Neuronal projections from V1 to V2 in amblyopia.

The mechanism of amblyopia in children with congenital cataract is not understood fully, but studies in macaques have shown that geniculate synapses are lost in striate cortex (V1). To search for other projection abnormalities in amblyopia, the pathway from V1 to V2 was examined using a triple-label technique in three animals raised with monocular suture. [(3)H]proline was injected into one eye to label the ocular dominance columns. Cholera toxin B subunit conjugated to gold (CTB-Au) was injected into V2 to label V1 projection neurons. Alternate sections were processed for cytochrome oxidase (CO) and CTB-Au, or dipped for autoradiography. Eight fields of CTB-Au-labeled cells in V1 opposite injection sites were plotted in layers 2/3 or 4B. After thin stripe injection, labeled cells were concentrated in CO patches. Despite column shrinkage, cells in deprived and normal columns were equal in size and density in both layers 2/3 and 4B. After pale or thick stripe injection, labeled cells were concentrated in interpatches. Only 23% of projection neurons originated from deprived columns. This reduction exceeded the degree of column shrinkage, a result explained by the fact that column shrinkage causes disproportionate loss of interpatch territory. These data indicate that early monocular form deprivation does not alter the segregation of patch and interpatch pathways to V2 stripes or cause selective loss or atrophy of V1 projection neurons. The effect of shrinkage of geniculocortical afferents in layer 4C following visual deprivation is not amplified further by attenuation of the amblyopic eye's projections from V1 to V2.

A watertight acrylic-free titanium recording chamber for electrophysiology in behaving monkeys.

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.

V1 interpatch projections to v2 thick stripes and pale stripes.

Cytochrome oxidase (CO) reveals two compartments in V1 (patches and interpatches) and three compartments in V2 (thin, pale, and thick stripes). Previously, it was shown that thin stripes receive input predominantly from patches. Here we examined the projections to thick and pale stripes in macaques, revealed by retrograde tracer injections. After thick stripe injection, cells were distributed in layer 2/3 (67%), layer 4A (7%), layer 4B (23%), and layer 5/6 (2%). Except in layer 5/6, cells were concentrated in interpatches, with a stronger bias in layer 2/3 (84%) than in layer 4B (75%). After pale stripe injection, cells were found in layer 2/3 (87%), layer 4A (2%), layer 4B (10%), and layer 5/6 (2%). As for thick stripes, cells were located preferentially in interpatches in layer 2/3 (84%) and layer 4B (72%) but not in layer 5/6. Thick stripes received a higher proportion of their input from layer 4B, compared with pale stripes, consistent with reports that thick stripe neurons exhibit a pronounced layer 4B influence. This difference aside, both stripe types receive similar inputs from V1, at least in terms of cortical layer and CO compartment. This finding was bolstered by injecting different tracers into pale and thick stripes; 10-27% of cells were double labeled, with most located in interpatches. These results suggest that the distinctive receptive field properties of neurons in thick and pale stripes are generated by local V2 circuits, or by other specific projections, rather than by differing sources of laminar and compartmental input from V1.

Ocular motor behavior in macaques with surgical exotropia.

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.

A biocompatible titanium headpost for stabilizing behaving monkeys.

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.

Neurons in V1 patch columns project to V2 thin stripes.

In the primate, connections between primary visual cortex (V1) and the second visual area (V2) are segregated according to the characteristic pattern of cytochrome oxidase (CO) activity in each of these cortical areas. Patches supply thin stripes, whereas interpatches supply pale stripes and thick stripes. Previously, the projection from patches to thin stripes was reported to arise exclusively from layer 2/3. In this present report, we made injections of a retrograde tracer, cholera toxin-B (CTB-Au), into macaque V2 thin stripes to re-examine the laminar origin of their input from V1. While the great majority of cells indeed resided in layer 2/3, small populations were also present in layers 4A, 4B, and 5/6. The location of CTB-filled cells in each layer was analyzed to determine the relationship with CO patches. Cells in layers 2/3, 4A, and 4B were aggregated into patches, forming columns that project to thin stripes. Surprisingly, cells in layer 5/6 were scattered, seemingly at random. These findings confirm that the main V1 projection to V2 stripes emanates from patches in layer 2/3. However, multiple V1 layers innervate V2 thin stripes, and the projection from layer 5/6 does not respect the patch/interpatch dichotomy.