Molecular Classification and Comparative Taxonomics of Foveal and Peripheral Cells in Primate Retina

We studied the relationship between the morphology of ganglion cells and the spatial density of photoreceptors in the retina of two Old World primates, human and macaque monkey; the diurnal New World marmoset Callithrix jacchus; and the cat. Ganglion cells in macaque and marmoset were labelled by intracellular injection with Neurobiotin or by DiI diffusion labelling in fixed tissue. Cone photoreceptor densities were measured from the same retinas. Supplemental data for macaque and data for human and cat were taken from published studies. For the primates studied, the central retina is characterised by a constant numerical convergence of cones to ganglion cells. Midget ganglion cells derive their input, via a midget bipolar cell, from a single cone. Parasol cells derive their input from 40-140 cones. Outside the central retina, the convergence increases with eccentricity. The convergence to beta cells in the cat retina is very close to that for parasol cells in primate retina. The convergence of rod photoreceptors to ganglion cells is similar in human, macaque, and marmoset, with parasol cells receiving input from 10-15 times more rods than midget cells. The low convergence of cones to midget cells in human and macaque retinas is associated with distinctive dendritic "clusters" in midget cells' dendritic fields. Convergence in marmoset is higher, and the clusters are absent. We conclude that the complementary changes in photoreceptor density and ganglion cell morphology should be considered when forming linking hypotheses between dendritic field, receptive field, and psychophysical properties of primate vision.

In this paper, we extend our previous light microscopic (LM) study of substance P (SP)-containing amacrine and ganglion cell types of the human retina (Cuenca et al. [1995] J. Comp. Neurol. 356:491-504) to an electron microscopic (EM) and confocal-imaging study in order to reveal synaptic circuitry and putative input and output neurons. SP-immunoreactive (-IR) amacrine cells in primate retina are typically wide-field cells with large cell bodies occurring in normal or displaced positions relative to the inner plexiform layer (IPL). Their main dendrites bear many spines and are monostratified in stratum 3 (S3) of the IPL. Axon-like processes arise from dendrites close to the cell body and run for hundreds of microns at the same level as the dendrites, thus forming a relatively dense plexus in S3 of the IPL. SP-IR axon processes also climb to S1 to surround some amacrine cell bodies, and others pass into the outer plexiform layer (OPL). Still other axons run down to the ganglion cell layer, where they encircle SP-IR ganglion cells and pass on to end in the nerve fiber layer. The SP-IR ganglion cell types have large cell bodies (20-22 microm diameter) and dendrites that costratify in S3 among the SP-IR amacrine cell processes. Double immunostaining and study by confocal microscopy reveals that SP-IR amacrine cells in the monkey colocalize gamma-aminobutyric acid (GABA). Their main plexus of dendrites in S3 of the IPL is skirted on the S2/S3 border by cone bipolar axons that stain for calbindin but intermingles primarily with glycinergic bipolar cell types of S3 and S3-S4. Strongly GABA-IR/weakly glycine-IR amacrine cell bodies, in addition to the SP-IR large-bodied ganglion cell type, are targets of encircling SP-IR axon processes. EM study of the human SP-IR amacrine cell indicates that input synapses to its dendrites are from bipolar cell axons of the S2/S3 border, S3, and the S3/S4 border of the IPL neuropil (33% of the synaptic input) and from amacrine cell processes (67% of the synaptic input). The input amacrine cells are of at least two distinct types based on cytological criteria. Synaptic output from the SP-IR amacrine cell dendrites is to bipolar cell axons as reciprocal synapses (31%), to amacrine cells (40%), and to ganglion cell profiles, primarily in S3 (29%) of the IPL. The SP-IR axons synapse upon SP-IR ganglion cell bodies and axons, upon normally placed and displaced amacrine cell bodies, and upon bipolar cell dendrites in the OPL. In addition, they appear to synapse among themselves. We shall discuss a wiring diagram and the possible role of SP-IR amacrine cells in the primate retina.

ABSTRACTThe human retina is a complex neural tissue that detects light and sends visual information to the brain. However, the molecular and cellular processes that underlie aging primate retina remain unclear. Here, we provide a comprehensive transcriptomic atlas based on 119 520 single cells of the foveal and peripheral retina of humans and macaques covering different ages. The molecular features of retinal cells differed between the two species, suggesting distinct regional and species specializations of the human and macaque retinae. In addition, human retinal aging occurred in a region- and cell-type-specific manner. Aging of human retina exhibited a foveal to peripheral gradient. MYO9A− rods and a horizontal cell subtype were greatly reduced in aging retina, indicating their vulnerability to aging. Moreover, we generated a dataset showing the cell-type- and region-specific gene expression associated with 55 types of human retinal disease, which provides a foundation to understanding of the molecular and cellular mechanisms underlying human retinal diseases. Such datasets are valuable to understanding of the molecular characteristics of primate retina, as well as molecular regulation of aging progression and related diseases.

Primate retinas contain two major ganglion cell types. Midget (or P type) cells have relatively sustained responses to light; the amplitude and polarity of these responses vary with stimulus wavelength. Parasol (or M type) cells are more sensitive to stimulus contrast and respond more transiently but are not selective for color. Both types can be further subdivided into a and b subtypes, according to the level of their dendritic stratification in the inner plexiform layer. To determine whether differences in receptors for amino acid transmitters are the basis for any differences in ganglion cell light responses, we made whole-cell, patch-clamp recordings from identified ganglion cells in slice preparations of macaque and baboon retinas. We found that midget and parasol cells of both a and b types had similar responses to excitatory amino acids, including kainate, alpha-amino-3-hydroxy-5-methylisoxalzole-4-propionic acid, and N-methyl-D-aspartate, with reversal potentials near the equilibrium potential for cations. Kainate responses were blocked by 6,7-dinitroquinoxaline, and N-methyl-D-aspartate responses were blocked by D-(-)-2-amino-7-phosphonoheptanoic acid. The four types of ganglion cells also had similar responses to bath-applied inhibitory amino acids. All cells had both gamma-aminobutyric acid and glycine receptors with reversal potentials near the equilibrium potential for Cl-, and the relative amplitudes of the responses to excitatory and inhibitory amino acids were similar among the various cell types. These results suggest that the differences in response properties of the different classes of ganglion cells in primate retina may be determined, to a significant degree, by the properties of the amacrine and bipolar cells that provide their input rather than by the nature of their postsynaptic receptors.

Center surround receptive field structure of cone bipolar cells in primate retina

Evidence that Circuits for Spatial and Color Vision Segregate at the First Retinal Synapse

We examined the functional microcircuitry of cone inputs to blue-ON/yellow-OFF (BY) ganglion cells in the macaque retina using multielectrode recording. BY cells were identified by their ON responses to blue light and OFF responses to red or green light. Cone-isolating stimulation indicated that ON responses originated in short (S) wavelength-sensitive cones, whereas OFF responses originated in both long (L) and middle (M) wavelength-sensitive cones. Stimulation with fine spatial patterns revealed locations of individual S cones in BY cell receptive fields. Neighboring BY cells received common but unequal inputs from one or more S cones. Inputs from individual S cones differed in strength, indicating different synaptic weights, and summed approximately linearly to control BY cell firing.

The midget pathway of the primate retina provides the visual system with the foundations for high spatial resolution and color perception. An essential contributor to these properties is center-surround organization, in which responses from the central area of a cell's receptive field are antagonized by responses from a surrounding area. Two key questions about center-surround organization are unresolved. First, the surround is largely or completely due to negative feedback from horizontal cells to cones: how can this feedback be reconciled with the popular difference of Gaussians (DOG) model, which implies feedforward inhibition? Second, can the spatial extent of center and surround be predicted from the components-optics, horizontal cell receptive field, ganglion cell dendrites-that give rise to them? We address these questions with a computational model of midget pathway signal processing in macaque retina; model parameters are derived from published literature. We show that, contrary to the DOG model, the surround's effect is better treated as divisive. A simplified version of our model-a ratio of Gaussians (ROG) model-has practical advantages over the DOG, such as accounting for spatiotemporal interactions and pulse responses. The ROG model also shows that both center and surround radii can be calculated from a sum of squared radii of their components. Finally, chromatic antagonism between center and surround in the full model predicts cone opponency as a function of eccentricity. We suggest that a signal-processing model gives new insight into retinal function.NEW & NOTEWORTHY We simulated signal processing from cones to midget ganglion cells in the monkey retina and found that: 1) center/surround structure is better described as a ratio of Gaussian functions than as the traditional difference of Gaussians; 2) ganglion cell center and surround radii can be calculated from a sum of squares of radii in upstream stages; 3) the model can predict chromatic dominance in the center and surround mechanisms as a function of eccentricity.

Orientation sensitivity of ganglion cells in primate retina

Visual information is conveyed to the brain by the retinal ganglion cells. Midget ganglion cells serve fine spatial vision by summing excitation from a receptive field 'centre', receiving input from a single cone in the central retina, with lateral inhibition from a receptive field 'surround', receiving input from many surrounding cones. Midget ganglion cells are also thought to serve colour opponent vision because the centre excitation is from a cone of one spectral type, while the surround inhibition is from cones of the other type. The two major cone types, middle(M)- and long-(L)wavelength sensitive, are equally numerous and randomly distributed in the primate central retina, so a spectrally homogeneous surround requires that the cells mediating lateral interactions (horizontal or amacrine cells) receive selective input from only one cone type. Horizontal cells cannot do this because they receive input indiscriminately from M and L cones. Here we report that the amacrine cells connected to midget ganglion cells are similarly indiscriminate. The absence of spectral specificity in the inhibitory wiring raises doubt about the involvement of midget ganglion cells in colour vision and suggest that colour opponency may instead be conveyed by a different type of ganglion cell.

The primate visual system consists of parallel pathways initiated by distinct cell types in the retina that encode different features of the visual scene. Small bistratified cells (SBCs), which form a major projection to the thalamus, exhibit blue-ON/yellow-OFF [S-ON/(L+M)-OFF] light responses thought to be important for high-acuity color vision. However, the spatial processing properties of individual SBCs and their spatial arrangement across the visual field are poorly understood. The present study of peripheral primate retina reveals that contrary to previous suggestions, SBCs exhibit center-surround spatial structure, with the (L+M)-OFF component of the receptive field approximately 50% larger in diameter than the S-ON component. Analysis of response kinetics shows that the (L+M)-OFF response in SBCs is slower than the S-ON response and significantly less transient than that of simultaneously recorded OFF-parasol cells. The (L+M)-OFF response in SBCs was eliminated by bath application of the metabotropic glutamate receptor agonist L-APB. These observations indicate that the (L+M)-OFF response of SBCs is not formed by OFF-bipolar cell input as has been suspected and suggest that it arises from horizontal cell feedback. Finally, the receptive fields of SBCs form orderly mosaics, with overlap and regularity similar to those of ON-parasol cells. Thus, despite their distinctive morphology and chromatic properties, SBCs exhibit two features of other retinal ganglion cell types: center-surround antagonism and regular mosaic sampling of visual space.

The major inhibitory neurotransmitter glycine is used by about half of the amacrine cells in the retina. Amacrine cells provide synaptic output to bipolar, ganglion, and other amacrine cells. The present study investigated whether different bipolar and amacrine cell types in the primate retina differ with respect to the expression of glycine receptor (GlyR) subtypes. Antibodies specific for the alpha1, alpha2, and alpha3 subunits of the GlyR were combined with immunohistochemical markers for bipolar and amacrine cells and applied to vertical sections of macaque (Macaca fascicularis) and marmoset (Callithrix jacchus) retinae. For all subunits, punctate immunoreactivity was expressed in the inner plexiform layer. The GlyRalpha2 immunoreactive (IR) puncta occur at the highest density, followed by GlyR(alpha)3 and GlyR(alpha)1 IR puncta. Postembedding electron microscopy showed the postsynaptic location of all subunits. Double immunofluorescence demonstrated that the three alpha subunits are clustered at different postsynaptic sites. Two OFF cone bipolar cell types (flat midget and diffuse bipolar DB3), are predominantly associated with the alpha1 subunit. Two ON bipolar cell types, the DB6 and the rod bipolar cell, are predominantly associated with the alpha2 subunit. The glycinergic AII amacrine cell is presynaptic to the alpha1 subunit in the OFF-sublamina, and postsynaptic to the alpha2 subunit in the ON-sublamina. Another putative glycinergic cell, the vesicular glutamate transporter 3 cell, is predominantly presynaptic to the alpha2 subunit. The dopaminergic amacrine cell expresses the alpha3 subunit at a low density.

Visual abilities change over the visual field. For example, our ability to detect movement is better in peripheral vision than in foveal vision, but colour discrimination is markedly worse. The deterioration of colour vision has been attributed to reduced colour specificity in cells of the midget, parvocellular (PC) visual pathway in the peripheral retina. We have measured the colour specificity (red-green chromatic modulation sensitivity) of PC cells at eccentricities between 20 and 50 degrees in the macaque retina. Here we show that most peripheral PC cells have red-green modulation sensitivity close to that of foveal PC cells. This result is incompatible with the view that PC pathway cells in peripheral retina make indiscriminate connections ('random wiring') with retinal circuits devoted to different spectral types of cone photoreceptors. We show that selective cone connections can be maintained by dendritic field anisotropy, consistent with the morphology of PC cell dendritic fields in peripheral retina. Our results also imply that postretinal mechanisms contribute to the psychophysically demonstrated deterioration of colour discrimination in the peripheral visual field.

Transcriptomic and Chromatin Accessibility Analysis of the Human Macular and Peripheral Retinal Pigment Epithelium at the Single-Cell Level

The collective representation of visual space in high resolution visual pathways was explored by simultaneously measuring the receptive fields of hundreds of ON and OFF midget and parasol ganglion cells in isolated primate retina. As expected, the receptive fields of all four cell types formed regular mosaics uniformly tiling the visual scene. Surprisingly, comparison of all four mosaics revealed that the overlap of neighboring receptive fields was nearly identical, for both the excitatory center and inhibitory surround components of the receptive field. These observations contrast sharply with the large differences in the dendritic overlap between the parasol and midget cell populations, revealing an unexpected relationship between the anatomical and functional architecture in the dominant circuits of the primate retina.