Number and distribution of neurons in the retinal ganglion cell layer in relation to foraging behaviors of tyrant flycatchers

The tyrant flycatchers represent a monophyletic radiation of predominantly insectivorous passerine birds that exhibit a plethora of stereotyped prey capture techniques. However, little is known about their retinal organization. Using retinal wholemounts, we estimated the total number and topography of neurons in the ganglion cell layer in the generalist yellow‐bellied elaenia (Elaenia flavogaster) and the up‐hover‐gleaner mouse‐colored tyrannulet (Phaeomyias murina) with the optical fractionator method. The mean estimated total number of neurons in the ganglion cell layer was 4,152,416 ± 189,310 in E. flavogaster and 2,965,132 ± 354,359 in P. murina. Topographic maps of isocounting lines revealed a similar distribution for both species: a central fovea and a temporal area surrounded by a poorly defined horizontal streak. In addition, both species had increased numbers of giant ganglion cells in the dorsotemporal retina forming an area giganto cellularis. In E. flavogaster, these giant ganglion cells were also distributed across the nasal and ventral retinal peripheries, which is in agreement with the generalist habits of this species. However, in P. murina these cells were rarely seen along the nasal and ventral peripheries, possibly reflecting a lesser need to perceive movement because this species captures stationary insects resting on foliage. Thus, we suggest that the retinas of the tyrant flycatchers in the present study show a general common pattern of neuron distribution in the ganglion cell layer irrespective of their foraging habits. We also suggest that the distribution of giant ganglion cells is indicative of the visual requirements of the species. J. Comp. Neurol. 514:66–73, 2009. © 2009 Wiley‐Liss, Inc.     

Neurogenesis and cell death in the ganglion cell layer of vertebrate retina

The correct formation of all central nervous system tissues depends on the proper balance of neurogenesis and developmental cell death. A model system for studying these programs is the ganglion cell layer (GCL) of the vertebrate retina because of its simple and well-described structure and amenability to experimental manipulations. The GCL contains approximately equal numbers of ganglion cells and displaced amacrine cells. Ganglion cells are the first or among the first cells born in the retina in all the studied vertebrates. Neurogenesis and cell death have been studied extensively in the GCL of various amniotes (rodents, chicks, and monkeys) and anamniotes (fish and frogs), and the two processes highlight developmental differences between the groups. In amniotes, neurogenesis occurs during a defined period prior to birth/hatch or the opening of the eyes, whereas in anamniotes, neurogenesis extends past hatching into adulthood-sometimes for years. Roughly half of GCL neurons die during development in amniotes, whereas developmental cell death does not occur in the GCL neurons of anamniotes. This review discusses the spatial and temporal patterns of neurogenesis, cell death, and possible explanation of cell death in the GCL. It also examines markers widely used to distinguish between ganglion cells and displaced amacrine cells, and methods employed to birth date neurons.     

Variations in cell density in the ganglion cell layer of the retina as a function of ocular pigmentation

Ocular melanin regulates retinal development, including cell density gradients in the central retina, a region essential for normal visual acuity. In albinos this region is underdeveloped and peak cell numbers are reduced. It is not known whether there is a dosage relationship between pigmentation and the degree of this underdevelopment, as studies of the retinal effects of albinism have commonly used rodents. These have poorly developed central regions even in the wild type. Rabbits, however, have a unique, highly specialized, visual streak in the central retina where cell density gradients are very steep and these are reduced in albinos. Here, cell densities in the ganglion cell layer of separate groups of rabbits, with different levels of ocular pigmentation and known mutations of the tyrosinase gene coding sequence, were examined. These revealed reductions in peak cell densities and/or in the regions over which high cell densities were maintained in all hypopigmented phenotypes. There was no dosage relationship between levels of pigmentation and deficits in the ganglion cell layer as animals with relatively small reductions in retinal pigment had deficits comparable to those found in albinos. The greatest variability between pigmentation phenotypes was between the two completely unpigmented strains. Consequently, although pigment may regulate the development of the central retina, this study failed to show that it does so in a dose-dependent manner.     

Amacrine cells in the ganglion cell layer of the cat retina

Following transection of the optic nerve, ganglion cells in the cat retina undergo retrograde degeneration. However, many small profiles (less than or equal to 10 micron) survive in the ganglion cell layer. Previously considered to be neuroglia, there is now substantial evidence that they are displaced amacrine cells. Their density increases from approximately 1,000 cells/mm2 in peripheral retina to 7,000 cells/mm2 in the central area. Their total number was found to be 850,000, which is five times the number of ganglion cells and also five times the number of astrocytes. Uptake of 3H-muscimol followed by autoradiography labelled 75% of the displaced amacrine cells; hence, the majority seem to be GABAergic. Immunocytochemistry with an antibody directed against choline-acetyl-transferase labelled approximately 10% of the displaced amacrines in the peripheral retina and 17% in the central area. Uptake of serotonin (5-HT) followed by immunocytochemistry was found in 25-30% of displaced amacrines. NADPH diaphorase histochemistry labelled approximately 5% of displaced amacrine cells. The sum of the various percentages make colocalization likely. Intracellular injection of Lucifer Yellow under microscopic control revealed that displaced amacrine cells constitute several morphological types.     

Asymmetry in the structural organization of the ganglion layer of the retina in the frog

Silver-impregnated retinal preparations were used to study the distribution density and topographic features of small and large ganglionic cells (GC) of Rana ridibunda and Rana temporaria. For both species the increased density of GC (a streak) stretched higher than the naso-temporal axis passing through the optic disk. Beyond the streak the density of small GC was maximal in the central zone of the retina and decreased towards its periphery. For the upper quadrants of the retina the density of small GC was higher than that for the lower ones by 26% on the average. On the contrary, the density of large GC was higher in the lower part of the retina as compared to the upper one, the difference being more pronounced for R. temporaria. The density of large GC was also asymmetric with respect to the dorso-ventral axis being higher in nasal quadrants than in temporal ones by 40-55%. The highest density of large GC was found in the middle zone of the retina. The found structural asymmetry in the retinal output raster may bear an adaptively ecological meaning and may condition the particularities of the formation of the visually guided prey-catching and avoidance reactions.     

Development of the human retina: patterns of cell distribution and redistribution in the ganglion cell layer

Neurogenesis in the ventricular layer and the development of cell topography in the ganglion cell layer have been studied in whole-mounts of human fetal retinae. At the end of the embryonic period mitotic figures were seen over the entire outer surface of the retina. By about 14 weeks gestation mitosis had ceased in central retina and differentiation of photoreceptor nuclei was evident within a well-defined area which constituted about 2% of total retina area. This area was approximately centered on the site of the putative fovea, identified by the exclusive development of cone nuclei at that location. The area of retina in which mitosis had ceased increased as gestation progressed. By mid-gestation mitosis in the ventricular layer occupied about 77% of the outer surface of the retina and by about 30 weeks gestation mitosis in the ventricular layer had ceased. Cell density distributions in the ganglion cell layer were nonuniform at all stages studied (14-40 weeks). Densities were highest at about 17 weeks gestation, and by mid-gestation the adult pattern of cell topography was present with maps showing elevated cell densities in posterior retina and along the horizontal meridian. Cell densities generally declined throughout the remainder of the gestation period, except in the posterior retina, where densities in the perifoveal ganglion cell layer remained high during the second half of gestation. There is a rapid decline in cell density in the foveal ganglion cell layer toward the end of gestation, and it is suggested that the persistence of high densities in the perifoveal region may be related to migration of cells away from the developing fovea. The total population of cells in the ganglion cell layer was highest (2.2-2.5 million cells) between about weeks 18 and 30 of gestation. After this the cell population declined rapidly to 1.5-1.7 million cells. It is suggested that naturally occurring neuronal death is largely responsible for this decline.     

Patterns of cell death in the ganglion cell layer of the human fetal retina

The distribution of dying cells in the ganglion cell layer (GCL) of retinae from human fetuses has been analysed. Both whole-mounted and sectioned retinae have been studied. Results suggest that cells are lost from the GCL between weeks 14 and 30 of the gestation period, approximately. This period corresponds to the period during which axons are lost from the developing optic nerve. Cell loss is greatest between weeks 16 and 21 of the gestation period. The pattern of cell loss is nonuniform, and between weeks 16 and 24, the relative frequency of pyknotic cells (pyknotic cells:viable cells) in peripheral retina is considerably higher than in central retina. This pattern of cell loss predominates during the period in which a distinct centroperipheral gradient of cell densities emerges in the GCL of the human fetal retina (between 18 and 23 weeks gestation). It is suggested that the regional loss of ganglion cells may contribute to the formation of the cell density gradient.     

The pigeon retina: Quantitative aspects of the optic nerve and ganglion cell layer

Studies of electron micrographs of pigeon optic nerve showed numerous unmyelinated fibers (29% of total) and more than twice as many total fibers as previously reported. The 2.4 million axons had a mean diameter below 1 μ, close to the limit of resolution of light microscopy. This probably accounts for the large discrepancy between light and electron microscopic counts. Studies of retinal whole mounts showed an irregular distribution of cells in the ganglion cell layer including a minor posterior fovea and an area of increased density in the posterior-superior quadrant. Sample counts produced an estimate of over 4.8 million cell bodies in this layer. Cytological investigation indicated that over 85% of these cells were neuronal. The considerable excess of neuron cell bodies over axons in thought to be explained by the presence of large numbers of amacrine cells, cells which do not project axons into the optic nerve.     

Isolation and characterization of seventeen polymorphic microsatellite DNA markers from Elaenia ruficeps (Aves: Tyrannidae)

Seventeen new polymorphic microsatellites were isolated and characterized for the South American Rufous-crowned Elaenia, (Elaenia ruficeps). E. ruficeps is a bird commonly found in white sand vegetation, a naturally fragmented and threatened Amazonian habitat. The number of alleles per locus ranged from 2 to 20, whilst the observed and expected heterozygosities varied from 0.074 to 0.967 and from 0.173 to 0.919, respectively. Seven loci significantly deviated from Hardy–Weinberg equilibrium. Eleven microsatellite loci were cross-amplified in at least one additional Tyrannidae species. These microsatellite markers will be useful tools for further studies of avian population structure and genetic variability in this unique habitat.     

A regional specialization in the opossum's retina: Quantitative analysis of the ganglion cell layer

The distribution of ganglion cells in the opossum's retina was determined from flat-mounted preparations stained with cresyl-violet. The retinal area is 109 mm² (SD = 16 mm²). Maps of ganglion cell density were made from retinae of seven animals. in all maps iso-density lines were approximately concentric, showing a slight elongation towards the nasal region. Cell density varied from 400 cells/mm² at the extreme periphery to 2,900 cells/mm² in the region of highest count, the are centralis. The center of this region lies 1.85 mm (26.3°) temporal to the center of the optic nerve head. The average total number of ganglion cells is 77,384 (SD = 10,173). Based upon soma diameter histograms ganglion cells were classified into three groups, showing at area centralis peaks at 7 μm, 12 μm and 15 μm respectively. Cell soma diameter ranged from 6 μm to 21 μm, larger values being observed at the periphery.     

Unusual topographic specializations of retinal ganglion cell density and spatial resolution in a cliff-dwelling artiodactyl,the Nubian ibex (Capra nubiana)

The Nubian ibex (Capra nubiana) occurs in information-rich visual habitats including the edges of cliffs and escarpments. In addition to needing enhanced spatial resolution to find food and detect predators, enhanced visual sampling of the lower visual field would be advantageous for the control of locomotion in such precarious terrains. Using retinal wholemounts and stereology, we sought to measure how the ganglion cell density varies across the retina of the Nubian ibex to reveal which portions of its surroundings are sampled with high resolution. We estimated a total of ~1 million ganglion cells in the Nubian ibex retinal ganglion cell layer. Topographic variations of ganglion cell density reveal a temporal area, a horizontal streak, and a dorsotemporal extension, which are topographic retinal features also found in other artiodactyls. In contrast to savannah-dwelling artiodactyls, the horizontal streak of the Nubian ibex appears loosely organized possibly reflecting a reduced predation risk in mountainous habitats. Estimates of spatial resolving power (~17 cycles/degree) for the temporal area would be reasonable to facilitate foraging in the frontal visual field. Embedded in the dorsotemporal extension, we also found an unusual dorsotemporal area not yet reported in any other mammal. Given its location and spatial resolving power (~6 cycles/degree), this specialization enhances visual sampling toward the lower visual field, which would be advantageous for visually guided locomotion. This study expands our understanding of the retinal organization in artiodactyls and offers insights on the importance of vision for the Nubian ibex ecology.     

The SCG10-related gene family in the developing rat retina: persistent expression of SCLIP and stathmin in mature ganglion cell layer

Neuronal growth-associated proteins (GAPs), such as GAP-43 and SCG10, are thought to play crucial roles in both axonal and dendritic outgrowth during neural development and regeneration, although the underlying mechanisms remain largely unknown. The recent finding that SCG10 is a microtubule regulator and also the identification of RB3 and SCLIP as two new SCG10-related members prompted us to investigate the roles of SCG10-related family in neural development, using the retina as a model system. We determined the temporal expression and the spatial distribution of SCG10-related mRNAs in the developing rat retina. Semiquantitative analysis by RT-PCR revealed that in prenatal retina, levels of SCG10 and stathmin mRNAs were higher than those of RB3 and SCLIP. In the postnatal retina, the level of SCLIP increased, whereas the level of RB3 remained low. In situ hybridization revealed that GAP-43 and all of the SCG10-related family mRNAs were present in the retinal ganglion cells (RGCs) at all stages of retinal development, and that stathmin mRNA was present in mitotic neuroblastic cells. Differential expression of SCG10 and other members of the family became more evident as retinal development proceeded; SCG10 and RB3 expression were relatively specific in the RGCs and amacrine cells, whereas SCLIP was also evident in bipolar and horizontal cells. Stathmin mRNA was highly expressed both in the RGCs and other interneurons. These results indicate that multiple SCG10-related proteins are expressed in single neurons including RGCs, and suggest that these nGAPs play similar but distinct roles in differentiation and functional maintenance of retinal neurons.     

Peak density, size and regional distribution of ganglion cells in the retina of the fur seal Callorhinus ursinus.

The total number, size, topographic distribution and peak density of ganglion cells were studied in retinal wholemounts of the fur seal, Callorhinus ursinus. The cell distribution showed a distinct zone of high ganglion cell density. It was located in the temporal retinal quadrant, near the horizontal meridian, 10-12 mm (25-31 degrees) from the optic disk. The peak cell density in this zone was 812-1332 cells/mm2 (mean 1053 cells/mm2), i.e. 125-205 cells/deg2 (mean 162 cells/deg2). These data predict a retinal resolution of 5.6-7.1 cycle/deg. The ganglion cell soma size ranged from 10 to 50 microns. Cell size histograms were bimodal in shape with modes below and above 30 microns.     

The ON-alpha ganglion cell of the cat retina and its presynaptic cell types

Anatomical circuits converging onto the ON-alpha (Y) ganglion cell were studied by computer-assisted reconstruction of substantial portions of 2 alpha cells from electron micrographs of serial sections. The alpha cells in the area centralis were labeled by a Golgi-like retrograde filling with horseradish peroxidase, and certain presynaptic amacrine processes were labeled by uptake of 3H-glycine. About 4400 synapses contacted the alpha cell. Eighty-six percent were from amacrine cells; the rest were from bipolar cells. About one-quarter of the amacrine synapses were specifically labeled by 3H-glycine and probably belong to the A4 amacrine. The bipolar inputs were provided by several types: cone bipolar CBb1 (85%), cone bipolar CBb5 (2%), the rod bipolar (5%), and some unidentified cone bipolars (11%). Contacts from each type occurred in specific strata, with the consequence that they tended to form spots or annulli over the alpha dendritic field. The CBb1 bipolars formed a moderately dense array (8000/mm2), with a nearest-neighbor distance of 8.6 +/- 1.3 microns. Most members of the array (84%) contacted the alpha cell, providing 1-7 synapses (average, 2.7 +/- 1.6). The placement of contacts from an individual CBb1 followed certain rules: they were restricted to a parent branch of the alpha arbor or to 2 daughter branches, but almost never crossed a branch of the alpha arbor. The synaptic territory of an individual CBb1 was not shared with other b1s (or cone bipolars of any sort), although it was shared with amacrine contacts. Rod bipolar cells also formed a very dense array (54,500/mm2) in the alpha dendritic field, but only a few of these (3%) contacted the alpha cell. The concentric receptive field of the CBb1, combined with the spatial organization of its array, is used to predict the CBb1 contribution to the alpha cell receptive field; this contribution resembles the spatial and temporal organization of the alpha receptive field itself.     

The organization of neuronal somata in the first sacral spinal ganglion of the cat.

The basis of organization of neuronal somata within a spinal ganglion has been studied using histological techniques. The positions of the largest neuronal cell bodies, which are thought to innervate muscle receptors, were plotted for unoperated and sham operated animals. These ganglion cells were found in all sectors of the ganglion and their distribution generally reflected that of the total ganglion cell population in the respective sectors. The positions of neuronal somata undergoing the axonal reaction following various nerve lesions were plotted. Altered neurons were not preferentially accumulated in any sector of the ganglion after lesions of cutaneous nerves, nerves to muscle, or nerves innervating the distal part of the hind limb, but they often occurred in clusters which were found in all parts of the ganglion. Following a lesion of either the pudendal nerve, the S1 dorsal primary ramus, or the S1 ventral root, altered neurons tended to be grouped in one or more sectors of the ganglion. These data suggest that ganglion cells innervating skin or muscle often occur in small groups distributed throughout the ganglion, while processes of afferent neurons which branch from the spinal nerve and do not travel through a limb plexus tend to have their cell bodies restricted to a particular region of the ganglion.     

The organization of the cardiac ganglion of the axolotl (Ambystoma mexicanum)

The heart of the axolotl Ambystoma mexicanum was studied with histochemical methods to determine the distribution of neurons containing acetylcholine esterase, catecholamines and 5-hydroxytryptamine. The cardiac ganglion is made up of cholinergic nerve fibers and somata, and of catecholaminergic fibers. Small intensely fluorescent cells were found along blood vessels in the pericardial wall at the base of the heart, but not in the heart itself, except, in a few instances, in the region bordering the pericardial wall. Both the cholinergic and the catecholaminergic innervation of the heart were poorly developed at hatching and reached their mature state after a few months. Cholinesterase staining fibers appeared several weeks before catecholaminergic fibers. The number of postganglionic cholinergic neurons in the heart increased several-fold during the first month after hatching. Histofluorescence studies of organ cultures suggested that all the catecholamine present in the heart are of extrinsic origin. Liquid chromatography with electrochemical detection demonstrated that the dominant catecholamine in the heart is norepinephrine. No neurons containing 5-hydroxytryptamine were found.     

Reduced glial cell density and neuronal size in the anterior cingulate cortex in major depressive disorder

BACKGROUND: Glial cells are more numerous than neurons in the cortex and are crucial to neuronal function. There is evidence for reduced neuronal size in schizophrenia, with suggestive evidence for reduced glial cell density in mood disorders. In this investigation, we have simultaneously assessed glial cell density and neuronal density and size in the anterior cingulate cortex in schizophrenia, major depressive disorder, and bipolar disorder. METHODS: We examined tissue from area 24b of the supracallosal anterior cingulate cortex in 60 postmortem brain specimens from 4 groups of 15 subjects, as follows: major depressive disorder, schizophrenia, bipolar disorder, and normal controls. Glial cell density and neuronal size and density were examined in all subjects using the nucleator and the optical disector. RESULTS: Glial cell density (22%) (P =.004) and neuronal size (23%) (P =.01) were reduced in layer 6 in major depressive disorder compared with controls. There was some evidence for reduced glial density in layer 6 (20%) (P =.02) in schizophrenia compared with controls, before adjusting for multiple layerwise comparisons, but there were no significant changes in neuronal size. There was no evidence for differences in glial density or neuronal size in bipolar disorder compared with controls. Neuronal density was similar in all groups to that found in controls. CONCLUSION: These findings suggest that there is reduced frontal cortical glial cell density and neuronal size in major depressive disorder.     

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

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

The distribution of ganglion cells in the retina of the north american opossum (Didelphis virginiana)

The distribution of ganglion cells in the retina of the opossum was determined from whole-mounted retinae stained with cresyl violet. Isodensity lines were approximately circular with a peak density of 2,000 to 2,700 cells/mm² in superior temporal retina (area centralis). The total number of retinal ganglion cells was estimated to be 72,000 to 135,000 (mean 101,026) in retinae ranging from 125 to 187 mm² in total area. Three groups of ganglion cells were distinguished on the basis of soma size and retinal topography. Large cells (24 to 32 μm diameter) were fairly evenly distributed across the retina. Medium cells (12 to 23 μm diameter) were more numerous in the superior temporal quadrant than in other regions of the retina. Small cells (7 to 11 μm diameter) were prominent in all retinal regions, but particularly in nasal and inferior retina. An analysis of topographical differences in soma size distribution suggests that the medium size cells can be further subdivided into small-medium and large-medium groups.