Monkey V1 Research Articles

Stimulus optimisation in primary visual cortex

A computational method is introduced for finding effective stimuli for sensory neurons. In single unit recording experiments, it maximises simultaneously recorded responses by changing stimuli along an estimated gradient with respect to stimuli. The estimate is the correlation between noise added to an evolving base stimulus and the response. Pixel optimisation for monkey V1 rapidly produces stimuli consistent with conventionally determined tuning even for complex cells. For the same complex cell, repeated runs of the optimisation gave solutions with different phase. Unlike reverse correlation, this method is applicable to non-linear, context-sensitive cells, possibly also in higher sensory areas.

Layer-Specific Input to Distinct Cell Types in Layer 6 of Monkey Primary Visual Cortex

Layer 6 of monkey V1 contains a physiologically and anatomically diverse population of excitatory pyramidal neurons. Distinctive arborization patterns of axons and dendrites within the functionally specialized cortical layers define eight types of layer 6 pyramidal neurons and suggest unique information processing roles for each cell type. To address how input sources contribute to cellular function, we examined the laminar sources of functional excitatory input onto individual layer 6 pyramidal neurons using scanning laser photostimulation. We find that excitatory input sources correlate with cell type. Class I neurons with axonal arbors selectively targeting magnocellular (M) recipient layer 4Calpha receive input from M-dominated layer 4B, whereas class I neurons whose axonal arbors target parvocellular (P) recipient layer 4Cbeta receive input from P-dominated layer 2/3. Surprisingly, these neuronal types do not differ significantly in the inputs they receive directly from layers 4Calpha or 4Cbeta. Class II cells, which lack dense axonal arbors within layer 4C, receive excitatory input from layers targeted by their local axons. Specifically, type IIA cells project axons to and receive input from the deep but not superficial layers. Type IIB neurons project to and receive input from the deepest and most superficial, but not middle layers. Type IIC neurons arborize throughout the cortical layers and tend to receive inputs from all cortical layers. These observations have implications for the functional roles of different layer 6 cell types in visual information processing.

Light-induced Down-regulation of the Rat Class 1 Dynein-associated Protein robl/LC7-like Gene in Visual Cortex

Light-induced Down-regulation of the Rat Class 1 Dynein-associated Protein robl/LC7-like Gene in Visual Cortex

Contour decouples gamma activity across texture representation in monkey striate cortex.

Previous work on figure-ground coding in monkey V1 revealed enhanced spike rates within an object's surface representation, synchronization of gamma oscillations (gamma = 35-90 Hz) in object and background regions, but no decrease in signal correlation across the representation of a contour. The latter observation seems to contradict previous statements on the role of gamma-synchronization for scene segmentation. We re-examine these findings by analyzing different coupling measures and frequency ranges of population activities potentially contributing to figure-ground segregation. Multiple unit activity (MUA) and local field potentials (LFPs) were recorded by parallel mu-electrodes in monkey V1 during stimulation by a grating in which an object was defined by a shifted rectangle. In contradiction to the conclusions in previous work, we find strong decoupling of population activity between figure and ground representations compared to the situation in which the object is absent. In particular, coherence of late gamma-LFPs is strongly reduced, while reduction is absent during the early epochs of high-amplitude transients for LFP- and MUA-coherence at all frequencies, and at low frequencies also in the subsequent epochs. Our results of decoupling in late LFP gamma-components among figure and ground representations suggest that these signals may support figure-ground segregation.

The connections of layer 4 subdivisions in the primary visual cortex (V1) of the owl monkey.

The primary visual cortex (V1) of primates receives signals from parallel lateral geniculate nucleus (LGN) channels. These signals are utilized by the laminar and compartmental [i.e. cytochrome oxidase (CO) blob and interblob] circuitry of V1 to synthesize new output pathways appropriate for the next steps of analysis. Within this framework, this study had two objectives: (i) to analyze the con- nections between primary input and output layers and compartments of V1; and (ii) to determine differences in connection patterns that might be related to species differences in physiological properties in an effort to link specific pathways to visual functions. In this study we examined the intrinsic interlaminar connections of V1 in the owl monkey, a nocturnal New World monkey, with a special emphasis on the projections from layer 4 to layer 3. Interlaminar connections were labeled via small iontophoretic or pressure injections of tracers [horseradish peroxidase, biocytin, biotinylated dextrine amine (BDA) or cholera toxin subunit B conjugated to colloidal gold particles]. Our most significant finding was that layer 4 (4C of Brodmann) can be divided into three tiers based upon projections to the superficial layers. Specifically, we find that 4alpha (4Calpha), 4beta (4Cbeta) and 4ctr send primary projections to layers 3C (4B), 3Bbeta (4A) and 3Balpha (3B), respectively. Examination of laminar structure with Nissl staining supports a tripartite organization of layer 4. The cortical output layer above layer 3Balpha (3B) (e.g. layer 3A) does not appear to receive any direct connections from layer 4 but receives heavy input from layers 3Balpha (3B) and 3C (4B). Some connectional differences also were observed between the subdivisions of layer 3 and the infragranular layers. No consistent differences in connections were observed that distinguished CO blobs from interblobs or that could be correlated with differences in visual lifestyle (nocturnal versus diurnal) when compared with connectional data in other primates. Re-examination of data from previous studies in squirrel and macaque monkeys suggests that the tripartite organization of layer 4 and the unique projection pattern of layer 4ctr are not restricted to owl monkeys, but are common to a number of primate species.

Light-induced down-regulation of the rat class 1 dynein-associated protein robl/LC7-like gene in visual cortex.

Dynein and kinesin are the main microtubule-dependent motors that mediate intracellular movement in eukaryotic organisms. We have cloned a full-length cDNA encoding rat dynein light chain protein, robl/LC7-like (class 1), from visual cortex. We found that rat robl/LC7-like gene is highly expressed in neocortex and displays the unusual feature of being rapidly down-regulated by sensory stimulation. This effect was seen at both mRNA and protein levels in visual cortex, being detectable in as little as 45 min after the onset of visual stimulation. Down-regulation by sensory stimulation was also found within ocular dominance columns of area V1 in monocularly deprived monkeys. Our results suggest a high turnover rate of the robl/LC7-like protein and the presence of a repressor mechanism in neurons that is tightly coupled to synaptic stimulation.

The distribution of NADPH diaphorase and nitric oxide synthetase (NOS) in relation to the functional compartments of areas V1 and V2 of primate visual cortex.

The primary visual cortex (V1) of primates receives visual signals from cells in the koniocellular (K), magnocellular (M) and parvocellular (P) layers of the lateral geniculate nucleus (LGN). The functional role of the K pathway is unknown, but one proposal is that it modulates visual activity locally via release of nitric oxide (NO). One goal of this study was to examine the distribution of nitric oxide synthetase (NOS), the enzyme that produces NO, using immunocytochemistry for brain NOS (bNOS) or histochemistry for nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity in the V1 target cells of the K pathway and within the LGN itself. A second goal was to examine bNOS and NADPH diaphorase activity within proposed functional compartments in the second visual area (V2). We examined the LGN, V1 and V2 in squirrel monkeys, owl monkeys and bushbabies. In V1 and V2, we found that dense neuropil staining for NADPH diaphorase mirrored the pattern of high metabolic activity shown with cytochrome oxidase (CO) staining but did not necessarily mirror the pattern of immunolabeling seen with antibodies against NOS. The smooth stellate cells stained for NADPH diaphorase or bNOS were sparse and did not colocalize with LGN recipient zones in V1 or with the CO compartments in V2. LGN cells projecting to V1, including K, M and P cells, were negative for bNOS and NADPH diaphorase. Therefore, high levels of NOS are not limited to the K pathway. Instead, dense NOS activity is present in interneurons and within the neuropil of V1 and V2 that exhibit high metabolic demand.

Specificity of color connectivity between primate V1 and V2.

To examine the functional interactions between the color and form pathways in the primate visual cortex, we have examined the functional connectivity between pairs of color oriented and nonoriented V1 and V2 neurons in Macaque monkeys. Optical imaging maps for color selectivity, orientation preference, and ocular dominance were used to identify specific functional compartments within V1 and V2 (blobs and thin stripes). These sites then were targeted with multiple electrodes, single neurons isolated, and their receptive fields characterized for orientation selectivity and color selectivity. Functional interactions between pairs of V1 and V2 neurons were inferred by cross-correlation analysis of spike firing. Three types of color interactions were studied: nonoriented V1/nonoriented V2 cell pairs, nonoriented V1/oriented V2 cell pairs, and oriented V1/nonoriented V2 cell pairs. In general, interactions between V1 and V2 neurons are highly dependent on color matching. Different cell pairs exhibited differing dependencies on spatial overlap. Interactions between nonoriented color cells in V1 and V2 are dependent on color matching but not on receptive field overlap, suggesting a role for these interactions in coding of color surfaces. In contrast, interactions between nonoriented V1 and oriented V2 color cells exhibit a strong dependency on receptive field overlap, suggesting a separate pathway for processing of color contour information. Yet another pattern of connectivity was observed between oriented V1 and nonoriented V2 cells; these cells exhibited interactions only when receptive fields were far apart and failed to interact when spatially overlapped. Such interactions may underlie the induction of color and brightness percepts from border contrasts. Our findings thus suggest the presence of separate color pathways between V1 and V2, each with differing patterns of convergence and divergence and distinct roles in color and form vision.

Probing the human stereoscopic system with reverse correlation.

Our two eyes obtain slightly different views of the world. The resulting differences in the two retinal images, called binocular disparities, provide us with a stereoscopic sense of depth. The primary visual cortex (V1) contains neurons that are selective for the disparity of individual elements in an image, but this information must be further analysed to complete the stereoscopic process. Here we apply the psychophysical technique of reverse correlation to investigate disparity processing in human vision. Observers viewed binocular random-dot patterns, with 'signal' dots in a specific depth plane plus 'noise' dots with randomly assigned disparities. By examining the correlation between the observers' ability to detect the plane and the particular sample of 'noise' disparities presented on each trial, we revealed detection 'filters', whose disparity selectivity was remarkably similar to that of individual neurons in monkey V1. Moreover, if the noise dots were of opposite contrast in the two eyes, the tuning inverted, just like the response patterns of V1 neurons. Reverse correlation appears to probe disparity processing at the earliest stages of binocular combination, prior to the generation of a full stereoscopic depth percept.

Cytochrome-oxidase blobs and intrinsic horizontal connections of layer 2/3 pyramidal neurons in primate V1.

Pyramidal neurons in superficial layers of cerebral cortex have extensive horizontal axons that provide a substrate for lateral interactions across cortical columns. These connections are believed to link functionally similar regions, as suggested by the observation that cytochrome-oxidase blobs in the monkey primary visual cortex (V1) are preferentially connected to blobs and interblobs to interblobs. To better understand the precise relationship between horizontal connections and blobs, we intracellularly labeled 20 layer 2/3 pyramidal neurons in tangential living brain slices from V1 of macaque monkeys. The locations of each cell body and the cell's synaptic boutons relative to blobs were quantitatively analyzed. We found evidence for two cell types located at characteristic distances from blob centers: (1) neurons lacking long-distance, clustered axons (somata 130-200 microm from blob centers) and (2) cells with clustered, long-distance axon collaterals (somata < 130 microm or >200 microm from blob centers). For all cells, synaptic boutons close to the cell body were located at similar distances from blob centers as the cell body. The majority of boutons from cells lacking distal axon clusters were close to their cell bodies. Cells located more than 200 microm from blob centers were in interblobs and had long-distance clustered axon collaterals selectively targeting distant interblob regions. Cells located less than 130 microm from blob centers were found within both blobs and interblobs, but many were close to traditionally defined borders. The distant synaptic boutons from these cells were generally located relatively near to blob centers, but the neurons closest to blob centers had synaptic boutons closer to blob centers than those farther away. There was not a sharp transition that would suggest specificity for blobs and interblobs as discrete, binary entities. Instead they appear to be extremes along a continuum. These observations have important implications for the function of lateral interactions within V1.

Cortical connections of the dorsomedial visual area in new world owl monkeys (Aotus trivirgatus) and squirrel monkeys (Saimiri sciureus).

The dorsomedial visual area (DM) is an extrastriate area that was originally described in owl monkeys as a complete representation of the visual hemifield in a heavily myelinated wedge of cortex just rostral to dorsomedial visual area V2. More recently, connections of DM in owl monkeys have been described (Krubitzer and Kaas [1993] J. Comp. Neurol 334:497-528). As part of an effort to determine whether DM exists in other primates, we compared the architecture, connections, and visual topography of DM in owl monkeys and the presumptive DM in squirrel monkeys. In both species of New World monkeys, the DM region was more heavily myelinated than adjacent cortex, and this region was connected with the first and second visual areas, the middle temporal area (MT), the medial area, the ventral posterior parietal area, the dorsointermediate area, the dorsolateral area, the ventral posterior and ventral anterior areas, the medial superior temporal area, the fundal area of the superior temporal sulcus, the inferior temporal cortex, and frontal cortex in or near the frontal eye field. In squirrel monkeys, both blob and interblob regions of V1 contributed equally to DM, whereas the blob regions provided most of the projections to V1 in owl monkeys. In squirrel monkeys, connections were also found with cortex on the ventral surface in the ventral occipital temporal sulcus. In owl monkeys and squirrel monkeys, connections were with both the upper and lower visual field representations in V1, V2, and MT, demonstrating that DM contains a complete representation of the visual field. These similarities in architecture, connections, and retinotopy argue that DM is a visual area of both owl and squirrel monkeys.

Extrastriate feedback to primary visual cortex in primates: a quantitative analysis of connectivity.

Knowledge-based or top-down influences on primary visual cortex (area V1) are believed to originate from information conveyed by extrastriate feedback axon connections. Understanding how this information is communicated to area V1 neurons relies in part on elucidating the quantitative as well as the qualitative nature of extrastriate pathway connectivity. A quantitative analysis of the connectivity based on anatomical data regarding the feedback pathway from extrastriate area V2 to area V1 in macaque monkey suggests (i) a total of around ten million or more area V2 axons project to area V1; (ii) the mean number of synaptic inputs from area V2 per upper-layer pyramidal cell in area V1 is less than 6% of all excitatory inputs; and (iii) the mean degree of convergence of area V2 afferents may be high, perhaps more than 100 afferent axons per cell. These results are consistent with empirical observations of the density of radial myelinated axons present in the upper layers in macaque area V1 and the proportion of excitatory extrastriate feedback synaptic inputs onto upper-layer neurons in rat visual cortex. Thus, in primate area V1, extrastriate feedback synapses onto upper-layer cells may, like geniculocortical afferent synapses onto layer IVC neurons, form only a small percentage of the total excitatory synaptic input.

Neural mechanisms underlying stereoscopic vision

The progressive frontalization of both eyes in mammals causes overlap of the left and right visual fields, having as a consequence a region of binocular field with single vision and stereopsis. The horizontal separation of the eyes makes the retinal images of the objects lying in this binocular field have slight horizontal and vertical differences, termed disparities. Horizontal disparities are the main cue for stereopsis. In the past decades numerous physiological studies made on monkeys, which have in many aspects a similar visual system to humans, showed that a population of visual cells are capable of encoding the amplitude and sign of horizontal disparity. Such disparity detectors were found in cortical visual areas V1, V2, V3, V3A, VP, MT (V5) and MST of monkeys and in the superior colliculus of the cat and opossum. According to their disparity tuning function, these cells were first grouped into tuned excitatory, tuned inhibitory, near and far sub-groups. Subsequent studies added two more categories, tuned near and tuned far cells. Asymmetries between left and right receptive field position, on and off regions, and intra-receptive field wiring are believed to be the neural mechanisms of disparity detection. Because horizontal disparity alone is insufficient to compute reliable stereopsis, additional information about fixation distance and angle of gaze is required. Thus, while there is unequivocal evidence of cells capable of detecting horizontal disparities, it is not known how horizontal disparity is calibrated. Sensitivity to vertical disparity and information about the vergence angle or eye position may be the source of this additional information.

Analysis of the postnatal growth of visual cortex

Development and growth of V1 begins during embryogenesis and continues postnatally. The growth of V1 has direct implications on the organization of features such as the retinotopic map and the pattern of visual cortical columns. We have examined the postnatal growth and two-dimensional shape of V1 in macaque monkeys, cats, and rats. The perimeter, area, and anterior-posterior length of V1 were measured from unfolded and flattened sections from neonatal and adult animals from each of these species. Although there were substantial differences in the overall amount of postnatal growth, from 18% in macaque monkeys to more than 100% in cats, in all three species the shape of V1 did not change during development. Thus, growth of the mammalian visual cortex is well described as an isotropic expansion, so the layout of the global features, such as the arrangement of ocular dominance columns and the retinotopic map, does not need to change during development. Furthermore, quantification of the shape confirms the observations that there is a similar, egg-like oval shape to the visual cortex of these mammalian species.

Neural activity in areas V1, V2 and V4 during free viewing of natural scenes compared to controlled viewing.

Under natural viewing conditions primates make frequent exploratory eye movements across complex scenes. We recorded neural activity of 62 cells in visual areas V1, V2 and V4 in an awake behaving monkey that freely viewed natural images. About half of the cells studied showed a modulation in firing rate following some of the eye movements made during free viewing, though the proportions showing a discernible modulation varied across areas. These cells were also examined under controlled viewing conditions in which gratings or natural image patches were flashed in and around the classical receptive field while the animal performed a fixation task. Activity rates were generally highest with flashed gratings and lowest during free viewing. Flashed natural image patches evoked responses between these two extremes, and the responses were higher when the patches were confined to the classical receptive field than when they extended into the non-classical surround. Thus the reduction of activity during free viewing relative to that obtained with flashed gratings is partly attributable to natural images being less effective stimuli and partly to suppressive spatio-temporal neural mechanisms that are important during natural vision.

Coding strategies in monkey V1 and inferior temporal cortices.

We would like to know whether the statistics of neuronal responses vary across cortical areas. We examined stimulus-elicited spike count response distributions in V1 and inferior temporal (IT) cortices of awake monkeys. In both areas, the distribution of spike counts for each stimulus was well described by a Gaussian distribution, with the log of the variance in the spike count linearly related to the log of the mean spike count. Two significant differences in response characteristics were found: both the range of spike counts and the slope of the log(variance) versus log(mean) regression were larger in V1 than in IT. However, neurons in the two areas transmitted approximately the same amount of information about the stimuli and had about the same channel capacity (the maximum possible transmitted information given noise in the responses). These results suggest that neurons in V1 use more variable signals over a larger dynamic range than IT neurons, which use less variable signals over a smaller dynamic range. The two coding strategies are approximately as effective in transmitting information.

Layer-specific differential regulation of transcription factors Zif268 and jun-D in visual cortex V1 and V2 of macaque monkeys

To investigate intracellular mechanisms of cortical layer-specific gene regulation, we quantitatively examined the expression of two transcription factors, Zif268 and JunD, and compared their expression levels in each layer of the primary visual cortex (V1) and visual area 2 (V2) of macaque monkeys ( Macaca fuscata). The brain sections were immunohistochemically stained for determination of the percentage of Zif268- or JunD-expressing neurons in the total neuronal population. We found area- and layer-specific expression of these transcription factors; Zif268 tended to be expressed at high levels in layers on the parvocellular pathway in V1, whereas JunD did not show such an expression pattern. In V1, many Zif268-immunopositive neurons were observed in layers II/III, IVC β and VI. The percentage of Zif268-immunopositive neurons was highest in layer IVC β and lowest in layer IVCα. JunD-immunopositive neurons were fewest in layer IVC β and most abundant in layer VI. In V2, the levels of expression of Zif268 was almost the same as that of JunD in layer II/III. However, layer IV of V2 tended to contain more Zif268-immunopositive neurons than JunD-immunopositive neurons, whereas layer VI contained more JunD-immunopositive neurons than Zif268-immunopositive neurons. Although it has been reported that the same extracellular signals induce both Zif268 and JunD, the present results indicate that the expression of these transcription factors is differentially controlled in each layer of the primate visual cortical areas. Furthermore, the present results suggest that these transcription factors contribute to area- and layer-specific gene regulation by mediating transmission of extracellular signals to the nucleus via different intracellular signalling pathways.

Phase correlation of cortical rhythms at different frequencies: higher-order spectral analysis of multiple-microelectrode recordings from cat and monkey visual cortex

In classical EEG analysis rhythms with different frequencies occuring at separable regions and states of the brain are analysed. Rhythms in different frequency bands have often been assumed to be independent and their occurrence was interpreted as a sign of different functional operations. Independence has scarcely proved because of conceptual and computational difficulties. It is, on the other hand, probable that different rhythmic brain processes are coupled because of the broad recurrent connectivity among brain structures. We therefore set out to find interactions among rhythmic signals at different frequencies. We were particularly interested in interactions between lower frequency bands and γ-activities (30–90 Hz), because the latter have been analysed in our laboratory in great detail and had properties suggesting their involvement in perceptual feature linking. Fast oscillations occurred synchronized in a stimulus-specific way in the visual cortex of cat and monkey. Their presence was often accompanied by lower frequency components at considerable power. Such multiple spectral peaks are known from many cortical and subcortical structures. Despite their well known occurrence, coupling among different frequencies has not been established, apart from harmonic components. For the present investigation we extended existing analytical tools to detect non-linear correlations among signal pairs at any frequency (including incommensurate ones). These methods were applied to multiple microelectrode recordings from visual cortical areas 17 and 18 of anesthetized cats and V1 of awake monkeys. In particular, we assessed non-linear correlations by means of higher order spectral analysis of multi-unit spike activities (MUA) and local slow wave field potentials (LFP, 1–120 Hz) recorded with microelectrodes. Non-linear correlations among signal components at different frequencies were investigated in the following steps. First, the frequency content of short (≈250 ms) sliding window signal epochs was analyzed for simultaneously occurring rhythms of significant power at different frequencies. This was done by a newly developed method derived from the trispectrum using separate averaging of the products of short-epoch power spectra for any possible combination of frequency pairs. Second, non-linear (quadratic) phase coupling between different frequencies was assessed by the methods of bispectrum and bicoherence. We found phase correlations at different frequencies in the visual cortex of the cat and monkey. These couplings were significant in about 60% of the investigated MUA and LFP recordings, including several cases of coupling among incommensurate (i.e. non-harmonic) frequencies. Significant phase correlations were present: (1) within the γ-frequency range; (2) between γ- and low frequency ranges (1–30 Hz, including α- and β-rhythms); and (3) within the low frequency range. Phase correlations depended, in most cases, on specific visual stimulation. We discuss the possible functional significance of phase correlations among high and low frequencies by including proposals from previous work about potential roles of single-frequency rhythms of the EEG. Our suggestions include: (1) visual feature linking across different temporal and spatial scales provided by coherent oscillations at high and low frequencies; (2) linking of visual cortical representations (high frequencies) to subcortical centers (low frequencies) like the thalamus and hippocampus; and (3) temporal segmentation of the sustained stream of incoming visual information into separate frames at different temporal resolutions in order to prevent perceptual smearing due to shifting retinal images. These proposals are, at present, merely speculative. However, they can, in principle, be proved by microelectrode recordings from trained behaving animals. © 1997 Elsevier Science B.V.

The role of temporal cortical areas in perceptual organization.

The visual areas of the temporal lobe of the primate are thought to be essential for the representation of visual objects. To examine the role of these areas in the visual awareness of a stimulus, we recorded the activity of single neurons in monkeys trained to report their percepts when viewing ambiguous stimuli. Visual ambiguity was induced by presenting incongruent images to the two eyes, a stimulation condition known to instigate binocular rivalry, during which one image is seen at a given time while the other is perceptually suppressed. Previous recordings in areas V1, V2, V4, and MT of monkeys experiencing binocular rivalry showed that only a small proportion of striate and early extrastriate neurons discharge exclusively when the driving stimulus is seen. In contrast, the activity of almost all neurons in the inferior temporal cortex and the visual areas of the cortex of superior temporal sulcus was found to be contingent upon the perceptual dominance of an effective visual stimulus. These areas thus appear to represent a stage of processing beyond the resolution of ambiguities--and thus beyond the processes of perceptual grouping and image segmentation--where neural activity reflects the brain's internal view of objects, rather than the effects of the retinal stimulus on cells encoding simple visual features or shape primitives.

Myelin patterns in V1 and V2 of normal and monocularly enucleated monkeys.

A pattern of alternating light and dark columns was observed in wet, unstained sections of macaque striate cortex after monocular enucleation. The columns were clearest in layer IV, but could be detected through the full thickness of the cortex. Subsequent processing for cytochrome oxidase (CO) showed that the light columns in wet sections viewed under darkfield illumination matched the ocular dominance columns serving the enucleated eye. These columns labeled preferentially with an antibody to myelin basic protein, suggesting that greater myelin content accounted for their brighter appearance. However, when sections were counterstained with luxol fast blue, Gallyas and Woelcke myelin techniques, the enucleated eye's columns appeared pale. It is unclear why classical myelin stains and myelin basic protein immunohistochemistry yielded opposite results. Discrepant patterns of myelin distribution were also found in normal animals using different myelin stains. Luxol fast blue showed homogeneous staining in layer IVc of macaque striate cortex, but the Gallyas stain revealed a pattern of thin pale bands alternating with wide dark bands, matching the pattern seen with the Liesegang stain. The CO patches in layers II and III fit in register with the wide dark myelin bands. In layers II and III of striate cortex, the Gallyas and luxol fast blue methods both labeled the CO patches. However, in squirrel monkey V2 the Gallyas stain labeled the pale CO stripes, whereas luxol fast blue labeled the dark CO stripes. These results indicate that pattern of myelin staining in visual cortex can vary according to the choice of technique, and may not reflect the true distribution of myelin. Studies of myeloarchitecture should employ a variety of myelin techniques, including examination of unstained sections, to obtain the most accurate picture of cortical myelin content.