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The orientation of cell division influences cell-fate choice in the developing mammalian retina.

Asymmetric segregation of cell-fate determinants during cell division plays an important part in generating cell diversity in invertebrates. We showed previously that cells in the neonatal rat retina divide at various orientations and that some dividing cells asymmetrically distribute the cell-fate determinant Numb to the two daughter cells. Here, we test the possibility that such asymmetric divisions contribute to retinal cell diversification. We have used long-term videomicroscopy of green-fluorescent-protein (GFP)-labeled retinal explants from neonatal rats to visualize the plane of cell division and follow the differentiation of the daughter cells. We found that cells that divided with a horizontal mitotic spindle, where both daughter cells should inherit Numb, tended to produce daughters that became the same cell type, whereas cells that divided with a vertical mitotic spindle, where only one daughter cell should inherit Numb, tended to produce daughters that became different. Moreover, overexpression of Numb in the dividing cells promoted the development of photoreceptor cells at the expense of interneurons and Müller glial cells. These findings indicate that the plane of cell division influences cell-fate choice in the neonatal rat retina and support the hypothesis that the asymmetric segregation of Numb normally influences some of these choices.

Animals↗

Gap junctions in the inner plexiform layer of the goldfish retina.

Oblique thin sections of the goldfish inner plexiform layer (IPL) were examined to characterize the processes engaging in gap junctions. Omission of en bloc uranyl acetate staining enhanced the relative visibility of gap junctions, thus facilitating surveys. Three general categories were identified: (1) extensive gap junctions between large caliber amacrine cell dendrites presynaptic to ganglion cells and other amacrine cells in distal and proximal IPL; (2) moderate to small gap junctions between amacrine cell synaptic terminals or small amacrine cell-like processes of similar cytoplasmic appearance; and (3) gap junctions between the telodendria/terminals of bipolar cells. Most instances of gap junctions in the goldfish IPL seem to represent cases of homologous coupling. We have not found unambiguous cases of heterologous coupling as in the mammalian retina, nor have we yet found mixed synapses. Taken together with previous physiological and anatomical findings, homologous coupling occurs between pairs of rods, cones, horizontal cells, bipolar cells and amacrine cells in the goldfish retina, and this in turn implies that direct intercellular communication among like cell types is a common theme in retinal circuitry.

Animals↗

The action spectra of cone photoreceptors in the turtle (Mauremys caspica) retina.

Cone photoreceptors in the turtle retina are involved in intricate neuronal interactions with other retinal neurons that modify the responses of the cones to photons absorbed in their outer segments. Therefore, the action spectra of cones strongly depend upon the conditions of measurements. This study describes an attempt to derive the action spectra of turtle cones which are the least distorted by neuronal interactions. To achieve this goal, the photoresponses of cones and horizontal cells were recorded from the turtle retina under different conditions of adaptation using different patterns of the stimulating test flashes. The sensitivity action spectra, derived from small-amplitude (<1 mV) photoresponses, were strongly affected by the recording conditions indicating the contributions of multiple neuronal inputs. Action spectra, constructed from large criterion photoresponses, were less distorted by neuronal interactions and better described the spectral properties of the "isolated" cones. The action spectra of the hyperpolarizing inputs to chromaticity-type horizontal cells were derived by stimulating these cells with mixtures of a saturating red light and a monochromatic light of different wavelength and intensity. The action spectra were constructed from the intensity of the addend component needed to "pull down" the depolarizing response to the red component by a fixed criterion. These spectra, measured in red/green and yellow/blue C-type horizontal cells, are suggested to best represent the "isolated" M-cones and S-cones, respectively.

Absorption↗

Anatomical evidence for the overlapped distribution of ipsilaterally and contralaterally projecting ganglion cells to the lateral geniculate nucleus in the cat retina: a morphologic study with fluorescent tracers.

The question of the nasotemporal overlap of ganglion cells in the central retina, which project to the ipsilateral and contralateral lateral geniculate nucleus, was reexamined in the cat by means of a retrograde fluorescent double-labeling technique. The horizontal separation between the ipsilateral and contralateral decussation lines, which corresponded to the width of the median strip of overlap, was approximately 0.2 mm. This is the first direct confirmation of the extent of such overlap in the cat retinal ganglion cells, which had been previously studied by indirect methods.

Animals↗

The retina of tyrant flycatchers: topographic organization of neuronal density and size in the ganglion cell layer of the great kiskadee Pitangus sulphuratus and the rusty margined flycatcher Myiozetetes cayanensis (Aves: Tyrannidae).

Tyrant flycatchers comprise the largest group of passerine birds of the Neotropical region but their retinal organization is unknown. The great kiskadee, Pitangus sulphuratus, is categorized as a supreme generalist and utilizes a variety of foraging strategies. The rusty margined flycatcher, Myiozetetes cayanensis, is partially frugivorous and captures insects in the air. Using retinal wholemounts, we described the topographic distribution of density and size of neurons lying in the retinal ganglion cell layer in those two species of tyrant flycatchers. Maps of neuron distribution showing isodensity contours revealed the presence of a pronounced central fovea and a temporal area in both species. Both retinal specializations were circumscribed by an inconspicuous horizontal visual streak. The highest foveal densities ranged from 48,000 to 55,000 cells/mm(2) for Pitangus sulphuratus and between 62,000 and 65,000 cells/mm(2) for Myiozetetes cayanensis. The peak density in the temporal area was around 40,000 cells/mm(2) for Pitangus sulphuratus and 46,000 cells/mm(2) for Myiozetetes cayanensis. At central, mid-peripheral and peripheral eccentricities, perikaryon size varied quite similarly in both species. A cohort of giant retinal ganglion cells with perikaryon size > 300 microm(2) was observed at the temporal periphery and defines an 'area giganto cellularis' described previously in procellariiform seabirds. This specialization is thought to be involved in movement detection and could aid the tyrant flycatchers to capture moving prey. Functionally, the presence of a fovea associated with a temporal area would allow high spatial resolution for capturing insects by the tyrant flycatchers. Nonetheless, even though both species exhibit different foraging strategies, they shared a similar topographic arrangement of neuronal density in the ganglion cell layer. This suggests that the retinal topography did not accompany changes in the foraging ecology throughout evolutionary history for these species of tyrant flycatchers.

Animals↗

Morphological types of horizontal cell in the retina of the domestic cat.

Two morphologically distinct types of horizontal cell are described from Golgi-stained whole mounts of the cat retina. They are referred to as A-type and B-type cells. The two types differ in their dendritic branching pattern, their overall size and the absence or presence of an axon. At every retinal position the dendrites of B-type cells branch more densely and overlap each other more frequently than do the dendrites of A-type cells. At equivalent retinal positions the dendritic field size of A-type cells is greater than that of B-type cells by a factor of about 1.5. Only B-type cells have an axon, which branches at the end into a large axon terminal system. The axons have no preferred direction of orientation. The stain-ability of horizontal cells by different Golgi methods is discussed.

Animals↗

Localization of natriuretic peptides and their activation of particulate guanylate cyclase and nitric oxide synthase in the retina.

In the vertebrate retina, cyclic guanosine monophosphate (cGMP) mediates photoreceptor signal transduction and modulates ion channel and gap junction conductivity. Although most previous studies have focused on its synthesis by nitric oxide (NO)-sensitive soluble guanylate cyclase, cGMP is also synthesized by NO-insensitive particulate guanylate cyclases (pGC). Natriuretic peptides and their associated pGC-coupled receptors have been reported in retina, but few studies have localized these natriuretic peptides or pGCs to specific retinal cells or demonstrated that activation of pGCs by natriuretic peptides increases cGMP synthesis. In this study, we immunocytochemically localized atrial, brain, and C-type natriuretic peptide-like immunoreactivity (ANP-LI, BNP-LI, and CNP-LI, respectively) in turtle retina by using isoform specific antisera, and determined the ability of each natriuretic peptide isoform to increase cGMP-like immunoreactivity (cGMP-LI) in retinal cells. ANP-LI and CNP-LI were localized in sparsely distributed amacrine cells with thin, intermittently varicose processes in the inner plexiform layer. BNP-LI was localized to abundant somata in the inner nuclear and ganglion cell layers and in specific amacrine and horizontal cells. Stimulation of turtle eyecups with each of these natriuretic peptides increased cGMP-LI in multistratified amacrine cells by means of NO-independent mechanisms in the central retina, and in select amacrine and bipolar cells in the peripheral retina by a nitric oxide-dependent mechanism. These results indicate that natriuretic peptides can modulate the synthesis of cGMP in select retinal neurons by two distinct signal transduction pathways in a regionally specific manner.

Animals↗

The ocular morphology of the southern hemisphere lamprey Mordacia mordax Richardson with special reference to a single class of photoreceptor and a retinal tapetum.

A description is given of the morphological and ultrastructural characteristics of the eye of Mordacia mordax, one of the two parasitic species of the southern hemisphere lamprey family Mordaciidae. The avascular retina is well-differentiated, containing two ganglion cell sublaminae, two layers of horizontal cells and a large population of displaced bipolar cells. Intermediate filaments, which stain positively for glial fibrillary acidic protein (GFAP), lie in perpendicularly-arranged bundles between the two layers of horizontal cells. M. mordax contains only a single type of rod-like photoreceptor, in contrast to representatives in the other two lamprey families Geotriidae and Petromyzontidae, which possess three (two cone-like and one rod-like) and two (one cone and one rod) types of photoreceptor, respectively. The large photoreceptors of M. mordax possess a large mitochondrial ellipsosome, a cylindrical outer segment with incisures, and a nucleus immediately abutting the outer limiting membrane. The photoreceptors are arranged in an hexagonal array with their outer segments surrounded by a retinal tapetum which elicits a yellow fundal eyeshine. The tapetum contains both diffuse (vitread spheroidal granules) and specular (sclerad needles) reflectors. The characteristics of the eye of M. mordax are related to the diurnal burrowing of this species during its upstream migration, a feature not found in the other two extant families of lampreys. The absence of cone-like photoreceptors and the presence of a tapetum suggest that although photopic vision is probably greatly diminished, characteristics have been evolved by the eye to maximize light capture and increase sensitivity in the low light intensities encountered during this species' nocturnal migration.

Animals↗

Depth perception and cortical physiology in normal and innate microstrabismic cats.

Evidence is presented that innate microstrabismus and abnormal cortical visual receptive-field properties can occur also in cats without any apparent involvement of the Siamese or albino genetic abnormalities in their visual system. A possible cause for microstrabismus in these cats may be sought in an abnormally large horizontal distance between blind spot and area centralis indicated by a temporal displacement of the most central receptive fields on both retinae. Depth perception was found to be impaired in cats with innate microstrabismus. Behavioral measurements using a Y-maze revealed in four such cats that the performance in recognizing the nearer of two random-dot patterns did not improve when they were allowed to use both eyes instead of only one. The ability of microstrabismic cats to perceive depth under binocular viewing conditions only corresponded to the monocular performance of five normal cats. Electrophysiological recordings were performed in the visual cortex (areas 17 and 18) of four awake cats, two normal, and two innate microstrabismic animals. Ocular dominance and orientation tuning of single neurons in area 17 and 18 were analyzed quantitatively. The percentage of neurons in area 17 and 18 which could be activated through either eye was significantly reduced to 49.7% in the microstrabismic animals when compared to the normal cats (74.8%). "True binocular cells," which can only be activated by simultaneous stimulation of both eyes, were significantly less frequent (1.6%) in microstrabismic cats than in normal animals (10.4%). However, subthreshold binocular interactions were identical in both groups of animals. In the strabismic animals, long-term binocular stimulation of monocular neurons did not give a clear indication of alternating use of one or the other eye. The range of stimulus orientations leading to discharge rates above 50% of the maximal response, i.e. the half-width of the orientation tuning curves, was the same in the two groups of cats. However, orientation sensitivity, i.e. the alternation in discharge rate per degree change in stimulus orientation, was higher in cortical cells of normal cats than in those of microstrabismic cats. In normal and microstrabismic cats, no clear sign of an "oblique effect," i.e. the preference of cortical neurons for vertical and horizontal orientations compared to oblique orientations, could be found neither in the incidence of cells with horizontal or vertical preferred orientation nor in the sharpness of orientation tuning and sensitivity of these neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Two systems of branching axons in monkey's retina.

Several monkey retinae were stained, by using the reduced silver technique, in order to analyse long-distance intraretinal connections. Long, bifurcating processes covering very large areas were identified. Morphological investigation of these processes suggest that they are members of two different systems of branching axons. The first population of these processes originates as axon collaterals from cell in the ganglion cell layer. These cells have a relatively large, elongated soma and straight, sparsely branching dendrites, stratified in the vitreal half of the inner plexiform layer. The main axon (0.6 microns average diameter) passes along the optic fibre bundles, disappearing into the optic disk, whilst its collaterals run mainly in the inner plexiform layer. A cell showing similar morphology has also been found in the ganglion cell layer of a cat retina. The second population of processes consists of very thick fibres (2.1 microns average diameter) apparently originating from the optic disk. The main branches run in the space between the optic fibre layer and the ganglion cell layer, with short, secondary processes crossing the ganglion cel layer orthogonally. Many higher-order processes originate from the second-order branches; these run almost horizontally in the inner plexiform layer. The ganglion cells generating axon collaterals may constitute an intraretinal firing synchronization system, or they may be a residual feature of retinal development. The centrifugal fibres may be related to the sensitivity control during retinal dark adaptation.

Animals↗

Two-photon imaging reveals somatodendritic chloride gradient in retinal ON-type bipolar cells expressing the biosensor Clomeleon.

A somatodendritic gradient of Cl(-) concentration ([Cl(-)](i)) has been postulated to generate GABA-evoked responses of different polarity in retinal bipolar cells, hyperpolarizing in OFF cells with low dendritic [Cl(-)](i), and depolarizing in ON cells with high dendritic [Cl(-)](i). As glutamate released by the photoreceptors depolarizes OFF cells and hyperpolarizes ON cells, the bipolars' antagonistic receptive field (RF) could be computed by simply integrating glutamatergic inputs from the RF center and GABAergic inputs from horizontal cells in the RF surround. Using ratiometric two-photon imaging of Clomeleon, a Cl(-) indicator transgenically expressed in ON bipolar cells, we found that dendritic [Cl(-)](i) exceeds somatic [Cl(-)](i) by up to 20 mM and that GABA application can lead to Cl(-) efflux (depolarization) in these dendrites. Blockers of Cl(-) transporters reduced the somatodendritic [Cl(-)](i) gradient. Hence, our results support the idea that ON bipolar cells employ a somatodendritic [Cl(-)](i) gradient to invert GABAergic horizontal cell input.

Animals↗

Frequency characteristics of retinal neurons in the carp.

Frequency characteristics of various retinal neurons in the carp were studied using sinusoidally modulated light as an input. They were affected by both intensity and pattern of illumination. In the horizontal cells, in which the effect of light intensity was studied most extensively, an increase in the light intensity brought about a decrease of the gain, which was more marked at lower frequencies, resulting in a shift of cutoff frequency towards higher frequencies and in a slight low frequency attenuation. A decrease in the area illuminated had an effect similar to a decrease in the light intensity. In the receptor, the low frequency attenuation was not apparent even at high light intensities. The adaptation process in receptors was not sufficient to explain the low frequency attenuation in the horizontal cells, and a possible contribution of negative feedback from horizontal cells to receptors was suggested. In the bipolar cell, the lateral interaction played an important role. An increase in an area resulted in the suppression of the response at low frequencies where the phases of the center and the surround responses were opposed, but in the augmentation near 5 Hz where the two responses were in phase. In amacrine cells, a low frequency attenuation and a phase advance at low frequencies were very prominent, and were considered to be due mainly to a process designated here as the neural adaptation.

Adaptation, Ocular↗

Lamina formation in the Mongolian gerbil retina (Meriones unguiculatus).

Retinae of nocturnal rodents, such as mice and rats, are almost exclusively rod-dominated. The gerbil, in contrast, shows active periods during day and night and uses both rod- and cone-based vision. However, its retina has not been studied in detail, except for one developmental study analysing its prenatal period (Wikler et al. 1989). Here, the formation of the laminar structure of the gerbil retina was studied from birth until late adult stages. At birth, the retina consisted of a wide neuroblastic layer, with 30% of cells still dividing, a rate decreasing to nearly zero by P6. Shortly after birth, segregation of a ganglion cell layer began. All retinal layers reached their final size around P20, as determined from DAPI-stained cryosections. Muller glial cells developed their typical structure from P1 onwards, e.g. announcing an outer plexiform layer (OPL) at P5, as analysed by the Ret-G7 and glutamine synthetase antibodies. The analyses of the inner retina were performed by antibodies to calretinin (CR) and calbindin (CB). CR is expressed in ganglion cells followed by amacrine cells from P1 onwards; their processes formed four subbands in the inner plexiform layer (IPL) and appeared sequentially after P5 until P20. CB stained a subtype of horizontal cells with their processes into the OPL from P14 onwards. The rod-specific antibody rho4D2 announced photoreceptors at P4, showing signs of outer segments from P10 onwards. The study shows that the formation of all retinal layers in the gerbil occurs postnatally. This and the fact that the gerbil retina is not exclusively rod-dominated could render the gerbil a valuable model for in vitro studies of retinogenesis in rodents.

Aging↗

Time course of appearance of GABA and GABA receptors during retinal regeneration in the adult newt.

Appearance and maturation of the GABA (gamma-aminobutyric acid) system during newt retinal regeneration were studied by electrophysiological, immunohistochemical, and biochemical techniques. (1) Responses to GABA appeared in neurons dissociated from regenerating retinae before the segregation of the plexiform layers; whereas (2) GABA immunoreactivity appeared at sites of the presumptive horizontal cell and amacrine cell layers at the beginning of the segregation of these layers. During subsequent regeneration, GABA-immunoreactive cells at the amacrine cell layer increased in number and extended lateral processes, forming a GABA-immunoreactive inner plexiform layer. Also GABA immunoreactivity increased in the region of the outer plexiform layer, but not their somata which showed decreased GABA immunoreactivity. (3) GABA synthesis in the retina increased significantly at the beginning of the segregation of the plexiform layers. These results suggest that the increase of GABA synthesis during retinal regeneration correlates well with the development of GABA-immunoreactive cells and that functional GABA receptors appear earlier than increased GABA synthesis.

Animals↗

Horizontal cells reveal cone type-specific adaptation in primate retina.

The human cone visual system maintains contrast sensitivity over a wide range of ambient illumination, a property known as light adaptation. The first stage in light adaptation is believed to take place at the first neural step in vision, within the long, middle, and short wavelength sensitive cone photoreceptors. To determine the properties of adaptation in primate outer retina, we measured cone signals in second-order interneurons, the horizontal cells, of the macaque monkey. Horizontal cells provide a unique site for studying early adaptational mechanisms; they are but one synapse away from the photoreceptors, and each horizontal cell receives excitatory inputs from many cones. Light adaptation occurred over the entire range of light levels evaluated, a luminance range of 15-1,850 trolands. Adaptation was demonstrated to be independent in each cone type and to be spatially restricted. Thus, in primates, a major source of sensitivity regulation occurs before summation of cone signals in the horizontal cell.

Adaptation, Physiological↗

Retinal ganglion cells in two teleost species, Sebastiscus marmoratus and Navodon modestus.

Distribution patterns of ganglion cells in the retina were examined in Nissl-stained retinal whole mounts of Sebastiscus and Navodon. The existence of area centralis in the temporal retina in both species suggests binocular vision. In Navodon, another high density area was found in the nasal retina, and a dense band of ganglion cells was observed along the horizontal axis between the two high-density areas. There is an obvious trend for the ganglion cell size to increase as the density decreases. The total number of ganglion cells was estimated to be about 45 X 10(4) in Sebastiscus and 87 X 10(4) in Navodon, whereas the total number of optic nerve fibers was about 35 X 10(4) and 70 X 10(4), respectively. The retinal ganglion cells labeled with HRP were classified into six types according to such morphological characteristics as size, shape, and location of the soma as well as dendritic arborization pattern. Type I cells have a small round or oval soma in the ganglion cell layer and a small dendritic field in the inner plexiform layer. Type II cells are similar to type I cells, but the dendrites arborize more closely to the ganglion cell layer in the innermost region of the inner plexiform layer. Type III cells have a medium-sized round soma in the ganglion cell layer, and the dendrites extend in an extremely wide area in the inner plexiform layer with few branches. Type IV cells have a large soma which is located in the ganglion cell layer. Dendrites emanate from the soma in all directions, branching out several times within a rather small region in the innermost part of the inner plexiform layer. Type V cells have large somata of various shapes, usually dislocated to the inner plexiform or granular layer. The dendrites extend in every direction and occupy an extremely large area in the inner plexiform layer. Type VI cells have the largest somata, which are also dislocated to the inner plexiform or granular layer. Type VI cells have a characteristic triangular or fan-shaped dendritic field. Soma size and the axon diameter are intimately linked, that is, small somata of type I and II cells give off thin axons, and large somata of type V and VI give off thick axons.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Color induction via non-opponent lateral interactions in the human retina.

Retinal connections causing colors in Benham's top (pattern induced flicker colors, PIFCs) are investigated by psychophysical experiments. PIFCs are still seen when stimuli to different cones are demodulated selectively, indicating the involvement of non-opponent channels. PIFCs also occur on retinal areas next to those affected by modulated stimuli; further, both monochromat and dark-adapted trichromats perceive PIFCs which are achromatic. These additional findings point to horizontal cells as neuronal mediators of modulated excitation leading to PIFCs. The unspecifity of the postulated connection with respect to cone types agrees with anatomic findings of Boycott, B. B., Hopkins, J. M. and Sperling, H. G. (1987, Proceedings of the Royal Society of London, B, 229, 345-379) on horizontal cells.

Color Perception↗

Pre- and post-synaptic effects of manipulating surface charge with divalent cations at the photoreceptor synapse.

Persistence of horizontal cell (HC) light responses in extracellular solutions containing low Ca2+ plus divalent cations to block Ca2+ currents (ICa) has been attributed to Ca2+-independent neurotransmission. Using a retinal slice preparation to record both ICa and light responses, we demonstrate that persistence of HC responses in low [Ca2+]o can instead be explained by a paradoxical increase of Ca2+ influx into photoreceptor terminals arising from surface charge-mediated shifts in ICa activation. Consistent with this explanation, application of Zn2+ or Ni2+ caused a hyperpolarizing block of HC light responses that was relieved by lowering [Ca2+]o. The same concentrations of Zn2+ and Ni2+ reduced the amplitude of ICa at the rod dark potential and this reduction was relieved by a hyperpolarizing shift in voltage dependence induced by lowering [Ca2+]o. Block of ICa by Mg2+, which has weak surface charge effects, was not relieved by low [Ca2+]o. Recovery of HC responses in low [Ca2+]o was assisted by enhancement of rod light responses. To bypass light stimulation, OFF bipolar cells were stimulated by steps to -40 mV applied to presynaptic rods during simultaneous paired recordings. Consistent with surface charge theory, the post-synaptic current was inhibited by Zn2+ and this inhibition was relieved by lowering [Ca2+]o. Nominally divalent-free media produced inversion of HC light responses even though rod light responses remained hyperpolarizing; HC response inversion can be explained by surface charge-mediated shifts in ICa. In summary, HC light responses modifications induced by low divalent cation solutions can be explained by effects on photoreceptor light responses and membrane surface charge without necessitating Ca2+-independent neurotransmission. Furthermore, these results suggest that surface charge effects accompanying physiological changing divalent cation levels in the synaptic cleft may provide a means for modulating synaptic output from photoreceptors.

Animals↗