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Abstract Background Although it has been suggested that alterations of nerve fiber layer vasculature may be involved in the etiology of eye diseases, including glaucoma, it has not been possible to examine this vasculature in-vivo. This report describes a novel imaging method, fluorescence adaptive optics (FAO) scanning laser ophthalmoscopy (SLO), that makes possible for the first time in-vivo imaging of this vasculature in the living macaque, comparing in-vivo and ex-vivo imaging of this vascular bed. Methods We injected sodium fluorescein intravenously in two macaque monkeys while imaging the retina with an FAO-SLO. An argon laser provided the 488 nm excitation source for fluorescence imaging. Reflectance images, obtained simultaneously with near infrared light, permitted precise surface registration of individual frames of the fluorescence imaging. In-vivo imaging was then compared to ex-vivo confocal microscopy of the same tissue. Results Superficial focus (innermost retina) at all depths within the NFL revealed a vasculature with extremely long capillaries, thin walls, little variation in caliber and parallel-linked structure oriented parallel to the NFL axons, typical of the radial peripapillary capillaries (RPCs). However, at a deeper focus beneath the NFL, (toward outer retina) the polygonal pattern typical of the ganglion cell layer (inner) and outer retinal vasculature was seen. These distinguishing patterns were also seen on histological examination of the same retinas. Furthermore, the thickness of the RPC beds and the caliber of individual RPCs determined by imaging closely matched that measured in histological sections. Conclusion This robust method demonstrates in-vivo, high-resolution, confocal imaging of the vasculature through the full thickness of the NFL in the living macaque, in precise agreement with histology. FAO provides a new tool to examine possible primary or secondary role of the nerve fiber layer vasculature in retinal vascular disorders and other eye diseases, such as glaucoma.

The ability to resolve single cells noninvasively in the living retina has important applications for the study of normal retina, diseased retina, and the efficacy of therapies for retinal disease. We describe a new instrument for high-resolution, in vivo imaging of the mammalian retina that combines the benefits of confocal detection, adaptive optics, multispectral, and fluorescence imaging. The instrument is capable of imaging single ganglion cells and their axons through retrograde transport in ganglion cells of fluorescent dyes injected into the monkey lateral geniculate nucleus (LGN). In addition, we demonstrate a method involving simultaneous imaging in two spectral bands that allows the integration of very weak signals across many frames despite inter-frame movement of the eye. With this method, we are also able to resolve the smallest retinal capillaries in fluorescein angiography and the mosaic of retinal pigment epithelium (RPE) cells with lipofuscin autofluorescence.

This study examined the question of which features of a complex grouping discrimination make it vulnerable to permanent elimination by V4 lesions. We first verified that the line element grouping discrimination, which we previously reported to be devastated by V4 lesions, was similarly affected in the monkeys of this study. The permanence of the deficit was established by mapping its visual field distribution and then testing this discrimination for an extended period at a locus on the border of the deficit. Also, a staircase procedure was used to provide the monkey with within session instruction in the grouping discrimination, but this did not improve V4 lesion performance. Grouping was then compared with several discriminations that shared some features with it, but which were found not to be permanently eliminated by V4 lesions. This comparison suggested that grouping (rather than segmentation or response to a single element) was one feature that made the discrimination vulnerable, a second was the similarity in shape of the texture elements to be grouped. Finally, we tested visual crowding, a property of peripheral vision that is thought to reflect neuronal interactions early in visual cortex, possibly in area V1, and found no effect of V4 lesions. A control experiment with human observers tested whether the elimination of grouping by V4 lesions might be due to an alteration of attention, but found no evidence to support this hypothesis. These results show that severe disruption of texture discriminations by V4 lesions depends on both the nature of the discrimination and the type of texture elements involved, but does not necessarily involve the disruption of attention.

Although human temporal cortex is known to be important for short- and long-term memory, its role in visual perception is not well understood. In this study, we compared the performance of three patients with unilateral temporal lobectomies to that of normal controls on both "simple" and "complex" visual discriminations that did not involve explicit memory components. Two types of complex tasks were tested that involved discriminations secondary to texture segmentation. These were contrasted with simple discriminations using luminance-defined stimuli. Patients showed impaired thresholds only on tasks involving texture segmentation, performing as well as controls when the targets were defined by luminance rather than texture. The minimum stimulus presentation times for threshold performance were also measured for all tasks and found to be elevated in temporal lobectomy patients relative to controls. Although the magnitude of the deficits observed was substantial, loss was equivalent in ipsi- and contra-lesional regions of the visual field. Additional control experiments showed that the patients' perceptual deficits were not due, even in part, to disturbances of basic visual capacities such as acuity and contrast sensitivity. Our results indicate that temporal lobe damage disrupts complex, but not simple, visual discriminations throughout the visual field.

The role of cortical area V4 in complex shape discriminations was studied by testing the effects of V4 lesions in macaques on the ability to visually discriminate between images of three-dimensional (3D) objects from different viewpoints. Stimuli were presented in pairs in the lower left or lower right visual field quadrants about 4 deg from the fovea, and the monkeys judged on each trial whether the two views were of the same or of different objects. Object similarity was varied to determine a threshold shape difference. V4 lesions caused profound, retinotopic, and apparently permanent disruptions of discrimination, regardless of whether the images represented single or multiple viewpoints. In V4 lesioned portions of the visual field, monkeys could discriminate objects only when they differed much more grossly in shape than was true in control locations. These effects of the lesion were virtually identical for discriminations that had been learned before lesions were placed and for those learned afterwards. As in previous studies, V4 lesions elevated contrast thresholds by approximately a factor of two, but control observations showed that this was not the basis of the disruption of shape discrimination. Manipulation of cues to shape showed that in control locations, monkeys maintained excellent shape discrimination despite a variety of stimulus alterations, whereas in V4 lesioned areas their performance was easily disrupted. This finding suggests that V4 may support visual shape discriminations by facilitating the use of multiple visual cues. However, the fact that single-viewpoint and multiple-viewpoint discriminations were similarly affected indicates that the disruption was not specific to 3D shape discrimination, but may apply to a variety of subtle discriminations.

This study shows the existence in humans of independent neural processing streams in early visual cortex, which had previously been demonstrated in macaque monkeys. This evidence was obtained by controlled fixation testing of a subject who had suffered a small stroke in the right fusiform gyrus. The patient showed a severe disruption of color perception, shape discrimination and contrast sensitivity for stationary gratings in the upper left quadrant of his visual field. However, motion perception and contrast sensitivity for drifting gratings were relatively preserved. These results support the view that there are independent visual processing streams early in human visual cortex, and that these streams may subserve such functions as motion and color/form perception.

Ibotenic acid lesions were made in four macaque monkeys in a region of cortical area V4 that corresponds to the lower quadrant of one hemifield. For visual testing, fixation locus was monitored with scleral search coils and controlled behaviorally to place test stimuli either in the lesioned quadrant or in a control location in the opposite hemifield. Some basic visual capacities were slightly altered by the lesions; there was a two-fold reduction of luminance contrast sensitivity as well as red-green chromatic contrast sensitivity, both tested with stationary gratings. On the other hand, little or no loss was found when contrast sensitivity for detection or direction discrimination was tested with 10-Hz drifting gratings nor was there a reliable change in visual acuity. Hue and luminance matching were tested with a spatially more complex matching-to-sample task, but monkeys could not learn this task in the visual field locus of a V4 lesion. If previously trained at this locus, performance was not affected by the lesion. In contrast to the small effects on basic visual capabilities, performance on two form discrimination tasks was devastated by V4 lesions. The first involved discriminating the orientation of colinear groups of dots on a background of randomly placed dots. The second involved discriminating the orientation of a group of three line segments surrounded by differently oriented line segments. Some selectivity of the deficits for form discrimination was shown by the lack of an effect of the lesions on a global motion discrimination. These results show that while V4 lesions cause only slight disruptions of basic visual capacities, they profoundly disrupt form discriminations.

Ibotenic acid lesions were placed in two monkeys in a portion of cortical area V2 that corresponds to a lower quadrant of the visual field extending approximately 3-7 degrees from the fovea. For purposes of comparison, another lesion was placed in area V1 in one animal. A wide range of visual capacities were then measured, using a discrimination between vertical and horizontal orientation, in and near the affected regions of the visual field. Visual acuity declined sharply as the test stimulus approached the visual field location corresponding to the V1 lesion, and no threshold could be measured at its center. In contrast, lesions of area V2 caused no measurable decrease in acuity, nor was there any substantial effect on several measures of contrast sensitivity. Subsequently, two types of more complex visual discriminations were measured (also using a vertical-horizontal discrimination), and these discriminations were severely disrupted by V2 lesions. The first discrimination was of the orientation of two parallel lines of five colinear dots each. We measured the number of background dots that would bring the discrimination to threshold, and this number of dots was greatly decreased by a V2 lesion. The second discrimination was of the orientation of a group of three distinctive texture elements embedded in a six by six element texture. This task could not be done in the visual field region affected by the V2 lesion when the distinctive elements differed in orientation from the others. Control experiments showed that the discrimination could be done when the three distinctive elements differed in size or color. These results suggest that cortical area V2 is not needed for some low-level discriminations, but may be essential for tasks involving complex spatial discriminations.

Four indices of visual performance were measured in control macaques and in macaques that had been exposed to monomeric acrylamide, a neurotoxicant that preferentially damages P retinal ganglion cells. Morphological examination of the retina and visual pathways of these monkeys showed virtually complete loss of P ganglion cells over a region extending to at least 40 deg from the fovea, and relative sparing of M ganglion cells. The four tests examined visual functions for which the visual pathway from P ganglion cells might be of great importance: visual acuity, contrast discrimination, hyperacuity, and shape discrimination. In the acrylamide-dosed monkeys, visual acuity was reduced slightly more than fourfold, a somewhat larger reduction than that seen previously after ibotenic-acid lesions of the P pathway in the geniculate. The residual acuity was in good agreement with the Nyquist frequency calculated from the density of ON or OFF M ganglion cells. Contrast increment thresholds were elevated for the dosed monkeys only in one of the two conditions tested. The elevation was found only under those spatiotemporal conditions for which we have previously shown that contrast thresholds are increased by acrylamide exposure, and was most marked at low background contrasts. Vernier acuity was elevated in one dosed monkey, but not affected in a second monkey that also had severe loss of P ganglion cells. Finally, we found no effect of acrylamide exposure on the number of training trials required to learn simple or complex shape discriminations. These results support previous findings in showing that the P pathway mediates visual acuity, and they show that several other important aspects of visual perception are not exclusively dependent on the P pathway.

This study examined the importance of the primate magnocellular retinocortical pathway in the perception of moving stimuli. A portion of the magnocellular pathway was permanently and selectively interrupted by ibotenic acid injections in the LGN of macaque monkeys. We then tested contrast sensitivity for detecting moving stimuli, as well as two indices of motion perception, contrast sensitivity for opposite direction discrimination and speed difference thresholds, in the affected portion of the visual field. Magnocellular lesions greatly reduced detection contrast sensitivity at high temporal and low spatial frequencies and had a similar effect on contrast sensitivity for opposite direction discrimination under these same stimulus conditions. Consequently, opposite direction discriminations could be made at contrast threshold, suggesting that magnocellular lesions reduced the visibility of stimuli used to test direction perception, but did not act directly on direction perception. Magnocellular lesions also elevated speed difference thresholds under some stimulus conditions. However, this deficit was reduced or eliminated by raising the contrast of the test stimulus. Together, these findings suggest that magnocellular lesions reduce the visibility of stimuli used to test motion perception but that they do not appear to alter motion perception otherwise.

The effects of ablating the visual pathway that passes through the parvocellular (dorsal) LGN were tested in 2 macaque monkeys by measuring acuity and both luminance and chromatic contrast sensitivity. Thresholds were tested monocularly before and after ibotenic acid was used to lesion parvocellular layers 4 and 6 of the contralateral geniculate. The injections were centered at the representation of 6 degrees in the temporal field on the horizontal meridian, and vision was tested with localized stimuli at this location. In addition, in one of the monkeys, a lesion was made in magnocellular layer 1 of the opposite geniculate, and the same thresholds were tested. Physiological and anatomical reconstructions demonstrated complete destruction of the target layers in 1 parvocellular lesions and in the magnocellular lesion, and sparing of the nontarget layers in the tested region. Parvocellular lesions caused a 3-4-fold reduction in visual acuity within the affected part of the visual field, while the magnocellular lesion did not affect acuity. Both luminance and chromatic contrast sensitivity, tested with stationary gratings of 2 c/degree, were severely reduced by parvocellular lesions, but not affected by the magnocellular lesion. However, when luminance contrast sensitivity was tested with 1 c/degree gratings, counterphase modulated at 10 Hz, it was reduced by both parvocellular and magnocellular lesions. This study demonstrates that the parvocellular pathway dominates chromatic vision, acuity, and contrast detection at low temporal and high spatial frequencies, while the magnocellular pathway may mediate contrast detection at higher temporal and lower spatial frequencies.

Ibotenic-acid lesions of the magnocellular portion of the macaque lateral geniculate nucleus were used to examine the role of the M-cell pathway in spatio-temporal contrast sensitivity. A lesion was placed in layer 1 of the lateral geniculate of each of two monkeys. Physiological mapping in one animal demonstrated that the visual-field locus of the lesion was on the horizontal meridian, approximately 6 deg in the temporal field. Visual thresholds were tested monocularly in the contralateral eye, and fixation locus was monitored with a scleral search coil to control the retinal location of the test target. Three threshold measures were clearly disrupted by the magnocellular lesions. Contrast sensitivity for a 1 cycle/deg grating that drifted at 10 Hz was reduced from about twofold greater than, to about the same as, that for 10-Hz counterphase modulated gratings. Sensitivity for a very low spatial frequency (Gaussian blob), 10-Hz flickering stimulus was reduced so severely that no threshold could be measured. In addition, flicker resolution was greatly reduced at lower modulation depths (0.22), but not at higher depths (1.0). Two of the measured thresholds were unaffected by the lesions. Contrast sensitivity for 2 cycle/deg stationary gratings remained intact, and little or no effect on sensitivity was found for 1 cycle/deg, 10-Hz counterphase modulated gratings. Together, these results suggest that the magnocellular pathway makes little contribution to visual sensitivity at low to moderate temporal frequencies. On the other hand, some contribution to detection sensitivity is evident at lower spatial and high temporal frequencies, especially for drifting stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)

Grating acuity was measured as a function of eccentricity from the fovea in two macaques. A vertical-horizontal orientation discrimination was used to determine acuity, and the retinal locus of the test grating was controlled by training them to fixate a spot placed at various distances from the stimulus. Their head was fixed in place and fixation was monitored with a scleral search coil. The acuity of monkeys across the retina was similar to that previously measured in human subjects, reaching a peak of about 38 c/deg at the fovea, and decreasing about 10-fold by 30 deg eccentricity. Acuity was slightly higher in the temporal than in the nasal visual field. The shape of the acuity-eccentricity function suggested a dependence on cone density near the fovea, and on the density of P ganglion cells at eccentricities beyond 10 deg. Existing physiological data suggest the possibility that macaque acuity may also be limited in part by spatial averaging across the receptive field of retinal ganglion cells.

Spatial contrast and temporal modulation sensitivity of two macaque monkeys were measured at three luminance levels before and after binocular laser coagulation of the fovea. The radius of the lesions ranged from 1.6 to 2.2 deg from the center of the fovea. After placement of the lesions, the visibility of high spatial frequencies was greatly reduced, although sensitivity at middle and low spatial frequencies was unaffected. No loss of spatial resolution was found at the lowest luminance tested. When temporal modulation sensitivity was tested with 4 deg targets, foveal lesions had no effect at any temporal frequency or luminance. However, with a 0.57 deg target, sensitivity to lower frequencies was impaired. Thus visual loss after destruction of the fovea is limited to high luminance, small targets, and the resolution of fine detail.

We examined the morphological effects of carbon disulfide exposure on neurons and vasculature of the visual system of macaque monkeys. Five monkeys were exposed to 256 ppm carbon disulfide (CS2) by inhalation for 6 hr a day, 5 days a week. One monkey, sacrificed immediately after exposure, had numerous axonal swellings in the distal optic tract. Four other monkeys survived the exposure period for at least 1 year and were found to have suffered marked degeneration of central retinal ganglion cells, with little or no effect on other neurons in the retina. No evidence was found for arteriosclerotic or aneurysmal changes, suggesting that visual system injury in primates induced by carbon disulfide exposure is not dependent on the occurrence of structural changes in retinal blood vessels.

P beta (color opponent) retinal ganglion cells in macaques were found to degenerate as a result of oral administration of acrylamide. Histological examination, wheat germ agglutinin-horseradish peroxidase transport and cytochrome oxidase histochemistry indicate that other retinal ganglion cells and other neurons in the visual pathways were spared.