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PubMed2004

Bipolar cells of the mouse retina: a gene gun, morphological study.

One of the key elements concerning our understanding of the organization of the mouse retina is the complete classification of the various types of bipolar cells. With the present study, we tried to contribute to this important issue. Unfortunately, most of the antibodies that stain specifically bipolar cells in the retina of other mammals hardly work for the retina of the mouse. We succeeded in overcoming this limitation by using a relatively novel technique based on the gene gun transfer of fluorescent dyes to cells. Hence, we were able to stain a considerable number of bipolar cells that could be characterized according to morphological and comparative criteria. We also performed a complete morphometric analysis of a subset of bipolar cells stained by anti-neurokinin-3 receptor antibodies. We found nine types of cone bipolar cells and one type of rod bipolar cell; these data are consistent with the findings of previous studies on the retinas of other mammals, such as rabbits, rats, and monkeys and with a recent study based on the mouse retina (Ghosh et al. [2004] J Comp Neurol 469:70-82). Our results also confirm the existence of a common structural similarity among mammalian retinas. It remains to be elucidated what is exactly the functional role of the various types of cone bipolar cells and what is the specific contribution they provide to the perception of a given visual stimulus. Most probably, each bipolar cell type constitutes a specialized channel for the computation of a selected component of the visual stimulus. More complex signal coding, involving the coordinated activity of various types of bipolar cells, could also be postulated, as it has been shown for ganglion cells (Meister [1996] Proc Natl Acad Sci U S A 93:609-614).

Animals↗
PubMed1979

Rods and cones in the mouse retina. II. Autoradiographic analysis of cell generation using tritiated thymidine.

The period of cell genesis of rod and cone photoreceptor cells has been determined in the retinas of C57BL/6J mice. Embryonic mice were exposed to a single dose of 3H-thymidine at embryonic day (E) 10--18 by injecting pregnant mice intraperitoneally. Animals at postnatal ages were injected subcutaneously once between postnatal day (P) 0--10. The eyes were removed at one to three months of age. After fixation, they were embedded in glycol methacrylate, sectioned at 1.5 micrometers and prepared for autoradiographic analysis. All of the cone cells are generated over a relatively short time interval during the fetal period. In the posterior retina, the peak of cone cell genesis occurs at E13-E14, and no cones are generated after E16. The rods, by contrast, are generated later and over a longer time period. They first begin to be generated in the posterior retina on E13, but the peak of cell genesis is not reached until the day of birth, and some rods are generated as late as P5. For both rods and cones the peaks of cell genesis in the peripheral retina occur two to three days later than in the posterior retina. The findings demonstrate that rods and cones are developmentally distinct cell types in the mouse retina.

Animals↗
PubMed1981

Regional specialization of the chick retina as revealed by the size and density of neurons in the ganglion cell layer.

On the basis of both morphological criteria and survival after ganglion cell axotomy, three populations of cells can be recognized in the ganglion cell layer of the chick retina. These are: 1) Irregularly shaped cells, which are not affected by axotomy and lack Nissl stain. They comprise about 5% of the total cell population and are probably glial cells. 2) Small cells which have a distinctive teardrop-shaped perikarya, stain for Nissl substance, and are not affected by axotomy. They occur with a uniform density of about 4,000 cells/mm2 across the entire retina and comprise 30-35% of the total number of cells in the ganglion cell layer. It is suggested that these cells are displaced amacrine cells. 3) Cells which stain strongly for Nissl substance and disappear after axotomy. These cells comprise 60-65% of the population of cells in this layer. The density of the latter cells varies throughout the retina. A high-density region in the central area extends into the superior-temporal retina. This high-density region corresponds to a position in the lateral visual field extending into the infero-frontal field. The total number of these cells agrees with the reported number of the optic nerve fibres in the chick, (Rager and Rager, '78); therefore they are presumably ganglion cells. The size distribution of the presumptive ganglion cells varies in different parts of the retina. The only previous study of the distribution and size of cells in the ganglion cell layer of the avian retina did not distinguish between ganglion cells and displaced amacrine cells (Binggeli and Paule, '69). The present results are therefore likely to be a more accurate description of the total number of ganglion cells, the regional variations in their density, and the characteristics of their size than previously reported.

Animals↗
PubMed1985

Parasol and midget ganglion cells of the human retina.

Golgi-impregnated ganglion cells were studied in two flat-mounted human retinas. A number of different morphologic forms were observed, and those showing a thickly branching dendritic field with terminals that stratified within a narrow zone of the inner plexiform layer were selected for further investigation. When the dendritic field diameter of these cells was plotted against distance from the fovea, the scatter diagram showed two distinct clusters. At any given eccentricity, there was no overlap between the cell group with large dendritic fields and the group with small dendritic fields. Those with the larger dendritic fields also tended to have larger somas and thicker axons than the group with the smaller dendritic fields. The dendritic fields of both groups tended to be elongated, and the orientation and degree of this elongation were quantified by determining the best-fitting ellipse for each dendritic field. The degree of elongation was independent of eccentricity. The orientation of the dendritic field (major axis of the ellipse) of a cell did not appear to be independent of its position on the retina. To test whether the major axes tended to be directed toward any particular point on the retina, the positions of the cells on the retinal flat mount were transformed to relative positions on the retinal hemisphere, and the orientations of the dendritic fields were expressed in a spherical coordinate system for this hemisphere. A search was made for the position on the hemisphere which minimized the mean square deviation of the orientations from this point. The group with the large dendritic fields showed a significant tendency to be radially oriented toward a specific location on the retinal hemisphere, and that location lay within a few degrees of the fovea. Leventhal and Schall ('83) have reported a similar finding for ganglion cells of the cat retina. For the group with small dendritic fields, the retinal location that minimized the mean square deviation was also near the fovea; however, the set of orientations showed no greater tendency for mutual alignment than did a randomized set. The cell group with the large dendritic fields appears to correspond to Dogiel's (1891) type II cells, to Polyak's ('41) parasol cells, to the A cells of the monkey retina described by Leventhal et al. ('81), observed following HRP injection to the magnocellular layer of the LGN, and to the P alpha cells of the monkey retina, observed by Perry and Cowey ('81), following HRP uptake by cut axons of the optic nerve.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1986

Choline acetyltransferase immunoreactivity suggests that ganglion cells in the goldfish retina are not cholinergic.

Published evidence that ganglion cells in the retinae of nonmammalian species are cholinergic is strong but indirect. In this paper we report results of attempts to demonstrate choline acetyltransferase immunoreactivity in ganglion cells of goldfish retina using two different antisera against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme. We obtained ChAT-immunoreactive staining of amacrine and displaced amacrine cells in the retina and type XIV cells in the tectum, but we obtained no direct immunocytochemical evidence that ganglion cells in the goldfish retina are cholinergic. Thus, ganglion cells identified by retrograde transport of propidium iodide were never ChAT-immunoreactive. Intraocular injections of colchicine did not result in the appearance of a population of ChAT-immunoreactive neurons in the ganglion cell layer. ChAT-immunoreactive axons were not observed in intact, ligated, or transected optic nerves. And finally, the ChAT immunoreactivity of cells and fibers in the optic tectum was unaffected by deafferentation. These experiments provide no positive evidence that any ganglion cells in goldfish retina contain the acetylcholine-synthesizing enzyme, ChAT. While it is possible that our method is too insensitive to detect the enzyme in ganglion cell somata or too specific to recognize the form of ChAT present in these cells, the fact that we can stain putatively cholinergic retinal amacrine cells and tectal neurons makes these alternative explanations improbable. We conclude that it is unlikely that any of the ganglion cells in the retina are cholinergic and that alternative explanations should be sought for previously published results that suggest that they are.

Acetylcholine↗
PubMed1989

Serotoninergic neurons in the retina of Xenopus laevis: selective staining, identification, development, and content.

Uptake of 3H-serotonin followed by autoradiography, and uptake of the serotonin analog 5,7-dihydroxytryptamine (5,7-DHT), with subsequent staining, were each used to define a unique set of neurons in the retina of the African clawed frog, Xenopus laevis. Both techniques demonstrated the same population of neurons, on the basis of perikaryal size, shape, and position within the retina. Two classes of amacrine cells accumulated 5,7-DHT at the proximal (vitread) margin of the inner nuclear layer; the two classes were distinguished by the size of their perikarya. Two similar populations of cells, observed in the ganglion cell layer with lower frequency, may represent "displaced" counterparts of these two amacrine cell types. A class of bipolar cells whose perikarya were located in middle-to-distal regions of the inner nuclear layer also accumulated 5,7-DHT and 3H-serotonin. Processes of these cells contributed to a dense plexus of fine fibers that appeared evenly distributed throughout the inner plexiform layer. 3H-Serotonin-accumulating cells first appeared in the developing retina at stage 35/36, a time immediately after retinal stratification but before elaboration of either plexiform layer. Electron microscopic analysis permitted an identification of 3H-serotonin-accumulating terminals in the inner plexiform layer. Serotonin-labeled terminals containing conventional contacts, suggestive of amacrine cells, were presynaptic to unidentified processes and postsynaptic to bipolar cells. Labeled terminals containing ribbon contacts, indicative of bipolar cells, were postsynaptic to amacrine cells. The amount of serotonin contained in isolated retinas was 15 pmol/mg protein as measured by HPLC with electrochemical detection. We attempted to stimulate the release of accumulated 3H-serotonin from mature retinas by increasing the K+-concentration in the bathing medium. Although preloaded glycine is readily released from 14C-glycine-accumulating neurons, from the same retinas there was no calcium-dependent, K+-stimulated release of 3H-serotonin. This finding suggests that serotonin and glycine are processed differently by retinal neurons, the consequence of which results in differing responses to 40 mM K+.

5,7-Dihydroxytryptamine↗
PubMed1989

Enzyme-histochemical demonstration of microglial cells in the adult and postnatal rabbit retina.

Enzyme-histochemical methods for thiamine pyrophosphatase (TPPase) and nucleoside diphosphatase (NDPase) were applied to wholemounted rabbit retinae to demonstrate the shape and distribution of microglial cells in early postnatal and adult animals. At birth, microglial cells were already present in the entire retina. They acquired their adult "resting shape" during the first 3 postnatal weeks. Early postnatally labeled microglial cells were scattered throughout the nerve fiber layer, the inner plexiform layer, and the outer plexiform layer (OPL); at adulthood, they were not detected in the OPL. Nissl-stained retinae revealed that the number of microglial cells continuously increased during postnatal development. The same Nissl-stained preparations were used to evaluate the topography of degenerating cells in the developing postnatal retina of the rabbit. Large numbers of degenerating pyknotic cells were observed throughout the entire retinal ganglion cell layer during the first postnatal week. Later their number decreased, and from the third postnatal week onward degenerating cells were rare. Also discussed is that the emergence of microglial cells during development may be related to cell death, whereas at adulthood the function(s) of microglial cells remains obscure. Evidence for the blood-derived origin of microglia was not obtained in this study. It is argued here that if this mode of development, which has been demonstrated for other species, is also applied to the rabbit retina, then microglia would have to migrate over considerable distances, since, postnatally, the rabbit retina is avascular for more than 1 week.

Acid Anhydride Hydrolases↗
PubMed1989

Topography of cones and rods in the tree shrew retina.

The topographical distribution of cones and rods in the tree shrew retina was analysed quantitatively in whole-mounted retinae and horizontal semithin sections stained with cresyl violet or toluidine blue. The outer nuclear layer consists of a single layer of photoreceptor nuclei with the rod nuclei slightly displaced towards the outer plexiform layer. This facilitated quantification of the photoreceptor populations. The density of cones ranges from 12,000/mm2 in the peripheral retina to a maximum of 36,000/mm2 in the inferior retina. Unlike ganglion cell density, the density of cones does not peak in the temporal retina. Rod density, between 500/mm2 and 3,500/mm2, also peaks in the inferior retina, but not in the same region as cone density. Rods constitute from 1 to 14% of the photoreceptor population, depending on retinal location, and have a local minimum at the central area. Amongst the cones a regularly arrayed subpopulation of presumed blue-sensitive cones is distinguished by its special staining properties. These cones constitute between 4 and 10% of the cone population depending on retinal location. A second, irregularly spaced, subpopulation of possibly pathological cones is also described.

Animals↗
PubMed1989

Development of microglia in the albino rabbit retina.

In this study the development of ameboid microglia and resting microglia in the retina of the albino rabbit has been examined by means of a lectin derived from Griffonia simplicifolia. Ameboid microglia are present in the retina as early as E12, when the optic fissure is in the process of closure, and appear to be concentrated initially at the vitreal surface. At E14 many ameboid microglia can be seen to extend processes to the ventricular surface of the cytoblast layer, but in subsequent ages these cells are rare and ameboid microglia are largely confined to the ganglion cell layer, inner plexiform layer, and occasionally the developing inner nuclear layer. By adult life, mature (or resting) microglia are confined to the inner plexiform and ganglion cell layers. Numbers of microglia increase steadily throughout fetal life from a mean of 400 at E14, the earliest age quantified, to a peak of 28,600 at E30. There is a small postnatal drop in numbers to 17,150 at P9. Microglia could only be labelled faintly in animals older than P11, but analysis of two adult (P130) retinas with adequate labelling suggested that numbers rise to a value of about 23,800 at this age. Ameboid microglia thus appear in the retina 11 days prior to the onset of axon loss in the optic nerve (about E23) and 14 days prior to the beginning of the period of reduction of retinal ganglion cell numbers (about E26). The present findings indicate that while some microglial precursors may enter the retina in response to debris generated during the natural retinal ganglion cell death period, most enter the retina well before this period. Also, microglia present a uniform density distribution with apparently regular spacing as early as E16, so the uniform regular distribution cannot simply be the consequence of regularly distributed pyknotic figures as previously suggested.

Animals↗
PubMed1989

Localization of L-glutamic acid decarboxylase mRNA in monkey and human retina by in situ hybridization.

Immunocytochemical studies with gamma-aminobutyric acid (GABA) antibodies and glutamic acid decarboxylase antibodies have shown that the primate retina contains GABAergic amacrine, interplexiform, and displaced amacrine cells. In addition, subpopulations of photoreceptors and horizontal cells have also been suggested to be GABAergic in this retina. In the present study, we have used in situ hybridization to localize GABAergic neurons in human and monkey retinas. In situ hybridizations were carried out with 35S-labeled DNA and RNA probes derived from human and cat glutamic acid decarboxylase cDNA clones. In the monkey retina, labeled cells were present in the inner nuclear and ganglion cells layers. The outer nuclear layer or the inner segment layer had only background levels of labeling. In the inner nuclear layer, all labeled somata were located in the vitread-half bordering the inner nuclear layer/inner plexiform layer boundary. These cells constituted approximately 83% of all labeled cells. Labeled cells were also seen in the ganglion cell layer. In the human retina, labeled somata were observed only in the inner nuclear and the ganglion cell layers. In the inner nuclear layer, the majority of labeled cells were located close to the inner nuclear layer/inner plexiform layer boundary although a minor population of labeled somata (approximately 20%) were found deeper in the inner nuclear layer. The distribution of glutamic acid decarboxylase mRNA-containing cells we observed is in good agreement with the known location of GABAergic neurons. We, however, did not find glutamic acid decarboxylase mRNA in either horizontal cells or photoreceptors in monkey and human retina.

Animals↗
PubMed1989

Parasol and midget ganglion cells of the primate retina.

We have intracellularly filled the dendritic arbors of 996 midget and parasol ganglion cells with horseradish peroxidase (HRP) in macaque and baboon retinas. Only minor differences in the properties of these cell groups were found between species. Ninety of these cells were cut from their retinas, embedded in methacrylate, and transversely sectioned. According to their depth of stratification, there are two types of parasol cells (termed a-parasol and b-parasol), and two types of midget ganglion cells (a-midget and b-midget). Each of these four types stratifies at a different level within the IPL. The dendritic fields of midget ganglion cells lie either near the border of the ganglion cell layer (GCL) or near the border of the amacrine cell layer (ACL). The dendrites of the two types of parasol cells stratify closer to the center of the IPL, where they divide it into three approximately equal parts. There was no vertical overlap in the dendritic fields of a-parasols and b-parasols; they were always separated by at least 1 micron. The border between the a- and b-sublaminae of the IPL, defined in terms of this narrow gap between the stratification of the two parasol cell types, lies approximately at the center of the IPL. The dendritic-field thickness for each of these types, on average, is no greater than 30% of the IPL thickness. At a similar location, there is no significant difference between the dendritic-field diameters of the two parasol types or between those of the two midget types. As previously reported (Perry et al.: Neuroscience 12:1101-1123, '84) the dendritic fields of both parasol and midget ganglion cells are smaller in the nasal retina than at a position in the temporal retina equidistant from the fovea. Because dendritic-field diameters prove to depend upon local ganglion-cell density, the scatter in these diameters as a function of retinal eccentricity is due in part to the asymmetric distribution of ganglion cells. We have devised a measure, termed equivalent eccentricity, that allows data points of cells from regions having the same local ganglion-cell density to be plotted at the same position on this scale. The use of this measure, rather than eccentricity per se, significantly reduces the scatter of dendritic-field diameters. The dendritic-field diameters of parasol cells within the nasal quadrant of the retina are not fully brought into line with those of cells lying elsewhere in the retina.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Photoreceptor mosaic: number and distribution of rods and cones in the rhesus monkey retina.

Video-enhanced differential interference contrast optics was used to determine the number and distribution of photoreceptors across the entire retinal surface of 9 eyes obtained from 7 adult rhesus monkeys. We found that the retina of this primate contains an average of 3,100,000 cones (+/- 130,000) and 61,000,000 rods (+/- 7,500,000). Variation among animals in the number of rods and cones cannot be accounted for by differences in sex, age, or retinal surface area, nor is there a correlation between the number of rods and cones (a retina with a high number of rods does not typically have a high number of cones). Cone density peaks at 141,000 cones/mm2 in the foveola and decreases about 100-fold toward the periphery. Rod density in a central annulus around the fovea is 130,000/mm2 and decreases 6-8-fold toward the periphery. In all 9 retinae, we found that an area 4-5 mm dorsal to the fovea had the highest rod density at 184,000 rods/mm2. The functional significance of this area, which we term the dorsal rod peak (DRP), may be related to high sensitivity vision under scotopic conditions. Outside of the DRP, rod density is symmetrical around the major axes of the retina, whereas cone density is elevated in nasal retina. Among animals, both rods and cones display a 2-fold individual difference in receptor density at any given eccentricity. Although rods and cones differ in absolute number, the location and magnitude of their peak densities, and their central to peripheral density gradients, the ratio of the density of rods to cones (15-30:1) is remarkably stable from 3 mm to 15 mm eccentricity. The relative consistency in the proportion of rods and cones in extrafoveal retina may be related to mechanisms of retinal development and functional interactions between scotopic and photopic systems.

Animals↗
PubMed1990

Microglial cell responses in the rabbit retina following transection of the optic nerve.

The response of retinal microglial cells, which accompanies retrograde degeneration of ganglion cell axons and perikarya (induced by transection of the optic nerve), was studied in whole-mounted rabbit retinae labeled enzyme-histochemically for nucleoside diphosphatase (NDPase), which is a microglial cell marker. A few days after transection, the number of microglial cells/mm2, as well as their staining intensity, began to increase in the inner plexiform layer. The mosaic-like distribution of the star-shaped microglial cells present in the inner plexiform layer of a normal rabbit retina was preserved during ganglion cell degeneration. As in the normal retina, processes of individual cells never overlapped with those of neighboring cells in the inner plexiform layer because individual cells in the "degenerating" retina acquired shorter processes, i.e., the cells occupied a smaller territory compared to the normal retina. In the nerve fiber layer the number and staining intensity of NDPase-labeled microglial cell processes (most of which are aligned in parallel with degenerating ganglion cell axons) transiently increased and returned to normal values by 5 months post-transection. Microglial cells that are not detectably NDPase labeled in the outer plexiform layer of a normal rabbit retina acquire intense staining a few days after the nerve is cut. The functional significance of the increased NDPase activity in the plasma membrane of microglial cells during degeneration remains to be determined.

Acid Anhydride Hydrolases↗
PubMed1991

Factors determining the migration of astrocytes into the developing retina: migration does not depend on intact axons or patent vessels.

Astrocytes migrate into the cat retina from the optic nerve, beginning from embryonic day (E) 52. Once they have entered the retina they concentrate along major axon bundles and fail to enter regions of the retina with high densities of neurones, in particular the area centralis region of the ganglion cell layer. These nonuniformities appear as the astrocytes spread over the retina during development, and in this study we have examined factors that might control their spread. First we examined astrocytes in a retina in which the axon bundles had degenerated following an optic nerve lesion at birth. The area over which astrocytes had spread was normal, suggesting that their spread does not depend on the presence of intact axons. Second, we noted that, despite the degeneration of all ganglion cells following the nerve lesion, astrocytes still did not spread over the area centralis. Their spread is apparently not inhibited by concentrations of neurones. Third, we examined astrocytes in retinas of animals raised in an atmosphere containing 70-80% oxygen, which prevents the formation of retinal vessels. Again, the area over which the astrocytes had spread was normal, suggesting that their spread does not depend on the presence of patent blood vessels. These negative findings led us to compare the distribution of spindle cells (precursors of retinal vasculature) and astrocytes in the cat during development. The close correspondence in their topographical distribution and the earlier spread of the spindle cells lead us to suggest that spindle cells provide a basal lamina component that may guide the migration of astrocytes.

Animals↗
PubMed1991

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 Wässle, '74). An antiserum to parvalbumin was used to visualize the dendrites (Röhrenbeck and Wässle, '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.

Animals↗
PubMed1991

Development of the rabbit retina: II. Müller cells.

Müller (glial) cells of the rabbit retina were stained with antibodies against the intermediate filament protein vimentin in retinal wholemounts from various developmental stages. Both the density of stained profiles and the mean diameter of these profiles were measured, with the microscope focus in the inner plexiform layer of the retinae. Within this retinal layer, every Müller cell possesses one stout vitread process; thus counts of the stained profiles allow an estimation of their number. After postnatal day (P) 9, the total number of stained cells was slightly above 4 million per retina; for the adult rabbit retina, this agrees well with earlier data obtained by our group based on another method, as well as with published data from other groups. We suggest that after P 9, only Müller cells are stained, and this population is numerically stable. In contrast, neonatal retinae contained significantly more stained profiles. This indicates that either the total number of Müller cells is reduced by "physiological cell death" or that additional cells are stained neonatally. We discuss why we favour the second possibility. After P 9, two peculiarities occur in the Müller cell population: (1) their density decreases gradually, to a greater extent in the retinal periphery than in the center (i.e., in the "visual streak"), and (2) Müller cell diameters increase, again more in the periphery than in the center. We argue that differential retinal expansion leads to dispersion of the pre-existing cell population and allows for widening of the Müller cell processes. We conclude that Müller cells can be used postnatally in the rabbit retina as "landmarks" of expansion.

Aging↗
PubMed1991

Glutamate, GABA, and glycine in the human retina: an immunocytochemical investigation.

The distribution of the neuroactive amino acids glutamate, GABA, and glycine in the human retina was examined in consecutive semithin sections treated with antisera specific for fixed glutamate, GABA, and glycine, respectively. Glutamate immunoreactivity was conspicuous in all photoreceptor cells (rods more strongly labelled than cones), and in a majority (85-89%) of the cells in the inner nuclear layer (INL). Rod spherules and cone pedicles showed a greater enrichment of glutamate immunoreactivity than the parent cell bodies and inner segments. Also, structures of the inner plexiform layer (IPL) were labelled. A large majority (83-91%) of cells in the ganglion cell layer (GCL) was strongly stained, as were most axons in the nerve fibre layer. Müller cell processes appeared unstained. GABA immunoreactivity was present in presumed amacrine but not in bipolar-like cells. The stained cells were restricted to the inner 1/3 of the INL and were more frequent in central than in peripheral retina (40% and 26% of all cells in the inner 1/2 of INL, respectively). GABA positive cell processes, probably originating from interplexiform cells, appeared to traverse the INL and end in the outer plexiform layer. Dense immunolabeling was found in the IPL. GABA immunoreactive cells (some also stained for glutamate) comprised 23% of all GCL cells in the peripheral retina, but only 5% in the central retina. Most of them were localized adjacent to the IPL. A few GABA positive (possibly ganglion) cells extended a single fibre toward the nerve fibre layer. Solitary GABA positive fibres were seen in this layer and in the optic nerve. Glycine immunoreactivity was observed in cells with the location typical of amacrine and bipolar (peripheral retina) cells, as well as in punctate structures of the IPL. In contrast to the GABA positive cells, the glycine positive cells were more frequent in the peripheral than in the central retina (42% and 23% of all cells in inner 1/3 of INL, respectively). A few cells in the GCL (0.5-1.5%) were glycine positive. Glutamate colocalized with GABA or glycine in a majority of the cells stained for either of these inhibitory transmitters (90-95% of the GABA positive cells, and 80-86% of the glycine positive cells, in the INL). Some bipolar cells were stained for both glutamate and glycine. Colocalization of GABA and glycine occurred in a subpopulation (3-4%) of presumed amacrine cells, about half of which was also glutamate positive.

Glutamates↗
PubMed1991

Calmodulin and calbindin localization in retina from six vertebrate species.

Calmodulin is abundant in the central nervous system, including the retina. However, the localization of calmodulin in the retina has not been described in detail. We therefore decided to investigate calmodulin localization in retinae from six vertebrate species, by using immunohistochemical labeling with four different rabbit polyclonal antibodies against calmodulin. The localization of calbindin-D28k, another calcium-binding protein already well described in retina, was compared. We found that calmodulin distribution is more highly conserved among species, contrasting with calbindin variability. The most striking result emerging is that calmodulin could not be detected in photoreceptors although other layers are intensely calmodulin-immunoreactive, casting doubt about a direct role of calmodulin in phototransduction. Horizontal cells are weakly calmodulin-immunoreactive, bipolar cells are calmodulin-immunoreactive except in turtle retina, numerous amacrine and ganglion cells are labeled in all species, and the fiber layer is always labeled. These data demonstrate that, while the calmodulin distribution in retina is similar among vertebrate species, selective differences in localization can be detected not only among the same cell types in different species but also among different cell types in the same species. The results showing differences in calmodulin immunoreactivity among cell types also provide further evidence that calmodulin expression in eukaryotes is not constitutive, in the sense that not every cell expresses similar levels of calmodulin.

Animals↗