OFF Pathways Research Articles

Spatial summation of S-cone ON and OFF signals: Effects of retinal eccentricity

We studied spatial summation for S-cone ON and OFF signals as a function of retinal eccentricity in human subjects. S-cone isolation was obtained by the two-colour threshold method of Stiles, modified by adding blue light to the yellow background. Test stimuli were blue light increments or decrements within a circular area of variable size. These were presented for 100 ms at 0 to 20 deg along the horizontal temporal retinal meridian. Ricco’s area of complete spatial summation was measured from the threshold vs. area curves. This was nearly constant and approximately the same for both types of stimuli within the 0–5 deg range and increased beyond this range. The decremental area increased faster, suggesting that separate mechanisms, presumably ON and OFF, integrate S-cone increments and decrements. The results appear to provide new evidence for the existence of separate S-cone ON and OFF pathways. We compare the data with known morphology of primate retina and assume that, if S-cone decrements are detected via separate OFF cells, these should differ in density and dendritic field size from the S-cone ON cells, but only in the retinal periphery.

Expression of vesicular glutamate transporter 1 in the mouse retina reveals temporal ordering in development of rod vs. cone and ON vs. OFF circuits.

Glutamatergic transmission is crucial to the segregation of ON and OFF pathways in the developing retina. The temporal sequence of maturation of vesicular glutamatergic transmission in rod and cone photoreceptor and ON and OFF bipolar cell terminals is currently unknown. Vesicular glutamate transporters (VGLUTs) that load glutamate into synaptic vesicles are necessary for vesicular glutamatergic transmission. To understand better the formation and maturation of glutamatergic transmission in the rod vs. cone and ON vs. OFF pathways of the retina, we examined the developmental expression of VGLUT1 and VGLUT2 immunocytochemically in the mouse retina. Photoreceptor and bipolar cell terminals showed only VGLUT1-immunoreactivity (-IR); no VGLUT2-IR was present in any synapses of the developing or adult retina. VGLUT1-IR was first detected in cone photoreceptor terminals at postnatal day 2 (P2), several days before initiation of ribbon synapse formation at P4-P5. Rod terminals showed VGLUT1-IR by P8, when they invade the outer plexiform layer (OPL) and initiate synaptogenesis. Developing OFF bipolar cell terminals showed VGLUT1-IR around P8, 2-3 days after bipolar terminals were first identified in the inner plexiform layer (IPL) by labeling for the photoreceptor and bipolar cell terminal marker, synaptic vesicle protein 2B. Although terminals of ON bipolar cells were present in the IPL by P6-P8, most did not show VGLUT1-IR until P8-P10 and increased dramatically from P12. These data suggest a hierarchical development of glutamatergic transmission in which cone circuits form prior to rod circuits in both the OPL and IPL, and OFF circuits form prior to ON circuits in the IPL.

Visual Stimulation Is Required for Refinement of ON and OFF Pathways in Postnatal Retina

ON and OFF pathways separately relay increment and decrement luminance signals from retinal bipolar cells to cortex. ON-OFF retinal ganglion cells (RGCs) are activated via synaptic inputs onto bistratified dendrites localized in the ON and OFF regions of the inner plexiform layer. Postnatal maturational processes convert bistratifying ON-OFF RGCs to monostratifying ON and OFF RGCs. Although visual deprivation influences refinement of higher visual centers, no previous studies suggest that light regulates either the development of the visual-evoked signaling in retinal ON and OFF pathways, nor pruning of bistratified RGC dendrites. We find that dark rearing blocks both the maturational loss of ON-OFF responsive RGCs and the pruning of dendrites. Thus, in retina, there is a previously unrecognized, pathway-specific maturation that is profoundly affected by visual deprivation.

Modeling reverse-phi motion-selective neurons in cortex: double synaptic-veto mechanism.

Reverse-phi motion is the illusory reversal of perceived direction of movement when the stimulus contrast is reversed in successive frames. Livingstone, Tsao, and Conway (2000) showed that direction-selective cells in striate cortex of the alert macaque monkey showed reversed excitatory and inhibitory regions when two different contrast bars were flashed sequentially during a two-bar interaction analysis. While correlation or motion energy models predict the reverse-phi response, it is unclear how neurons can accomplish this. We carried out detailed biophysical simulations of a direction-selective cell model implementing a synaptic shunting scheme. Our results suggest that a simple synaptic-veto mechanism with strong direction selectivity for normal motion cannot account for the observed reverse-phi motion effect. Given the nature of reverse-phi motion, a direct interaction between the ON and OFF pathway, missing in the original shunting-inhibition model, it is essential to account for the reversal of response. We here propose a double synaptic-veto mechanism in which ON excitatory synapses are gated by both delayed ON inhibition at their null side and delayed OFF inhibition at their preferred side. The converse applies to OFF excitatory synapses. Mapping this scheme onto the dendrites of a direction-selective neuron permits the model to respond best to normal motion in its preferred direction and to reverse-phi motion in its null direction. Two-bar interaction maps showed reversed excitation and inhibition regions when two different contrast bars are presented.

Morphological and functional organization of ON and OFF pathways in the adult newt retina.

Morphological and functional organization of ON and OFF pathways in the adult newt retina were examined by intracellular recording and staining techniques and immunohistochemistry. Synaptotagmin immunoreactivity discriminated three broad bands within the IPL: the distal band (sublamina I), the middle band (sublamina II) consisting of two dense punctate bands (sublaminae II(a) and II(b)), and proximal band (sublamina III). The Lucifer-yellow labeled OFF amacrine and ganglion cells send their processes mainly in sublamina I and/or II(a) where OFF bipolar cells extend their axon terminals, while ON amacrine and ganglion cells send their processes in sublamina III and/or II(b) where ON bipolar cells extend their axon terminals. Processes of ON-OFF amacrine and ganglion cells ramify broadly in the whole thickness of the IPL. Many bipolar cells responded to light spot with a transient hyperpolarization at both light onset and offset. They are probably subtypes of ON bipolar cells, because their axon terminals branch mainly in sublaminae III and/or II(b), although a few cells ramified the axon at both sublaminae II(a) and III. Two immunohistochemical markers for bipolar cells, PKC and RB-1, identified axon terminals in sublaminae III and/or II(b). From the ramification pattern of axon terminal, they are probably subtypes of ON bipolar cells. ChAT-ir amacrine cells ramified their dendrites in either sublamina I or II(b). Altogether, present studies support the general idea of segregation of ON and OFF pathways in sublaminae a and b of the IPL.

Adaptation of spatiotemporal mechanisms by increment and decrement stimuli.

Sawtooth modulation has been used in the past to examine visual sensitivity to luminance increments and decrements. The threshold elevation caused by adaptation depends on the spatial profile of the stimulus field and the polarities of the adaptation and test stimuli. We hypothesized that the adaptation effects reflect a change in the sensitivity of the spatiotemporal channels that detect the stimuli. We used a 2-deg disk centered in a larger surround field. Five levels of contrast between the test field and surround were investigated: equiluminant, three intermediate levels, and dark. At each contrast, observers adapted for 5 s to 2-Hz sawtooth modulation (rapid-on or rapid-off). Immediately after adaptation, thresholds were measured for detection of a single cycle of either a rapid-on or a rapid-off waveform. Varying the contrast of the surround affected observers' sensitivity to the polarity of the sawtooth stimulus to the extent that the pattern of sensitivity with the equiluminant surround was the opposite of that with the dark surround. To examine temporal factors, we measured thresholds for slow (500-ms ramps) and fast (8.3-ms pulses) test stimuli. The adaptation effect was preserved with the ramp stimuli but not with the pulse stimuli. Blurring the edge between the test and surround fields in the equiluminant surround condition raised thresholds for all sawtooth test stimuli, suggesting that spatiotemporal channels sensitive to high spatial frequencies and low temporal frequencies facilitate detection in that condition. These findings suggest that adaptation to sawtooth modulation can differentially effect the sensitivity of ON and OFF pathways, but the relative desensitization of each pathway depends on an interaction with the adaptation state of spatiotemporal channels that are involved in detection.

Functional asymmetries in ON and OFF ganglion cells of primate retina.

Functional asymmetries in the ON and OFF pathways of the primate visual system were examined using simultaneous multi-electrode recordings from dozens of retinal ganglion cells (RGCs) in vitro. Light responses of RGCs were characterized using white noise stimulation. Two distinct functional types of cells frequently encountered, one ON and one OFF, had non-opponent spectral sensitivity, relatively high response gain, transient light responses, and large receptive fields (RFs) that tiled the region of retina recorded, suggesting that they belonged to the same morphological cell class, most likely parasol. Three principal functional asymmetries were observed. (1) Receptive fields of ON cells were 20% larger in diameter than those of OFF cells, resulting in higher full-field sensitivity. (2) ON cells had faster response kinetics than OFF cells, with a 10-20% shorter time to peak, trough and zero crossing in the biphasic temporal impulse response. (3) ON cells had more nearly linear light responses and were capable of signaling decrements, whereas OFF cells had more strongly rectifying responses that provided little information about increments. These findings suggest specific mechanistic asymmetries in retinal ON and OFF circuits and differences in visual performance on the basis of the activity of ON and OFF parasol cells.

Functional Asymmetries in ON and OFF Ganglion Cells of Primate Retina

Functional asymmetries in the ON and OFF pathways of the primate visual system were examined using simultaneous multi-electrode recordings from dozens of retinal ganglion cells (RGCs)in vitro. Light responses of RGCs were characterized using white noise stimulation. Two distinct functional types of cells frequently encountered, one ON and one OFF, had non-opponent spectral sensitivity, relatively high response gain, transient light responses, and large receptive fields (RFs) that tiled the region of retina recorded, suggesting that they belonged to the same morphological cell class, most likely parasol. Three principal functional asymmetries were observed. (1) Receptive fields of ON cells were 20% larger in diameter than those of OFF cells, resulting in higher full-field sensitivity. (2) ON cells had faster response kinetics than OFF cells, with a 10–20% shorter time to peak, trough and zero crossing in the biphasic temporal impulse response. (3) ON cells had more nearly linear light responses and were capable of signaling decrements, whereas OFF cells had more strongly rectifying responses that provided little information about increments. These findings suggest specific mechanistic asymmetries in retinal ON and OFF circuits and differences in visual performance on the basis of the activity of ON and OFF parasol cells.

Abnormalities of the long flash ERG in congenital stationary night blindness of the Schubert–Bornschein type

We investigated abnormalities of the retinal cone ON- and OFF-pathways in 24 males with Schubert–Bornschein congenital stationary night blindness (CSNB). Substantial differences were found between both CSNB types. In incomplete type, a-, b- and d-waves were reduced and delayed, whereas in complete type only the b-wave showed significant changes. Oscillatory potentials (OPs) were not discernible from noise in incomplete CSNB and showed significant peak alterations of the ON-OPs only in complete CSNB. In the complete CSNB type, the ON pathway appeared to be mainly affected. In the incomplete CSNB form marked involvement of both the ON and the OFF pathways was noted.

On bipolar cells: following in the footsteps of phototransduction.

The electrical signals resulting from phototransduction are decomposed by bipolar cells and then encoded into spike trains by ganglion cells. The signal decomposition by bipolar cells includes formation of ON and OFF pathways and separation of tonic and phasic signals. The decomposition is accomplished by post-synaptic receptors in the ON and OFF bipolar cells. This chapter focuses on these phenomena in ON bipolar cells and the role of metabotropic glutamate receptors in these processes.

Effects of APB, PDA, and TTX on ERG responses recorded using both multifocal and conventional methods in monkey. Effects of APB, PDA, and TTX on monkey ERG responses.

Results from studies of human subjects suggest that the multifocal ERG technique developed by Erich Sutter and colleagues has considerable potential for assessment of retinal function in both the clinic and laboratory. While the utility of this measure depends to a large extent upon an understanding of the physiological origin for the different response components, relatively little is known in this regard. For the experiments described in this report, we made ERG recordings using both multifocal and conventional methods. Intravitreal injections of APB, PDA, and TTX were used to identify contributions from activity in ON pathway, OFF pathway, and third order retinal neurons, respectively. The results show that photoreceptor activity makes a small direct contribution to 1st and 2nd order multifocal photopic luminance responses. TTX-sensitive activity in third order retinal neurons contributes to both 1st and 2nd order responses with relatively greater contribution to the 2nd order response. Blockade of TTX-sensitive activity in third order cells produces effects on the 2nd order response which are very similar to changes observed in eyes suffering selective loss of retinal ganglion cells resulting from experimental glaucoma. Effects of these intravitreally injected test agents were also determined, in the same recording session, for flash, 30 Hz flicker, and oscillatory potential responses.

Cone-dominated ERG luminance-response function: the Photopic Hill revisited.

In response to progressively brighter stimuli, the b-wave of the photopic ERG gradually augments in amplitude, reaches a plateau for a narrow range of intensities and then rapidly decreases with further increments in the luminance of the flash. This unique luminance-response function was originally introduced as the Photopic Hill. The purpose of this study was to further characterize this unique feature of the cone ERG, investigate if it was only limited to b-wave measurements and if it could be obtained under different photopic background luminances. Photopic ERGs and oscillatory potentials were generated in response to flashes of light ranging from 0.5 to 16 cd m(-2) s in intensity and presented against photopic backgrounds varying from 18 to 525 cd m(-2) in luminance. All but the brightest background yielded a clear Photopic Hill like luminance-response function which could only be evidenced with the b-wave, the i-wave and OP4 amplitude measurements. Interestingly, the maximal amplitude reached remained almost identical irrespective of the background luminance. Our results suggest that the retinal mechanisms at the origin of the Photopic Hill effect could represent a voltage limitation mechanism, intimately tied to the OFF pathway. The latter would however be intrinsic to the cone system only and not to the entire retinal network since significantly higher peak amplitudes are reached with dark adaptation.

Analysis of photoreceptor function and inner retinal activity in juvenile X-linked retinoschisis

Thirteen retinoschisis males with genotyped XLRS1 gene mutations were examined by electroretinogram (ERG) techniques to determine photoreceptor involvement and ON-pathway and OFF-pathway sites of dysfunction. Parameters R max and log S determined by fitting the mathematical model of the activation phase of phototransduction to the scotopic and photopic a-wave responses, were not significantly different from normal. However, the XLRS photopic a-wave amplitudes were significantly lower than normal across all intensities, consistent with defective signaling in the OFF pathway. Long flash (150 ms) ON–OFF photopic responses showed reduced b-wave amplitude but normal d-wave amplitude, giving a reduced b/d ratio of <1.32 Hz photopic flicker ERG fundamental frequency responses showed reduced amplitude and delayed phase, consistent with abnormal signaling by both the ON- and OFF-pathway components. These results indicate that the XLRS1 protein appears not to affect photoreceptor function directly for most XLRS males, and that ERG signaling abnormalities occur in both the ON- and OFF-pathway components that originate in the proximal retina.

ON and OFF Pathways Segregated at the Auditory Thalamus of the Guinea Pig

ON and OFF auditory responses were examined on the medial geniculate body (MGB) of the guinea pig. Single-unit and multiunit recordings were performed on 13 anesthetized subjects while either noise-burst or pure-tone stimuli were applied to the ear contralateral to the recorded hemisphere. Of 2187 uneven-spaced samples, 1142 were ON neurons, 386 were OFF neurons, and 488 were ON-OFF neurons. Neither ON nor OFF neurons changed their response patterns when the stimulus was changed from a noise burst of 60 dB sound pressure level intensity (expressed in decibels per 20 mPa) to pure tones or noise bursts of other intensities. However, most of the ON-OFF neurons changed to either ON or OFF responses with some stimuli. OFF neurons formed clusters. With anatomical confirmation, we found that OFF neuron clusters are always segregated from ON neuron clusters and form OFF sheets in various divisions of the MGB. In the ventral division of the MGB (MGv), the neurons showed mainly ON responses in its core and OFF responses in its periphery or on its boundary with other divisions. The MGv was partially surrounded by an OFF sheet dorsolaterally at the caudal part and medioventrally at the rostral part and was almost completely surrounded by an OFF sheet at the central level rostrocaudally.

Alterations in NMDA receptor expression during retinal degeneration in the RCS rat.

To determine how a progressive loss of photoreceptor cells and the concomitant loss of glutamatergic input to second-order neurons can affect inner-retinal signaling, glutamate receptor expression was analyzed in the Royal College of Surgeons (RCS) rat, an animal model of retinitis pigmentosa. Immunohistochemistry was performed on retinal sections of RCS rats and congenic controls between postnatal (P) day 3 and the aged adult (up to P350) using specific antibodies against N-methyl-D-aspartate (NMDA) subunits. All NMDA subunits (NR1, NR2A-2D) were expressed in control and dystrophic retinas at all ages, and distinct patterns of labeling were found in horizontal cells, subpopulations of amacrine cells and ganglion cells, as well as in the outer and inner plexiform layer (IPL). NRI immunoreactivity in the inner plexiform layer of adult control retinas was concentrated in two distinct bands, indicating a synaptic localization of NMDA receptors in the OFF and ON signal pathways. In the RCS retina, these bands of NRI immunoreactivity in the IPL were much weaker in animals older than P40. In parallel, NR2B immunoreactivity in the outer plexiform layer (OPL) of RCS rats was always reduced compared to controls and vanished between P40 and P120. The most striking alteration observed in the degenerating retina, however, was a strong expression of NRI immunoreactivity in Müller cell processes in the inner retina which was not observed in control animals and which was present prior to any visible sign of photoreceptor degeneration. The results suggest functional changes in glutamatergic receptor signaling in the dystrophic retina and a possible involvement of Müller cells in early processes of this disease.

Interocular interactions reveal the opponent structure of motion mechanisms

Interactions between motion sensors tuned to the same and to opposite directions were probed by means of measuring summation indexes for sensitivities ( d′) to contrast increments and/or decrements applied to drifting gratings presented in binocular and in dichoptic vision. The data confirm a phenomenon described by Stromeyer, Kronauer, Madsen & Klein (1984, J. Opt. Soc. Am. A 1, 876–884), whereby opposite polarity contrast changes applied to binocular gratings drifting in opposite directions yield sensitivities significantly higher than same sign changes for which performance complies with probability summation (PS). The effect disappears in dichoptic vision where opposite sign contrast changes yield a performance close to, or below PS, whether they are applied to same or to opposite direction stimuli. Same sign changes in dichoptic drifting stimuli yield a performance higher than PS independently of their relative directions and close to the performances obtained when these same sign changes are applied to dichoptic, static ±45° gratings. Opposite sign changes applied to such static gratings yield PS. The data support the view according to which: (i) motion direction is extracted at the monocular site; (ii) motion sensors exhibit mutual inhibition within each eye when tuned to opposite directions; and (iii) binocular summation when tuned to the same direction. The data also suggest that (iv) independently of their directional tuning, all motion sensors converge on a binocular, motion non-specific (‘flicker’) unit; and that (v) signals carried by ON and OFF pathways are slightly inhibitory to each other.

Immunocytochemical localization of kainate-selective glutamate receptor subunits GluR5, GluR6, and GluR7 in the cat retina

Localizations of the kainate-selective glutamate receptor subunits GluR5, 6, and 7 were studied in the cat retina by light and electron microscopic immunocytochemistry. GluR5 immunoreactivity was observed in the cell bodies and dendrites of numerous cone bipolar cells and ganglion cells. The labeled cone bipolar cells make basal or flat contacts with cone pedicles in the outer plexiform layer, leading to their identification as OFF-center bipolar cells. Reaction product within the inner plexiform layer was observed in processes of ganglion cells at their sites of input from cone bipolar cells. Staining for GluR6 was localized to A- and B-type horizontal cells, numerous amacrine cells, and an occasional cone bipolar cell. The larger ganglion cells were also immunoreactive. As with other GluR molecules, labeling was usually confined to one of the two postsynaptic elements at a cone bipolar dyad contact. Immunoreactivity for GluR7 was very limited and was seen only in a few amacrine and displaced amacrine cells. Findings of this study are consistent with a major role for kainate receptors in mediating OFF pathways in the outer retina with participation in both OFF and ON pathways in the inner retina.

Bipolar or rectified chromatic detection mechanisms?

It is widely accepted that human color vision is based on two types of cone-opponent mechanism, one differencing L and M cone types (loosely termed "red-green"), and the other differencing S with the L and M cones (loosely termed "blue-yellow"). The traditional view of the early processing of human color vision suggests that each of these cone-opponent mechanisms respond in a bipolar fashion to signal two opponent colors (red vs. green, blue vs. yellow). An alternative possibility is that each cone-opponent response, as well as the luminance response, is rectified, so producing separable signals for each pole (red, green, blue, yellow, light, and dark). In this study, we use psychophysical noise masking to determine whether the rectified model applies to detection by the postreceptoral mechanisms. We measured the contrast-detection thresholds of six test stimuli (red, green, blue, yellow, light, and dark), corresponding to the two poles of each of the three postreceptoral mechanisms. For each test, we determined whether noise presented to the cross pole had the same masking effect as noise presented to the same pole (e.g. comparing masking of luminance increments by luminance decrement noise (cross pole) and luminance increment noise (same pole)). To avoid stimulus cancellation, the test and mask were presented asynchronously in a "sandwich" arrangement (mask-test-mask). For the six test stimuli, we observed that noise masks presented to the cross pole did not raise the detection thresholds of the test, whereas noise presented to the same pole produced a substantial masking. This result suggests that each color signal (red, green, blue, and yellow) and luminance signal, (light and dark) is subserved by a separable mechanism. We suggest that the cone-opponent and luminance mechanisms have similar physiological bases, since a functional separation of the processing of cone increments and cone decrements could underlie both the separation of the luminance system into ON and OFF pathways as well as the splitting of the cone-opponent mechanisms into separable color poles.

Spatial structure, contrast polarity and motion integration

It has been shown that in the initial stages of motion processing, the ON and OFF pathways stay more or less separated. There is evidence that this distinction between motion signals from opposite contrast polarities remains at least partly intact in the integration stage of local motion information. At the same time, interactions between the two systems are also apparent. Here we constructed stimuli that contained a constant number of moving checks. The checks were either assigned only one contrast polarity, or contrast polarity was distributed across the checks either randomly or evenly. We investigated how the spatial configuration of the moving stimulus affected direction discrimination thresholds for the different polarity distributions. Our results provide new evidence for contrast-sign-specific integration of local motion signals within areas of limited size, and inhibitory interactions between these separate ON and OFF motion sensor pools.

Interaction of ON and OFF pathways for visual contrast measurement.

We propose a novel model of visual contrast measurement based on segregated On and Off pathways. Two driving forces have shaped our investigation: (1) establishing a mechanism selective for sharp local transitions in the luminance distribution; (2) generating a robust scheme of oriented contrast detection. Our starting point was the architecture of early stages in the mammalian visual system. We show that the circuit behaves as a soft AND-gate and analyze the scale-space selectivity properties of the model in detail. The theoretical analysis is supplemented by computer simulations in which we selectively investigate key functionalities of the proposed contrast detection scheme. We demonstrate that the model is capable of successfully processing synthetic as well as natural images, thus illustrating the potential of the method for computer vision applications.