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The superior colliculus receives a direct projection from retinal ganglion cells. In primates, it remains unknown if the same ganglion cells also supply the lateral geniculate nucleus. To address this issue, a double-label experiment was performed in two male macaques. The animals fixated a target while injection sites were scouted in the superior colliculus by recording and stimulating with a tetrode. Once suitable sites were identified, cholera toxin subunit B-Alexa Fluor 488 was injected via an adjacent micropipette. In a subsequent acute experiment, cholera toxin subunit B-Alexa Fluor 555 was injected into the lateral geniculate nucleus at matching retinotopic locations. After a brief survival period, ganglion cells were examined in retinal flatmounts. The percentage of double-labeled cells varied locally, depending on the relative efficiency of retrograde transport by each tracer and the precision of retinotopic overlap of injection sites in each target nucleus. In counting boxes with extensive overlap, 76-98% of ganglion cells projecting to the superior colliculus were double labeled. Cells projecting to the superior colliculus constituted 4.0-6.7% of the labeled ganglion cell population. In one particularly large zone, there were 5,746 cells labeled only by CTB-AF555, 561cells double labeled by CTB-AF555 and CTB-AF488, but no cell labeled only by CTB-AF488. These data indicate that retinal input to the macaque superior colliculus arises from a collateral axonal branch supplied by ∼5% of the ganglion cells that project to the lateral geniculate nucleus. Surprisingly, there exist no ganglion cells that project exclusively to the SC., Competing Interests: The authors declare no competing financial interests., (Copyright © 2024 the authors.)

Adeno-associated virus (AAV) is widely used as a vector for delivery of gene therapy. Long term therapeutic benefit depends on perpetual expression of the wild-type gene after transduction of host cells by AAV. To address this issue in a mass population of identified single cells, 4 rats received an injection of a 1:1 mixture of rAAV2-retro-hSyn-EGFP and rAAV2-retro-hSyn-mCherry into each superior colliculus. After the virus was transported retrogradely to both retinas, serial fundus imaging was performed at days 14, 45, 211, and 375 to visualize individual fluorescent ganglion cells. The location of each cell was plotted to compare labeling at each time point. In 12/16 comparisons, 97% or more of the cells identified in the initial baseline fundus image were still labeled at day 375. In 4 cases the percentage was lower, but in these cases the apparent reduction in the number of labeled cells at day 375 was attributable to the lower quality of follow-up fundus images, rather than true loss of transgene expression. These data indicate that retinal ganglion cells transduced by rAAV2-retro are transduced permanently., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Purpose: To explore the utility of eye tracking glasses in patients with intermittent exotropia as a means for quantifying the occurrence of exotropia, defined as the percentage of time that the eyes are misaligned., Design: Prospective observational study., Methods: Eye tracking glasses were used to obtain 68 recordings in 44 ambulatory patients with a history of intermittent exotropia. Vergence angle was monitored for up to 12 hours to document the occurrence of exotropia., Results: Intermittent exotropia was present in 31 of 44 patients. They had a mean exotropia of 19.3 ± 5.3° and a mean occurrence of 40% (range 3-99%). There was a moderate correlation between the magnitude of exotropia and its occurrence (r = 0.59). In 13 patients the occurrence of exotropia was <1%; they were deemed to have an exophoria only. In 35 of 44 cases, families reported an occurrence of intermittent exotropia greater than that measured by the eye tracking glasses., Conclusions: Eye tracking glasses may be a useful tool for quantifying the severity of intermittent exotropia and for defining more precisely its clinical features., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: The axons of ganglion cells in the nasal retina decussate at the optic chiasm. It is unclear why tumours cause more injury to crossing nasal fibres, thereby giving rise to temporal visual field loss in each eye. To address this issue, the course of fibres through the optic chiasm was examined following injection of a different fluorescent tracer into each eye of a monkey., Methods: Under general anaesthesia, cholera toxin subunit B-Alexa Fluor 488 was injected into the right eye and cholera toxin subunit B-Alexa Fluor 594 was injected into the left eye of a single normal adult male rhesus monkey. After a week's survival for anterograde transport, serial coronal sections through the primary optic pathway were examined., Results: A zone within the core of the anterior and mid portions of the optic chiasm was comprised entirely of crossing fibres. This zone of decussation was delineated by segregated, interwoven sheets of green (right eye) and red (left eye) fibres. It expanded steadily to fill more of the optic chiasm as fibres coursed posteriorly towards the optic tracts. Eventually, crossed fibres became completely intermingled with uncrossed fibres, so that ocular separation was lost., Conclusions: A distinct, central compartment located within the anterior two-thirds of the optic chiasm contains only crossing fibres. Sellar tumours focus their compressive force on this portion of the structure, explaining why they so often produce visual field loss in the temporal fields., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: Wearable eye trackers record gaze position as ambulatory subjects navigate their environment. Tobii Pro Glasses 3 were tested to assess their accuracy and precision in the measurement of vergence angle., Methods: Four subjects wore the eye tracking glasses, with their head stabilized, while fixating at a series of distances corresponding to vergence demands of: 0.25, 0.50, 1, 2, 4, 8, 16, and 32°. After these laboratory trials were completed, 10 subjects wore the glasses for a prolonged period while carrying out their customary daily pursuits. A vergence profile was compiled for each subject and compared with interpupillary distance., Results: In the laboratory, the eye tracking glasses were comparable in accuracy to remote video eye trackers, outputting a mean vergence value within 1° of demand at all angles except 32°. In ambulatory subjects, the glasses were less accurate, due to tracking interruptions and measurement errors, partly mitigated by the application of data filters. Nonetheless, a useful record of vergence behavior was obtained in every subject. Vergence profiles often had a bimodal distribution, reflecting a preponderance of activities at near (mobile phone and computer) or far (driving and walking). As expected, vergence angle correlated with interpupillary distance., Conclusions: Wearable eye tracking glasses make it possible to compile a nearly continuous record of vergence angle over hours, which can be correlated with the corresponding visual scene viewed by ambulatory subjects., Translational Relevance: This technology provides new insight into the diversity of human ocular motor behavior and may become useful for the diagnosis of disorders that affect vergence function such as: convergence insufficiency, Parkinson disease, and strabismus.

Abstract: The primate lateral geniculate nucleus has long been a favorite structure among anatomists because of its striking lamination. It has been shown that each lamina receives input from a different eye using various single label techniques but never by double labeling. Here, we illustrate the organization of retinal inputs to the lateral geniculate nucleus by injection of cholera toxin-B conjugated to Alexa Fluor-488 into the right eye and cholera toxin-B conjugated to Alexa Fluor-594 into the left eye., Competing Interests: The authors report no conflicts of interest., (Copyright © 2022 by North American Neuro-Ophthalmology Society.)

Access of adeno-associated virus (AAV) to ganglion cells following intravitreal injection for gene therapy is impeded by the internal limiting membrane of the retina. As an alternative, one could transduce ganglion cells via retrograde transport after virus injection into a retinal target nucleus. It is unknown if recombinant AAV2-retro (rAAV2-retro), a variant of AAV2 developed specifically for retrograde transport, is capable of transducing retinal ganglion cells. To address this issue, equal volumes of rAAV2-retro-hSyn-EGFP and rAAV2-retro-hSyn-mCherry were mixed in a micropipette and injected into the rat superior colliculus. The time-course of viral transduction was tracked by performing serial in vivo fundus imaging. Cells that were labeled by the fluorophores within the first week remained consistent in distribution and relative signal strength on follow-up imaging. Most transduced cells were double-labeled, but some were labeled by only EGFP or mCherry. Fundus images were later aligned with retinal wholemounts. Ganglion cells in the wholemounts matched precisely the cells imaged by fundus photography. As seen in the fundus images, ganglion cells in wholemounts were sometimes labeled by only EGFP or mCherry. Overall, there was detectable label in 32-41% of ganglion cells. Analysis of the number of cells labeled by 0, 1, or 2 fluorophores, based on Poisson statistics, yielded an average of 0.66 virions transducing each ganglion cell. Although this represents a low number relative to the quantity of virus injected into the superior colliculus, the ganglion cells showed sustained and robust fluorescent labeling. In the primate, injection of rAAV2-retro into the lateral geniculate nucleus might provide a viable approach for the transduction of ganglion cells, bypassing the obstacles that have prevented effective gene delivery via intravitreal injection., (Copyright © 2022. Published by Elsevier Ltd.)

Purpose: After the lateral geniculate nucleus, the superior colliculus is the richest target of retinal projections in primates. Hubel et al. used tritium autoradiography to show that axon terminals emanating from one eye form irregular columns in the stratum griseum superficiale. Unlabeled gaps were thought to be filled by the other eye, but this assumption was never tested directly., Methods: Experiments were performed in two normal macaques. In monkey 1, [3H]proline was injected into the left eye and the pattern of radiolabeling was examined in serial cross-sections through the entire superior colliculus. In monkey 2, cholera toxin subunit B conjugated to Alexa 488 was injected into the right eye and cholera toxin subunit B - Alexa 594 was injected into the left eye. The two fluorescent labels were compared in a reconstruction of the superior colliculus prepared from serial sections., Results: In monkey 1, irregular columns of axon terminals were present in the superficial grey. The projection from the peripheral retina was stronger than the projection from the macula. In monkey 2, the two fluorescent Alexa tracers mainly interdigitated: a conspicuous gap in one label was usually filled by a clump of the other label. There was also partial laminar segregation of ocular inputs. In the far peripheral field representation, the contralateral eye's input generally terminated closer to the tectal surface. In the midperiphery the eyes switched, bringing the ipsilateral input nearer the surface., Conclusions: Direct retinal input to the macaque superior colliculus is segregated into alternating columns and strata, despite the fact that tectal cells respond robustly to stimulation of either eye.

Background: The purpose of this study was to investigate which portions of the visual scene are perceived by each eye in an exotropic subject with acquired hemianopia. The pattern of suppression is predictable from knowledge of how suppression scotomas are organized in exotropic subjects with intact visual fields., Methods: Dichoptic perimetry was performed by having a subject wear red/blue goggles while fixating a cross that was either red or blue. Red, blue, or purple spots were presented briefly at peripheral locations. The subject's identification of the spot color revealed which eye was perceptually engaged at any given location in the visual fields., Results: A 17-year-old female with a history of exotropia was evaluated after rupture of a right parietal arteriovenous malformation. Dichoptic perimetry showed a left homonymous hemianopia. All stimuli to the right of the right fovea's projection point were perceived via the right eye. Stimuli between the foveal projection points, which were separated horizontally by the 20° exotropia, were perceived by the left eye., Conclusions: Perception of the visual scene is shared by the eyes in hemianopia and exotropia. Suppression occurs only in the peripheral temporal retina of the eye contralateral to the brain lesion, regardless of which eye is engaged in fixation. Although exotropia expands the binocular field of vision in hemianopia, it is probably not an adaptive response, even when it develops after hemianopia., (Copyright © 2021. Published by Elsevier Inc.)

Diplopia (double vision) in strabismus is prevented by suppression of the image emanating from one eye. In a recent study conducted in two macaques raised with exotropia (an outward ocular deviation) but having normal acuity in each eye, simultaneous display of stimuli to each eye did not induce suppression in V1 neurons. Puzzled by this negative result, we have modified our protocol to display stimuli in a staggered sequence, rather than simultaneously. Additional recordings were made in the same two macaques, following two paradigms. In trial type 1 , the receptive field in one eye was stimulated with a sine-wave grating while the other eye was occluded. After 5 s, the occluder was removed and the neuron was stimulated for another 5 s. The effect of uncovering the eye, which potentially exposed the animal to diplopia, was quantified by the peripheral retinal interaction index (PRII). In trial type 2 , the receptive field in the fixating eye was stimulated with a grating during binocular viewing. After 5 s, a second grating appeared in the receptive field of the nonfixating eye. The impact of the second grating, which had the potential to generate visual confusion, was quantified by the receptive field interaction index (RFII). For 82 units, the mean PRII was 0.48 ± 0.05 (0.50 = no suppression) and the mean RFII was 0.46 ± 0.08 (0.50 = no suppression). These values suggest mild suppression, but the modest decline in spike rate registered during the second epoch of visual stimulation might have been due to neuronal adaptation, rather than interocular suppression. In a few instances neurons showed unequivocal suppression, but overall, these recordings did not support the contention that staggered stimulus presentation is more effective than simultaneous stimulus presentation at evoking interocular suppression in V1 neurons. NEW & NOTEWORTHY In strabismus, double vision is prevented by interocular suppression. It has been reported that inhibition of neuronal firing in the primary visual cortex occurs only when stimuli are presented sequentially, rather than simultaneously. However, these recordings in alert macaques raised with exotropia showed, with rare exceptions, little evidence to support the concept that staggered stimulus presentation is more effective at inducing interocular suppression of V1 neurons.

Patients with homonymous hemianopia sometimes show preservation of the central visual fields, ranging up to 10°. This phenomenon, known as macular sparing, has sparked perpetual controversy. Two main theories have been offered to explain it. The first theory proposes a dual representation of the macula in each hemisphere. After loss of one occipital lobe, the back-up representation in the remaining occipital lobe is postulated to sustain ipsilateral central vision in the blind hemifield. This theory is supported by studies showing that some midline retinal ganglion cells project to the wrong hemisphere, presumably driving neurons in striate cortex that have ipsilateral receptive fields. However, more recent electrophysiological recordings and neuroimaging studies have cast doubt on this theory by showing only a minuscule ipsilateral field representation in early visual cortical areas. The second theory holds that macular sparing arises because the occipital pole, where the macula is represented, remains perfused after occlusion of the posterior cerebral artery because it receives collateral flow from the middle cerebral artery. An objection to this theory is that it cannot account for reports of macular sparing in patients after loss of an entire occipital lobe. On close scrutiny, such reports turn out to be erroneous, arising from inadequate control of fixation during visual field testing. Patients seem able to detect test stimuli on their blind side within the macula or along the vertical meridian because they make surveillance saccades. A purported treatment for hemianopia, called vision restoration therapy, is based on this error. The dual perfusion theory is supported by anatomical studies showing that the middle cerebral artery perfuses the occipital pole in many individuals.In patients with hemianopia from stroke, neuroimaging shows preservation of the occipital pole when macular sparing is present. The frontier dividing the infarcted territory of the posterior cerebral artery and the preserved territory of the middle cerebral artery is variable, but always falls within the representation of the macula, because the macula is so highly magnified. For physicians, macular sparing was an important neurological sign in acute hemianopia because it signified a posterior cerebral artery occlusion. Modern neuroimaging has supplanted the importance of that clinical sign but at the same time confirmed its validity. For patients, macular sparing remains important because it mitigates the impact of hemianopia and preserves the ability to read fluently.

Purpose: The most common form of strabismus, intermittent exotropia, is thought to become manifest when the drive to fuse is overcome by excessive divergent muscle tone. This principle is tested by examining the alignment of the eyes in the absence of vision. We compare the ocular deviation in patients with intermittent exotropia under conditions of monocular versus binocular occlusion., Methods: This prospective study of a patient cohort referred to our laboratory enrolled 18 patients with typical findings of well-controlled intermittent exotropia. Eye positions were recorded with video eye trackers while patients looked at a fixation spot at a distance of 57 cm. One eye was occluded, and the resulting ocular deviation was measured. Both eyes were then occluded, and the ocular deviation was re-measured., Results: The majority of patients (11/18) had a smaller deviation when both eyes were covered. Occlusion of one eye resulted in a mean exotropia of 13.5° ± 4.7°. Occlusion of both eyes reduced the mean exotropia to 6.0° ± 6.5° (paired t-test, P < 0.001), corresponding to a 56% reduction in the ocular deviation. This reduction persisted during prolonged bilateral occlusion but reversed as soon as vision was restored., Conclusions: Bilateral occlusion reveals a fixation-free state of alignment that is different from orthotropia and usually less than the exotropia that occurs spontaneously during binocular viewing. This finding demonstrates that the deviation angle in patients with intermittent exotropia is actively mediated by visual feedback, which the fixating eye is capable of providing alone.

Patient with exotropia frequently alternate fixation, looking at something with one eye and then switching their attention to acquire a new target with the other eye. Which eye informs the brain about the location of the new target? To address this issue, we presented targets dichoptically to 16 exotropes that were visible to the fixating eye, the deviated eye, or to both eyes. We then compared the subjects' choice of eye for target acquisition with the organization of their suppression scotomas. There was a correspondence between suppression scotoma maps and the eye used to acquire peripheral targets. In other words, a target perceived via an eye was also fixated by it. These studies reveal how patients with alternating strabismus, despite eye misalignment, manage to localize and fixate efficiently visual targets in their environment., Competing Interests: Conflict of interest: Yes

Purpose: In patients with early ocular misalignment and nystagmus, vertical optokinetic stimulation reportedly increases the horizontal component of the nystagmus present during fixation, resulting in diagonal eye movements. We tested patients with infantile nystagmus syndrome but normal ocular alignment to determine if this crosstalk depends on strabismus., Methods: Eye movements were recorded in seven patients with infantile nystagmus. All but one patient had normal ocular alignment with high-grade stereopsis. Nystagmus during interleaved trials of right, left, up, and down optokinetic stimulation was compared with waveforms recorded during fixation. Six patients with strabismus but no nystagmus were also tested., Results: In infantile nystagmus syndrome, horizontal motion evoked a mostly jerk nystagmus with virtually no vertical component. A vertical optokinetic pattern produced nystagmus with a diagonal trajectory. It was not simply a combination of a vertical component from optokinetic stimulation and a horizontal component from the subject's congenital nystagmus, rather in six of seven patients, the slow-phase velocity of the horizontal component during vertical optokinetic stimulation differed from that recorded during fixation. In the six strabismus patients without nystagmus, responses to vertical optokinetic stimulation were normal., Conclusions: In patients with congenital motor nystagmus, a vertical noise pattern drives a diagonal nystagmus. This appears to arise because of crosstalk between the vertical and horizontal components of the optokinetic system. This abnormal response to vertical stimulation is not caused by strabismus because it occurs in patients with infantile nystagmus without strabismus. Moreover, it is absent in patients with strabismus and no spontaneous nystagmus.

Purpose: To assess the outcome of free tenotomy of the medial rectus muscle in post-natal monkeys., Methods: The medial rectus muscle was disinserted in both eyes of 6 macaques at age 4 weeks to induce an alternating exotropia. After the impact on the visual cortex and superior colliculus was investigated, the animals were examined post-mortem to assess the anatomy of the medial rectus muscles., Results: After tenotomy, the monkeys eventually recovered partial adduction. Necropsy revealed that all 12 medial rectus muscles had reattached to the globe. They were firmly connected via an abnormally long tendon, but at the native insertion site., Conclusions: Medial rectus muscles are able to reattach spontaneously to the eye following free tenotomy in post-natal macaques. The early timing of surgery and the large size of the globe relative to the orbit may explain why reinsertion occurs more readily in monkeys than in children with a lost muscle after strabismus surgery. [J Pediatr Ophthalmol Strabismus. 2018;55(5):335-338.]., (Copyright 2018, SLACK Incorporated.)

In subjects with alternating strabismus, either eye can be used to saccade to visual targets. The brain must calculate the correct vector for each saccade, which will depend on the eye chosen to make it. The superior colliculus, a major midbrain center for saccade generation, was examined to determine whether the maps serving each eye were shifted to compensate for strabismus. Alternating exotropia was induced in two male macaques at age 1 month by sectioning the tendons of the medial recti. Once the animals grew to maturity, they were trained to fixate targets with either eye. Receptive fields were mapped in the superior colliculus using a sparse noise stimulus while the monkeys alternated fixation. For some neurons, sparse noise was presented dichoptically to probe for anomalous retinal correspondence. After recordings, microstimulation was applied to compare sensory and motor maps. The data showed that receptive fields were offset in position by the ocular deviation, but otherwise remained aligned. In one animal, the left eye's coordinates were rotated ∼20° clockwise with respect to those of the right eye. This was explained by a corresponding cyclorotation of the ocular fundi, which produced an A-pattern deviation. Microstimulation drove the eyes accurately to the site of receptive fields, as in normal animals. Single-cell recordings uncovered no evidence for anomalous retinal correspondence. Despite strabismus, neurons remained responsive to stimulation of either eye. Misalignment of the eyes early in life does not alter the organization of topographic maps or disrupt binocular convergence in the superior colliculus. SIGNIFICANCE STATEMENT Patients with strabismus are able to make rapid eye movements, known as saccades, toward visual targets almost as gracefully as subjects with normal binocular alignment. They can even exercise the option of using the right eye or the left eye. It is unknown how the brain measures the degree of ocular misalignment and uses it to compute the appropriate saccade for either eye. The obvious place to investigate is the superior colliculus, a midbrain oculomotor center responsible for the generation of saccades. Here, we report the first experiments in the superior colliculus of awake primates with strabismus using a combination of single-cell recordings and microstimulation to explore the organization of its topographic maps., (Copyright © 2018 the authors 0270-6474/18/380173-10$15.00/0.)

Purpose: To determine if the deviation angle changes in subjects with intermittent exotropia as they alternate fixation between the right and left eye in primary gaze., Methods: In this prospective observational cohort study, 37 subjects with intermittent exotropia were tested for evidence of incomitance. The position of each eye was recorded with a video tracker during fixation on a small central target. A cover-uncover test was performed by occluding one eye with a shutter that passed infrared light, allowing continuous tracking of both eyes. The deviation angle was measured during periods of right eye and left eye fixation. Incomitance was assessed as a function of eye preference, fixation stability, and exotropia variability., Results: The mean exotropia was 18.2° ± 8.1°. A difference between right exotropia and left exotropia was detectable in 16/37 subjects. Allowing for potential tracking error, the incomitance had a mean amplitude of 1.7°. It was not related to a difference in accommodative effort, eye preference, fixation stability, or variability in deviation., Conclusions: Comitance is regarded as a feature that distinguishes strabismus from paralytic or restrictive processes. Unexpectedly, eye tracking during the cover-uncover test showed that incomitance is present in approximately 40% of subjects with intermittent exotropia. It averages 10% of the exotropia, and can equal up to 5°. When substantial, it may be worth considering when planning surgical correction.

Purpose: To characterize eye movements made by patients with intermittent exotropia when fusion loss occurs spontaneously and to compare them with those induced by covering 1 eye and with strategies used to recover fusion., Design: Prospective study of a patient cohort referred to our laboratory., Participants: Thirteen patients with typical findings of intermittent exotropia who experienced frequent spontaneous loss of fusion., Methods: The position of each eye was recorded with a video eye tracker under infrared illumination while fixating on a small central near target., Main Outcome Measures: Eye position and peak velocity measured during spontaneous loss of fusion, shutter-induced loss of fusion, and recovery of fusion., Results: In 10 of 13 subjects, the eye movement made after spontaneous loss of fusion was indistinguishable from that induced by covering 1 eye. It reached 90% of full amplitude in a mean of 1.75 seconds. Peak velocity of the deviating eye's movement was highly correlated for spontaneous and shutter-induced events. Peak velocity was also proportional to exotropia amplitude. Recovery of fusion was more rapid than loss of fusion, and often was accompanied by interjection of a disconjugate saccade., Conclusions: Loss of fusion in intermittent exotropia is not influenced by visual feedback. Excessive divergence tone may be responsible, but breakdown of alignment occurs via a unique, pathological type of eye movement that differs from a normal, physiological divergence eye movement., (Copyright © 2017 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

The superior colliculus is a major brain stem structure for the production of saccadic eye movements. Electrical stimulation at any given point in the motor map generates saccades of defined amplitude and direction. It is unknown how this saccade map is affected by strabismus. Three macaques were raised with exotropia, an outwards ocular deviation, by detaching the medial rectus tendon in each eye at age 1 mo. The animals were able to make saccades to targets with either eye and appeared to alternate fixation freely. To probe the organization of the superior colliculus, microstimulation was applied at multiple sites, with the animals either free-viewing or fixating a target. On average, microstimulation drove nearly conjugate saccades, similar in both amplitude and direction but separated by the ocular deviation. Two monkeys showed a pattern deviation, characterized by a systematic change in the relative position of the two eyes with certain changes in gaze angle. These animals' saccades were slightly different for the right eye and left eye in their amplitude or direction. The differences were consistent with the animals' underlying pattern deviation, measured during static fixation and smooth pursuit. The tectal map for saccade generation appears to be normal in strabismus, but saccades may be affected by changes in the strabismic deviation that occur with different gaze angles., (Copyright © 2016 the American Physiological Society.)

Purpose: To determine how formation of an acquired myopic crescent adjacent to the optic disc affects metabolic activity in the primary visual cortex., Design: Laboratory animal study., Participants: Three macaque monkeys., Methods: The blind spot region in the primary visual cortex was labeled by cytochrome oxidase (CO) histochemistry analysis or [(3)H]proline autoradiography., Main Outcome Measures: Visualization of the representation of the blind spot and myopic peripapillary crescent in the visual cortex., Results: In high myopia, a region resembling the myopic peripapillary crescent was visible in cortical sections processed for CO. In this region, metabolic activity was reduced in ocular dominance columns that normally would be driven by input from retina corresponding to the myopic peripapillary crescent., Conclusions: The formation of a myopic crescent is accompanied by loss of metabolic activity in the cortex supplied by the affected retina. This observation confirms that retinal tissue is damaged by the development of a myopic crescent, rather than simply translocated in a temporal direction. The cortical defect matches the myopic peripapillary crescent in size and shape, indicating that fill-in of the retinotopic map by healthy, surrounding retina does not occur., (Copyright © 2016 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Importance: In strabismus, the fixating eye conveys the direction of gaze while the fellow eye points at a peripheral location in space. The stability of the eyes may be reduced by the absence of a common target., Objective: To quantify the stability of eye position in strabismus and to measure variability in the ocular deviation., Design, Setting, and Participants: From 2010 to 2014, a prospective comparative case study of 25 patients with alternating exotropia with normal visual acuity in each eye and 25 control individuals was conducted in a laboratory at a tertiary eye center. A video eye tracker was used to measure the position of each eye while participants alternated fixation on the center of a cross under dichoptic conditions or scanned pictures of natural scenes., Main Outcomes and Measures: Spatial and temporal variability in the position of the fixating eye and the nonfixating eye in patients with strabismus and control individuals, quantified by the log area of ellipses containing 95% of eye positions or mean SDs of eye position., Results: In the 25 patients with strabismus, the mean (SD) age was 28 (14) years (range, 8-55 years) and the mean (SD) ocular deviation was 14.2° (5.9°) (range, 4.4°-22.4°). In the patients with strabismus, the mean position variability (1.80 log units; 95% CI, 1.66-1.93) for the deviating eye was greater than for the fixating eye (1.26 log units; 95% CI, 1.17-1.35) (P < .001). The fixating eye of patients with strabismus was more variable in position than the fixating eye of individuals without strabismus (0.98 log units; 95% CI, 0.88-1.08) (P < .005)., Conclusions and Relevance: In patients with strabismus, even without amblyopia, the deviated eye is more variable in position than the fixating eye. Both eyes are less stable in position than the eyes of control individuals, which indicates that strabismus impairs the ability to fixate targets steadily. Saccades contribute to variability of the deviation angle because they are less conjugate in patients with strabismus.

The vascular supply to layers and columns was compared in macaque primary visual cortex (V1) by labeling red blood cells via their endogenous peroxidase activity. Alternate sections were processed for cytochrome oxidase to reveal "patches" or "blobs," which anchor the interdigitated column systems of striate cortex. More densely populated cell layers received the most profuse blood supply. In the superficial layers the blood supply was organized into microvascular lobules, consisting of a central venule surrounded by arterioles. Each vessel was identified as an arteriole or venule by matching it with the penetration site where it entered the cortex from a parent arteriole or venule in the pial circulation. Although microvascular lobules and cytochrome oxidase patches had a similar periodicity, they bore no mutual relationship. The size and density of penetrating arterioles and venules did not differ between patches and interpatches. The red blood cell labeling in patches and interpatches was equal. Moreover, patches and interpatches were supplied by an anastomotic pial arteriole system, with no segregation of blood supply to the two compartments. Often a focal constriction was present at the origin of pial arterial branches, indicating that local control of cortical perfusion may be accomplished by vascular sphincters., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

To probe the mechanism of visual suppression, we have raised macaques with strabismus by disinserting the medial rectus muscle in each eye at 1 mo of age. Typically, this operation produces a comitant, alternating exotropia with normal acuity in each eye. Here we describe an unusual occurrence: the development of severe amblyopia in one eye of a monkey after induction of exotropia. Shortly after surgery, the animal demonstrated a strong fixation preference for the left eye, with apparent suppression of the right eye. Later, behavioral testing showed inability to track or to saccade to targets with the right eye. With the left eye occluded, the animal demonstrated no visually guided behavior. Optokinetic nystagmus was absent in the right eye. Metabolic activity in striate cortex was assessed by processing the tissue for cytochrome oxidase (CO). Amblyopia caused loss of CO in one eye's rows of patches, presumably those serving the blind eye. Layers 4A and 4B showed columns of reduced CO, in register with pale rows of patches in layer 2/3. Layers 4C, 5, and 6 also showed columns of CO activity, but remarkably, comparison with more superficial layers showed a reversal in contrast. In other words, pale CO staining in layers 2/3, 4A, and 4B was aligned with dark CO staining in layers 4C, 5, and 6. No experimental intervention or deprivation paradigm has been reported previously to produce opposite effects on metabolic activity in layers 2/3, 4A, and 4B vs. layers 4C, 5, and 6 within a given eye's columns., (Copyright © 2015 the American Physiological Society.)

The patches in primary visual cortex constitute hot spots of metabolic activity, manifested by enhanced levels of cytochrome oxidase (CO) activity. They are also labeled preferentially by immunostaining for glutamic acid decarboxylase (GAD), γ-aminobutyric acid (GABA), and parvalbumin. However, calbindin shows stronger immunoreactivity outside patches. In light of this discrepancy, the distribution of the vesicular GABA transporter (VGAT) was examined in striate cortex of two normal macaques. VGAT immunoreactivity was strongest in layers 4B, 4Cα, and 5. In tangential sections, the distribution of CO, GAD, and VGAT was compared in layer 2/3. There was a close match between all three labels. This finding indicates that GABA synthesis is enriched in patches, and that inhibitory synapses are more active in patches than interpatches.

In strabismus, potentially either eye can inform the brain about the location of a target so that an accurate saccade can be made. Sixteen human subjects with alternating exotropia were tested dichoptically while viewing stimuli on a tangent screen. Each trial began with a fixation cross visible to only one eye. After the subject fixated the cross, a peripheral target visible to only one eye flashed briefly. The subject's task was to look at it. As a rule, the eye to which the target was presented was the eye that acquired the target. However, when stimuli were presented in the far nasal visual field, subjects occasionally performed a "crossover" saccade by placing the other eye on the target. This strategy avoided the need to make a large adducting saccade. In such cases, information about target location was obtained by one eye and used to program a saccade for the other eye, with a corresponding latency increase. In 10/16 subjects, targets were presented on some trials to both eyes. Binocular sensory maps were also compiled to delineate the portions of the visual scene perceived with each eye. These maps were compared with subjects' pattern of eye choice for target acquisition. There was a correspondence between suppression scotoma maps and the eye used to acquire peripheral targets. In other words, targets were fixated by the eye used to perceive them. These studies reveal how patients with alternating strabismus, despite eye misalignment, manage to localize and capture visual targets in their environment., (Copyright © 2014 the authors 0270-6474/14/3414578-11$15.00/0.)

When an eye becomes deviated in early childhood, a person does not experience double vision, although the globes are aimed at different targets. The extra image is prevented from reaching perception in subjects with alternating exotropia by suppression of each eye's peripheral temporal retina. To test the impact of visual suppression on neuronal activity in primary (striate) visual cortex, the pattern of cytochrome oxidase (CO) staining was examined in four macaques raised with exotropia by disinserting the medial rectus muscles shortly following birth. No ocular dominance columns were visible in opercular cortex, where the central visual field is represented, indicating that signals coming from the central retina in each eye were perceived. However, the border strips at the edges of ocular dominance columns appeared pale, reflecting a loss of activity in binocular cells from disruption of fusion. In calcarine cortex, where the peripheral visual field is represented, there were alternating pale and dark bands resembling ocular dominance columns. To interpret the CO staining pattern, [(3)H]proline was injected into the right eye in two monkeys. In the right calcarine cortex, the pale CO columns matched the labeled proline columns of the right eye. In the left calcarine cortex, the pale CO columns overlapped the unlabeled columns of the left eye in the autoradiograph. Therefore, metabolic activity was reduced in the ipsilateral eye's ocular dominance columns which serve peripheral temporal retina, in a fashion consistent with the topographic organization of suppression scotomas in humans with exotropia.

Misalignment of the eyes can lead to double vision and visual confusion. However, these sensations are rare when strabismus is acquired early in life, because the extra image is suppressed. To explore the mechanism of perceptual suppression in strabismus, the visual fields were mapped binocularly in 14 human subjects with exotropia. Subjects wore red/blue filter glasses to permit dichoptic stimulation while fixating a central target on a tangent screen. A purple stimulus was flashed at a peripheral location; its reported color ("red" or "blue") revealed which eye's image was perceived at that locus. The maps showed a vertical border between the center of gaze for each eye, splitting the visual field into two separate regions. In each region, perception was mediated by only one eye, with suppression of the other eye. Unexpectedly, stimuli falling on the fovea of the deviated eye were seen in all subjects. However, they were perceived in a location shifted by the angle of ocular deviation. This plasticity in the coding of visual direction allows accurate localization of objects everywhere in the visual scene, despite the presence of strabismus.

The mechanism of diplopia from enophthalmos is not well understood. We describe a 55-year-old man who underwent a left transorbital craniotomy for clipping of a basilar aneurysm. The lateral orbital wall was not reconstructed properly, resulting in 8 mm of left enophthalmos. Months after surgery the patient developed diplopia with ocular excursions, although he remained orthotropic in primary gaze. The left eye was limited in elevation, adduction, and abduction. These findings were confirmed by eye movement recordings, which showed ocular separation increasing with gaze eccentricity. A CT scan demonstrated a defect in the sphenoid and frontal bones, profound enophthalmos, and shortening of the rectus muscles. Slack in the extraocular muscles reduced the force generated by each muscle, causing diplopia with ocular rotation. This case underscores the value of careful orbital wall reconstruction after orbitotomy and suggests a mechanism for diplopia produced by postoperative enophthalmos.

The abundant concentration of cytochrome oxidase in patches or blobs of primate striate cortex has never been explained. Patches are thought to contain unoriented, color-opponent neurons. Lacking orientation selectivity, these cells might endow patches with high metabolic activity because they respond to all contours in visual scenes. To test this idea, we measured orientation tuning in layer 2/3 of macaque cortical area V1 using acutely implanted 100-electrode arrays. Each electrode recording site was identified and assigned to the patch or interpatch compartment. The mean orientation bandwidth of cells was 28.4° in patches and 25.8° in interpatches. Neurons in patches were indeed less orientation selective, but the difference was subtle, indicating that the processing of form and color is not strictly segregated in V1. The most conspicuous finding was that patch cells had a 49% greater overall firing rate. This global difference in neuronal responsiveness, rather than an absence of orientation tuning, may account for the rich mitochondrial enzyme activity that defines patches.

Neurophysiological recording in alert monkeys requires the creation of a permanent aperture in the skull for repeated insertion of microelectrodes. Most laboratories use polymethyl methacrylate to attach a recording chamber over the skull opening. Here, we describe a titanium chamber that fastens to the skull with screws, using no polymethyl methacrylate. The gap between the base of the chamber and the skull is filled with hydroxyapatite, forming a watertight gasket. As the chamber base osseointegates with the skull, the hydroxyapatite is replaced with bone. Rather than having a finite lifetime, the recording chamber becomes more firmly anchored the longer it is in place. It has a small footprint, low profile, and needs little maintenance to control infection. Toilette consists of occasional application of betadine to clean the scalp margin, followed by application of neomycin, polymyxin, and bacitracin ointment. Antibiotic is also placed inside the chamber to suppress bacterial proliferation. Thickening of the dura within the chamber can be prevented by regular application of mitocycin C and/or bevacizumab, an antibody against vascular endothelial growth factor. By conducting an e-mail survey, this protocol for chamber maintenance was compared with procedures used in 37 other vision research laboratories. Refinement of appliances and techniques used for recordings in awake monkeys promises to increase the pace of scientific discovery and to benefit animal welfare.

To provide an animal model of human exotropia, a free tenotomy of the medial recti was performed in two infant macaques. When the animals were old enough to record eye movements with video eye trackers, we measured their ductions, ocular alignment, comitance, smooth pursuit, fixation preference, and gaze stability. Partial recovery of adduction occurred in each monkey from spontaneous re-attachment of the medial rectus muscle to the eye. However, each animal was left with a relatively comitant, large angle exotropia. The magnitude of the exotropia was not affected by covering one eye. There was no dissociated vertical deviation or any significant "A" or "V" pattern to the horizontal misalignment. Smooth pursuit was more accurate when tracking nasally compared with temporally in both animals. Compensatory catch-up saccades in the tracking eye were always accompanied by conjugate movements in the deviated eye. Despite tenotomy of the medial recti, the velocity of adducting saccades was normal. Both monkeys alternated fixation, preferring to use the left eye for targets on the left side and the right eye for targets on the right. Each animal was capable of switching fixation while making accurate saccades. One of the monkeys developed a vertical pendular nystagmus, which was most prominent in the deviated eye. Macaques with ocular misalignment from medial rectus tenotomy exhibit features that are present in humans with alternating exotropia. These animals will be valuable for probing the cortical mechanisms that underlie visual suppression in strabismus.

Many neurophysiological experiments involving monkeys require that the head be stabilized while the animal performs a task. Often a post is attached to the skull to accomplish this goal, using a headcap formed from dental acrylic. We describe a new headpost, developed by refinement of several prototypes, and supply an AutoCAD file to aid in machine shop production. This headpost is fabricated from a single piece of commercially pure titanium. It has a footplate consisting of four limbs arranged in the configuration of a "K." These are bent during surgery to match the curvature of the skull and attached with specialized titanium bone screws. Headposts were implanted in seven rhesus monkeys ranging in age from 2 yr to adult. None has been rejected after up to 17 mo of regular use. They require little or no daily toilette and create only a 0.80-cm(2) defect in the scalp. Computed tomography after implantation showed that the skull undergoes remodeling to embed the footplate in bone. This finding was confirmed by necropsy in two subjects. The outer table of the skull had grown over the titanium footplate, whereas the inner table had thickened to bury the tips of the titanium screws. The remarkable strength of the skull/implant bond was demonstrated by applying increasing amounts of torque to the headpost. At 26.3 Nm, the headpost tore from its metal footplate, but no screws came loose. The excellent performance of this implant is explained by integration of biocompatible titanium into remodeled bone tissue. The headpost is simpler to implant, more securely anchored, easier to maintain, and less obtrusive than devices attached with acrylic.

Retinal spikes impinging on relay neurons in the lateral geniculate nucleus (LGN) generate synaptic potentials, which sometimes produce spikes sent to visual cortex. We examined how signal transmission is regulated in the macaque LGN by recording the retinal input to a single LGN neuron while stimulating the receptive field center with a naturalistic luminance sequence. After extracting the EPSPs, which are often partially merged with spike waveforms, we found that >95% of spikes were associated with an EPSP from a single retinal ganglion cell. Each spike within a "burst" train was generated by an EPSP, indicating that LGN bursts are inherited from retinal bursts. LGN neurons rarely fired unless at least two EPSPs summated within 40 ms. This facilitation in EPSP efficacy was followed by depression. If a spike was generated by the first EPSP in a pair, it did not alter the efficacy of the second EPSP. Hence, the timing of EPSPs arising from the primary retinal driver governs synaptic efficacy and provides the basis for successful retinogeniculate transmission.

Primate striate cortex contains a prominent system of columns referred to as cytochrome oxidase (CO) patches. Studies directed at the morphology and interconnections of cells in patches would be facilitated by a method that revealed the location of patches in the intact cortex. In three adult rhesus monkeys we prepared flatmounts of striate cortex [Horton JC, Hocking DR. Intrinsic variability of ocular dominance column periodicity in normal macaque monkeys. J Neurosci 1996;16:7228-39; Sincich LC, Adams DL, Horton JC. Complete flatmounting of the macaque cerebral cortex. Vis Neurosci 2003;20:663-86]. The flattened specimens were then reacted for CO activity prior to sectioning. Transillumination of the intact cortical sheet revealed an extensive pattern of dark ovals. It was confirmed that this pattern corresponded to the CO patches by subsequently cutting tangential sections and comparing them with images from the intact block. In vitro labeling of CO patches in the intact striate cortex may prove useful for directing injections of anatomical tracers such as Lucifer Yellow or DiI into identified patch and interpatch compartments.

The dorsolateral pontine nucleus (DLPN) and nucleus reticularis tegmenti pontis (NRTP) comprise obligatory links in the cortico-ponto-cerebellar system supporting smooth pursuit eye movements. We examined the response properties of DLPN and rNRTP neurons during step-ramp smooth pursuit of a small target moving across a dark background. Our neurophysiological studies were conducted in awake, behaving juvenile macaques (Macaca mulatta). We used multiple linear-regression modeling to estimate the relative sensitivities of neurons to eye parameters (position, velocity, and acceleration) and retinal-error parameters (position, velocity, and acceleration). We found that a large proportion of pursuit-related DLPN neurons primarily code eye-velocity information, whereas a large proportion of rNRTP neurons primarily code eye-acceleration information. We calculated the relative decrease in variance found when using a six-component model that included both eye- and retinal-error parameters compared with three-component models that include either eye or retinal error. These comparisons show that a majority of DLPN (14/20) and rNRTP (17/19) neurons have larger contributions from eye compared with retinal-error parameters (P < 0.001, paired t-test). Even though eye-motion parameters provide the strongest contributions in a given model, a significant contribution from retinal error was often present (i.e., >20% reduction in variance in 6-component model compared with 3-component models). Thus our results indicate that the DLPN plays a larger role in maintaining steady-state smooth pursuit eye velocity, whereas rNRTP contributes to both the initiation and maintenance of smooth pursuit.

We recorded from single units in the pretectal nucleus of the optic tract (NOT) of the nonhuman primate. Specifically, we examined units that are modulated during smooth tracking of a small laser spot against a dark background. We used a nonlinear optimization procedure to determine whether the unit responses of these parafoveal cells are better described by a model that incorporates retinal error motion parameters or by a model that incorporates eye motion parameters. Our main finding was that all the cells in our sample group were better fit with a three-component model that incorporated retinal error motion parameters of position, velocity and acceleration (average coefficient of determination = 0.84) than a model that used position, velocity and acceleration components of eye motion (average coefficient of determination = 0.68). Other analyses involved comparison of goodness of fit between the three-component retinal error model and two-component retinal error models that excluded position or acceleration related terms. We found that there was a statistically significant degradation in the fit when position and acceleration related terms were dropped from the retinal error based model (P<0.05). Unit data from experiments in which the laser spot was extinguished for a brief period of time during tracking showed that the unit response was decreased following the target blink. We conclude on the basis of this and previous experimental data and our dynamic modeling approach that the parafoveal cells in the NOT primarily encode retinal error motion. Further they encode position, velocity and acceleration components of retinal error that could be used by other downstream structures for synthesis of a smooth-pursuit eye movement.