Evidence for an amacrine cell system in the ganglion cell layer of the rat retina

In the ganglion cell layer of the rat retina approx 50% of the cells with the Nissl morphology of neurons survive optic nerve section in infant and adult rats and cannot be retrogradely labelled with horseradish peroxidase. The number of neurons which can be retrogradely labelled with horseradish peroxidase from subcortical visual centres is similar to the number of axons in the optic nerve, and it is concluded that the small neurons do not send an axon into the optic nerve. The dendritic tree of the cells which have axons was demonstrated by filling the cells with horseradish peroxidase from the optic nerve. The dendritic structure of the cells which survive optic nerve section was shown by injecting horseradish peroxidase into the retina or impregnating with the Golgi method the cells which survive optic nerve section. A variety of amacrine cells were found in the ganglion cell layer which form branches in the lower part of the inner plexiform layer.It can be concluded that amacrine cells form a substantial number of the neurons in the ganglion cell layer.     

Is Thy-1 expressed only by ganglion cells and their axons in the retina and optic nerve?

Summary The distribution of Thy-1 in the retina and optic nerve has been examined immunohistochemically, and compared to that of the astrocytic marker glial fibrillary acidic protein. The axons and cell bodies of ganglion cells were found to be Thy-1 positive as were processes within the inner plexiform layer. Transection of the optic nerve in the neonatal rat results in the rapid degeneration of the ganglion cells but some Thy-1 staining remains in the inner plexiform layer. We have estimated using an immunoassay of normal and optic nerve transected retinae that about 70% of the Thy-1 in the retina is on ganglion cells and their axons and the remainder is on cells which contribute processes to the inner plexiform layer, presumably amacrine, bipolar or Müller cells.In the optic nerve the Thy-1 was found to be limited to the fascicles of optic nerve fibres and the intrafascicular spaces, containing astrocytes and their processes, were not stained. Axotomy of the adult nerve, which produced axonal degeneration and astrocytic proliferation, led to a loss of over 95% of the Thy-1 from the nerve. We found no evidence that the astrocytes of the retina or optic nerve were Thy-1 positive in normal animals or during degeneration.     

Specific transcellular carbocyanine-labelling of rat retinal microglia during injury-induced neuronal degeneration

The present work employed a new technique for labelling phagocytizing microglia in the axotomized retinal of adult rats. Transection axotomy was performed within the intraorbital segment of the optic nerve, and the fast-transported, vital fluorescent carbocyanine dyes DiI and 4Di-10ASP were deposited at the ocular stump of the nerve in order to retrogradely prelabel the ganglion cells which were destined to die. Optic nerve transection resulted in progressive degradation of ganglion cell axons, perikarya and dendrites within the retina and in release of fluorescent material which was then incorporated into cells identified as microglia but not into other cells of the retina. Incorporation of labelled material into microglia occurred only when the ganglion cells degenerated and not when the non-lesioned ganglion cells were labelled from the superior colliculus. Double-staining of microglia with both dyes helped to compare the pattern of labelling for each dye. After progression of ganglion cell degeneration, microglia displayed a staggered, bilaminated distribution within the ganglion cell layer and within the inner plexiform layer. Fluorescent microglia were not found within the deeper layers of the retina indicating that transneuronal degeneration and subsequent labelling of microglial cells do not occur. The results show that one major function of microglia within the ganglion cell and inner plexiform layers of the lesioned retina is to remove debris produced after degradation of neurons.     

An immunocytochemical marker for hamster retinal ganglion cells

Summary We examined the specificity and developmental time course of the labelling of retinal ganglion cells in Syrian hamsters by a monoclonal antibody AB5. In adult hamsters, AB5 selectively labelled somata in the ganglion cell layer, dendrites in the inner plexiform layer and axons in the nerve fibre layer. When retinal ganglion cells were retrogradely labelled with Dil prior to AB5 immunocytochemistry, all of the retrogradely labelled retinal ganglion cells in the ganglion cell layer were AB5 immunoreactive, indicating that AB5 labels all classes of ganglion cell in that layer. In retinae depleted of retinal ganglion cells by neonatal optic nerve transections, AB5 did not label any somata or processes, indicating that AB5 specifically labels retinal ganglion cells. During development, AB5 labelling first appeared as a weak staining of cell bodies in the ganglion cell layer on postnatal day 12 (P12; PO=first 24 h following birth) and acquired the staining pattern seen in the adult by postnatal day 14. From the onset of AB5 immunoreactivity, AB5-labelled somata of varying sizes were present across the entire retinal surface. Although AB5 labelled retinal ganglion cell axons in the nerve fibre layer of the retina it did not label the optic nerve or retinal ganglion cell axons in the brain at any age examined. AB5 labelling was also found to be compatible with bromodeoxyuridine immunocytochemistry and, therefore, useful for determining the time of generation of hamster retinal ganglion cells.     

大鼠视神经切断后视网膜双极细胞PKC-α和recoverin的表达

为了探讨视神经切断后视网膜内部是否存在突触可塑性改变,本实验采用大鼠视神经切断模型,通过免疫组织化学方法检测视神经切断后视网膜双极细胞PKC-α和recoverin的表达变化。结果显示:正常视网膜中,PKC-α和recoverin阳性产物主要见于视网膜内核层、内网层及节细胞层,另外外核层也可见少量recoverin阳性细胞。视神经切断后3d,大鼠视网膜内网层高倍镜下可见PKC-α和recoverin免疫阳性终末的数量开始增加,14d时增至最高,21d、28d呈现逐渐减少的趋势。本研究结果提示视神经切断后视网膜双极细胞与节细胞之间的突触可能存在早期增生,后期溃变的可塑性变化。     

Changes in NADPH diaphorase expression in the fish visual system during optic nerve regeneration and retinal development

The various functions of nitric oxide (NO) in the nervous system are not fully understood, including its role in neuronal regeneration. The goldfish can regenerate its optic nerve after transection, making it a useful model for studying central nervous regeneration in response to injury. Therefore, we have studied the pattern of NO expression in the retina and optic tectum after optic nerve transection, using NADPH diaphorase histochemistry. NO synthesis was transiently up-regulated in the ganglion cell bodies, peaking during the period when retinal axons reach the tectum, between 20–45 days after optic nerve transection. Enzyme activity in the tectum was transiently down-regulated and then returned to control levels at 60 days after optic nerve transection, during synaptic refinement. To compare NO expression in the developing and regenerating retina, we have looked at NO expression in the developing zebrafish retina. In the developing zebrafish retina the pattern of staining roughly followed the pattern of development with the inner plexiform layer and horizontal cells having the strongest pattern of staining. These results suggest that NO may be involved in the survival of ganglion cells in the regenerating retina, and that it plays a different role in the developing retina. In the tectum, NO may be involved in synaptic refinement.     

Antidromic responses of orthodromically identified ganglion cells in monkey retina

1. Conduction velocities of two types of on-centre monkey ganglion cells, called phasic and tonic, have been measured by stimulating their axons in the optic tract while recording from their cell bodies in the retina.2. The average conduction velocity of twenty-two phasic and twenty-seven tonic cells is 3.8+/-S.D. 0.6 and 1.8+/-S.D. 0.4 m/sec respectively. Since the latter, but not the former, show opponent-colour responses, retinal signals carrying information about colour appear to be travelling in smaller axons than those not handling such information.3. Stimulation of the optic tract elicits several graded intraretinal potentials, which are negative in the optic nerve fibre layer and positive in the inner plexiform layer. One of these potentials, which is largest near the fovea, occurs simultaneously with the antidromic impulses of tonic ganglion cells and is considered to result from extracellular current generated by these cells.4. Stimulation of the optic tract suppresses the orthodromic responses of ganglion cells, more for phasic than for tonic ones. This suppression is only observed after a cell is antidromically driven and is considered most likely due to a transient hyperpolarization of the cell's membrane potential following an impulse.     

Histology of the ferret retina

The retina of the adult ferret, Mustelo furo, was studied with light and transmission electron microscopy to provide an anatomical basis for use of the ferret as a model for retinal research. The pigment epithelium is a simple cuboidal layer of cells characterized by a zone of basal folds, apical microvilli, and pigment granules at various stages of maturation. The distinction between rod and cone photoreceptor cells is based on their location, morphology, heterochromatin pattern and the electron density of their inner segments. The round, light-staining cone cell nuclei occupy the layer of perikarya along the apical border of the outer nuclear layer. The remainder of the outer nuclear layer consists of oblong, deeply-stained rod cell nuclei. Ribbon type synaptic complexes involving photoreceptor cell axons, horizontal cell processes, and bipolar cell dendrites characterize the outer plexiform layer. The inner nuclear layer is comprised of horizontal, bipolar, and amacrine cell perikarya as well as the perikarya of the Müller cells. The light-staining horizontal cell nuclei are prominent along the apical border of the inner nuclear layer. The light-staining amacrine cell nuclei form a more or less continuous layer along the basal border of the inner nuclear layer. Both conventional and ribbon-type synapses characterize the inner plexiform layer. The ganglion cells form a single cell layer. The optic fiber layer contains bundles of axons surrounded by Müller cell processes. Small blood vessels and capillaries are present in the basal portion of the retina throughout the region extending from the internal limiting membrane to the outer plexiform layer. The adult one-year-old retina is compared with the retina at the time of eye opening.     

Displaced retinal ganglion cells in normal frogs and those with regenerated optic nerves

Summary We have analysed the number and spatial distribution of displaced retinal ganglion cells in the frog Litoria (Hyla) moorei. A series of normal animals was compared with one in which the optic nerve was crushed and allowed to regenerate. Ganglion cells were labelled with horseradish peroxidase (HRP) applied to the optic nerve, and retinae were examined as sections or whole mounts. We analysed separately ganglion cells with somata displaced to the inner nuclear (Dogiel cells, DGCs) and to the inner plexiform layer (IPLGCs). These findings were related to data for the orthotopic ganglion cells (OGCs). The mean number of DGCs in the normal series was 2,550 (±281) and fell to 1,630 (±321) after regeneration, representing a mean loss of 36%. This reduction was not significantly different from the mean loss of 43% from the OGC population in which mean values fell from 474,700 (±47,136) to 268,700 (±54,395). In both the normal and the regenerate series, DGCs were estimated to represent means of only 0.6% of the OGC population. Densities of DGCs were highest in the nasoventral and temporo-dorsal peripheries; densities of both DGCs and OGCs were lower after optic nerve regeneration. We conclude that the factors which affect ganglion cell death during optic nerve regeneration, do so to similar extents amongst the DGC and the OGC populations. The IPLGCs were very rare in normal animals with a mean of 420 (±95). However, their numbers increased after regeneration to a mean of 3,350 (±690), estimated to be 1.2% of the OGC population. These cells normally favoured peripheral retina but became pan-retinal after regeneration. The primary dendrites of the majority of IPLGCs were oriented in the same direction as those of OGCs. We conclude that most IPLGCs were OGCs which had relocated their somata to the inner plexiform layer.     

Early expression of recoverin in a unique population of neurons in the human retina

 The calcium-binding protein recoverin has been reported as present in photoreceptors, cone bipolar cells and sparse cells in the ganglion cell layer in the adult retinae of various vertebrate species. The present study was undertaken to clarify the developmental pattern of recoverin-immunoreactive cells in the human retina with particular attention to the cells in the inner retinal layers. In the adult human retina, small populations of recoverin-containing cells are present in the ganglion cell and nerve fiber layers. However, the precursors of these cells are quite numerous on the inner and outer borders of the nerve fiber layer in the fetal retina. By 13 weeks of gestation these cells express recoverin very intensely. By 24 weeks they are mature-looking with relatively large soma sizes (mean=118 μm²) and appear round, oval or multipolar in shape, with varying numbers of short processes. There follows a noticeable reduction of the mean soma size, but little change in morphology and process number during the remaining gestational stages up to and after birth. The mean numerical density of the recoverin-positive cells in the fetal inner retinal layers is gradually reduced from the high level at 13 weeks until birth, when there is a great drop to the adult level. The recoverin-immunoreactive cells in the ganglion cell layer demonstrate distinctively different developmental and morphological features from the principle neurons and glial cells in the retina. They are probably the neurons derived from the marginal zone of the retinal primordium that reside in the inner and outer borders of the nerve fiber layer due to the invasion of ganglion cell axons. The expression of recoverin in the neurons may be significant in maintaining an inside-out and centro-peripheral gradient of calcium concentration in the premature retina, thereby playing a role in determining the polarity of the differentiating ganglion cells and the growth of their axons in a centrifugal spatiotemporal order. Accepted: 3 June 1996     

Identification of amacrine and ganglion cells in the carp retina.

1. Amacrine and ganglion cells in the carp retina were identified from such criteria as photoresponses, intracellular dye staining, responses to optic nerve stimulation and behaviour to a synapse blocking agent. 2. Responses of ganglion cells were accompanied by spike discharges., either facilitated or suppressed by photic stimulation. The cells were also invaded by antidromic impulses, which survived after chemical synapses had been blocked by application of atomized CoCl2 solution. In subsequent histology of the Procion-stained neurones, the cell bodies were found in the ganglion cell layer and the axons were often traced. 3. Amacrine cells were subdivided into two types. The first type gave rise to transient depolarizations at both on- and offsets of spot and annulus illuminations, usually being associated with spike discharges of which the amplitudes varied in different cells. In histology, the cell bodies of this type were situated in the inner nuclear layer and dendrites ramified in two or more discrete sublayers of the inner plexiform layer (the stratified amacrine cell of Cajal). 4. The second type of amacrine cells produced sustained responses during illumination, being associated with no spike but with small oscillatory wavelets. The cell bodies were situated in the inner nuclear layer and the dendrites ramified in a single sublayer of the inner plexiform layer (the monolayered amacrine cell). 5. An attempt was made to see the effect of activation of centrifugal fibres on amacrine cells, but almost all of about 200 cells examined did not respond to optic nerve stimulation. Only two cells produced, with long latency, a small post-synaptic depolarization which disappeared after chemical synapses in the retina had been blocked. It is considered that the physiological role of the centrifugal system is insignificant in the carp retina.     

高血压大鼠视网膜溶酶体酶组织化学研究

目的：探讨溶酶体酶在高血压视网膜网变发生过程中的作用。方法：应用光镜定量酶组织化学方法对ＷＫＹ大鼠和自发性高血压大鼠视网膜原酸性磷酸的分布和活性变化进行定量观察。结果：视网膜各层酸性磷酸酶活性岂强到弱依次是（Ｆ检验，Ｐ〈０．０５）；（１）色素上皮层；（２）视杆维层内节和外网层（两层间活性无显著性差异）；（３）内网层；（４）节细胞层和神经纤维层，（５）外核层和内核层（两层间活性无显著性差异）杆锥层外     

Regeneration of axons in the optic nerve of the adult Browman-Wyse (BW) mutant rat

Summary We have studied the regeneration of axons in the optic nerves of the BW rat in which both oligodendrocytes and CNS myelin are absent from a variable length of the proximal (retinal) end of the nerve. In the optic nerves of some of these animals, Schwann cells are present. Axons failed to regenerate in the exclusively astrocytic environment of the unmyelinated segment of BW optic nerves but readily regrew in the presence of Schwann cells even across the junctional zone and into the myelin debris filled distal segment. In the latter animals, the essential condition for regeneration was that the lesion was sited in a region of the nerve in which Schwann cells were resident. Regenerating fibres appeared to be sequestered within Schwann cell tubes although fibres traversed the neuropil intervening between the ends of discontinuous bundles of Schwann cell tubes, in both the proximal unmyelinated and myelin debris laden distal segments of the BW optic nerve. Regenerating axons never grew beyond the distal point of termination of the tubes. These observations demonstrate that central myelin is not an absolute requirement for regenerative failure, and that important contributing factors might include inhibition of astrocytes and/or absence of trophic factors. Regeneration presumably occurs in the BW optic nerve because trophic molecules are provided by resident Schwann cells, even in the presence of central myelin, oligodendrocytes and astrocytes. All the above experimental BW animals also have Schwann cells in their retinae which myelinate retinal ganglion cell axons in the fibre layer. Control animals comprised normal Long Evans Hooded rats, BW rats in which both retina and optic nerve were normal, and BW rats with Schwann cells in the retina but with normal, i.e. CNS myelinated, optic nerves. Regeneration was not observed in any of the control groups, demonstrating that, although the presence of Schwann cells in the retina may enhance the survival of retinal ganglion cells after crush, concomitant regrowth of axons cut in the optic nerve does not take place.     

Intermediate filament complement of the normal and gliotic canine retina.

Intermediate filament expression of various cell types in the adult canine normal and gliotic retina was determined by an immunoperoxidase method of using monoclonal antibodies on aldehyde-fixed tissues. In the normal retina, vimentin was present in astrocytes in the nerve fibre layer, horizontal cell processes, and Müller cell fibres from the internal limiting membrane to the outer nuclear layer. Neurofilamentous axons were noted in the nerve fibre, inner plexiform layer, and outer plexiform layer, although the degree of staining intensity varied among the three molecular weight neurofilament antisera used. Glial fibrillary acidic protein (GFAP) staining was confined to the nerve fibre and ganglion cell layer; this was interpreted as representing fibrous astrocytes. Astrocyte density varied according to retinal topography with an increased number around retinal blood vessels and in the peripapillary retina. Quantitative, but not qualitative differences in staining for vimentin and the neurofilaments were noted in degenerative, gliotic retinas. In common with several other mammalian species previously studied, the canine Müller cells accumulate or express GFAP under pathological conditions involving a gliotic response.     

Evidence that migratory oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells are kept out of the rat retina by a barrier at the eye-end of the optic nerve

Summary There is evidence that oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells migrate along the developing rat optic nerve from the chiasm toward the eye before differentiating into oligodendrocytes that myelinate the retinal ganglion cell axons in the nerve. Why, then, do these progenitor cells not migrate into the eye, differentiate into oligodendrocytes and myelinate the nerve fibre layer of the retina? Myelination would opacify the neural retina and thereby severely impair vision. Here we provide evidence that there is a barrier at the eye-end of the rat optic nerve that prevents the migration of O-2A progenitor cells into the retina. Our findings in the rat support a previous hypothesis that such a barrier keeps myelin-forming glial cells out of the human retina.     

Transplanted olfactory ensheathing cells incorporated into the optic nerve head ensheathe retinal ganglion cell axons: possible relevance to glaucoma

A mixture of olfactory ensheathing cells and fibroblasts cultured from the adult rat olfactory mucosa was transplanted through a scleral incision into the retina. A major stream of transplanted cells migrated through the stratum opticum and penetrated for up to about 0.5mm into the optic nerve head. This stream of transplanted cells consisted of a mixture of bipolar olfactory ensheathing cells with long processes which give rise to a non-myelinating ensheathment of single retinal ganglion cell axons, and olfactory nerve fibroblasts embedded in a dense fibronectin-positive extracellular matrix. A second stream of ovoid olfactory ensheathing cells with tufted processes and unaccompanied by fibroblasts or matrix migrate into the internal plexiform layer. The incorporation of olfactory ensheathing cells in the optic nerve head may suggest future possibilities for protection of the axons in this vulnerable region from mechanical damage, as in the raised intraocular pressure of glaucoma.     

Chronotopographical distribution patterns of cell death and of lectin‐positive macrophages/microglial cells during the visual system ontogeny of the small‐spotted catshark Scyliorhinus canicula

The patterns of distribution of TUNEL‐positive bodies and of lectin‐positive phagocytes were investigated in the developing visual system of the small‐spotted catshark Scyliorhinus canicula, from the optic vesicle stage to adulthood. During early stages of development, TUNEL‐staining was mainly found in the protruding dorsal part of the optic cup and in the presumptive optic chiasm. Furthermore, TUNEL‐positive bodies were also detected during detachment of the embryonic lens. Coinciding with the developmental period during which ganglion cells began to differentiate, an area of programmed cell death occurred in the distal optic stalk and in the retinal pigment epithelium that surrounds the optic nerve head. The topographical distribution of TUNEL‐positive bodies in the differentiating retina recapitulated the sequence of maturation of the various layers and cell types following a vitreal‐to‐scleral gradient. Lectin‐positive cells apparently entered the retina by the optic nerve head when the retinal layering was almost complete. As development proceeded, these labelled cells migrated parallel to the axon fascicles of the optic fiber layer and then reached more external layers by radial migration. In the mature retina, lectin‐positive cells were confined to the optic fiber layer, ganglion cell layer and inner plexiform layer. No evident correlation was found between the chronotopographical pattern of distribution of TUNEL‐positive bodies and the pattern of distribution of lectin‐labelled macrophages/microglial cells during the shark′s visual system ontogeny.     

人胚胎视网膜内神经丝蛋白的出现和分布

用免疫组织化学ＡＢＣ法研究了２５０例４－３８周人胚胎视网膜中神经丝蛋白（ＮＦ）的出现和分布。发现第３２周胎儿视网膜神经纤维层出现ＮＦ阳性纤维。至出生时，神经纤维层卫性纤维增多、增粗，但仍未达成人的水平。人胎视网膜ＮＦ主要分布于节细胞轴突，成人除节细胞胞体和轴突外，内网层也有少数ＮＦ阳性纤维。     

Protein gene product 9.5-immunoreactive neurons in the retina of striped dolphin (Stenella coeruleoalba) and Fraser dolphin (Lagenodelphis hosei).

This study demonstrates immunocytochemically that protein gene product 9.5 (PGP 9.5), a neuronal marker, is expressed by various populations of retinal cells in Stenella coeruleoalba (striped dolphin) and Lagenodelphis hosei (Fraser dolphin): one in the retinal ganglion cells and the other in the inner nuclear layer, resembling horizontal and amacrine cells. The specific distribution of PGP 9.5 in a dolphin closely resembles that in rodents and carnivores; however, some differences arise among these animals. In a dolphin's retina, for example, only a few of giant ganglion cells are immunoreacted while almost all the small ganglion cells are stained strongly. The processes of horizontal cells, identified according to their localization, appear not to connect entirely in a dolphin. Instead, PGP 9.5 positive cells are widely distributed in the small to moderate ganglion cells and have distinct processes which are ramified extensively in the outer plexiform layer in rodents and carnivores. The high levels of PGP 9.5 expressing in the inner part of dolphin retina, including ganglion cells and their axons as well as distinct sublamination in the inner plexiform layer, indicate that this molecule markedly influences the retinal system, possibly in visual connection. Although mammals have various visual behavior, i.e., living marine vs. terrestrial environment, and active during daytime vs. in the night, the retina is a common model to characterize the neurochemical properties.