Subgroup of alpha ganglion cells in the adult cat retina is immunoreactive for somatostatin.

We have previously shown that two types of cells in the ganglion cell layer of the adult cat retina are immunoreactive for somatostatin (White et al., '90). One of the types was identified by morphological criteria as a wide-field amacrine cell. The other cell type had a large, angular soma that resembled the alpha ganglion cell, but evidence was not available to identify it definitively as a ganglion cell. Both cell types were distributed preferentially in the inferior retina. In this report, we demonstrate that the two types of cell are, indeed, displaced amacrine cells and alpha ganglion cells. First, when retrograde tracers were injected into central visual targets, the immunoreactive large cells but not the displaced amacrine cells were found to be labeled. Second, after unilateral section of the optic nerve, the immunoreactive large cells disappeared from the retina on the lesioned side, but the displaced amacrine cells occurred in the same numbers in both retinae. In the periphery, the large cells ranged in diameter from 33 to 47 microns, comparable only to alpha ganglion cells (Boycott and W?ssle, '74). An antiserum to parvalbumin was used to visualize the dendrites (R?hrenbeck and W?ssle, '88) of somatostatin-immunoreactive large cells. Based on dendritic stratification within the inner plexiform layer (Famiglietti and Kolb, '76), the somatostatin-immunoreactive large cells were found to include both on-center cells and off-center cells, but were predominantly of the off-center type. Within a local region, they were found to be arrayed with greater regularity than the overall population of alpha ganglion cells. These results indicate that alpha ganglion cells of the cat retina can be subdivided on the basis of their immunoreactive staining for somatostatin and suggest that the diversity of ganglion cells in the cat retina may be greater than has been recognized on the basis of morphological criteria alone.     

Somatostatin-immunoreactive cells in the adult cat retina

Peptides have been found in the retinas of all mammalian species studied to date, but little is known about their localization and function in the cat. Using two mouse monoclonal antibodies directed to somatostatin 14, we have observed two sparse groups of somatostatin-immunoreactive neurons in the cat, both distributed preferentially in the inferior retina. The more numerous cell type is characterized by a small- to medium-sized soma (mean diameter = 16.3 +/- 9.0 microns; n = 186) with sparsely branching, far-reaching varicose processes that ramify mainly in the inner plexiform layer. The majority of these cells are located in the ganglion cell layer, with the remainder in the proximal inner nuclear layer and the inner plexiform layer. They are in especially high density at the retinal margin. In morphology and soma size, these cells resemble wide-field amacrine cells. The second cell type has a large, granular-staining soma (mean diameter = 29.7 +/- 14.8 microns; n = 145) with poorly stained primary processes and is found only in the ganglion cell layer. Cells of this type are most similar in their size and morphology to alpha ganglion cells. In contrast to the location of somatostatin-immunoreactive somata, a dense meshwork of immunoreactive processes was observed at all eccentricities within the inner plexiform layer, adjacent to the inner nuclear layer and to the ganglion cell layer. Labeled processes arising from the inner plexiform layer were also occasionally detected in the outer plexiform layer and the nerve fiber layer. Additional processes of unknown origin were observed in the nerve fiber layer and the optic nerve head. The extensive distribution of immunoreactive processes suggests that somatostatin-immunoreactive somata located preferentially in the inferior half of the retina have a widespread influence on neural activity.     

Somatostatin-like immunoreactive material in the rabbit retina: immunohistochemical staining using monoclonal antibodies

Two mouse monoclonal antibodies to somatostatin-14 were used with avidin-biotin-peroxidase immunohistochemical technique to examine the rabbit retina. In agreement with a previous study using a polyclonal anti-serum, a sparse population (about 1,000 per retina) of neurons in the ganglion cell layer are immunoreactive for somatostatin; the vast majority of these cells are inferior to the myelinated fiber bundle. In addition, the monoclonal antibodies disclose a second neuronal population that forms a circumferential band of immunoreactive neurons around the extreme periphery of the retina. The cells in the body of the inferior retina have dendrites that ramify in the inner plexiform layer. Both the circumferential band of cells and the cells in the body of the inferior retina give off axonlike processes that run in the inner plexiform layer and do not enter the optic nerve. These long, straight varicose fibers form a meshwork that covers the entire retina. The superior retina, which contains only rare immunoreactive cell bodies, has a plexus of stained fibers comparable to that of the inferior retina. The circumferential band of cells is relatively resistant to the neurotoxin kainic acid, explaining a previously reported observation that this toxin depletes only about 50% of the content of somatostatin-like immunoreactivity from the rabbit retina. Moreover, the somatostatin immunoreactive neurons are not labeled by the intraocular injection of the fluorescent dye DAPI, which labels the cholinergic displaced amacrine cells of the rabbit retina. These observations imply that somatostatin-like immunoreactivity is localized to two populations of associational ganglion cells, neurons with cell bodies in the ganglion cell layer, the axons of which remain within the retina.     

GABA-like immunoreactivity in the macaque monkey retina: a light and electron microscopic study

The distribution of GABA-like immunoreactivity in the macaque monkey retina was studied by using postembedding techniques on semithin and ultrathin sections. At the light microscopic level, both inner and outer plexiform layers showed strong GABA-like immunoreactivity in the central retina. All the horizontal cells, some bipolar cells, 30-40% of amacrine cells, occasional interplexiform cells, and practically all displaced amacrine cells were labeled. In the peripheral retina (beyond 5 mm eccentricity), the outer plexiform layer and the horizontal cells were not labeled, but all other cell types showed the same labeling pattern as in the central retina. Synapses of the inner plexiform layer involving a pre- or postsynaptic GABA-labeled process were studied electron microscopically. Synapses involving a GABA-labeled presynaptic amacrine cell process made up 80% of the synapses observed. These GABA-labeled amacrine processes synapsed onto amacrine, bipolar, and ganglion cell processes as well as onto amacrine and ganglion cell bodies. Synapses involving a postsynaptic GABA-labeled process made up 20% of the synapses studied. The GABA-like immunoreactive processes were postsynaptic to bipolar cells at the dyads and to amacrine cells at conventional synapses.     

Differential expression of phospholipase D1 in the developing retina

Expression patterns of phospholipase D1 (PLD1) in the developing rat retina were investigated using immunocytochemistry and Western blot analysis and compared with the expression patterns of glutamine synthetase. PLD1 immunoreactivity appeared first in a few neuroblasts in the middle of the mantle zone of the primitive retina by embryonic (E) day 13. PLD1-immunoreactive primitive ganglion cells were characterized in the ganglion cell layer by E17. Faint immunoreactivity at E17 profiled radially orientated cells and this pattern appeared up to postnatal (P) day 7. In the ganglion cell layer at P3, displaced amacrine cells and ganglion cells were classified. At P5, presumptive horizontal cells and amacrine cells were identified. By P7, a thin outermost layer of newly formed segments of the photoreceptor cells was also PLD1 immunoreactive. PLD1 immunoreactivity at P8 was limited to radial Müller cells and the outer segment layer of the photoreceptor cells, and the expression pattern was conserved to adulthood. Western blot analysis showed relatively high amounts of PLD1 protein at E17 and P3, a decrease at P7, and moderate amounts from P8 onward. Co-expression of PLD1 with glutamine synthetase in the retina appeared first after birth in differentiating neurons and in Müller cells by P8; thereafter the pattern was maintained. The expression pattern of the PLD1 during development of the retina suggests that PLD1 plays important roles in glutamate-associated differentiation of both specific neurons and radial glial cells, and in glutamate-mediated cellular signalling in Müller cells.     

Distribution and synaptic connectivity of neuropeptide Y-immunoreactive amacrine cells in the rat retina

Neuropeptide Y (NPY) is a potent bioactive peptide that is widely expressed in the nervous system, including the retina. Here we show that specific NPY immunoreactivity was localized to amacrine and displaced amacrine cells in the rat retina. Immunoreactive cells had a regular distribution across the retina and an overall cell density of 280 cells/mm(2) in the inner nuclear layer (INL) and 90 cells/mm(2) in the ganglion cell layer (GCL). In the INL, most immunoreactive cells were characterized by small cell bodies and fine processes that appeared to ramify primarily in stratum 1 of the inner plexiform layer (IPL). A few cells in the INL also ramified in stratum 3 of the IPL. In the GCL, small to medium immunoreactive cells appeared to ramify primarily in stratum 5 of the IPL. A few immunoreactive processes, originating from somata in the INL and processes in the IPL, ramified in the OPL. NPY-immunoreactive cells contained GABA immunoreactivity, and some amacrine cells also contained tyrosine hydroxylase immunoreactivity. NPY-immunostained processes were most frequently presynaptic to nonimmunostained amacrine and ganglion cell processes and postsynaptic to nonimmunostained amacrine cell processes and cone bipolar cell axonal terminals. These findings indicate that NPY immunoreactivity is present in two populations of amacrine cells, one located in the INL and the other in the GCL, and that these cells mainly form synaptic contacts with other amacrine cells. These observations suggest that NPY-immunoreactive cells participate in multiple circuits mediating visual information processing in the inner retina.     

Substance P-like immunoreactive amacrine cells in the cat retina

Substance P-like immunoreactivity was localized by immunocytochemical techniques to two subpopulations of amacrine cells in the cat retina. One cell was a unistratified amacrine with processes ramifying within stratum 4 of the inner plexiform layer. The other cell type was a bistratified cell with processes in both stratum 1 (s1) and stratum 4 (s4). Both cell types were seen with their somas displaced to the ganglion cell layer as well as in the conventional amacrine location in the inner nuclear layer. Substance P cells were present in the greatest density within the area centralis and decreased in number toward the periphery. The ratio of amacrine to displaced amacrine cells also decreased peripherally. However, the coverage by immunoreactive fibers in s4 remained three times that seen in s1. Computer-assisted analysis confirmed the location of substance P-containing processes at 5-15% (s1) and 50-70% (s4) depth levels in the inner plexiform layer. A comparison of substance P-like immunoreactivity in light- and dark-adapted cat retinas showed no apparent differences in the distribution of immunoreactivity due to lighting conditions.     

Localization of two calcium binding proteins, calbindin (28 kD) and parvalbumin (12 kD), in the vertebrate retina

We used immunocytochemistry to locate two calcium binding proteins, calbindin (CaB) and parvalbumin (PV), in the retina of goldfish, frog, chick, rat, guinea pig, dog, and man. The location of CaB depended on the type of dominant photoreceptor cells in birds and mammals. In cone-dominant retinas such as those of the chick, CaB-like immunoreactivity was found in the cones, cone bipolars, and ganglion cells. Amacrine cells 5-12 microns across were also labeled. In rod-dominant retinas, such as those of the rat, guinea pig, and dog, horizontal cells, small amacrine cells (about 6 microns across), and cells in the ganglion cell layer were labeled. In the human retina, which has both cones and rods in abundance, cones, cone bipolars, ganglion cells, horizontal cells, and small and large amacrine cells were labeled. In the frog and goldfish, the level of CaB-like immunoreactivity was low. In the frog, a few cones, amacrine cells, and cells in the ganglion cell layer were labeled. No immunoreactive structures were seen in the goldfish retina. PV-like immunoreactivity was found in chicks, rats, and dogs. No such immunoreactive structures were seen in the other species. In the chick, only amacrine cells were labeled. In the rat, amacrine cells and several displaced amacrine cells were labeled. In the dog, in addition to amacrine cells and displaced amacrine cells, horizontal cells were strongly labeled. Thus, PV-like immunoreactivity was found in those elements relating to the modulation of the main pathway of the visual transmission system.     

Characterization of the cell death promoter, Bad, in the developing rat retina and forebrain.

Neuronal programmed cell death, or apoptosis, occurs during development, following injury or in certain disease processes, and is regulated by members of the B-cell leukemia-2 (Bcl-2) protein family. These molecules include both positive and negative regulators of cell death and act by selective dimerization that results in permissive or inhibitory effects on a cascade of cellular events, including mitochondrial release of cytochrome c, stimulation of cysteine protease activity and subsequent cellular deterioration. Here, we have characterized the expression of the cell death agonist, Bad, in the postnatal rat retina and forebrain. Isolation, subsequent amplification by RT-PCR and DNA sequence analysis revealed that retinal Bad was identical to Bad expressed in the developing and adult rat brain. Using a polyclonal antibody to Bad, we determined that, in the retina, on the day of birth (postnatal day-0, PND-0) Bad immunoreactivity was expressed primarily by retinal ganglion cells, some cells in the inner neuroblastic layer (NBL) and an indistinct plexus of processes in the inner plexiform layer (IPL). On PND-7, Bad immunoreactivity was observed in most cells in the ganglion cell layer (GCL), numerous cells scattered throughout the inner nuclear layer (INL), a lightly stained IPL and in a distinct band of immunostained fibers in the forming outer plexiform layer (OPL). By PND-15, Bad immunoreactivity was present in cells in the GCL, in some cells in the proximal INL and in horizontal cell processes in the OPL. The IPL was only faintly labeled. In the adult retina, specific Bad immunostaining was confined to large cells in the ganglion cell layer (presumed ganglion cells), occasional lightly stained horizontal cells and their processes in the OPL and to occasional small, lightly stained cells in the proximal INL (presumed amacrine cells) and GCL (presumed displaced amacrine cells). Again, the interposed IPL was faintly labeled. In the brain, Bad immunoreactive cells were scattered throughout the forebrain parenchyma but were particularly concentrated in neurons of the cerebral cortex, hippocampus and amygdala. Bad immunoreactivity was heaviest in these cells at PND-7, distinctly weaker at PND-10 and absent by PND-24. At all time points examined, Bad immunoreactivity was present in epithelial cells of the choroid plexus, as previously reported in the adult rat brain. These data suggest that Bad is transiently expressed by various cell types in the perinatal retina, particularly ganglion cells, and in discrete forebrain regions. In the context of corroborative observations, Bad expression may be regulated in response to acute ischemia and may act as a control point for retinal neuronal apoptosis.     

Plasmalemmal and vesicular gamma-aminobutyric acid transporter expression in the developing mouse retina

Plasmalemmal and vesicular gamma-aminobutyric acid (GABA) transporters influence neurotransmission by regulating high-affinity GABA uptake and GABA release into the synaptic cleft and extracellular space. Postnatal expression of the plasmalemmal GABA transporter-1 (GAT-1), GAT-3, and the vesicular GABA/glycine transporter (VGAT) were evaluated in the developing mouse retina by using immunohistochemistry with affinity-purified antibodies. Weak transporter immunoreactivity was observed in the inner retina at postnatal day 0 (P0). GAT-1 immunostaining at P0 and at older ages was in amacrine and displaced amacrine cells in the inner nuclear layer (INL) and ganglion cell layer (GCL), respectively, and in their processes in the inner plexiform layer (IPL). At P10, weak GAT-1 immunostaining was in Müller cell processes. GAT-3 immunostaining at P0 and older ages was in amacrine cells and their processes, as well as in Müller cells and their processes that extended radially across the retina. At P10, Müller cell somata were observed in the middle of the INL. VGAT immunostaining was present at P0 and older ages in amacrine cells in the INL as well as processes in the IPL. At P5, weak VGAT immunostaining was also observed in horizontal cell somata and processes. By P15, the GAT and VGAT immunostaining patterns appear similar to the adult immunostaining patterns; they reached adult levels by about P20. These findings demonstrate that GABA uptake and release are initially established in the inner retina during the first postnatal week and that these systems subsequently mature in the outer retina during the second postnatal week.     

Unique Distribution of Somatostatin-immunoreactive Cells in the Retina of the Tree Shrew (Tupaia belangeri)

Somatostatin-like immunoreactive cells in the tree shrew retina were studied with the monoclonal antibody S8 against the neuropeptide somatostatin 14. As in some other mammals, immunoreactive somata are exclusively found in the ganglion cell layer. Immunoreactive processes form a sparse main plexus in the inner plexiform layer near the border of the inner nuclear layer; fewer additional processes are found closer to the ganglion cell layer. With retrograde labelling of retinal ganglion cells by injections of the tracer Fast Blue into the superior colliculus and lateral geniculate body and counterstaining of the retinae with S8, ˜5% of the immunoreactive somata were double-labelled at any retinal location. The vast majority of somatostatin-like immunoreactive cells are thus displaced amacrine cells. Their somata are distributed over the entire retina. Their population density is highest in the temporal retina, with peak densities of ˜5000 cells/mm² near the central area and a steep density gradient. In the remaining retina densities are 200–400 cells/mm², falling to ˜100 cells/mm² at the retinal margins. This is in stark contrast to the somatostatin-like immunoreactive cells in other mammalian retinae which have densities of 10–40 cells/mm² and are confined to restricted retinal regions (inferior retina and/or retinal margin).     

Shifting of parvalbumin expression in the rat retina in experimentally induced diabetes

The AII amacrine cell, a unique rod signal integrator passing through the cone bipolar cell to ganglion cells, uses parvalbumin as a transducer of cytosolic calcium ion signals in the mammalian retina. For clarification of whether AII amacrine cell network contributes to the early neuropathogenesis of diabetic retinopathy, this study first analyzed alteration of parvalbumin expression in experimental diabetic retinas using immunohistochemical methods. Parvalbumin immunoreactivity was found in AII amacrine cells, some amacrine cells of a wide-field type, and displaced amacrine cells of the normal rat retina. During diabetes, cell density of each parvalbumin immunoreactive amacrine cell type showed no large changes despite decrease in immunoreactivity especially in AII amacrine cells. In addition to these parvalbumin immunoreactive amacrine cell types, a type of cone bipolar cells co-expressing glutamate transporter 1b and connecting electrically with AII amacrine cells appeared clearly by 4 weeks of diabetes, and thereafter sharply increased in number to that of AII amacrine cells. Protein levels of parvalbumin throughout the diabetic retinas also showed no large changes, except a transitional slight increase at 4 weeks of diabetes. These results suggest that the parvalbumin expression propagates from AII amacrine cells to a type of cone bipolar cell through electrical synapses due to dysfunction of biased mechanism in calcium ion buffering, caused by diabetic injury, and thus AII amacrine cells are closely involved in neuropathogenesis of ongoing diabetic retinopathy.     

Immunocytochemical localization of the amino acid neurotransmitters in the chicken retina

Postembedding immunocytochemistry was used to determine the cellular localization of the amino acid neurotransmitters glutamate, aspartate, gamma-aminobutyric acid (GABA), and glycine in the avian retina. The through retinal pathway was glutamatergic, with all photoreceptors, bipolar cells, and ganglion cells being immunoreactive for glutamate. Bipolar cells displayed the highest level of glutamate immunoreactivity, with the cell bodies terminating just below the middle of the inner nuclear layer. All lateral elements, horizontal cells, amacrine cells, and interplexiform cells were immunoreactive for glycine or GABA. The GABAergic neurons consisted of two classes of horizontal cells and amacrine cells located in the lower part of the inner nuclear layer. GABA was also localized in displaced amacrine cells in the ganglion cell layer, and a population of ganglion cells that co-localize glutamate and GABA. Both the horizontal cells and GABAergic amacrine cells had high levels of glutamate immunoreactivity, which probably reflects a metabolic pool. At least two types of horizontal cells in the avian retina could be discriminated on the basis of the presence of aspartate immunoreactivity in the H2 horizontal cells. Glycine was contained in a subclass of amacrine cells, with their cell bodies located between the bipolar cells and GABAergic amacrine cells, two subclasses of bipolar cells, displaced amacrine cells in the ganglion cell layer, and ganglion cells that colocalize glutamate and glycine. Glycinergic amacrine cells had low levels of glutamate. We have also identified a new class of glycinergic interplexiform cell, with its stellate cell body located in the middle of the inner nuclear layer among the cell bodies of bipolar cells. Neurochemical signatures obtained by analyzing data from serial sections allowed the classification of subclasses of horizontal cells, bipolar cells, amacrine cells, and ganglion cells. © 1993 Wiley-Liss, Inc.     

Immunocytochemical localization of kainate-selective glutamate receptor subunits GluR5, GluR6, and GluR7 in the cat retina

Localizations of the kainate-selective glutamate receptor subunits GluR5, 6, and 7 were studied in the cat retina by light and electron microscopic immunocytochemistry. GluR5 immunoreactivity was observed in the cell bodies and dendrites of numerous cone bipolar cells and ganglion cells. The labeled cone bipolar cells make basal or flat contacts with cone pedicles in the outer plexiform layer, leading to their identification as OFF-center bipolar cells. Reaction product within the inner plexiform layer was observed in processes of ganglion cells at their sites of input from cone bipolar cells. Staining for GluR6 was localized to A- and B-type horizontal cells, numerous amacrine cells, and an occasional cone bipolar cell. The larger ganglion cells were also immunoreactive. As with other GluR molecules, labeling was usually confined to one of the two postsynaptic elements at a cone bipolar dyad contact. Immunoreactivity for GluR7 was very limited and was seen only in a few amacrine and displaced amacrine cells. Findings of this study are consistent with a major role for kainate receptors in mediating OFF pathways in the outer retina with participation in both OFF and ON pathways in the inner retina.     

Cellular localization of cyclooxygenase-1 and cyclooxygenase-2 in the normal mouse,rat, and human retina

Prostaglandins, synthesized by cyclooxygenase (COX), regulate diverse neurophysiological actions such as regulation of autonomic responses, transmission of pain, generation of fever, control of sleep-wake cycle, synaptic signaling, and cross-talk between neurons and glia in the central nervous system. Although prostaglandins have been widely studied in the anterior segment tissues of the eye, relatively little is known about prostaglandins in the neural retina. By using immunohistochemistry, we have compared the cellular expression and localization of COX-1 and COX-2 in the normal mouse, rat, and human retina. In the normal mouse retina, COX-1 immunoreactivity is present in the outer segments of photoreceptor cells, horizontal cells, microglia, retinal ganglion cells, and displaced amacrine cells. In the normal rat retina, COX-1 immunoreactivity is present in microglia, retinal ganglion cells, and displaced amacrine cells. In the normal human retina, COX-1 immunoreactivity is present in microglia, astrocytes, retinal ganglion cells, and displaced amacrine cells. In the normal mouse and rat retina, COX-2 immunoreactivity is present in processes of the outer plexiform layer and in certain amacrine cells and retinal ganglion cells. In the normal human retina, COX-2 immunoreactivity is only present in processes of the outer plexiform layer. These results suggest that prostaglandins, synthesized by COX-1 or COX-2, may contribute to normal physiological and homeostatic functions in the retina.     

Vanilloid receptor like 1 (VRL1) immunoreactivity in mammalian retina: colocalization with somatostatin and purinergic P2X1 receptors

The distribution of vanilloid receptor like1 immunoreactivity (VRL1-IR) in the retinas of rat, cat, and monkey was studied by single- and double-labeling immunocytochemistry. The patterns were similar for all three species in that VRL1-IR was most prominent in the inner plexiform layer, with scattered compact projections to the outer plexiform layer (OPL). VRL1-immunoreactive cell bodies were present throughout the rat retina, represented by amacrine cells in the inner nuclear layer and ganglion cell layer (GCL). In cat and monkey retinas, VRL1-immunoreactive cell bodies were restricted to the GCL in the inferior retina. Occasional cell bodies were associated with retinal blood vessels, but their identity as pericytes, glia, or neurons is uncertain. All VRL1-immunoreactive cells and processes colocalized with somatostatin and purinergic P2X1 receptor-IR but not with tyrosine hydroxylase-IR. VRL1-immunoreactive processes in the OPL did not label with antisera against synaptic vesicle 2 (SV2), suggesting that they were dendritic and did not derive from interplexiform cells. However, VRL1-immunoreactive processes in the far periphery toward the pars plana labeled for SV2, suggesting that these processes were presynaptic. The VRL1-immunoreactive cell bodies in the monkey GCL were not calbindin-immunoreactive, demonstrating that they were not displaced H2 horizontal cells. The VRL1-immunoreactive cells in cat and monkey could represent biplexiform and/or associational ganglion cells that receive input in the OPL throughout the retina and direct output to the far periphery. The presence of P2X1 receptors and vanilloid receptor like 1 protein on somatostatin-containing neurons in mammalian retina adds to the growing complexity regarding the chemical control of retinal function that is likely to include the microcirculation.     

Role of cell death in the topogenesis of neuronal distributions in the developing cat retinal ganglion cell layer

The neurons of the developing and adult ganglion cell layer of the cat retina may be morphologically divided into two major populations. One population, the classic neurons, is mainly composed of ganglion cells, and of a small percentage of displaced amacrines, the bar cells. The remaining neurons are microneurons, which make up the majority of the displaced amacrine population. The loss of ganglion cells during the development has been attributed to cell death. It has alternatively been suggested that some ganglion cells may lose their axon and be transformed into displaced amacrine cells, without degeneration of the cell soma. Reexamination of foetal and postnatal cat retinas confirms the presence of degenerating cells in the ganglion cell layer. Their number appears to be at a maximum on embryonic day (E) 57 but declines rapidly until birth. The peak of cell death thus coincides with the decline in optic nerve fibre counts and classical neuron or ganglion cell numbers. Some cells in early stages of degeneration resemble classical neurons, but the original morphology of those advanced stages of degeneration could not be identified, nor was it possible to identify pyknotic microneurons at any stage. Substantial degeneration of the microneurons is not suggested but if it occurs, it is masked by an overall increase in the population of these cells before birth. Cell death in the microneuron population thus cannot yet be ruled out. It has been argued in the literature that fragments of degenerating cells in developing neural tissue are cleared by microglia within 10-14 hours. In order to test the hypothesis that operation of cell death can alone account for the observed loss of classical neurons in the foetal cat retina, we have modelled the effect of various presumed clearance times on corresponding neuronal population magnitudes. It is found that a constant clearance time of 10-24 hours would be consistent with the observed loss of classical neurons before birth. If this is true, then no ganglion cells would remain for transformation into amacrine cells. The absolute density of degenerating or pyknotic cells is found to be relatively constant across the retina. However their density expressed as a percentage of the local population of classical neurons is markedly higher in peripheral than central retina. In the former region, they compose more than 10% of classical neurons at stage E57. On the same day, the percentage distribution maps define an elongated central area containing only 3-5% pyknotic profiles. This region corresponds to the location of the future visual streak.(ABSTRACT TRUNCATED AT 400 WORDS)     

AII amacrine cells in the mammalian retina show disabled-1 immunoreactivity

Disabled 1 (Dab1) is an adapter molecule in a signaling pathway, stimulated by Reelin, which controls cell positioning in the developing brain. It has been localized to AII amacrine cells in the mouse and guinea pig retinas. This study was conducted to identify whether Dab1 is commonly localized to AII amacrine cells in the retinas of other mammals. We investigated Dab1-labeled cells in human, rat, rabbit, and cat retinas in detail by immunocytochemistry with antisera against Dab1. Dab1 immunoreactivity was found in certain populations of amacrine cells, with lobular appendages in the outer half of the inner plexiform layer (IPL) and a bushy, smooth dendritic tree in the inner half of the IPL. Double-labeling experiments demonstrated that all Dab1-immunoreactive amacrine cells were immunoreactive to antisera against calretinin or parvalbumin (i.e., other markers for AII amacrine cells in the mammalian retina) and that they made contacts with the axon terminals of the rod bipolar cells in the IPL close to the ganglion cell layer. Furthermore, all Dab1-labeled amacrine cells showed glycine transporter-1 immunoreactivity, indicating that they are glycinergic. The peak density was relatively high in the human and rat retinas, moderate in the cat retina, and low in the rabbit retina. Together, these morphological and histochemical observations clearly indicate that Dab1 is commonly localized to AII amacrine cells and that antiserum against Dab1 is a reliable and specific marker for AII amacrine cells of diverse mammals.     

Development of ganglion cell topography in ferret retina

The adult ferret has approximately 90,000 retinal ganglion cells, arranged in a prominent area centralis and visual streak. The role of differential cell generation, cell death, and retinal growth in the control of adult retinal ganglion cell number and distribution was evaluated by examining basic aspects of retinogenesis, including growth in retinal area, developmental changes in the number, size, and distribution of retinal ganglion cells (identification aided by retrograde transport of HRP), and the incidence of degenerating cells in the ganglion cell layer. Retinal development in the ferret was also compared to retinal development in the cat (which has an even more differentiated area centralis) to determine what alterations of developmental parameters are most closely associated with this species difference in adult morphology. The area of the retina increases linearly from birth (12 mm2) to postnatal day 24 (54 mm2), reaching an eventual adult value of 64 mm2. Ganglion cell numbers peak at 155,000 (approximately twice the adult number) on postnatal day 3, and fall to adult numbers by postnatal day 6. The remaining cells of the ganglion cell layer, principally displaced amacrine cells, reach their peak number on postnatal day 10 (approximately 280,000), falling to 200,000 by adulthood. Degenerating cells are abundant in the ganglion cell layer in the immediate postnatal period. A difference in the incidence of degenerating cells in the presumptive area centralis versus that in the retinal periphery was not observed postnatally, though there were other striking spatial nonuniformities, suggesting that differential cell loss might contribute to other features of retinal topographic organization. Ganglion cell density is virtually uniform across the retina at birth. Cell density is first reduced in the dorsal retina, resulting in a dorsal-to-ventral gradient in cell density that persists until day 10, when ganglion cell number has stabilized. By postnatal day 24, an area centralis and visual streak has emerged, but not of adult magnitude. Because ganglion cell number has stabilized long before the area centralis and visual streak emerge, we conclude that differential retinal growth is the principal mechanism producing this feature of retinal topography. Comparison with the cat suggests that the proportionately greater nonuniform growth of the cat's eye accounts for the greater differentiation of its area centralis.     

Neuropeptide Y- and substance P-like immunoreactive amacrine cells in the retina of the developing Xenopus laevis.

The development of neuropeptide Y-like (NPY-LI) and substance P-like (SP-LI) immunoreactive neurons was studied in retinas of Xenopus laevis from young tadpole through to adult animals. In adult retina these neuropeptides are present in wide-field amacrine cells located in the inner nuclear layer and the ganglion cell layer of the retina. Retinal wholemount preparations and sectioned material showed that immunoreactive cells appeared during early larval life and NPY-LI occurred earlier than SP-LI cells. The primary dendritic branching of NPY-LI neurons appeared from early larval life whilst SP-LI was evident in dendrites from mid-larval stages. In postmetamorphic animals the numbers of immunoreactive cells increased in proportion to retinal area growth with a relatively constant cell density of about 35 cells/mm2 for SP-LI and 45 cells/mm2 for NPY-LI. The maturation of dendritic morphology of both NPY- and SP-LI amacrine cells appeared later in larval development than the appearance of immunoreactivity in cell somas. However, the sequence of expression of NPY- or SP-LI and their dendritic maturation was different for the two classes of amacrine cells. It is suggested that the maturation of dendritic fields of amacrine cells is complete just prior to metamorphosis, consistent with the postmetamorphic onset of electrophysiological features of ganglion cells attributed to amacrine cells.