Research Articles
Background contrast modulates kinetics and lateral spread of responses to superimposed stimuli in outer retina
- Eric S. Reifsnider, Daniel Tranchina
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- Published online by Cambridge University Press:
- 02 June 2009, pp. 1105-1126
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Surround enhancement (sensitization) is a poorly understood form of network adaptation in which the kinetics of the responses of retinal neurons to test stimuli become faster, and absolute sensitivity of the responses increases with increasing level of steady, surrounding light. Surround enhancement has been observed in all classes of retinal neurons in lower vertebrates except cones, in some primate retinal ganglion cells, and in human psychophysical studies. In theory, surround enhancement could be mediated by two broad classes of mechanisms, which are not mutually exclusive: one in which the kinetics of the transduction linking cone voltage to postsynaptic current in second-order neurons is modulated, and another in which the transformation of postsynaptic current to membrane voltage is modulated. We report here that both classes of mechanism play a role in surround enhancement measured in turtle horizontal cells (HCs). We stimulated the retina by modulating sinusoidally the illuminance of a bar placed at various positions in the HC receptive field. The bar was surrounded by either equally luminant or dim, steady light. Interpretation of responses in the context of a model for the cone-HC network led to the conclusion that the speeding up of response kinetics —due to selective increase in response gain at high temporal frequencies — by surround illuminance is almost completely accounted for by the change in the kinetics of the transduction linking cone membrane potential to HC postsynaptic current. However, surround illuminance also had an additional, surprising effect on the transformation between postsynaptic current and voltage: the space constant for signal spread in the HC network for the dim-surround condition was roughly twice as large as that for the bright-surround condition. Thus, increasing surround illuminance had analogous effects in the spatial and temporal domains: it restricted the time course and the spatial spread of signal. Both effects were dependent on the contrast between the mean bar illuminance and that of the surround, rather than on overall light level. When the stimulus with the bright surround was dimmed uniformly by a neutral density filter, the space constant did not increase, and response gain at high temporal frequencies did not decrease. Pharmacological experiments performed with dopamine and various agonists and antagonists indicated that, although exogenous dopamine can influence surround enhancement, endogenous dopamine does not play an important role in surround enhancement. We conclude that contrast in background light modulates the spatiotemporal properties of signal processing in the outer retina, and does so by a non-dopaminergic mechanism.
The morphology and physiology of a “mini-ommatidium” in the median optic nerve of Limulus polyphemus
- Faramarz H. Samie, Robert N. Jinks, William W. Weiner, Steven C. Chamberlain
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- 02 June 2009, pp. 69-76
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Examination of the Limulus median optic nerve with low-magnification light microscopy allows clear visualization of an ultraviolet-sensitive mini-ommatidium enshrouded by pigment cells, glial cells, and guanophores. Serial 1-μm sections of median optic nerves containing mini-ommatidia revealed the presence of a single, heavily pigmented photoreceptor (retinular) cell and a single, unpigmented arhabdomeric cell. Computer-assisted serial reconstructions from 1-μm sections confirmed the presence of two cells, each bearing a nucleus, and two axons leaving the mini-ommatidium. The retinular cell is morphologically similar to retinular cells from the median and lateral eyes. Its rhabdomere appears to be a continuous sheet of microvilli with much infolding. The structure of the arhabdomeric cell is nearly identical to those found in the median ocellus. As in other photoreceptors in Limulus, the retinular cell of the mini-ommatidium is innervated by efferent fibers from the brain. Each mini-ommatidium generates a single train of nerve impulses in response to light, presumably from the arhabdomeric cell. Measurement of the spectral sensitivity of the mini-ommatidium based upon a constant-response criterion indicated that the retinular cell is maximally sensitive to near ultraviolet light with λmax = 380 nm. Comparison of intensity-response functions revealed that those of the mini-ommatidium are significantly steeper than those of the ocellus almost certainly as the result of neural processing in the ocellus which is absent in the mini-ommatidium.
Development of the lateral eye of American horseshoe crabs: Visual field and dioptric array
- Michael J. Shih, William W. Weiner, Kathleen Kier Wheatley, Jennifer L. DePonceau, Mary Anne Sydlik, Steven C. Chamberlain
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- 02 June 2009, pp. 485-492
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We used a precision two-circle goniometer mounted to the stage of a compound microscope to determine the optical alignment and to measure the entrance aperture diameter of individual cuticular cones in the dioptric array of the lateral eye of juvenile horseshoe crabs in order to learn about the development of the visual field. Our results show that the extent of the visual field of juvenile horseshoe crabs with prosomal lengths about 20% ofadult size (14–21 mm) is about 70% that of the visual field of adult horseshoe crabs (prosomal lengths: 100+ mm). The visual field of such juvenile animals covers between 77 and 85 deg vertically and 140 and 145 deg horizontally. Assuming that the dioptric array is uniform and square packed, the average interommatidial angle of the juvenile animals is between 5.6 and 6.0 deg as compared to 4.6 deg for an adult animal. The diameter of the entrance aperture of individual cuticular cones increases markedly with increasing animal size. In addition, we noted a statistically significant trend for entrance aperture diameters to increase from anterior to posterior within the eye for animals of all sizes. There may be a slight trend for entrance aperture diameters to increase from dorsal to ventral within the eye. Our results indicate that the extent of the visual field and the resolution of the lateral eye approach adult values in advance of animals' reaching sexual maturity.
The pupillary response to light in the turtle
- A. M. Granda, J. R. Dearworth, Jr, C. A. Kittila, W. D. Boyd
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- 02 June 2009, pp. 1127-1133
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When intense adapting lights are turned off, the pupil of the turtle, Pseudemys scripta elegans, enlarges. The recovery functions for pupillary dilation have different time constants that are defined by red- and green-sensitive cones and rods as they are affected by prior light adaptation and time in the dark. Pupillary area related to dilation responds over at least a three- to four-fold range. Following white-light adaptation, the course of pupil dilation in the dark shows a three-legged curve of differing time constants. With spectral-light adaptations, the contributions of separate classes of photoreceptors can be isolated. Red- and green-sensitive cones contribute shorter time constants of 3.31 and 3.65 min to prior white-light adaptation—4.81 and 4.18 min to prior spectral-light adaptations. Rods contribute a much longer time constant of 6.69 min to prior white-light adaptation—7.60 min to prior spectral-light adaptation. The ratios are in keeping with the flash sensitivities of photoreceptors in this same animal, as well as with psychophysical visual threshold mechanisms of color sensitivity.
Retinotopic organization of the superior colliculus in relation to the retinal distribution of afferent ganglion cells
- D.M. Berson, J.J. Stein
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- 02 June 2009, pp. 671-686
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Sensory representations in the brain exhibit topographic variations in magnification. These variations have been thought to reflect regional differences in the density of innervation at the sensory receptor surface. In the primate visual cortex, for example, local magnification factors have been reported to be proportional to the corresponding densities of retinal ganglion cells. We sought to learn whether this principle also operates in a second major retinofugal pathway—the projection to the superior colliculus. In cats, we first used retrograde transport to determine the retinal distributions of the ganglion cells that project to the colliculus. Then, we compared the numbers of colliculopetal ganglion cells in selected retinal sectors to the areas of the corresponding collicular representations. Collicular areal magnification was not simply proportional to the density of afferent ganglion cells, being instead at least 5-fold greater than expected in the representation of the central visual field. These data imply that incoming retinal afferents are more widely spaced in the central regions of the tectal map than in the map's periphery. Such variations in afferent density appear to play as large a role as the distribution of ganglion cells in determining the metric of the collicular map.
Characterization and localization of an aldehyde dehydrogenase to amacrine cells of bovine retina
- John C. Saari, Robert J. Champer, Mary Ann Asson-Batres, Gregory G. Garwin, Jing Huang, John W. Crabb, Ann H. Milam
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- 02 June 2009, pp. 263-272
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An enzyme of bovine retina that catalyzes oxidation of retinaldehyde to retinoic acid was purified to homogeneity and a monoclonal antibody (mAb H-4) was generated. MAb H-4 recognized a single component (Mr = 55,000) in extracts of bovine retina and other bovine tissues. The antibody showed no cross-reactivity with extracts of rat, monkey, or human retinas. A 2067 bp cDNA was selected from a retina cDNA expression library using mAb H-4. The cDNA hybridized with a similarly sized, moderately abundant mRNA prepared from bovine retina. Nucleotide sequence analysis indicated that the cDNA contained a single open reading frame encoding 501 amino acids that have 88% sequence identity with the amino-acid sequence of human hepatic Class 1 aldehyde dehydrogenase. Amino-acid sequence analysis of purified enzyme demonstrated that the cDNA encodes the isolated enzyme. MAb H-4 specifically labeled the somata and processes of a subset of amacrine cells in bovine retinal sections. Labeled amacrine somata were located on both sides of the inner plexiform layer, and their processes ramified into two laminae within the inner plexiform layer. The inner radial processes of Müller (glial) cells were weakly reactive with mAb H-4. Weak immunostaining of amacrine cells was found in monkey retina with mAb H-4, but no signal was detected in rat or human retina. The results provide further evidence for metabolism and function of retinoids within cells of the inner retina and define a novel class of retinal amacrine cells.
The influence of center-surround antagonism on light adaptation in cones in the retina of the turtle
- Dwight A. Burkhardt
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- Published online by Cambridge University Press:
- 02 June 2009, pp. 877-885
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The influence of center-surround antagonism on light adaptation in cone photoreceptors was investigated by intracellular recording from red-sensitive cones in the retina of the turtle, Pseudemys scripta elegans. Test flashes of 0.15-mm diameter were applied at the center of background fields of 0.25-mm or 2.2-mm diameter. Immediately upon expanding the background from 0.25 to 2.2 mm, the membrane potential depolarized by about 1–4 mV. The test flash response was enhanced if the depolarization was primarily due to synaptic feedback from horizontal cells, whereas the response was attenuated if the prolonged depolarization, an intrinsic response of the cone, was the dominant source of the depolarization. After several seconds, however, only the synaptic depolarization was maintained so maintained illumination of the large background field produced an enhancement of the cone's incremental sensitivity. The enhancement was examined in detail in steady-state conditions by obtaining amplitude-intensity measurements for centered test flashes on steady background fields over a large range of intensity. The effect of the large background field at any fixed intensity was fairly well described as a vertical (upward) shift of the amplitude-intensity curve obtained on the small field. This operation constitutes a quasi-subtractive mechanism of light adaptation and might provide a basis for the sort of subtractive mechanisms inferred from psychophysical studies of human vision. The enhancement was quantified by measuring the incremental sensitivity over four decades of background illumination. The magnitude of the enhancement increased with background intensity and then tended to stabilize at higher background intensities. The maximum difference in incremental sensitivity obtained on the large vs. small background field averaged 0.46 log unit (±0.12 s.d.). At higher background intensities, incremental sensitivity conformed to Weber's Law behavior about equally well for flashes applied on either small or large background fields. In sum, the present results provide evidence for an additional mechanism of light adaptation in cone photoreceptors by showing that the incremental light sensitivity, initially set by mechanisms in the outer segment, can be modulated some three-fold by synaptic feedback at the inner segment of the cone.
Contrast coding by cells in the cat's striate cortex: Monocular vs. binocular detection
- Akiyuki Anzai, Marcus A. Bearse, Jr, Ralph D. Freeman, Daqing Cai
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- 02 June 2009, pp. 77-93
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Many psychophysical studies of various visual tasks show that performance is generally better for binocular than for monocular observation. To investigate the physiological basis of this binocular advantage, we have recorded, under monocular and binocular stimulation, contrast response functions for single cells in the striate cortex of anesthetized and paralyzed cats. We applied receiver operating characteristic analysis to our data to obtain monocular and binocular neurometric functions for each cell. A contrast threshold and a slope were extracted from each neurometric function and were compared for monocular and binocular stimulation. We found that contrast thresholds and slopes varied from cell to cell but, in general, binocular contrast thresholds were lower, and binocular slopes were steeper, than their monocular counterparts. The binocular advantage ratio, the ratio of monocular to binocular thresholds for individual cells, was, on average, slightly higher than the typical ratios reported in human psychophysics. No single rule appeared to account for the various degrees of binocular summation seen in individual cells. We also found that the proportion of cells likely to contribute to contrast detection increased with stimulus contrast. Less contrast was required under binocular than under monocular stimulation to obtain the same proportion of cells that contribute to contrast detection. Based on these results, we suggest that behavioral contrast detection is carried out by a small proportion of cells that are relatively sensitive to near-threshold contrasts. Contrast sensitivity functions (CSFs) for the cell population, estimated from this hypothesis, agree well with behavioral data in both the shape of the CSF and the ratio of binocular to monocular sensitivities. We conclude that binocular summation in behavioral contrast detection may be attributed to the binocular superiority in contrast sensitivity of a small proportion of cells which are responsible for threshold contrast detection.
The glial ensheathment of the soma and axon hillock of retinal ganglion cells
- Jonathan Stone, Felix Makarov, Horstmar Holländer
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- 02 June 2009, pp. 273-279
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We have studied the glial investment of ganglion cells of the cat's retina, orienting the sections taken for electron microscopy so that the investment could be traced from the soma along the axon. The soma of each ganglion cell is covered by a close-fitting, continuous sheath formed by Müller cells. The axon hillock and the first part of the initial segment are invested by an extension of the somal sheath, and are thus enclosed in the same glial compartment as the soma. The initial segment extends a few microns past the Müller cell sheath; this last length of the initial segment is contacted by numerous processes of astrocytes, which converge on it in a pattern found also on nodes of the same axons, in the optic nerve. Beyond the initial segment, the intraretinal lengths of the axons are invested by both Müller cells and astrocytes, but the investment is strikingly incomplete. Large areas of axonal membrane have no glial cover, and lie close to other axonal membranes. The sequential arrangement of these distinct forms of glial wrapping of the soma, initial segment, and axon is described here for the first time. It is suggested that this pattern of glial investment controls the flow of current between dendrite and initial segment of the ganglion cell, defines the site of initiation of action spikes, and controls the formation of synapses on the soma and initial segment.
Are the preferred directions of neurons in cat extrastriate cortex related to optic flow?
- Helen Sherk, Jong-Nam Kim, Kathleen Mulligan
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- Published online by Cambridge University Press:
- 02 June 2009, pp. 887-894
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It has been proposed that one area of extrastriate cortex in the cat, the lateral suprasylvian area (LS), plays an important role in visual analysis during locomotion (Rauschecker et al., 1987). Cells in LS reportedly tend to prefer directions along a trajectory originating at the center of gaze, and passing outward through the receptive-field center. Such directions coincide with the directions of image motion in an optic flow field, the pattern seen by locomoting observers when they fixate the point towards which they are heading (Gibson, 1950). We re-examined this issue for cells in LS with receptive fields in the lower visual field. Cells recorded posterior to Horsley-Clarke A2 showed a clear correlation between preferred direction and receptive-field location, but not that predicted: preferred directions were generally orthogonal to “optic flow” directions. Since these cells were all located posterior to those in studies showing a bias for “optic flow” directions, we hypothesized that there are two cell populations within LS, an anterior population that tends to prefer radial-outward directions, and a posterior population that tends to prefer directions orthogonal to radial. Data from earlier mapping experiments (Sherk & Mulligan, 1993) supported this idea.
Na+ - and Cl−-dependent neurotransmitter transporters in bovine retina: Identification and localization by in situ hybridization histochemistry
- Eugenia M. C. Jones
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- Published online by Cambridge University Press:
- 02 June 2009, pp. 1135-1142
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The physiological actions of biogenic amine and amino-acid neurotransmitters are terminated by their removal from the synaptic cleft by specific high-affinity transport proteins. The members of the Na+- and Cl−-dependent neurotransmitter transporter family expressed in bovine retina and responsible for the uptake of biogenic amine and amino-acid neurotransmitters were identified using a reverse transcriptase-polymerase chain reaction-based approach. cDNA clones encoding bovine homologues of glycine (GLYT-1), γ-aminobutyric acid (GAT-1) creatine (CreaT), and orphan (NTT4) transporters were identified using this strategy. The expression pattern of mRNAs encoding these proteins in the retina was determined by in situ hybridization histochemistry GLYT-1 CreaT NTT4 and GAT-1 mRNAs were expressed in the retina by cells in the inner nuclear inner plex, iform and ganglion cell layers They were not expressed at detectable levels in the photoreceptor cells whose cell bodies are in the outer nuclear layer and are the most abundant cell type in the retina GLYT-1 mRNA was present exclusively in the proximal inner nuclear layer GAT-1 mRNA was localized to both the inner nuclear and ganglion cell layers CreaT mRNA was expressed in all cell types in the retina except photoreceptors and NTT4 mRNA was expressed by a sub subpoulation of cells in the ganglion cell laver. Elucidation of the expression pattern of these neurotransmitter transporter mRNAs in the retina provides a basis for studies of the role of glycine γ-aminobutyric acid and creatine transporters in retinal function.
On the distribution of gamma cells in the cat retina
- J.J. Stein, D.M. Berson
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- Published online by Cambridge University Press:
- 02 June 2009, pp. 687-700
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Ganglion cells of the cat retina that are neither alpha nor beta cells are often lumped for convenience into a single anatomical group—the gamma cells (Boycott & Wässle, 1974; Stone, 1983; Wässle & Boycott, 1991). Defined in this way, gamma cells are the morphological counterpart to the physiological W-cell class, which includes all ganglion cells that are neither Y (alpha) nor X (beta) cells. We have estimated the retinal distribution of gamma cells by using retrograde transport to label ganglion cells innervating the superior colliculus and by assuming that these included virtually all gamma cells and no beta cells. We excluded labeled alpha cells on the basis of soma size. Our data suggest that gamma cells represent just under half of the ganglion cells in most of the nasal retina, but only about a third of those in the area centralis and temporal retina. Gamma cells do not appear to be more highly concentrated in the nasal visual streak than are other ganglion cells. In the temporal retina, gamma cells with crossed projections to the brain are apparently at least twice as common as those with uncrossed projections.
Asymmetric retinal growth in the adult teleost green sunfish (Lepomis cyanellus)
- David A. Cameron
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- 02 June 2009, pp. 95-102
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Previous studies on fish retina have suggested that a curved, non-fused embryonic fissure is associated with, and perhaps caused by, asymmetric growth along the retina's marginal germinal zone (where neurons and Miiller glia are added appositionally throughout life). In this report retinal growth was measured directly in adult green sunfish (Lepomis cyanellus), which has a curved, non-fused embryonic fissure. Growth was asymmetric in both small and large fish: ventral and nasal retina grew more than temporal and dorsal retina. This asymmetry was due to different net rates of cellular addition, rather than differential passive expansion. The absolute rates of retinal growth in the centroperipheral direction were roughly exponential functions of fish size—smaller fish grow faster than large fish—but the area of new retina added per unit time did not vary with fish size. Visual implications of asymmetric retinal growth are evaluated.
Nitric oxide synthesis in retinal photoreceptor cells
- Akiko Yoshida, Nikolay Pozdnyakov, Loan Dang, Stephen M. Orselli, Venkat N. Reddy, Ari Sitaramayya
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- 02 June 2009, pp. 493-500
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Nitric oxide (NO) is known to be synthesized in several tissues and to increase the formation of cyclic GMP through the activation of soluble guanylate cyclases. Since cyclic GMP plays an important role in visual transduction, we investigated the presence of nitric oxide synthesizing activity in retinal rod outer segments. Bovine rod outer segments were isolated intact and separated into membrane and cytosolic fractions. Nitric oxide synthase activity was assayed by measuring the conversion of L-arginine to L-citrulline. Both membrane and cytosolic fractions were active in the presence of calcium and calmodulin. The activity in both fractions was stimulated by the nitric oxide synthase cofactors FAD, FMN, and tetrahydrobiopterin and inhibited by the L-arginine analog, L-monomethyl arginine. The Km for L-arginine was similar, about 5 μM for the enzyme in both fractions. However, the two fractions differed in their calcium/calmodulin dependence: the membrane fraction exhibited basal activity even in the absence of added calcium and calmodulin while the cytosolic fraction was inactive. But the activity increased in both fractions when supplemented with calcium/calmodulin: in membranes from about 40 to 110 fmol/min/mg of protein and in the cytosol from near zero to about 350 fmol/min/mg of protein in assays carried out at 0.3 μM L-arginine. The two enzymes also responded differently to detergent: the activity of the membrane enzyme was doubled by Triton X-100 while that of the cytosolic enzyme was unaffected. These results show that NO is produced by cytosolic and membrane-associated enzymes with distinguishable properties. Investigations on the purity of isolated ROS showed that about 50% of the NOS activity is endogenous to the outer segments, and that the rest is due to membrane vesicles rich in Na, K-ATPase activity. If and how NO influences the rod outer segment physiology remains to be investigated.
Qualitative and quantitative features of axons projecting from caudal to rostral inferior temporal cortex of squirrel monkeys
- G.E. Steele, R.E. Weller
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- 02 June 2009, pp. 701-722
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On the basis of cortical and subcortical connections and architectonics, inferior temporal (IT) cortex of squirrel monkeys consists of a caudal region, ITC, with dorsal (ITCd) and ventral (ITCv) subdivisions; a rostral region, ITR; and possibly a third region intermediate to ITC and ITR, IT1 (Weller & Steele, 1992; Steele & Weller, 1993). The present study qualitatively and quantitatively examined the terminal arborizations of 26 axons in ITR and IT1 labeled by injections of biocytin or, in one case, horseradish peroxidase, in ITCv. The majority of axons gave rise to a single terminal arbor, with a small number branching into two overlapping or nearby arbors. Presumptive terminal specializations consisted of rounded, bead-like swellings, most often located en passant. All axons terminated in layer 4 of cortex, and most had additional terminations in layers 3 and 5. The total extent of each axon's terminal arbor was 125–750 μm dorsoventrally (mean = 360.6 μm) and 150–725 μm anteroposteriorly (mean = 328.1 μm; all values uncorrected for shrinkage). In most axons, especially those with larger terminal fields, boutons were not uniformly distributed, but formed 2–4 clumps (mean = 2.2), with a mean width of 149 μm, separated by narrower regions of fewer boutons. Based on a cluster analysis of characteristics of the 26 axons, axons projecting from caudal (ITCv) to rostral (ITR or IT1) IT cortex of squirrel monkeys comprised three groups that we called Type I, Type II, and Type III. Type I axons, the smallest in areal extent of terminal arbor, terminated predominantly in dorsal ITR. Type III axons, largest in areal extent, and Type II axons, intermediate in areal extent, terminated in ventral ITR and throughout IT1. The three classes of axons may correspond to different types of visual information entering rostral IT cortex. The clumping of boutons suggests that individual axons terminate in limited patches within their terminal fields.
Spatial structure of chromatically opponent receptive fields in the human visual system
- Pascal Girard, Maria Concetta Morrone
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- 02 June 2009, pp. 103-116
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This study investigates the receptive-field structure of mechanisms operating in human color vision, by recording visual evoked potentials (VEPs) to multiharmonic gratings modulated either in luminance or color (red-green). Varying the Fourier phase of the harmonics from 0 deg to 90 deg produced a family of stimulus profiles that varied from lines to edges. The stimuli were contrast reversed to elicit steady-state VEPS, and also randomly jittered (at a higher temporal frequency than the contrast reversal) to ensure that the evoked response resulted from the polarity reversal, rather than from local variation of luminance or color. Reliable VEPs were recorded from both luminance and chromatic stimuli at all phases, suggesting that the mechanisms sensitive to chromatic contrast and those sensitive to luminance contrast have both symmetric and asymmetric receptive fields. Contrast thresholds estimated by extrapolation of the contrast response curves were very similar to psychophysical thresholds for phase discrimination, suggesting that the VEP response is generated by mechanisms mediating phase discrimination. The results support the idea that human color mechanisms have receptive fields with a variety of spatial symmetries (including odd- and even-symmetric fields) and that these mechanisms may contribute to phase discrimination of chromatic stimuli in a similar way to what has been suggested for luminance vision.
The cat's pupillary light response under urethane anesthesia
- J.K. Oh, D.L. Bohnsack, J.B. Troy, Ch. Enroth-Cugell
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- 02 June 2009, pp. 281-284
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Pupillary area was measured in urethane-anesthetized cats as a function of retinal illuminance. When appropriate corrections are made for differences in experimental procedures, it was found that the pupillary response of the urethane-anesthetized cat's eyes to light was basically unchanged from that of the alert behaving cat. This preparation may therefore be a very satisfactory one in which to study the pupillary response pathway in a higher mammal.
Immunocytochemical evidence for an axonal localization of GABA in the optic nerves of rabbits, rats, and cats
- P. C. Rogers, D. V. Pow
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- 02 June 2009, pp. 1143-1149
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We have examined, by light-microscopic immunocytochemistry, the distribution of GABA in the optic nerves of adult rabbits, rats, and cats. Within the optic nerves, immunoreactivity for GABA was restricted to a small subset of axons; some axons were strongly labelled, others weakly labelled, whilst most axons were unlabelled. Glia and other non-neuronal elements were always unlabelled. Our ability to detect GABA in optic nerve axons of adult mammals contrasts with previous reports that indicate a lack of GABA immunoreactivity in such axons. We suggest that this discrepancy may be due to the sensitivity of our immunocytochemical techniques which enable us to detect low concentrations of GABA.
Temporal and spatial patterns of opsin gene expression in zebrafish (Danio rerio)
- Judith Robinson, Ellen A. Schmitt, John E. Dowling
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- Published online by Cambridge University Press:
- 02 June 2009, pp. 895-906
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In zebrafish, the first class of cone photoreceptor to become morphologically distinct is the ultraviolet-sensitive short single cone, at 4 days postfertilization, whereas the last class, the red- and green-sensitive double cone, becomes distinct at 10 days postfertilization. We have examined the time course of visual pigment gene expression in zebrafish using whole-mount in situ hybridization. Within the retina, opsins may be detected as early as 40 h postfertilization with the ultraviolet and rod visual pigments being expressed before the blue- (48 h) and red- (60 h) sensitive pigments. In the pineal, red-sensitive opsin is expressed at 48 h postfertilization. Visual pigment expression provides a useful tool for investigations of early cell fate in zebrafish.
Expression of glycine receptor subunits and gephyrin in single bipolar cells of the rat retina
- Ralf Enz, Joachim Bormann
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- 02 June 2009, pp. 501-507
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We studied the expression of glycine receptor (GlyR) subunits and gephyrin in the adult rat retina. Reverse transcribed RNA was amplified by polymerase chain reaction (RT-PCR) with primers designed to recognize GlyR α1, α2, α3, β subunits, and gephyrin. Using RNA isolated from the whole retina, signals for all four GlyR subunits and gephyrin could be observed. In rod bipolar cells, in contrast, we detected a subset of GlyR subunits, α1 and β, and no gephyrin. Patch-clamp recording employing two subtype-specific blockers of the GlyR, picrotoxinin and cyanotriphenylborate (CTB), indicated that the GlyR in rod bipolar cells is a heteromeric protein composed of the α1 and β subunit. Moreover, the absence of detectable amounts of gephyrin mRNA suggests that the anchor protein is not required for the function of GlyRs in rod bipolar cells.