Cone Inputs Research Articles

Temporal Sensitivity for Achromatic and Chromatic Flicker across the Visual Cortex.

The retinal ganglion cells (RGCs) receive different combinations of L, M, and S cone inputs and give rise to one achromatic and two chromatic postreceptoral channels. The goal of the current study was to determine temporal sensitivity across the three postreceptoral channels in subcortical and cortical regions involved in human vision. We measured functional magnetic resonance imaging (fMRI) responses at 7 T from three participants (two males, one female) viewing a high-contrast, flickering, spatially uniform wide field (∼140°). Stimulus flicker frequency varied logarithmically between 2 and 64 Hz and targeted the L + M + S, L - M, and S - (L + M) cone combinations. These measurements were used to create temporal sensitivity functions of the primary visual cortex (V1) across eccentricity and spatially averaged responses from the lateral geniculate nucleus (LGN), and the V2/V3, hV4, and V3A/B regions. fMRI responses reflected the known properties of the visual system, including higher peak temporal sensitivity to achromatic versus chromatic stimuli and low-pass filtering between the LGN and V1. Peak temporal sensitivity increased across levels of the cortical visual hierarchy. Unexpectedly, peak temporal sensitivity varied little across eccentricity within area V1. Measures of adaptation and distributed pattern activity revealed a subtle influence of 64 Hz achromatic flicker in area V1, despite this stimulus evoking only a minimal overall response. The comparison of measured cortical responses to a model of the integrated retinal output to our stimuli demonstrates that extensive filtering and amplification are applied to postretinal signals.

Cone-Opponent Ganglion Cells in the Primate Fovea Tuned to Noncardinal Color Directions.

A long-standing question in vision science is how the three cone photoreceptor types-long (L), medium (M), and short (S) wavelength sensitive-combine to generate our perception of color. Hue perception can be described along two opponent axes: red-green and blue-yellow. Psychophysical measurements of color appearance indicate that the cone inputs to the red-green and blue-yellow opponent axes are M vs. L + S and L vs. M + S, respectively. However, the "cardinal directions of color space" revealed by psychophysical measurements of color detection thresholds following adaptation are L vs. M and S vs. L + M. These cardinal directions match the most common cone-opponent retinal ganglion cells (RGCs) in the primate retina. Accordingly, the cone opponency necessary for color appearance is thought to be established in the cortex. While neurons with the appropriate M vs. L + S and L vs. M + S opponency have been reported in the retina and lateral geniculate nucleus, their existence continues to be debated. Resolving this long-standing debate is necessary because a complete account of the cone opponency in the retinal output is critical for understanding how downstream neural circuits process color. Here, we performed adaptive optics calcium imaging to noninvasively measure foveal RGC light responses in the living Macaca fascicularis eye. We confirm the presence of L vs. M + S and M vs. L + S neurons with noncardinal cone opponency and demonstrate that cone-opponent signals in the retinal output are more diverse than classically thought.

A dynamical adaptation model of visual spatiotemporal processing in cones and horizontal cells

In this work, we introduce a phenomenological model for the cone-horizontal cell assembly, including spatial integration and formation of receptive field-like structures. The model extends our previous dynamical adaptation description with gain control accounting for processes in single cones, valid in severe nonlinear regimes. Here, a spatially extended feedback mechanism is introduced from horizontal cells to cones to account for experimental evidence, contributing thus to the development of a center–surround receptive field in cones and downstream bipolar cells. Feedback gain is defined on different spatial scales by weighting spatial filters: a short scale accounting for cone input to the feedback mechanism and a large scale driven by the syncytium characteristics of horizontal cells. A third spatial scale improves the description, mimicking neighboring cone–cone coupling. This overall spatial integration couples to temporal signal processing, thus obtaining a spatiotemporal model of outer retina responses capable of reproducing nonlinear features in both dimensions (space and time). The model was tested and validated using measurements on horizontal cells from different studies, with excellent performance. By its phenomenological nature, signal processing properties are inferred from model parameters. The model can be used in arrays of processing units with more complex incoming patterns of visual stimuli.

Contributed Session III: In vivo calcium imaging of macaque foveolar retinal ganglion cells reveals spatiochromatic receptive field properties.

Here, we optically record responses to spatial and chromatic stimuli using a calcium indicator in the living macaque eye to characterize the receptive field (RF) properties of retinal ganglion cells (RGCs) serving the foveal center. GCaMP6s was expressed in three female macaques. Adaptive optics ophthalmoscopy was used to image fluorescence (488nm ex, 520/35nm em) from RGCs whose RF centers were driven by cones in the central 36 arcmin of the fovea and additional RGCs driven by cones in the central 6 arcmin of the foveola. Using cone isolating and luminance flicker (1.3deg, 0.15Hz, LED 420nm, 530nm, 660nm), we derived cone weights in over 250 RGCs. Using drifting gratings (1.9deg, 6Hz, 4-50c/deg, 561nm), we derived the spatial frequency responses of 15 L vs. M cone opponent RGCs at the foveolar center. Employing computational modeling (ISETbio), we inferred the full spatial difference of gaussians center and surround structure for those 15 cells. Of the 34 foveolar RGCs, 44% exhibited L vs. M cone opponency, 15% were L+M ON, 6% were -L-M OFF, and 35% showed only L or only M responses. The spatial frequency response functions of 12/15 L vs. M opponent cells peaked at high spatial frequencies (25-40c/deg) and had a strong bandpass characteristic. Our model indicates that the responses of all 15 L vs. M opponent cells are consistent with single cone input to their RF centers and that our data are consistent with extrafoveal data when the blurring of the optics is accounted for.

Functional and structural adaptations to lifelong lack of cone input and its implications for gene therapy outcomes

Functional and structural adaptations to lifelong lack of cone input and its implications for gene therapy outcomes

Dopamine differentially affects retinal circuits to shape the retinal code.

Dopamine has long been reported to enhance antagonistic surrounds of retinal ganglion cells (RGCs). Yet, the retina contains many different RGC subtypes and the effects of dopamine can be subtype-specific. Using multielectrode array (MEA) recordings we investigated how dopamine shapes the receptive fields of RGCs in the mouse retina. We found that the non-selective dopamine receptor agonist apomorphine can either increase or decrease RGCs' surround strength, depending on their subtype. We thenused two-photon targeted patch-clamp to target a specific RGC subtype, the transient-Off-αRGC. In line with our MEA recordings, apomorphine did not increase the antagonistic surround of transient-Off-αRGCs but enhanced their responses to Off stimuli in the centre receptive field. Both D1 - and D2 -like family receptor (D1 -R and D2 -R) blockers had the opposite effect and reduced centre-mediated responses, but differently affected transient-Off-αRGC's surround. While D2 -R blocker reduced surround antagonism, D1 -R blocker led to surround activation, revealing On responses to large stimuli. Using voltage-clamp recordings we separated excitatory inputs from Off cone bipolar cells and inhibitory inputs from the primary rod pathway. In control conditions, cone inputs displayed strong surround antagonism, while inputs from the primary rod pathway showed no surround. Yet, the surround activation in the D1 -R blockade originated from the primary rod pathway. Our findings demonstrate that dopamine differentially affects RGC subtypes via distinct pathways, suggesting that dopamine has a more complex role in shaping the retinal code than previously reported. KEY POINTS: Receptive fields of retinal ganglion cells (RGCs) have a centre-surround organisation, and previous work has shown that this organisation can be modulated by dopamine in a light-intensity-dependent manner. Dopamine is thought to enhance RGCs' antagonistic surround, but a detailed understanding of how different RGC subtypes are affected is missing. Using a multielectrode array recordings, clustering analysis and pharmacological manipulations, we found that dopamine can either enhance or weaken antagonistic surrounds, and also change response kinetics, of RGCs in a subtype-specific manner. We performed targeted patch-clamp recordings of one RGC subtype, the transient-Off-αRGC, and identified the underlying circuits by which dopamine shapes its receptive field. Our findings demonstrate that dopamine acts in a subtype-specific manner and can have complex effects, which has implications for other retinal computations that rely on receptive field structure.

Synaptotagmins 1 and 7 in vesicle release from rods of mouse retina.

Synaptotagmins are the primary Ca2+ sensors for synaptic exocytosis. Previous work suggested synaptotagmin-1 (Syt1) mediates evoked vesicle release from cone photoreceptor cells in the vertebrate retina whereas release from rods may involve another sensor in addition to Syt1. We found immunohistochemical evidence for syntaptotagmin-7 (Syt7) in mouse rod terminals and so performed electroretinograms (ERG) and single-cell recordings using mice in which Syt1 and/or Syt7 were conditionally removed from rods and/or cones. Synaptic release was measured in mouse rods by recording presynaptic anion currents activated during glutamate re-uptake and from exocytotic membrane capacitance changes. Deleting Syt1 from rods reduced glutamate release evoked by short depolarizing steps but not long steps whereas deleting Syt7 from rods reduced release evoked by long but not short steps. Deleting both sensors completely abolished depolarization-evoked release from rods. Effects of various intracellular Ca2+ buffers showed that Syt1-mediated release from rods involves vesicles close to ribbon-associated Ca2+ channels whereas Syt7-mediated release evoked by longer steps involves more distant release sites. Spontaneous release from rods was unaffected by eliminating Syt7. While whole animal knockout of Syt7 slightly reduced ERG b-waves and oscillatory potentials, selective elimination of Syt7 from rods had no effect on ERGs. Furthermore, eliminating Syt1 from rods and cones abolished ERG b-waves and additional elimination of Syt7 had no further effect. These results show that while Syt7 contributes to slow non-ribbon release from rods, Syt1 is the principal sensor shaping rod and cone inputs to bipolar cells in response to light flashes.

A hyperpolarizing rod bipolar cell in the sea lamprey, Petromyzon marinus.

Retinal bipolar cells receive direct input from rod and cone photoreceptors and send axons into the inner retina, synapsing onto amacrine and ganglion cells. Bipolar cell responses can be either depolarizing (ON) or hyperpolarizing (OFF); in lower vertebrates, bipolar cells receive mixed rod and cone input, whereas in mammals, input is mostly segregated into 14 classes of cone ON and OFF cells and a single rod ON bipolar cell. We show that lamprey, like mammals, have rod bipolar cells with little or no cone input, but these cells are OFF rather than ON. They have a characteristic morphology and a spectral sensitivity nearly indistinguishable from that of rod photoreceptors. In background light known to saturate rods, rod bipolar cells are also saturated and cannot respond to increment flashes. Our results suggest that early vertebrate progenitors of both agnathans and gnathostomes may have had a more fluid retinal organization than previously thought.

Inhibition, but not excitation, recovers from partial cone loss with greater spatiotemporal integration, synapse density, and frequency.

SUMMARYNeural circuits function in the face of changing inputs, either caused by normal variation in stimuli or by cell death. To maintain their ability to perform essential computations with partial inputs, neural circuits make modifications. Here, we study the retinal circuit’s responses to changes in light stimuli or in photoreceptor inputs by inducing partial cone death in the mature mouse retina. Can the retina withstand or recover from input loss? We find that the excitatory pathways exhibit functional loss commensurate with cone death and with some aspects predicted by partial light stimulation. However, inhibitory pathways recover functionally from lost input by increasing spatiotemporal integration in a way that is not recapitulated by partially stimulating the control retina. Anatomically, inhibitory synapses are upregulated on secondary bipolar cells and output ganglion cells. These findings demonstrate the greater capacity for inhibition, compared with excitation, to modify spatiotemporal processing with fewer cone inputs.

Structural changes to primary visual cortex in the congenital absence of cone input in achromatopsia

Autosomal recessive Achromatopsia (ACHM) is a rare inherited disorder associated with dysfunctional cone photoreceptors resulting in a congenital absence of cone input to visual cortex. This might lead to distinct changes in cortical architecture with a negative impact on the success of gene augmentation therapies. To investigate the status of the visual cortex in these patients, we performed a multi-centre study focusing on the cortical structure of regions that normally receive predominantly cone input. Using high-resolution T1-weighted MRI scans and surface-based morphometry, we compared cortical thickness, surface area and grey matter volume in foveal, parafoveal and paracentral representations of primary visual cortex in 15 individuals with ACHM and 42 normally sighted, healthy controls (HC). In ACHM, surface area was reduced in all tested representations, while thickening of the cortex was found highly localized to the most central representation. These results were comparable to more widespread changes in brain structure reported in congenitally blind individuals, suggesting similar developmental processes, i.e., irrespective of the underlying cause and extent of vision loss. The cortical differences we report here could limit the success of treatment of ACHM in adulthood. Interventions earlier in life when cortical structure is not different from normal would likely offer better visual outcomes for those with ACHM.

Structural Differences Across Multiple Visual Cortical Regions in the Absence of Cone Function in Congenital Achromatopsia.

Most individuals with congenital achromatopsia (ACHM) carry mutations that affect the retinal phototransduction pathway of cone photoreceptors, fundamental to both high acuity vision and colour perception. As the central fovea is occupied solely by cones, achromats have an absence of retinal input to the visual cortex and a small central area of blindness. Additionally, those with complete ACHM have no colour perception, and colour processing regions of the ventral cortex also lack typical chromatic signals from the cones. This study examined the cortical morphology (grey matter volume, cortical thickness, and cortical surface area) of multiple visual cortical regions in ACHM (n = 15) compared to normally sighted controls (n = 42) to determine the cortical changes that are associated with the retinal characteristics of ACHM. Surface-based morphometry was applied to T1-weighted MRI in atlas-defined early, ventral and dorsal visual regions of interest. Reduced grey matter volume in V1, V2, V3, and V4 was found in ACHM compared to controls, driven by a reduction in cortical surface area as there was no significant reduction in cortical thickness. Cortical surface area (but not thickness) was reduced in a wide range of areas (V1, V2, V3, TO1, V4, and LO1). Reduction in early visual areas with large foveal representations (V1, V2, and V3) suggests that the lack of foveal input to the visual cortex was a major driving factor in morphological changes in ACHM. However, the significant reduction in ventral area V4 coupled with the lack of difference in dorsal areas V3a and V3b suggest that deprivation of chromatic signals to visual cortex in ACHM may also contribute to changes in cortical morphology. This research shows that the congenital lack of cone input to the visual cortex can lead to widespread structural changes across multiple visual areas.

Melanopic stimulation does not alter psychophysical threshold sensitivity for luminance flicker

In addition to the rod and cone photoreceptors the retina contains intrinsically photosensitive retinal ganglion cells (ipRGCs). These cells express the photopigment melanopsin and are known to be involved in reflexive visual functions such as pupil response and photo-entrainment of the circadian rhythm. It is possible that the ipRGCs contribute to conscious visual perception, either by providing an independent signal to the geniculo-striate pathway, or by interacting with and thus modifying signals arising from “classical” retinal ganglion cells that combine and contrast cone input. Here, we tested for the existence of an interaction by asking if a 350% change in melanopsin stimulation alters psychophysical sensitivity for the detection of luminance flicker. In Experiment 1, we tested for a change in the threshold for detecting luminance flicker in three participants after they adapted to backgrounds with different degrees of tonic melanopsin stimulation. In Experiments 2 and 3, this test was repeated, but now for luminance flicker presented on a transient pedestal of melanopsin stimulation. Across the three experiments, no effect of melanopsin stimulation upon threshold flicker sensitivity was found. Our results suggest that even large changes in melanopsin stimulation do not affect near-threshold, cone-mediated visual perception.

Anatomical changes to primary visual cortex in the congenital absence of cone input

Anatomical changes to primary visual cortex in the congenital absence of cone input

Ganglion cells in larval zebrafish retina integrate inputs from multiple cone types.

We recently showed the presence of seven physiological cone opsins-R1 (575 nm), R2 (556 nm), G1 (460 nm), G3 (480 nm), B1 (415 nm), B2 (440 nm), and UV (358 nm)-in electroretinogram (ERG) recordings of larval zebrafish (Danio rerio) retina. Larval ganglion cells (GCs) are generally thought to integrate only four cone opsin signals (red, green, blue, and UV). We address the question as to whether they may integrate seven cone spectral signals. Here we examined the 127 possible combinations of seven cone signals to find the optimal representation, as based on impulse discharge data sets from GC axons in the larval optic nerve. We recorded four varieties of light-response waveform, sustained-ON, transient-ON, ON-OFF, and OFF, based on the time course of mean discharge rates to all stimulus wavelengths combined. Modeling of GC responses revealed that each received 1-6 cone opsin signals, with a mean of 3.8 ± 1.3 cone signals/GC. Most onset or offset responses were opponent (ON, 80%; OFF, 100%). The most common cone signals were UV (93%), R2 (50%), G3 (55%), and G1 (60%). Seventy-three percent of cone opsin signals were excitatory, and 27% were inhibitory. UV signals favored excitation, whereas G3 and B2 signals favored inhibition. R1/R2, G1/G3, and B1/B2 opsin signals were selectively associated along a nonsynergistic/opponent axis. Overall, these results suggest that larval zebrafish GC spectral responses are complex and use inputs from the seven expressed opsins.NEW & NOTEWORTHY Ganglion cells in larval zebrafish retina have complex spectral responses driven by seven different cone opsin types. UV cone inputs are significant and excitatory to ganglion cells, whereas green and blue cone inputs favor inhibition. Most dramatic are the pentachromatic cells. These responses were identified at 5-6 days after fertilization, reflecting an impressive level of color processing not seen in older fish or mammals.

Variations in photoreceptor throughput to mouse visual cortex and the unique effects on tuning

Visual input to primary visual cortex (V1) depends on highly adaptive filtering in the retina. In turn, isolation of V1 computations requires experimental control of retinal adaptation to infer its spatio-temporal-chromatic output. Here, we measure the balance of input to mouse V1, in the anesthetized setup, from the three main photoreceptor opsins—M-opsin, S-opsin, and rhodopsin—as a function of two stimulus dimensions. The first dimension is the level of light adaptation within the mesopic range, which governs the balance of rod and cone inputs to cortex. The second stimulus dimension is retinotopic position, which governs the balance of S- and M-cone opsin input due to the opsin expression gradient in the retina. The fitted model predicts opsin input under arbitrary lighting environments, which provides a much-needed handle on in-vivo studies of the mouse visual system. We use it here to reveal that V1 is rod-mediated in common laboratory settings yet cone-mediated in natural daylight. Next, we compare functional properties of V1 under rod and cone-mediated inputs. The results show that cone-mediated V1 responds to 2.5-fold higher temporal frequencies than rod-mediated V1. Furthermore, cone-mediated V1 has smaller receptive fields, yet similar spatial frequency tuning. V1 responses in rod-deficient (Gnat1−/−) mice confirm that the effects are due to differences in photoreceptor opsin contribution.

Rod and Cone Connections With Bipolar Cells in the Rabbit Retina.

Rod and cone pathways are segregated in the first stage of the retina: cones synapse with both ON- and OFF-cone bipolar cells while rods contact only rod bipolar cells. However, there is an exception to this specific wiring in that rods also contact certain OFF cone bipolar cells, providing a tertiary rod pathway. Recently, it has been proposed that there is even more crossover between rod and cone pathways. Physiological recordings suggested that rod bipolar cells receive input from cones, and ON cone bipolar cells can receive input from rods, in addition to the established pathways. To image their rod and cone contacts, we have dye-filled individual rod bipolar cells in the rabbit retina. We report that approximately half the rod bipolar cells receive one or two cone contacts. Dye-filling AII amacrine cells, combined with subtractive labeling, revealed most of the ON cone bipolar cells to which they were coupled, including the occasional blue cone bipolar cell, identified by its contacts with blue cones. Imaging the AII-coupled ON cone bipolar dendrites in this way showed that they contact cones exclusively. We conclude that there is some limited cone input to rod bipolar cells, but we could find no evidence for rod contacts with ON cone bipolar cells. The tertiary rod OFF pathway operates via direct contacts between rods and OFF cone bipolar cells. In contrast, our results do not support the presence of a tertiary rod ON pathway in the rabbit retina.

Inhibition, but Not Excitation, Recovers From Partial Cone Loss With Greater Spatiotemporal Integration, Synapse Density and Frequency

Neural circuits function in the face of changing inputs, either caused by normal variation in stimuli or cell death. To maintain their ability to perform essential computations with partial inputs, neural circuits make modifications. Here, we study the retinal circuit’s responses to changes in light stimuli or in photoreceptor inputs by inducing partial cone death in the mature mouse retina. Can the retina withstand or recover from input loss? We find that the excitatory pathways exhibit functional loss commensurate with cone death and with some aspects predicted by partial light stimulation. However, Inhibitory pathways recover functionally from lost input by increasing spatiotemporal integration in a way that is not recapitulated by partially stimulating the control retina. Anatomically, inhibitory synapses are upregulated on secondary bipolar cells and output ganglion cells. These findings demonstrate the greater capacity for inhibition, compared to excitation, to modify spatiotemporal processing with fewer cone inputs.

Influence of common lighting conditions and time-of-day on the effort-related cardiac response.

Melanopic stimuli trigger diverse non-image-forming effects. However, evidence of a melanopic contribution to acute effects on alertness and performance is inconclusive, especially under common lighting situations. Effects on cognitive performance are likely mediated by effort-related physiological changes. We assessed the acute effects of lighting in three scenarios, at two times of day, on effort-related changes to cardiac contraction as indexed by the cardiac pre-ejection period (PEP). In a within-subject design, twenty-seven participants performed a cognitive task thrice during a morning and a late-afternoon session. We set the lighting at 500 lux in all three lighting scenarios, measured horizontally at the desk level, but with 54 lux, 128 lux, or 241 lux melanopic equivalent daylight illuminance at the eye level. Impedance cardiography and electrocardiography measurements were used to calculate PEP, for the baseline and task period. A shorter PEP during the task represents a sympathetic heart activation and therefore increased effort. Data were analysed with linear mixed-effect models. PEP changes depended on both the light scene and time of day (p = 0.01 and p = 0.002, respectively). The highest change (sympathetic activation) occurred for the medium one of the three stimuli (128 lux) during the late-afternoon session. However, effect sizes for the singular effects were small, and only for the combined effect of light and time of day middle-sized. Performance scores or self-reported scores on alertness and task demand did not change with the light scene. In conclusion, participants reached the same performance most efficiently at both the highest and lowest melanopic setting, and during the morning session. The resulting U-shaped relation between melanopic stimulus intensity and PEP is likely not dependent solely on intrinsic ipRGC stimuli, and might be moderated by extrinsic cone input. Since lighting situations were modelled according to current integrative lighting strategies and real-life indoor light intensities, the result has implications for artificial lighting in a work environment.

Multiplicative modulations enhance diversity of hue-selective cells

There is still much to understand about the brain’s colour processing mechanisms and the transformation from cone-opponent representations to perceptual hues. Moreover, it is unclear which area(s) in the brain represent unique hues. We propose a hierarchical model inspired by the neuronal mechanisms in the brain for local hue representation, which reveals the contributions of each visual cortical area in hue representation. Hue encoding is achieved through incrementally increasing processing nonlinearities beginning with cone input. Besides employing nonlinear rectifications, we propose multiplicative modulations as a form of nonlinearity. Our simulation results indicate that multiplicative modulations have significant contributions in encoding of hues along intermediate directions in the MacLeod-Boynton diagram and that our model V2 neurons have the capacity to encode unique hues. Additionally, responses of our model neurons resemble those of biological colour cells, suggesting that our model provides a novel formulation of the brain’s colour processing pathway.

Homeostatic Plasticity Shapes the Retinal Response to Photoreceptor Degeneration.

Homeostatic plasticity stabilizes input and activity levels during neural development, but whether it can restore connectivity and preserve circuit function during neurodegeneration is unknown. Photoreceptor degeneration is the most common cause of blindness in the industrialized world. Visual deficits are dominated by cone loss, which progresses slowly, leaving a window during which rewiring of second-order neurons (i.e., bipolar cells) could preserve function. Here we establish a transgenic model to induce cone degeneration with precise control and analyze bipolar cell responses and their effects on vision through anatomical reconstructions, invivo electrophysiology, and behavioral assays. In young retinas, we find that three bipolar cell types precisely restore input synapse numbers when 50% of cones degenerate but one does not. Of the three bipolar cell types that rewire, two contact new cones within stable dendritic territories, whereas one expands its dendrite arbors to reach new partners. In mature retinas, only one of four bipolar cell types rewires homeostatically. This steep decline in homeostatic plasticity is accompanied by reduced light responses of bipolar cells and deficits in visual behaviors. By contrast, light responses and behavioral performance are preserved when cones degenerate in young mice. Our results reveal unexpected cell type specificity and a steep maturational decline of homeostatic plasticity. The effect of homeostatic plasticity on functional outcomes identify it as a promising therapeutic target for retinal and other neurodegenerative diseases.