Connectivity of neuronal populations within and between areas of primate somatosensory cortex.

Functions of the cerebral cortex emerge via interactions of horizontally distributed neuronal populations within and across areas. However, the connectional underpinning of these interactions is not well understood. The present study explores the circuitry of column-size cortical domains within the hierarchically organized somatosensory cortical areas 3b and 1 using tract tracing and optical intrinsic signal imaging (OIS). The anatomical findings reveal that feedforward connections exhibit high topographic specificity, while intrinsic and feedback connections have a more widespread distribution. Both intrinsic and inter-areal connections are topographically oriented across the finger representations. Compared to area 3b, the low clustering of connections and small cortical magnification factor supports that the circuitry of area 1 scaffolds a sparse functional representation that integrates peripheral information from a large area that is fed back to area 3b. Fast information exchange between areas is ensured by thick axons forming a topographically organized, reciprocal pathway. Moreover, the highest density of projecting neurons and groups of axon arborization patches corresponds well with the size and locations of the functional population response reported by OIS. The findings establish connectional motifs at the mesoscopic level that underpin the functional organization of the cerebral cortex.

A rapid topographic mapping and eye alignment method using optical imaging in Macaque visual cortex.

In optical imaging experiments, it is often advantageous to map the field of view and to converge the eyes without electrophysiological recording. This occurs when limited space precludes placement of an electrode or in chronic optical chambers in which one may not want to introduce an electrode each session or for determining eye position in studies of ocular disparity response in visual cortex of anesthetized animals. For these purposes, we have developed a spot imaging method that can be conducted rapidly and repeatedly throughout an experiment. Using small 0.2 degrees -0.5 degrees spots, the extent of the imaged field of view is mapped by imaging cortical response to single spots, placed at different positions (0.2 degrees steps) in either the horizontal or vertical axes. By shifting the relative positions of two spots, one presented to each eye, eye convergence can be assessed to within 0.1 degrees resolution. Once appropriate eye alignment is determined, stimuli for further optical imaging procedures (e.g. imaging random dot stimuli for study of disparity responses) can then be confidently placed. This procedure can be quickly repeated throughout the experiment to ensure maintained eye alignment.

Fine-scale organization of SI (area 3b) in the squirrel monkey revealed with intrinsic optical imaging.

Optical imaging of intrinsic cortical activity was used to study the somatotopic map and the representation of pressure, flutter, and vibration in area 3b of the squirrel monkey (Saimiri sciureus) cortex under pentothal or isoflurane anesthesia. The representation of the fingerpads in primary somatosensory cortex was investigated by stimulating the glabrous skin of distal fingerpads (D1-D5) with Teflon probes (3-mm diam) attached through an armature to force feedback-controlled torque motors. Under pentothal anesthesia, intrinsic signal maps in area 3b obtained in response to stimulation (trapezoidal indentation) of individual fingerpads showed focal activations. These activations (ranging from 0.5 to 1.0 mm) were discrete and exhibited minimal overlap between adjacent fingerpad representations. Consistent with previously published maps, a somatotopic representation of the fingerpads was observed with an orderly medial to lateral progression from the D5 to D1 fingerpads. Under isoflurane anesthesia, general topography was still maintained, but the representation of fingerpads on adjacent fingers had higher degrees of overlap than with pentothal anesthesia. Multi- and single-unit recordings in the activation zones confirmed the somatotopic maps. To examine preferential inputs from slowly adapting type I (SA) and rapidly adapting type I (RA) and type II (PC) mechanoreceptors, we applied stimuli consisting of sinusoidal indentations that produce sensations of pressure (1 Hz), flutter (30 Hz), and vibration (200 Hz). Under pentothal anesthesia, activation patterns to these different stimuli were focal and coincided on the cortex. Under isoflurane, activation zones from pressure, flutter, and vibratory stimuli differed in size and shape and often contained multiple foci, although overall topography was maintained. Subtraction and vector maps revealed cortical areas (approximate 250-microm diam) that were preferentially activated by the sensations of pressure, flutter, and vibration. Multi- and single-unit recordings aided in the interpretation of the imaging maps. In conclusion, the cortical signals observed with intrinsic signal optical imaging delineated a somatotopic organization of area 3b and revealed different topographical cortical activation patterns for pressure, flutter, and vibratory stimuli. These patterns were dependent on anesthesia type. Possible relationships of these anesthesia effects to somatosensory cortical plasticity are discussed.

Real and illusory contour processing in area V1 of the primate: a cortical balancing act.

It is known that neurons in area V2 (the second visual area) can signal the orientation of illusory contours in the primate. Whether area V1 (primary visual cortex) can signal illusory contour orientation is more controversial. While some electrophysiology studies have ruled out illusory signaling in V1, other reports suggest that V1 shows some illusory-specific response. Here, using optical imaging and single unit electrophysiology, we report that primate V1 does show an orientation-specific response to the 'abutting line grating' illusory contour. However, this response does not signal an illusory contour in the conventional sense. Rather, we find that illusory contour stimulation leads to an activation map that, after appropriate subtraction of real line signal, is inversely related to the real orientation map. The illusory contour orientation is thus negatively signaled or de-emphasized in V1. This 'activation reversal' is robust, is not due merely to presence of line ends, is not dependent on inducer orientation, and is not due to precise position of line end stimulation of V1 cells. These data suggest a resolution for previous apparently contradictory experimental findings. We propose that the de-emphasis of illusory contour orientation in V1 may be an important signal of contour identity and may, together with illusory signal from V2, provide a unique signature for illusory contour representation.

A hierarchy of the functional organization for color, form and disparity in primate visual area V2.

By combining optical imaging, single unit electrophysiology and cytochrome oxidase (CO) histology, we sought to reveal in greater detail the functional organization within the CO stripes of visual area V2 of primates. To visualize the disparity selective regions of V2, the imaging of binocular interaction was employed. These imaging maps guided single unit penetrations that then revealed a columnar organization for disparity. Our studies also showed a pattern of intermixing between the color and disparity pathways of V2, including the existence of single cells tuned for both color and disparity. While previous studies have suggested that the CO stripes of V2 constitute the fundamental organizational unit within V2, our results show a further level of organization consisting of functionally distinct subcompartments, 0.7-1.5 mm in diameter, within individual stripes. These subcompartments, which are not clearly revealed by CO histochemistry, lie within each of the thin, pale, and thick CO dense stripes in V2 and are specific for aspects of color, orientation and retinal disparity, respectively. The present results favor an architectural view of V2, not unlike that of V1, as a collection of functionally distinct subcompartments or modules situated within each of the V2 stripes. These modules also support the notion that for each cortical area (e.g. V1, V2, V4), there exists a stereotyped cortical module with a geometry that is characteristic for each area. These modules exist as a middle tier in a hierarchy of functional organization within V2.

Building surfaces from borders in Areas 17 and 18 of the cat.

Several brightness illusions indicate that borders can dramatically affect the perception of adjoining surfaces. In the Craik-O'Brien-Cornsweet illusion, in particular, two equiluminant surfaces can appear different in brightness due to the contrast border between them. Although the psychophysical nature of this phenomenon has been well characterized, the neural circuitry underlying this effect is unexplored. Here, we have asked whether there are cells in visual cortex which respond to edge-induced illusory brightness percepts such as the Cornsweet. Using optical imaging and single unit recordings methods, we have studied responses of the primary (Area 17) and second (Area 18) visual cortical areas of the anesthetized cat to both real luminance change and Cornsweet brightness change. We find that there are indeed cells whose responses are modulated in phase with the modulation of the Cornsweet stimulus. These cells are present in both Area 17 and Area 18, but are more prevalent in Area 18. These responses are generally weak and are found even when receptive fields are distant from the contrast border. Consistent with perception, cells which respond to the Cornsweet border are modulated in antiphase to the Narrow Real (another border-induced illusory brightness stimulus). Remarkably, we also find evidence of edge-induced responses to illusory brightness change using intrinsic signal optical imaging. Both real luminance change and edge-induced brightness change produces a greater imaged response in Area 18 than in Area 17. Thus, in the absence of direct luminance stimulation, cells in visual cortex can respond to modulation of distant border contrasts. We suggest that the perception of surface brightness was encoded in the early visual cortical pathway by both surface luminance contrast signals in Area 17 (Rossi, A. F., Rittenhouse, C. D., & Paradiso, M. A. (1996). The representation of brightness in primary visual cortex. Science, 273, 1104-7) and border-induced contrast signals that predominate in Area 18.

Spontaneous and stimulus-evoked intrinsic optical signals in primary auditory cortex of the cat.

Spontaneous and tone-evoked changes in light reflectance were recorded from primary auditory cortex (A1) of anesthetized cats (barbiturate induction, ketamine maintenance). Spontaneous 0.1-Hz oscillations of reflectance of 540- and 690-nm light were recorded in quiet. Stimulation with tone pips evoked localized reflectance decreases at 540 nm in 3/10 cats. The distribution of patches "activated" by tones of different frequencies reflected the known tonotopic organization of auditory cortex. Stimulus-evoked reflectance changes at 690 nm were observed in 9/10 cats but lacked stimulus-dependent topography. In two experiments, stimulus-evoked optical signals at 540 nm were compared with multiunit responses to the same stimuli recorded at multiple sites. A significant correlation (P < 0.05) between magnitude of reflectance decrease and multiunit response strength was evident in only one of five stimulus conditions in each experiment. There was no significant correlation when data were pooled across all stimulus conditions in either experiment. In one experiment, the spatial distribution of activated patches, evident in records of spontaneous activity at 540 nm, was similar to that of patches activated by tonal stimuli. These results suggest that local cerebral blood volume changes reflect the gross tonotopic organization of A1 but are not restricted to the sites of spiking neurons.

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.

Optical imaging of functional domains in the cortex of the awake and behaving monkey.

As demonstrated by anatomical and physiological studies, the cerebral cortex consists of groups of cortical modules, each comprising populations of neurons with similar functional properties. This functional modularity exists in both sensory and association neocortices. However, the role of such cortical modules in perceptual and cognitive behavior is unknown. To aid in the examination of this issue we have applied the high spatial resolution optical imaging methodology to the study of awake, behaving animals. In this paper, we report the optical imaging of orientation domains and blob structures, approximately 100-200 micrometer in size, in visual cortex of the awake and behaving monkey. By overcoming the spatial limitations of other existing imaging methods, optical imaging will permit the study of a wide variety of cortical functions at the columnar level, including motor and cognitive functions traditionally studied with positron-emission tomography or functional MRI techniques.

Visual topography in primate V2: multiple representation across functional stripes.

The second visual cortical area (V2) of the primate is composed of repeating thin, pale, and thick cytochrome oxidase stripes containing primarily color-selective, broad-band oriented, and disparity-selective cells, respectively. We have now examined topography in V2 with respect to these functional subdivisions. Our data suggest that there are multiple, interleaved visual maps in V2, one for each of the color, orientation, and disparity domains. The same region of visual space is re-represented by each stripe within a stripe cycle, resulting in discontinuities or "jumps back" in representation at stripe borders. Adjacent stripe cycles represent adjacent regions of space such that the visual map is continuous from one stripe to the next like stripe. Receptive field size and scatter are significantly larger for thin stripes than for thick stripes. Unexpectedly, our data suggest two types of pale stripes within each stripe cycle, one with scatter similar to thin stripes and another to thick stripes. Some evidence also suggests the presence of multiple maps within individual stripes in V2. Consistent with functional clustering within single stripes (Ts'o et al., 1990b), we have recorded re-representations and topographic discontinuities coincident with functional borders within single stripes. These results suggest that multiple and interleaved mapping may be a common organizational strategy for representing multiple functional domains within a single cortical area.

Early discordant binocular vision disrupts signal transfer in the lateral geniculate nucleus.

The mammalian lateral geniculate nucleus (LGN) is known to regulate signal transfer from the retina to the brain neocortex in a highly complex manner. Besides inputs from the brainstem, extraretinal inputs via corticogeniculate projections and local inhibitory neurons modulate signal transfer in the LGN. However, very little is known about whether the postnatal development of LGN signal-transfer mechanisms is influenced by early discordant binocular vision. By intraunit comparisons of responses between individual X-LGN cells and their direct retinal inputs, the efficiency of signal transfer was found permanently reduced due to an early interocular misalignment (strabismus). The contrast sensitivity and spatial resolution of cat LGN cells were significantly lower relative to their retinal inputs, and there was substantial decrease in signal-transfer speed. The observed physiological deficits were associated with immature X-retinogeniculate axon arbors. Thus, contrary to previous ideas, conflicting binocular inputs can produce neural deficits in subcortical visual structures.

Abnormal development of retinogeniculate X axons in strabismic cats: a possible substrate for visual dysfunction.

Rearing cats with surgically induced esotropia results in a number of structural and functional abnormalities in the visual system. In the present experiments, we have studied the morphology of individual retinogeniculate X axons in esotropic cats using intracellular recording and staining techniques. After serially reconstructing stained axonal arbors, we find that some X axon terminations in A-laminae of the lateral geniculate nucleus (LGN) have larger volumes and lower bouton densities than normal. This is true for X axons projecting from both the deviated and the non-deviated eye. It is possible that these abnormalities in morphological development contribute to the disruptions in signal transfer that has been observed in X-cells in the LGN of esotropic cats.

Experimentally induced visual projections to the auditory thalamus in ferrets: evidence for a W cell pathway.

We have previously reported that following specific neonatal brain lesions in ferrets, a retinal projection is induced into the auditory thalamus (Sur et al., Science 242:1437, '88). In these "rewired" ferrets, a novel visual pathway is established through auditory thalamus [the medial geniculate nucleus (MGN)] and primary auditory cortex (A1); cells in both MGN and A1 are visually responsive and exhibit properties similar to those of visual cells in the normal visual pathway. In this paper, we use three approaches--physiological, anatomical, and developmental--to examine which of the retinal ganglion cells project to the MGN in these rewired ferrets. We find that: 1) physiological response properties of postsynaptic visual cells in the MGN are W-like; 2) retinal ganglion cells back-filled from the MGN are small and similar to soma sizes of subsets of the normal retinal W cell population; and 3) subpopulations of these small cells can be preferentially rerouted to the MGN in response to different surgical manipulations at birth, consistent with differential W cell projection patterns in normal animals. These data suggest that retinal W cells come to project to the MGN in rewired animals. These findings not only provide a basis on which to interpret functional properties of this novel visual pathway, but also provide important information about the developmental capabilities of specific retinal ganglion cell classes and the regulation of their projections by target structures in the brain during development.

Visual projections routed to the auditory pathway in ferrets: receptive fields of visual neurons in primary auditory cortex.

How does cortex that normally processes inputs from one sensory modality respond when provided with input from a different modality? We have addressed such a question with an experimental preparation in which retinal input is routed to the auditory pathway in ferrets. Following neonatal surgical manipulations, a specific population of retinal ganglion cells is induced to innervate the auditory thalamus and provides visual input to cells in auditory cortex (Sur et al., 1988). We have now examined in detail the visual response properties of single cells in primary auditory cortex (A1) of these rewired animals and compared the responses to those in primary visual cortex (V1) of normal animals. Cells in A1 of rewired animals differed from cells in normal V1: they exhibited larger receptive field sizes and poorer visual responsivity, and responded with longer latencies to electrical stimulation of their inputs. However, striking similarities were also found. Like cells in normal V1, A1 cells in rewired animals exhibited orientation and direction selectivity and had simple and complex receptive field organizations. Furthermore, the degree of orientation and directional selectivity as well as the proportions of simple, complex, and nonoriented cells found in A1 and V1 were very similar. These results have significant implications for possible commonalities in intracortical processing circuits between sensory cortices, and for the role of inputs in specifying intracortical circuitry.

A map of visual space induced in primary auditory cortex.

Maps of sensory surfaces are a fundamental feature of sensory cortical areas of the brain. The relative roles of afferents and targets in forming neocortical maps in higher mammals can be examined in ferrets in which retinal inputs are directed into the auditory pathway. In these animals, the primary auditory cortex contains a systematic representation of the retina (and of visual space) rather than a representation of the cochlea (and of sound frequency). A representation of a two-dimensional sensory epithelium, the retina, in cortex that normally represents a one-dimensional epithelium, the cochlea, suggests that the same cortical area can support different types of maps. Topography in the visual map arises both from thalamocortical projections that are characteristic of the auditory pathway and from patterns of retinal activity that provide the input to the map.

Visual projections induced into the auditory pathway of ferrets. I. Novel inputs to primary auditory cortex (AI) from the LP/pulvinar complex and the topography of the MGN-AI projection.

The organization of cortical circuitry responsible for processing sensory information is a subject of intense examination. However, it is not known whether cortical cells in different sensory cortices process information in a way that is specific to the modality of their input, or whether there are commonalities in processing circuitry across different cortices. In our laboratory, this question has been investigated at the level of the geniculocortical pathway by routing information of one sensory modality into the processing circuitry of another modality. Appropriate early lesions cause growth of retinal axons into the auditory thalamus (MGN) (Sur et al., Science 242:1437, '88). Previously, we have established that the MGN carries the resulting visual information on to primary auditory cortex (AI), which thus contains visually responsive neurons and a topographic representation of the retina (Roe et al., Soc. Neurosci. Abstr. 14:460, '88; Sur et al., Science 242:1437, '88). In this paper, we describe anomalous projections from the dorsal part of the thalamus, specifically the lateral posterior/pulvinar complex, into AI. This result demonstrates that thalamic neurons belonging to one modality can be induced to project to cortex that is normally of a different modality. In addition, we have studied in detail the nature of the MGN to AI projection in these animals as compared to the normal projection. The MGN to AI projection appears to be unaltered by the lesions; the location and topography of labelled cells are similar to that in normal animals. Because the MGN to AI projection is still highly divergent along the "isofrequency" dimension when compared to the tonotopic dimension, our data suggest that visual topography in the cortical map is created within the auditory cortex, perhaps by activity-dependent sharpening of the retinal representation during development.

Cross-modal plasticity in cortical development: differentiation and specification of sensory neocortex.

Early developmental manipulations can induce sensory afferents of one modality to project to central targets of a different sensory modality. We and other investigators have used such cross-modal plasticity to examine the role of afferent inputs and their patterns of activity in the development of sensory neocortex. We suggest that the afferent rewiring can significantly influence the internal connectivity or microcircuitry of sensory cortex, aspects of which appear to be determined or specified relatively late in development, but that they cannot influence, or influence only to a minor extent, the laminar characteristics and external connectivity patterns of cortex, which appear to be specified earlier.

Three-dimensional autoradiographic localization of quench-corrected glycine receptor specific activity in the mouse brain using 3H-strychnine as the ligand.

The autoradiographic analysis of neurotransmitter receptor distribution is a powerful technique that provides extensive information on the localization of neurotransmitter systems. Computer methodologies are described for the analysis of autoradiographic material which include quench correction, 3-dimensional display, and quantification based on anatomical boundaries determined from the tissue sections. These methodologies are applied to the problem of the distribution of glycine receptors measured by 3H-strychnine binding in the mouse CNS. The most distinctive feature of this distribution is its marked caudorostral gradient. The highest densities of binding sites within this gradient were seen in somatic motor and sensory areas; high densities of binding were seen in branchial efferent and special sensory areas. Moderate levels were seen in nuclei related to visceral function. Densities within the reticular formation paralleled the overall gradient with high to moderate levels of binding. The colliculi had low and the diencephalon had very low levels of binding. No binding was seen in the cerebellum or the telencephalon with the exception of the amygdala, which had very low levels of specific binding. This distribution of glycine receptors correlates well with the known functional distribution of glycine synaptic function. These data are illustrated in 3 dimensions and discussed in terms of the significance of the analysis techniques on this type of data as well as the functional significance of the distribution of glycine receptors.

Effects of convergent strabismus on the development of physiologically identified retinogeniculate axons in cats.

We have studied the effects of surgically induced convergent strabismus (esotropia) on the morphological development of retinogeniculate X and Y axon arbors in cats. Single axons were recorded in the lateral geniculate nucleus or in the optic tract adjacent to the nucleus, classified physiologically, and injected intracellularly with horseradish peroxidase. The arbors of recovered axons were compared with X and Y axon arbors from normally reared adult cats. Our data demonstrate that while X axon arbors are relatively normal, the arbors of Y axons are profoundly affected by rearing with strabismus. Y axons, whether originating from the deviated or the nondeviated eye, have substantially smaller arbors and fewer boutons in the A-laminae of the lateral geniculate nucleus compared to Y axons in normal cats. The C-lamina terminations of contralaterally projecting Y axons in the strabismic cats are unaffected. These results suggest that the postnatal development of retinogeniculate Y axon arbors in the A-laminae is strongly influenced by abnormalities in postnatal visual experience. Furthermore, the present data suggest that, in addition to intraocular competitive interactions between X and Y axons previously proposed to account for the effects of other rearing conditions, interactions between afferents from the two eyes must also be involved in the development of at least Y axons.

Terminal arbors of single ON-center and OFF-center X and Y retinal ganglion cell axons within the ferret's lateral geniculate nucleus.

The lateral geniculate nucleus of the ferret contains not only eye-specific layers, but a further subdivision of layers A and A1 into inner and outer sublaminae that contain, respectively, ON-center and OFF-center cells (Stryker and Zahs, '83). To study how the arbors of single retinal ganglion cell axons correlate with these cellular divisions, we have examined the morphology of physiologically classified retinal axons in the ferret's lateral geniculate nucleus. As in cats, we could classify retinal axons as X or Y on the basis of a number of physiological criteria. X and Y axons have distinct patterns of termination in the lateral geniculate nucleus. Contralateral X axons innervate lamina A and ipsilateral axons lamina A1. X axons are further segregated in these laminae so that ON-center axons terminate in the inner sublamina, and OFF-center axons in the outer sublamina. We did not observe any branches of X axons innervating the medial interlaminar nucleus or the midbrain. Y axons have much larger terminal arbors and exhibit greater variation in their terminations. Generally, within layers A and A1, ON-center Y axons innervate the inner sublamina and OFF-center Y axons innervate the outer sublamina. However, they often innervate both sublaminae, and occasionally have a few boutons in the inappropriate lamina as well. Y axons also terminate in the dorsal C laminae, the interlaminar zones, and the media interlaminar nucleus; branches of these axons course toward the midbrain, presumably to innervate the superior colliculus. Thus, whereas the Y pathway in the ferret is one of high divergence, the X pathway appears to be the substrate for segregated ON and OFF channels through the lateral geniculate nucleus.