The development and activity-dependent expression of aggrecan in the cat visual cortex.

The Cat-301 monoclonal antibody identifies aggrecan, a chondroitin sulfate proteoglycan in the cat visual cortex and dorsal lateral geniculate nucleus (dLGN). During development, aggrecan expression increases in the dLGN with a time course that matches the decline in plasticity. Moreover, examination of tissue from selectively visually deprived cats shows that expression is activity dependent, suggesting a role for aggrecan in the termination of the sensitive period. Here, we demonstrate for the first time that the onset of aggrecan expression in area 17 also correlates with the decline in experience-dependent plasticity in visual cortex and that this expression is experience dependent. Dark rearing until 15 weeks of age dramatically reduced the density of aggrecan-positive neurons in the extragranular layers, but not in layer IV. This effect was reversible as dark-reared animals that were subsequently exposed to light showed normal numbers of Cat-301-positive cells. The reduction in aggrecan following certain early deprivation regimens is the first biochemical correlate of the functional changes to the γ-aminobutyric acidergic system that have been reported following early deprivation in cats.

Postnatal expression profile of OBCAM implies its involvement in visual cortex development and plasticity.

This study examined the expression of a neuron-specific cell adhesion molecule, OBCAM (opioid-binding cell adhesion molecule), at both the mRNA and protein levels in the cat primary visual cortex at various postnatal ages, using cDNA array analysis and immunocytochemistry. Results obtained using both methods showed that the expression level of OBCAM was high in young and low in older and adult visual cortex. OBCAM-immunoreactivities were associated predominantly with perikarya and dendrites of pyramidal neurons, and OBCAM-immunopositive neurons were present in all cortical layers. Immunostaining of OBCAM in adult visual cortex showed a reduced number of immunopositive neurons and neurites and relatively lower staining intensities as compared with younger animals. In addition, the number of OBCAM-immunopositive neurons was significantly higher in the visual cortex of 4-month-old animals dark-reared from birth than those in age-matched normally reared animals. These results suggest that OBCAM may play an important role in visual cortex development and plasticity.

Gene expression patterns during enhanced periods of visual cortex plasticity.

During a critical period in its postnatal development the mammalian visual cortex displays susceptibility to experience-dependent alterations of neuronal response properties. Plasticity represents an integrated set of developmental processes controlled by a transcriptional hierarchy that coordinates the action of many genes. To illuminate the expression of these critical genes, we examined gene expression patterns of 18371 non-redundant cDNAs in the visual cortex of cats at birth, at eye opening, at the peak of the critical period of eye dominance plasticity and in the adult cat using filter-based cDNA arrays and software-based hierarchical cluster analysis. We identified a small set of genes that were selectively expressed during the peak of the critical period for plasticity. We further examined the patterns of expression of these genes by analyzing the gene expression pattern of dark-reared chronologically older animals that are known to retain this ocular dominance plasticity beyond the chronologically defined critical period. This additional cluster assessment allowed us to separate age-related changes in the patterns of gene expression from plasticity-related changes, thus identifying a subset of genes that we define as plasticity candidate genes. Those plasticity candidate genes that have previously characterized functions include participants in second messenger systems, in cell adhesion, in transmitter recycling and cytokines, among others. Comparison of cDNA array quantitation with reverse transcription-polymerase chain reaction showed almost identical expression profiles for three genes that we examined. The expression pattern of one identified gene, opioid binding cell adhesion molecule, from the cDNA array analysis, is also in agreement with immunocytochemical results. We conclude that the approach of high-density cDNA array hybridization can be used as a useful tool for examining a complex phenomenon of developmental plasticity since it is amenable to multiple developmental stage gene expression comparisons.

Initial recovery of vision after early monocular deprivation in kittens is faster when both eyes are open.

A comparison was made of the speed of visual recovery in the deprived eye of kittens after a 6-day period of monocular deprivation imposed at 5-9 weeks of age in two postdeprivation conditions. In one condition, binocular recovery (BR), both eyes were open, whereas in the other condition, reverse lid-suture (RLS), the formerly nondeprived eye was closed to force the animal to use the originally deprived eye. In littermate pairs, BR kittens began to recover form vision 12 to 30 h before those subjected to RLS. The vision of the deprived eye of the BR animals remained superior to that of their RLS littermates for 4-8 days. Although this finding is difficult to reconcile with competitive mechanisms of synaptic plasticity, it supports a prediction of an alternative model of synaptic plasticity [Bienenstock, E. L., Cooper, L. N. & Munro, P. W. (1982) J. Neurosci. 2, 32-48] for slower initial recovery with RLS because of the time required to reset the modification threshold.

Development of cytochrome oxidase blobs in visual cortex of normal and visually deprived cats.

Cytochrome oxidase (CO) blobs are central to our understanding of the columnar organization and parallel pathways in primate and cat visual cortex. In primates, development of blobs and their relationship with other columnar features of the visual cortex begins pre-natally, before visual experience. In kittens, the supragranular layers differentiate post-natally, after eye opening, raising the possibility that visual experience may influence the development of blobs in cat V1. We have examined the development of blobs in unfolded and flattened sections through the visual cortex of normally reared, dark-reared, monocularly deprived and binocularly deprived kittens. Blobs were found in superficial layers of V1 of normally reared kittens as early as 2 weeks of age, although at this age the overall CO staining in V1 was lighter than in V2. By 6 weeks of age the blobs were adult-like. A patchy pattern of CO staining was also found in V2 of young kittens but not in adults. Visual experience was not necessary for expression of the blobs and monocularly deprived kittens had well developed blobs, indicating that strong Y cell drive is not necessary for the development of blobs in cat V1. CO blobs appear in kitten V1 very early in post-natal development and their expression is independent of visual experience, suggesting that they may be an intrinsic feature of V1 organization.

Visual recovery after monocular deprivation is driven by absolute, rather than relative, visually evoked activity levels.

It is now well established that the anatomical and functional development of the central visual pathways of a number of higher mammalian species is activity-dependent [1-3]. This dependence was revealed by the functional effects of an early period of monocular deprivation, where one eye of a young animal was deprived for a time of patterned visual input. Subsequently, most cells in the visual cortex (area 17) could be excited only by visual stimuli delivered to the non deprived eye [4-6] and the animal appeared blind through the deprived eye [7,8]. These effects have been attributed to a competitive activity-dependent mechanism in development, whereby the two eyes compete for control of cortical cells [9,10]. There are, however, suggestions that the substantial recovery that can occur after monocular deprivation may be mediated by a different mechanism. Here, insight into the nature of this mechanism has been provided by monitoring the speed of changes in the vision of the deprived eye of a kitten after 6 days of monocular deprivation. Although both eyes were open during the recovery period, the kitten was able to see with its deprived eye only 2 hours after visual input was restored to this eye. The visual acuity of this eye improved rapidly in the first 24 hours and continued in an orderly way for 6 weeks. In contrast to the effects during monocular deprivation, which depend upon a competitive activity-dependent process, we propose that the events that follow deprivation rely on a mechanism driven by the absolute level of visually evoked activity through the formerly deprived eye.

Stereodeficient subjects show substantial differences in interocular transfer of two motion adaptation aftereffects.

Interocular transfer (IOT) of two motion aftereffects was examined in subjects with normal and deficient stereopsis. Normal subjects showed complete (100%) IOT of motion adaptation on coherent motion thresholds, but only partial IOT of a conventional motion aftereffect, supporting suggestions that the latter aftereffect may be mediated at a lower level in the visual pathway than the extrastriate regions implicated in processing coherent motion. This idea was strengthened by an even greater dissociation between the extent of IOT of the two aftereffects among stereodeficient subjects who exhibited very low IOT of the conventional motion aftereffect, but high (> 87%) IOT of the coherence motion aftereffect.

Remodeling of lesioned kitten visual cortex after xenotransplantation of fetal mouse neopallium.

Remodeling of the mechanically injured cerebral cortex of kittens was studied in the presence of a neural xenograft taken from mouse fetuses. Solid neural tissue from the neopallium of a 14-day-old fetus was transferred into a cavity prepared in visual cortical area 18 of 33-day-old kittens. Injections of bromodeoxyuridine (BrdU) were used to monitor postoperative cell proliferation. Two months after transplantation, the presence of graft tissue in the recipient brain was assessed by Thy-1 immunohistochemistry. Antibodies specific for neurons, astrocytes, and oligodendrocytes and hematoxylin staining for endothelial cells were used for the characterization of proliferating (BrdU+) cells. The following were the major observations: 1) Of ten transplanted kittens, four had the cavity completely filled with neural tissue that resembled the intact cerebral cortex in its cytoarchitecture, whereas, in four other kittens, the cavity was partially closed. In two kittens, the cavity remained or became larger, which was also the case with all four sham-operated (lesioned, without graft) animals. 2) A substantial part of the remodeled tissue was of host origin. Only a few donor cells survived and dispersed widely in the host parenchyme. 3) In the remodeled region of transplanted animals, the densities of nerve, glial, and endothelial cells were similar to those in intact animals. 4) Cell proliferation increased after transplantation but only within a limited time, because, 2 months after the operation, the number of mitotic cells in the grafted cerebral cortex did not differ from that in intact controls. Our data suggest that the xenograft evokes repair processes in the kitten visual cortex that lead to structural recovery from a mechanical insult. The regeneration seems to rely on a complex interplay of many different mechanisms, including attenuation of necrosis, cell proliferation, and immigration of host cells into the wounded area.

Effects of early periods of monocular deprivation and reverse lid suture on the development of Cat-301 immunoreactivity in the dorsal lateral geniculate nucleus (dLGN) of the cat.

During certain sensitive periods early in postnatal life, the anatomical and physiological development of the central visual pathways of cats and monkeys can be affected by the nature of an animal's early visual experience. In the last few years, studies have been started on some of the molecular and biochemical events that underlie the many functional changes induced by early selected visual deprivation in the visual cortex of kittens. In this respect, the monoclonal antibody Cat-301 provides a potentially powerful tool, because it recognizes in the cat dorsal lateral geniculate nucleus (dLGN) a proteoglycan associated with the surface of a particular class of cells, namely Y cells. In the dLGN, the Cat-301 proteoglycan appears late in postnatal development, and it expression has been shown to be experience dependent in both the dLGN and visual cortex (M. Sur, D. Frost, and S. Hockfield, 1988, J. Neurosci. 8:874-882; A. Guimaraes, S. Zaremba, and S. Hockfield, 1990, J. Neurosci. 10:3014-3024). We have explored further the experience-dependent nature of Cat-301 expression in the dLGN with a view to establishing a biochemical correlate of the many functional changes induced by early monocular deprivation and its reversal in the kitten visual system. In addition to demonstrating differences in Cat-301 expression between deprived and nondeprived laminae of the dLGNs of kittens monocularly deprived to only 4 or 5 weeks of age, the magnitude of the laminar difference was found to increase as the period of deprivation was extended. Moreover, monocularly deprived kittens that subsequently received long periods of reverse lid suture exhibited a reversal of the pattern of immunoreactivity, so that the greatest immunoreactivity occurred in laminae innervated by the initially deprived eye. However, possibly the most surprising and important finding was the extremely low levels of immunoreactivity observed in both A laminae of monocularly deprived animals that had received relatively short periods of reverse lid suture. These data suggest that Y cell development can be drastically altered depending on the time of initiation of the period of reverse lid suture and its duration.

A method to study changes in eye-related columns in the visual cortex of kittens during and following early periods of monocular deprivation.

An immunohistochemical method that exploits the rapid light-evoked expression of Fos, the protein product of the immediate early gene, c-fos, to visualize eye-related columns in the visual cortex, has been used to provide preliminary data on the relative innervation of the cortex by the two eyes of monocularly deprived kittens and the speed of the changes that occur afterward during reverse occlusion. In contrast to conventional anatomical techniques, the method allows both cellular resolution and documentation of the dimensions of eye-related columns through the depth of the cortex. In kittens monocularly deprived from near birth, Fos-immunoreactive neurones were observed in oval or circular patches, the size of which decreased as the duration of deprivation was increased from 4 to 6 weeks. Following reverse occlusion at 5 weeks of age, the size of the patches increased rapidly so that after 4 days their area had approximately tripled. In addition to providing possible insights into the anatomical underpinnings of the puzzling behavioural effects that occur following termination of short periods of reverse occlusion, the method can be used to investigate the temporal order of the anatomical effects of monocular deprivation in different cortical layers.

Depth perception in monocularly deprived cats following part-time reverse occlusion.

The behavioural effects of an early period of monocular deprivation can be extremely profound. However, it is possible to achieve a high degree of recovery, even to normal levels of visual acuity, by prompt imposition of certain regimes of part-time reverse occlusion where the initially non-deprived eye is occluded for only part of each day in order to allow a daily period of binocular visual exposure. In this paper we report on the depth perception of five monocularly deprived cats that had recovered normal visual acuity in both eyes following imposition of certain of the above occlusion regimes. Although three of the animals exhibited five- to sevenfold superiority of binocular over monocular depth thresholds, subsequent tests made on two of the animals revealed that they were unable to make stereoscopic discriminations with random-dot stereograms. Despite the recovery of normal visual acuity in both eyes, we conclude that these animals recover at best only local stereopsis.

Comparison of receptive field properties of neurons in area 17 of normal and bilaterally amblyopic cats.

Receptive field properties of extracellularly recorded units in the visual cortex (area 17) of cats made bilaterally amblyopic by a variety of rearing conditions were measured and compared with the properties of units in normal cats. Properties studied included sensitivity to vernier offset, response facilitation to increasing bar length, receptive field size, responsiveness to moving and flashed stimuli, orientation tuning, the relation between mean firing rate and its variance, the amount of overlap of regions of on and off responsiveness in simple and complex cells, and, for flashed stimuli, latency to response onset, time to peak response, and response decay time constant. Behavioural testing of the amblyopic animals showed that spatial resolution was 2-4 times lower and vernier acuity thresholds 10-20 times greater than normal. Despite this, several neuronal response properties did not differ significantly from those in normal animals. These included peak responsiveness to moving stimuli, widths of orientation tuning curves, response variability, and latency to initial response for flashed stimuli. Other properties showed small but significant changes. Sensitivity to vernier offset (impulses per degree of offset) was reduced to nearly half its normal level; receptive field sizes increased by about 24% and an incomplete segregation of regions of on and off responsiveness was found in some cells, which made them hard to classify as simple or complex. Responses to flashed stimuli were smaller and more persistent. Their statistical significance notwithstanding, it seems unlikely that these relatively small response abnormalities in area 17 can fully account for the observed behavioural deficits.

Immunohistochemical study of the pattern of rapid expression of C-Fos protein in the visual cortex of dark-reared kittens following initial exposure to light.

Brief alterations to the nature of the visual input during critical periods in the early life of cats and monkeys can result in rapid anatomical and physiological changes in the central visual pathways. The immediate early genes (IEGs) represent a possible way in which these changes could be mediated since the protein products of a number of these genes have been shown to be induced rapidly in neurons in response to a variety of transsynaptic stimuli. Immunohistochemical methods were employed to examine the tempo and pattern of expression of Fos, the protein product of the c-fos gene, induced in the visual cortex of kittens dark-reared from birth to 30 days of age by brief periods of binocular visual exposure. In visual cortical area 17, the number of Fos immunoreactive cells increased rapidly from virtually zero in control kittens that received no visual exposure, to reach high levels in animals that received between 1 and 2 hours of visual experience. Immunoreactive cells were absent in the dorsal lateral geniculate nucleus, but were numerous in the ventral lateral geniculate nucleus, and in area 17, were most numerous in the extragranular layers (2, 3 and 6) but sparse in lower layer 4 and layer 5, and virtually absent in layer 1. Substantial constitutive Fos immunoreactivity was observed in area 17 of normal 30-day-old kittens but very few immunopositive cells were evident in adult animals. However, Fos immunoreactivity was observed in the visual cortex of a dark-reared (for 30 days) adult animal following a brief period of visual exposure, a finding that suggests that Fos might serve other roles in the visual cortex in addition to those it might play uniquely during development. It is suggested that Fos, in combination with the protein products of a select number of other IEGs, may mediate a variety of rapid changes in the visual cortex including those that underlie visual system plasticity during early postnatal life.

Patterns of Assimilate Production and Translocation in Muskmelon (Cucumis melo L.) : I. Diurnal Patterns.

Continuous monitoring of steady-state carbon dioxide exchange rates in mature muskmelon (Cucumis melo L.) leaves showed diurnal patterns of photosynthesis and respiration that were translated into distinct patterns of accumulation and phloem export of soluble sugars and amino acids. Leaf soluble sugar patterns in general followed the pattern of photosynthetic activity observed in the leaf, whereas starch accumulated steadily throughout the light period. Sugar and starch levels declined through the dark phase. Phloem exudate analysis revealed that diurnal levels of the major transport sugars (stachyose and sucrose) in the phloem did not appear to correlate directly with the photosynthetic activity of the leaf but instead were inversely correlated with leaf starch accumulation and degradation. The amino acid pool in leaf tissues remained constant throughout the diurnal period; however, the relative contribution of individual amino acids to the total pool varied with the diurnal photosynthetic and respiratory activity of the leaf. In contrast, the phloem sap amino acid pool size was substantially larger in the light than in the dark, a result primarily due to enhanced export of glutamine, glutamate, and citrulline during the light period. The results indicate that the sugar and amino acid composition of cucurbit phloem sap is not constant but varies throughout the diurnal cycle in response to the metabolic activities of the source leaf.

Patterns of Assimilate Production and Translocation in Muskmelon (Cucumis melo L.) : II. Low Temperature Effects.

Muskmelon (Cucumis melo L.) plants were exposed to a 10 degrees C chilling treatment for 72 hours, which induced leaf chilling injury symptoms (wilting, appearance of water-soaked areas, necrosis). Chilling caused an accumulation of starch, sucrose, hexoses (glucose and fructose), and certain amino acids (glutamate, aspartate, and citrulline) in source leaf tissues, but no accumulation of stachyose or other galactosyl-oligosaccharides occurred. Chilling also caused a general increase in sugar (stachyose, raffinose, sucrose) and amino acid content of the phloem sap, although rates of phloem transport were apparently reduced. Pretreatment of the leaves with a 20-milligram per liter abscisic acid (ABA) spray before chilling prevented the appearance of chilling injury symptoms. ABA pretreatment had little or no affect on sugar accumulation in leaf tissues but greatly reduced or eliminated the chilling-induced amino acid accumulation. Higher levels of aspartate and particularly of arginine were found in phloem saps from ABA-pretreated plants. The data indicate that changes in leaf metabolism caused by environmental stresses such as chilling may change the composition of cucurbit phloem sap. This raises the possibility that some of the deleterious effects of stress on sink tissues may, in part, be due to alterations in the nature of the assimilate supply.

Grating acuity of cats measured with detection and discrimination tasks.

In order to help resolve the widely discrepant claims for the grating acuity of the cat, behavioral measurements were made of the acuities of two normal adult cats using two different tasks. In one, a conventional detection task, the cats were required to discriminate between high contrast vertical gratings and a uniform field of the same space-averaged luminance. The second, less commonly-used task, required the cats to discriminate between vertical and horizontal gratings of the same spatial frequency. Both cats obtained thresholds of between 8.5 and 9 cycles/degree, with no difference between tasks. These results suggest that detection of aliased patterns is not a likely factor contributing to the wide range of published acuities, and they provide support for one of two competing models of beta-ganglion cell sampling in the retina.

The long-term effectiveness of different regimens of occlusion on recovery from early monocular deprivation in kittens.

Although the behavioural effects of an early period of monocular deprivation imposed on kittens can be very severe, resembling an extreme form of the human clinical condition deprivation amblyopia, they are not necessarily irreversible. Considerable behavioural as well as physiological recovery can occur if normal visual input is restored to the deprived eye sufficiently early, particularly if the other (initially non-deprived) eye is occluded at the same time (reverse occlusion). However, past work has shown that in many situations the improvement in the vision of the initially deprived eye that occurs during reverse occlusion is not retained following the subsequent introduction of binocular visual input. Furthermore, the vision of the other eye is often reduced as well, with the result that the eventual outcome is a condition of bilateral amblyopia. This study first examines the consequences of several periods of reverse occlusion whose onset and duration would be thought to maximize the opportunity for good and long-standing recovery of vision in the initially deprived eye. However, only in a very restricted set of exposure conditions did animals acquire good vision in one or both eyes; in most situations the final outcome was one of bilateral amblyopia. A second set of experiments examined the consequences of various regimens of part-time reverse occlusion, where the initially non-deprived eye was occluded for only part of each day to allow a period of binocular visual exposure, on kittens that had been monocularly deprived until 6, 8, 10 or 12 weeks of age. Whereas short or long daily periods of occlusion of the initially non-deprived eye resulted eventually in amblyopia in one, or usually both, eyes, certain intermediate occlusion times (3.5 or 5 h each day) resulted in recovery of normal acuities, contrast sensitivity and vernier acuity in both eyes, in animals that had been monocularly deprived until 6, 8 or 10 weeks of age, but not in animals deprived for longer periods. Experiments were done to establish some of the factors that contributed to the successful outcome associated with certain of the regimens of part-time reverse occlusion. It was established that recovery was just as good in animals in which the visual axes were vertically misaligned by means of prisms during the daily period of binocular visual exposure, thereby indicating that the visual input to the two eyes need not be concordant. However, animals that received equivalent visual exposure of the two eyes each day, but successively rather than simultaneously, all developed very severe bilateral amblyopia.(ABSTRACT TRUNCATED AT 400 WORDS)

The enzymes in cyclooxygenase and lipoxygenase pathways of arachidonic acid metabolism in human corpora lutea: dependence on luteal phase, cellular and subcellular distribution.

Eicosanoids synthesized within corpus luteum are presumed to regulate luteal function in women. However, the potential cellular source(s) of the eicosanoids, whether small and large luteal cells differ in eicosanoid synthesis and whether eicosanoids other than prostaglandin (PG)E2, PGF2 alpha and 6-keto-PGI1 alpha can be synthesized, have not been investigated. The present immunocytochemical studies were undertaken to answer these questions using mono and polyclonal antibodies to several enzymes in arachidonic acid metabolism by cyclooxygenase and lipoxygenase pathways. Human corpora lutea from early (n = 5), mid (n = 6) and late (n = 3) luteal phases were specifically immunostained for all the enzymes. All the enzymes were present in small and large luteal cells as well as in non luteal cells. However, small luteal cells contained more immunoreactive 5-lipoxygenase, PGD2 and PGF2 alpha synthases; large luteal cells contained more TXA2 synthase and 12-lipoxygenase; small and large luteal cells contained similar amounts of cyclooxygenase and PGI2 synthase. In all the cells, immunoreactive PGD2, PGI2 and TXA2 synthases increased from early to mid luteal phase and then declined in late luteal phase. Cyclooxygenase, 5- and 12-lipoxygenases and PGF2 alpha synthase, on the other hand, increased from early to mid and mid to late luteal phases. Immunoreactive cyclooxygenase and 5- and 12-lipoxygenases were present primarily in rough endoplasmic reticulum (ER) and/or smooth ER and cytoplasm. Quite unexpectedly, all three enzymes were also found in nuclear membranes, condensed chromatin and especially at the perimeter of condensed chromatin. Dispersed chromatin contained very little or no immunoreactive enzyme. These results indicate that regulation of human luteal function by eicosanoids synthesized within the corpus luteum is complex involving perhaps a) small and large luteal as well as non luteal cells, b) eicosanoids which have not been previously considered to play a role in luteal function and c) coordinate regulation of more than one enzyme in the pathways of arachidonic acid metabolism.

Vernier acuity of normal and visually deprived cats.

This study examines the relationship between grating and vernier acuity in cats that were either normally reared, unilaterally amblyopic as a result of a period of monocular deprivation, or bilaterally amblyopic resulting from a period of reverse occlusion followed by binocular visual experience. Vernier acuity was assessed on a jumping stand by use of a vernier-grating stimulus similar to that devised for use with human infants. The vernier thresholds for normal cats were 1.2-1.3 min arc, values that were approx. 6 times better than their grating acuity, and hence may represent a true hyperacuity. By contrast, the vernier acuity of the visually deprived cats were substantially below normal (19-83 min arc). The vernier thresholds for the deprived eye of the monocularly deprived cat and both eyes of the reverse occluded cats had fallen to the point where they were at best equal, and sometimes worse than the corresponding grating acuity. This pattern of results is similar to those observed in some types of human amblyopia, where vernier acuity also no longer represents a hyperacuity, and where in severe cases the thresholds may be worse than grating acuity.

Vernier acuity with opposite-contrast stimuli.

Vernier acuity has usually been tested with stimuli of the same contrast polarity (SC). This traditional vernier acuity was compared to that obtained with stimuli of opposite-contrast (OC) in which one target was brighter than the background and the other was darker. For both bar and dot targets vernier acuity with OC stimuli was about half as good as with SC stimuli. There were large individual differences in the size of the disadvantage with OC stimuli, although thresholds remained within the hyperacuity range. There were also individually-differing biases to see a dark vernier stimulus on one or the other side of a bright stimulus. Differences between OC and SC vernier acuities persisted over a wide range of interstimulus spacings, widths, and contrasts. At extremes of these spatial manipulations acuities became similar, but only because SC acuities were degraded to the level of OC acuities. Subjects showed little improvement in OC vernier acuity, even after 50,000 trials. It is concluded that finest judgements of spatial position arise in a level of the visual system at which light and dark stimuli are treated independently.