences are consistent with distinct functional roles for the two main afferent systems ascending to the cuneate nucleus. © 1995 Wiley-Liss, Inc.     

Topographical origin and ganglion cell type of the retino-pulvinar projection in the cat

In order to examine the pattern of the retino-pulvinar projection in the cat, the existence of which has been recently demonstrated using autoradiographic fiber tracing technique, a small amount of horseradish peroxidase (HRP) was injected into the lateral part of the pulvinar nucleus at various rostocaudal levels. The retrogradely labeled ganglion cells were analyzed in terms of their topographical location and cell size, as seen inretinal whole mounts. The results were compared with those obtained following injections into the lateral geniculate nucleus. Retrogradely labeled cells were found in the retina bilaterally after injections of HRP into the pulvinar nucleus. Pulvinar injections produced labeling of retinal cells in the nasal half of the retina contralaterally, and in the temporal half ipsilaterally. The labeled cells were diffusely distributed in a retinotopically organized fashion. The representation of the area centralis in the retino-pulvinar projection is displaced rostrally as compared with the retino-geniculate projection. All labeled cells after pulvinar injections were medium to small size and no large cells were encountered.     

Ganglion cells of the cat accessory optic system: Morphology and retinal topography

The ganglion cells of the cat's retina form classes that are distinct in their cell morphology, retinal distribution, central projections, and functional properties. By means of the retrograde transport of horseradish peroxidase injected into the accessory optic nuclei of the cat midbrain, we have characterized the retinal ganglion cells projecting to these nuclei. The retinal projection is virtually completely crossed to the medial terminal nucleus and to the lateral terminal nucleus. This appears to be true for the dorsal terminal nucleus as well, although difficulties of the technique limit our findings for this region. No differences were found in either the spatial distribution, or the somal size distribution, or the morphological characteristics of the ganglion cells projecting to these three nuclei. In spatial distribution, these cells are concentrated in the area centralis and visual streak and show no evidence of a nasotemporal divison. Morphologically, they have small to medium-sized somas and delicate, sparsely branching dendrites. They do not appear to belong to any of the morphological cell types thus far identified.     

Retinal ganglion cells projecting to the rabbit accessory optic system

Recent evidence from extracellular recording studies indicates that the medial terminal nucleus (MTN) of the rabbit accessory optic system receives inputs from a particular functional class of retinal ganglion cells—specifically, the on-type direction-selective cells. These ganglion cells have been selectively labeled by the retrograde transport of horseradish peroxidase (HRP) injected into the MTN. The number of labeled cells, their distribution over the retina, and their soma areas were determined. In one animal in which the HRP injection completely filled the nucleus, two thousand ganglion cells were labeled. This number agrees with previous estimates of the number of retinal axons terminating in the MTN. Unlike results in avians, none of the ganglion cells was displaced—i.e., they were not Dogiel cells. The density of labeled cells was highest in the visual streak and, overall, the distribution of labeled cells corresponded to the physiologically determined distribution of on-type direction-selective cells. Cells labeled by the HRP injection were among the 20% largest cells in the retina. This result, in conjunction with conclusions from other studies, leads to the prediction that on-type direction-selective cells can be characterized morphologically as cells with large cell bodies and a very extensive dendritic spread in which the dendrites ramify in the vitreal sublamina of the inner plexiform layer.     

Direct vagal input to neurons in the area postrema which project to the parabrachial nucleus: An electron microscopic-HRP study in the cat

This study in cat examines the synaptic relationship of vagal afferents to parabrachial projecting neurons in the area postrema (AP) using anterograde and retrograde transport of horseradish peroxidase (HRP). Wheat germ agglutinin-HRP injected into the parabrachial nucleus (PBN) produced retrograde neuronal labeling in the AP and in the nucleus of the tractus solitarius bilaterally, but with an ipsilateral predominance. Labeled neurons were confined mainly to the caudal 2/3's of the AP. Following injection of WGA-HRP into the PBN and HRP into the nodose ganglion in the same animal, examination of sections of the AP with the electron microscope revealed anterogradely labeled axon terminals in apposition to retrogradely labeled somata and dendrites. In some instances, labeled terminals were observed to form synaptic contacts with retrogradely labeled neurons. We conclude that in the cat a vagal input to neurons in the AP is monosynaptically relayed to the PBN.     

Lucifer yellow, retrograde tracers, and fractal analysis characterise adult ferret retinal ganglion cells.

The dendritic morphology of retinal ganglion cells in the ferret was studied by the intracellular injection of lucifer yellow in fixed tissue. Ganglion cells were identified by the retrograde transport of red or green fluorescent microspheres that had been injected into different target nuclei, usually the lateral geniculate nucleus or superior colliculus. This approach allows the comparison of dendritic morphologies of ganglion cells in the same retina with different central projections and also identifies cells with branching axons. The digitised images of dendritic arbors were analysed quantitatively by a variety of measures. Dendritic complexity was assessed by calculating the fractal dimension of each arbor. The ferret has distinct alpha, beta, and gamma morphological classes of cells similar to those found in the cat. The gamma cell class was morphologically diverse and could be subdivided into "sparse," "loose," and "tight" groups, reflecting increasing dendritic complexity. Whereas the beta cell projection was limited to the lateral geniculate nucleus alone, alpha and gamma cells could project to either or both nuclei. Retinal ganglion cells labelled from the pretectal nuclei formed a morphologically distinct class of retinal ganglion cells. The ipsilateral projection lacked alpha cells and the most complex, "tight" gamma cells. However, ipsilaterally projecting "loose" gamma cells overlapped alpha cells in both soma and dendritic dimensions. Different morphological classes of retinal ganglion cells hence show characteristic axon behaviour both in their decussation at the chiasm and in which targets they innervate. Fractal measures were used to contrast variation within and between these identified classes.     

Direct retinal projections to the hypothalamus,piriform cortex,and accessory optic nuclei in the golden hamster as demonstrated by a sensitive anterograde horseradish peroxidase technique

The central projections of the retinal ganglion cells of the golden hamster were examined using horseradish peroxidase (HRP) as the anterograde tracer molecule. Following monocular injections of HRP into the vitreous, retinfugal fibers were histochemically demonstrated using the chromagen tetramethylbenzidine. This procedure, being more sensitive than the ³H-amino acid radioautographic technique, provided a clear demonstration of previously controversial retinal projections, clearer definition of established projections, and the discovery of new retinal pathways. An inferior accessory optic system was shown to be unequivocally present in this species and to consist of both crossed and uncrossed components. A direct retinal projection to the suprachiasmatic nucleus (SCN) of the hypothalamus was confirmed in this study. But the distribution of terminals as seen by this procedure was substantially different than previously reported; both rostrocaudal and mediolateral asymmetries in the distribution of label between the ipsilateral and contralateral SCN were observed. Substantial differences in the retinal projection to the SCN in the hamster and the rat were also noted. It is suggested that these differences may reflect the different effects photic input has on the neuroendocrine-gonadal axis in these two species. Finally, labeled retinal axons were followed leaving the optic tract and coursing anteriorly through the plexiform layer of the piriform cortex; other labeled fibers were seen to enter the septal region. The physiological significance of these previously undescribed retinal projections is not known.     

Localization and morphology of cat extraocular muscle afferent neurones identified by retrograde transport of horseradish peroxidase

Afferent neurones that provide proprioceptive innervation of extraocular muscles of the cat have been identified by means of retrograde axonal transport of horseradish peroxidase (HRP). Discrete injections of HRP into the medial rectus, lateral rectus, or retractor bulbi muscles labeled pseudounipolar neurones that were localized exclusively to the ipsilateral semilunar ganglion. The distribution of labeled neurones within the ganglion was consistent with its somatotopic organization with the majority found within the ophthalmic subdivision. Cell counts indicating approximately 90 labeled neurones per horizontal rectus muscle correlated well with earlier quantitative observations regarding the percentage of afferent fibers in oculomotor nerves and the number of proprioceptive terminals in the extraocular muscles. Neither the trigeminal mesencephalic nucleus nor the contralateral semilunar ganglion contained labeled neurones following injections of HRP into extraocular muscles. Consistent with other studies of spinal and cranial ganglia the contingent of pseudounipolar neurones present in the cat semilunar ganglion included both light and dark cell types. Light and electron microscope analysis of HRP-labeled neurones in combination with acetylcholinesterase (AChE) histochemistry revealed that only one of the two neuronal types, the light cell, subserves extraocular muscle proprioception. Our data support the hypothesis that ganglion neurone type and, more specifically, soma diameter, are important determinants of functional status.     

The relationship between cell density and the nasotemporal division in the rat retina

Cell counts in the ganglion cell layer of the rat retina have been undertaken following unilateral injections of horseradish peroxidase into the ipsilateral thalamus. By retrograde transport, the tracer defined the location of the uncrossed retinal projection, making it possible to determine the relationship between the area of highest cell density and the nasotemporal division. Here it is demonstrated that unlike in the primate and cat, these two regions of retinal specialisation reside in different locations, with the nasotemporal division displaced temporally to the area of highest cell density.     

Extra-organ sources of parasympathetic innervation of small intestine in the region of ligament of Treitz]

Localization sites of neurons in the dorsal motor nucleus of cat vagus innervating the duodenal-jejunal region and upper part of the jejunum were investigated using the technique of the retrograde axonal transport of horseradish peroxidase. Most part of the neurons was located within the ventrolateral area of the nucleus from +1.0 to +2.7 mm (according to obex). Morphological features of the ganglion nodosum neurons which organize afferent innervation of the intestine were analyzed. It has been shown that the maximal amount of such neurons is concentrated in the medium and caudal parts of the ganglion.     

The organization of retinal projections to the diencephalon and pretectum in the cichlid fish, Haplochromis burtoni

The organization of retinofugal projections was studied in a cichlid fish by labelling small groups of retinal ganglion cell axons with either horseradish peroxidase or cobaltous lysine. Two major findings resulted from these experiments. First, optic tract axons show a greater degree of pathway diversity than was previously appreciated, and this pathway diversity is related to the target nuclei of groups of axons. The most striking example is the formation of the medial optic tract. Fibers that will become the medial optic tract move abruptly away from their neighbors, at about the level of the optic chiasm, and coalesce at the dorsomedial edge of the marginal optic tract. The medial optic tract projects to the thalamus, the dorsal pretectum, and the deep layer of the optic tectum. The axial optic tract is a group of fibers which segregates from the most medial portion of the marginal optic tract, at about the level of the optic chiasm. The axial tract stays medial to the marginal optic tract for a few hundred microns and then curves laterally to rejoin the marginal optic tract. At least some axial trat axons terminate in the suprachiasmatic nucleus. Within the marginal optic tract, retinal ganglion cell axons from a given retinal quadrant are always segregated into at least two groups. The smaller group projects to the superficial pretectal nucleus. The larger group projects to the superficial layer of the optic tectum. Second, each nontectal retinal termination site receives a unique pattern of retinal input. Within the pretectum the parvocellular superficial pretectal nucleus receives a highly retinotopically organized input from all retinal regions; the basal optic nucleus receives a roughly retinotopically organized input from all retinal regions; the dorsal pretectum receives an input from all retinal regions; and the central pretectal nucleus receives input only from the ventral hemiretina. Within the diencephalon the thalamus receives an input from all retinal regions, but this input is not retinotopically organized; the suprachiasmatic nucleus receives input from the region of central retina that lies just dorsal to the optic nerve head, via the axial optic tract. The accessory optic nucleus receives input from the dorsal hemiretina.