The primate small bistratified blue-ON ganglion cell responds to the onset of a short wavelength light and to the offset of a middle and long wavelength light. We hypothesize, based on the small bistratified cell's well described anatomy (Dacey, 1993), and extracellular recordings from blue-ON ganglion cells combined with pharmacology (Crook et al., 2009), that the blue-ON cell's basic wavelength opponent response is generated primarily by excitatory conductances originating from ON and OFF bipolar cell pathways. The blue-ON bipolar cell exclusively contacts short (S)-wavelength sensitive cone photoreceptors and relays to the small bistratified ganglion cell's inner dendrites; and OFF diffuse bipolar cells contact long (L)- and middle (M)-wavelength sensitive cone photoreceptors and synapse on the outer dendrites. Recording from macaque monkey blue-ON cells in vitro in whole-cell voltage-clamp, we show that light evoked ON and OFF responses, to S- and LM-cone modulation, respectively, are produced primarily by excitatory conductances. ON pathway block with the mGluR6 agonist, L-AP-4, knocks out the excitatory conductance to S-cone modulation, but spares the excitatory conductance to LM-cone modulation. Blocking inner retinal inhibitory pathways with the addition of GABAa/c receptor antagonists, TPMPA and GABAzine, and the glycine receptor antagonist strychnine, further isolates this conductance. These results are consistent with the blue-ON cells basic wavelength opponent response originating from ON and OFF bipolar pathways. Inhibition, from inner retinal amacrine cells, appears to shape the kinetics of the excitatory inputs. CrookJD J Neurosci 29( 26):8372–8387 (2009). DaceyDM Vis Neurosci 10( 6):1081–1098 (1993). YIA

Horizontal cells typical of the vertebrate retina are strongly coupled by gap junctions. The resulting horizontal cell network has extremely large receptive fields that extend well beyond the boundaries of a single dendritic tree. This network has been modeled as a syncytium of cytoplasm bounded by cell membrane (Lamb 1976; Naka & Rushton, 1967). Horizontal cells in the primate retina are also coupled by gap junctions, but their receptive fields are relatively small and in some cases may approximate the span of the dendritic tree of an individual cell (Packer & Dacey, 2002). The receptive field of the macaque H1 horizontal cell type has been modeled as the sum of two spatial components: a strong but small diameter excitatory center, and a weak but broad excitatory surround. Here we explore the hypothesis that the receptive field center of H1 cells derives from direct cone synaptic input and that the synergistic surround derives from gap-junctional coupling among H1 cell neighbors. We measured the receptive field structure of H1 cells in the presence of carbenoxolone, a gap junction blocker, to determine the effects of uncoupling center and surround components and compared these data to a neural simulation of the H1 network in which gap-junctional conductance could be manipulated. Carbenoxolone reduced the surround component and eliminated irregularities in spatial structure thought to be associated with the surround. The effects of carbenoxolone could be mimicked by manipulating gap-junctional conductance in an H1 cell network simulation. These results provide strong support for the two-component model of H1 receptive field structure. In addition, carbenoxolone eliminated a slow depolarization following light onset thought to be mediated by cone-H1 feedback (Kamermans & Spekreijse, 1999). Low concentrations of cobalt, a calcium channel blocker that spares gap junctions, had an effect similar to that of carbenoxolone but did not affect receptive field structure. These results are consistent with a calcium-mediated mechanism of feedback from H1 cells to cones that is independent of the synergistic two-component model of receptive field organization.

Although the center-surround receptive field is a fundamental property of retinal ganglion cells, the circuitry that mediates surround inhibition remains controversial. We examined the contribution of horizontal cells and amacrine cells to the surround of parasol ganglion cells of macaque and baboon retina by measuring receptive field structure before and during the application of drugs that have been shown previously to affect surrounds in a range of mammalian and nonmammalian species. Carbenoxolone and cobalt, thought to attenuate feedback from horizontal cells to cones, severely reduced the surround. Tetrodotoxin, which blocks sodium spiking in amacrine cells, and picrotoxin, which blocks the inhibitory action of GABA, only slightly reduced the surround. These data are consistent with the hypothesis that the surrounds of light-adapted parasol ganglion cells are generated primarily by non-GABAergic horizontal cell feedback in the outer retina, with a small contribution from GABAergic amacrine cells of the inner retina.

Analysis of cone inputs to primate parvocellular ganglion cells suggests that red–green spectral opponency results when connections segregate input from long wavelength (L) or middle wavelength (M) sensitive cones to receptive field centers and surrounds. However, selective circuitry is not an obvious retinal feature. Rather, cone receptive field surrounds and H1 horizontal cells get mixed L and M cone input, likely indiscriminately sampled from the randomly arranged cones of the photoreceptor mosaic. Red–green spectral opponency is consistent with random connections in central retina where the mixed cone ganglion cell surround is opposed by a single cone input to the receptive field center, but not in peripheral retina where centers get multiple cone inputs. The selective and random connection hypotheses might be reconciled if cone type selective circuitry existed in inner retina. If so, the segregation of L and M cone inputs to receptive field centers and surrounds would increase from horizontal to ganglion cell, and opponency would remain strong in peripheral retina. We measured the relative strengths of L and M cone inputs to H1 horizontal cells and parasol and midget ganglion cells by recording intracellular physiological responses from morphologically identified neurons in anin vitropreparation of the macaque monkey retina. The relative strength of L and M cone inputs to H1 and ganglion cells at the same locations matched closely. Peripheral midget cells were nonopponent. These results suggest that peripheral H1 and ganglion cells inherit their L and M cone inputs from the photoreceptor mosaic unmodified by selective circuitry.

In non-mammalian vertebrates, retinal bipolar cells show center-surround receptive field organization. In mammals, recordings from bipolar cells are rare and have not revealed a clear surround. Here we report center-surround receptive fields of identified cone bipolar cells in the macaque monkey retina. In the peripheral retina, cone bipolar cell nuclei were labeled in vitro with diamidino-phenylindole (DAPI), targeted for recording under microscopic control, and anatomically identified by intracellular staining. Identified cells included 'diffuse' bipolar cells, which contact multiple cones, and 'midget' bipolar cells, which contact a single cone. Responses to flickering spots and annuli revealed a clear surround: both hyperpolarizing (OFF) and depolarizing (ON) cells responded with reversed polarity to annular stimuli. Center and surround dimensions were calculated for 12 bipolar cells from the spatial frequency response to drifting, sinusoidal luminance modulated gratings. The frequency response was bandpass and well fit by a difference of Gaussians receptive field model. Center diameters were all two to three times larger than known dendritic tree diameters for both diffuse and midget bipolar cells in the retinal periphery. In one instance intracellular staining revealed tracer spread between a recorded cell and its nearest neighbors, suggesting that homotypic electrical coupling may contribute to receptive field center size. Surrounds were around ten times larger in diameter than centers and in most cases the ratio of center to surround strength was ∼1. We suggest that the center-surround receptive fields of the major primate ganglion cell types are established at the bipolar cell, probably by the circuitry of the outer retina.

We simultaneously measured the light being absorbed at every location across a patch of fresh excised primate retina. Rod and cone axial absorptances were 0.5 and 0.3 respectively, consistent with the specific density of photopigment and outer segment length. The photometric quantum efficiency of patches of peripheral retina was 0.12. There was little evidence for directional sensitivity in patches of rods. Great care must be taken when interpreting bright spots in images of excised retina.

We introduce a new technique for classifying many photoreceptors simultaneously in fresh, excised primate retina on the basis of their absorptance spectra. Primate retina is removed from the pigment epithelium and illuminated under a microscope from the same direction as in the intact eye. To facilitate the guiding of light into the receptor outer segments, the optical axes of the photoreceptors are oriented parallel to the optical axis of the microscope. Photoreceptor outer-segment tips are imaged on a charge-coupled device array, which provides radiometric measurements of the light passing through each photoreceptor. These images are acquired sequentially at three wavelengths chosen to maximize the absorptance differences among the three cone photopigments. After the photopigment is bleached, a second set of three images is acquired. The ratios of the images before and after bleaching at each wavelength are photopigment transmittance maps of the retina. These are combined into a single trichromatic image showing the distribution of photopigment if the retina could be viewed directly in white light without bleaching. We have found patches of receptors in peripheral macaque retina where the measured absorptance at the wavelength of maximum absorptance is consistent with the predicted axial absorptance of th photopigment. The cones in these patches cluster into two groups corresponding to the middle wavelength- sensitive (n = 53, mean absorptance = 0.28) and the long wavelength- sensitive (n = 63, mean absorptance = 0.30) cones. The mean absorptances of 273 macaque and 183 human rods were 0.51 and 0.41, respectively.

A fixation-and-flash technique has been developed to provide control over the retinal eccentricities of stimuli presented to infant subjects, to within a few degrees of visual angle. The technique is a variant of forced-choice preferential looking (FPL). An adult observer triggers presentation of the test stimulus when she judges that the infant is fixating a centrally located fixation target. The stimuli are short in relation to the infant's refixation latency. Auxiliary experiments confirmed that on most trials the stimuli fell within ± 4° of the designated eccentricity. Test fields of two sizes, 3.1 and 17°, were presented to 1-month-old infants at one of four retinal locations, 9, 18, 27 and 36° eccentric. The infants' data show a perfect area-intensity tradeoff at all four locations. Adult control subjects showed summation over areas of only 1–2°. The results are discussed in relation to other evidence of coarse spatial processing in human infants and other immature mammalian systems.