Area-specific laminar distribution of cortical feedback neurons projecting to cat area 17: Quantitative analysis in the adult and during ontogeny

Corticocortical pathways can be classified as feedback and feedforward, in part according to the laminar distribution of the parent cell bodies. Here, we have developed exhaustive sampling procedures to determine unambiguously this laminar distribution. This shows that individual extrastriate areas in the adult cat have highly stereotyped proportions of supragranular layer neurons with respect to the total population of neurons back-projecting to area 17. During development, these adult laminar patterns emerge from an initially uniform radial distribution through a process of selective reorganization, which is highly specific to each area. Injections of fluorescent retrograde tracers were made in area 17. In areas 19, 20, posteromedial lateral suprasylvian area, and anteromedial lateral suprasylvian area, we defined a projection zone as the region containing retrogradely labeled neurons. In the neonate, counts of labeled neurons throughout the projection zones show constant percentages of 40% in the supragranular layers. During development, there is an area-specific reduction in the percentage of supragranular labeled neurons generating the laminar distributions characteristic of each area. Numbers of labeled neurons were estimated at different eccentricities of the projection zone. This finding indicates that during development there is a relative decrease in the numbers of labeled neurons of the periphery of the projection zone in the supragranular layers but not in the infragranular layers. This decrease is accompanied by a relative decrease in the dimensions of the supragranular projection zone with respect to the infragranular projection zone. These findings suggest that each extrastriate area precisely adjusts the proportions of supragranular layer neurons back-projecting to striate cortex in part by developmental changes in the divergence-convergence values of individual neurons. This shaping of corticocortical connectivity occurs relatively late in postnatal development and could, therefore, be under epigenetic control. J. Comp. Neurol. 396:493–510, 1998. © 1998 Wiley-Liss, Inc.     

Development of projections from auditory to visual areas in the cat

In newborn kittens, cortical auditory areas (including AI and AII) send transitory projections to ipsi- and contralateral visual areas 17 and 18. These projections originate mainly from neurons in supragranular layers but also from a few in infragranular layers (Innocenti and Clarke: Dev. Brain Res. 14:143-148, '84; Clarke and Innocenti: J. Comp. Neurol. 251:1-22, '86). The postnatal development of these projections was studied with injections of anterograde tracers (wheat germ agglutinin-horseradish peroxidase [WGA-HRP]) in AI and AII and of retrograde tracers (WGA-HRP, fast blue, diamidino yellow, rhodamine-labeled latex beads) in areas 17 and 18. It was found that the projections are nearly completely eliminated in development, this, by the end of the first postnatal month. Until then, most of the transitory axons seem to remain confined to the white matter and the depth of layer VI; a few enter it further but do not appear to form terminal arbors. As for other transitory cortical projections the disappearance of the transitory axons seems not to involve death of their neurons of origin. In kittens older than 1 month and in normal adult cats, retrograde tracer injections restricted to, or including, areas 17 and 18 label only a few neurons in areas AI and AII. Unlike the situation in the kitten, nearly all of these are restricted to layers V and VI. A similar distribution of neurons projecting from auditory to visual areas is found in adult cats bilaterally enucleated at birth, which suggests that the postnatal elimination of the auditory-to-visual projection is independent of visual experience and more generally of information coming from the retina.     

Topography of the afferent connectivity of area 17 in the macaque monkey: a double-labelling study

The various structures afferent to area 17 (or V1) of the macaque monkey have widely differing retinotopic organizations. It is likely that these differences are reflected in the topographic organizations of the projections from these structures to area V1. We have investigated this issue by placing side-by-side injections of two retrograde fluorescent tracers, fast blue and diamidino yellow, in V1. By examining the extent of mixing of the two populations of singly labelled cells and the presence of doubly labelled cells, in different structures, we have characterized the topography of each projection in terms of the size of its axonal arborization and the amount of convergence and divergence. The afferents from the lateral geniculate nucleus (LGN) and from the pulvinar are organized in a point-to-point fashion. The maximum extent of axonal arborization of these afferents is 0.5 mm and these projections demonstrate little scatter (i.e., neighboring LGN neurons project to adjacent regions of V1). The other two subcortical structures examined, the claustrum and the intralaminar nuclei, demonstrate a much larger scatter and wider axonal arborizations in their projections to V1 than do the LGN and pulvinar. Two-dimensional reconstructions were made of the distribution of labelled neurons in extrastriate cortical areas. Using the separation between patches of labelled cells and transitions in myelin-stained sections, we have identified seven separate cortical regions containing labelled cells. Two of these can be identified as area V2 and the middle temporal visual area (MT). Three other regions correspond to areas V3, V3A and V4t. Finally, two more regions of labelling have been distinguished that belong to area V4. These results demonstrate that, at least within the central 6 degrees of visual field, all the presently known extrastriate visual cortical areas project to V1. This result is interesting in view of the fact that only a few extrastriate cortical areas are reported to receive afferents from V1. Three groups of cortical areas can be distinguished on the basis of the characteristics of their cortical connections to V1. The first group contains area V2, V3, and the posterior region of V4. These areas project to V1 with infra- as well as supragranular layer neurons and show limited axonal arborization and scatter in the projection. The second group consists of two regions of labelling in the superior temporal sulcus corresponding to V4t and MT and another on the annectant gyrus (V3A). These regions contain almost exclusively infragranular labelling and show wide axonal arborization and scatter in their projections to V1.(ABSTRACT TRUNCATED AT 400 WORDS)     

Postnatal development of afferent projections to the lateral suprasylvian visual area in the cat: an HRP study

The postnatal development of thalamic and cortical projections to the medial bank of the lateral suprasylvian area was studied in the cat by using the retrograde and orthograde HRP methods. Both projections are already present at birth. In both newborn kittens and adult cats, the thalamic projections arise from the same nuclei. By far the heaviest thalamic projection originates from a relatively lateral portion of the lateral posterior nucleus (the presumed LPl). The cortical laminar distribution of the afferents arising from the presumed LPl changes markedly with aging. In kittens younger than 1 week, the terminals are distributed densely in layer I and sparsely in layer IV. With age, the terminals in layer I become less dense while those in layer IV become denser. By 1 month of age, the terminal distribution is similar to that found in adult cats, in which the terminals are sparse in layer I and dense in depth--particularly, in layer IV. The terminal distribution of the corticocortical projections from areas 17 and 18 also changes with aging. The terminals in kittens younger than 2 weeks are distributed in both superficial and deep cortical layers, whereas those in kittens older than 1 month and in an adult cat are distributed only in deep layers.     

Organization of association projections from area 17 to areas 18 and 19 and to suprasylvian areas in the cat's visual cortex.

Cells in area 17 that are labelled by single, discrete injections of retrogradely transported tracers into extrastriate visual areas are discontinuously distributed in dense patches. In this study we made multiple, closely spaced injections of fluorescent dyes into extrastriate areas, to generate large deposits that would reveal whether the distributions of corticocortical cell bodies in area 17 are truly patchy or appear clustered only after small injections. By injecting a different tracer into each extrastriate area, or group of areas, we examined the spatial relationships between the populations of association cells. All deposits of tracers in areas 18, 19, or suprasylvian cortex, irrespective of size, label cells in a series of clusters in topographically related parts of area 17. We conclude that the complete populations of cells in area 17 that project to areas 18, 19, and the lateral suprasylvian cortex are all genuinely distributed in a patchy fashion. There appears to be a complex relationship between the sets of association cells projecting to different extrastriate regions: they do not completely overlap, only partially, and share some cortical zones but not others. In these experiments, only tiny percentages (2-5%) of labelled cells in the overlapping regions were filled with both tracers, suggesting that very few association cells in area 17 project to more than one of the extrastriate areas we studied. By comparing the dimensions of each injection site and of the labelled region in area 17, we estimated the extent of the convergence from area 17 to areas 18, 19, and posteromedial suprasylvian areas in retinotopic terms. The functional convergence was very similar in these pathways.     

Inactivation of lateral connections in cat area 17

Excitatory synapses arising from local neurons in the cat visual cortex are much more numerous than the thalamocortical synapses, which provide the primary sensory input. Many of these local circuit synapses are involved in the connections between cortical layers, but lateral connections within layers provide a major component of the local circuit synapses. We tested the influence of these lateral connections in the primary visual cortex of cats by inactivating small patches of cortex about 450 μm lateral from the recording pipette. By use of the neurotransmitter γ-aminobutyric acid (GABA), small patches of cortex were inhibited and released from inhibition in seconds. Orientation tuning curves derived from responses to oriented drifting gratings were obtained during short control periods interleaved with periods of GABA inactivation. About 30% of the cells (18/62, recorded in all layers) changed their orientation tuning when a small portion of their lateral input was silenced. There was no broadening of the orientation tuning curve during lateral inactivation. Instead, the recorded cells shifted their preferred orientation towards the orientation of the inactivated site. One explanation is that the GABA inactivation alters the balance of excitatory and inhibitory inputs to a cell, which results in a shift of the cell's preferred orientation.     

Non-mirror-symmetric patterns of callosal linkages in areas 17 and 18 in cat visual cortex

In the cat, callosal connections in area 17 are largely confined to a 5–6-mm-wide strip at the 17/18 border. It is commonly thought that callosal fibers extending from between the 17/18 border regions interconnect loci that are mirror-symmetric with respect to the midline of the brain, but this idea has not been tested experimentally. The present study examined the organization of callosal linkages in the 17/18 border region of normal adult cats by analyzing the patterns of connections revealed in one hemisphere after small injections of different fluorescent tracers into the opposite 17/18 callosal region. The location of the injection sites within areas 17 and 18 was assessed by examining architectonic data and by inspecting the labeling pattern in the ipsilateral visual thalamus. Area 17 and 18 were separated by a 1 –1.5-mm-wide zone of cytoarchitectonic transition rather than by a sharp border. The results show that, in general, callosal fibers interconnect loci that are not mirror-symmetric with respect to the midline. Thus, area 17 injections placed nearly 3 mm away from the 17/18 transition zone produced discrete labeled areas located preferentially within the contralateral 17/18 transition zone. However, when the injection site was within the 17/18 transition zone, labeled cells were found primarily medial and lateral to, but not within, the 17/18 transition zone in the contralateral hemisphere. Previous studies have indicated that the 17/18 transition zone contains a representation of a strip of the ipsilateral visual field. Comparison of the retinotopy of the 17/18 border region with the mirror-reversed pattern of callosal linkages found in the present study suggests that callosal fibers link points that are in retinotopic correspondence in both hemispheres. © 1996 Wiley-Liss, Inc.     

Area 3a in the cat. II. Projections to the motor cortex and their relations to other corticocortical connections.

It is well known that area 3a in the cat may monosynaptically influence the activity of neurons in the motor cortex. Much less information is available, however, on the anatomy of these connections. By using single or combined injections of different retrograde axonal tracers, we investigated the topography (horizontal and laminar) of area 3a neurons projecting to the motor cortex, and the anatomical relationships between these neurons and those projecting to other areas (2, 5, and SII) which, in turn, project to the motor cortex. Area 3a projects to all parts of area 4 gamma, to area 4 delta, and to the agranular area 6 in the lateral bank of the presylvian sulcus (area 6 alpha gamma), but not to other parts of areas 4 and 6. This projection exhibits a loose topographic organization along the mediolateral dimension of area 3a, and, in many cases, arises predominantly from the rostral half of this area. Although single small injections in the motor cortex produced two or more separate patches of retrograde labeling in 3a, after simultaneous injections of fluorochromes in two separate loci there often appeared in area 3a overlapping populations of neurons which were labeled retrogradely by each of the dyes, but with very few double-labeled neurons. In horseradish peroxidase (HRP) cases, 72% of area 3a neurons projecting to area 4 gamma were distributed in supragranular layers (mainly layer III), although the proportion of labeling in infragranular layers was larger when using fluorescent dyes. Double-labeled cells predominated in infragranular layers. These results have a bearing upon the functional roles that have been attributed to area 3a, as a cortical locus involved in muscle sensation, and a cortical relay to the motor cortex of rapid feedback information from muscle activity during movement.     

Reciprocal projections of cat extrastriate cortex: I. distribution and morphology of neurons projecting from posterior medial lateral suprasylvian sulcus to area 17

Reciprocal projections between cortical areas have been subdivided into two functionally distinct components, “feedforward” and “feedback” (for review, see Felleman and Van Essen [1991] Cereb. Cortex 1:1–47). Some anatomical evidence, such as differences in the laminar distribution of the neurons of origin and of the terminations of their axons, has supported this division. However, very little is actually known about the distribution and morphology of the neurons of the feedback projections. In order to contribute further to our understanding of these two components of the corticocortical projections, I studied the distribution and morphology of a feedback projection, the reciprocal projection from the posterior medial lateral suprasylvian sulcus (PMLS), to primary visual cortex (area 17). Retrograde transport of horseradish peroxidase and fluorescent tracers in vivo combined with intracellular dye injections in lightly fixed cortical slices revealed many similarities between the feedforward and feedback projections: 1) They both emanate from all layers but layer 1; 2) each layer of origin contains a wide variety of standard and/or inverted pyramidal neurons; and 3) all of these, with the exception of a rare, large layer 5 neuron, have dendritic fields restricted principally to their layers of origin. There was, however, one major difference between the feedforward and feedback projections: In contrast to the projection from area 17 to PMLS, the projection from PMLS had a dense projection from layer 6 that comprised a striking abundance of spiny fusiform and inverted pyramidal neurons. These were morphologically distinct from other layer 6 neurons that project to the thalamus. Taken together, these data suggest that the reciprocal projections between area 17 and area PMLS, although not completely equivalent, share essential features that form a distinct population of neurons differing in morphology from corticothalamic projection neurons. © 1996 Wiley-Liss, Inc.     

Afferent projections to the ventral lateral geniculate nucleus in the cat

Horseradish peroxidase (HRP) injections were made into the dorsal lateral geniculate nucleus (LGN_d) and ventral lateral geniculate nucleus (LGN_v) of the cat in order to define afferent projections to LGN_v. These were found from the superior colliculus, contralateral LGN_v, dorsal median raphe nucleus, locus coeruleus, ipsilateral pretectum, and various portions of visual cortex. While many cortical areas project to LGN_v (17, 18, 19, 21 and lateral suprasylvian), the heaviest input arises from areas 17 and 20. The cell bodies of origin are in layer 5 in contrast to layer 6 which projects to LGN_d.     

The number of neurons in the different laminae of the binocular and monocular regions of area 17 in the cat

The number of neurons in individual laminae of area 17 was determined separately for both the binocular and the monocular, regions in the left hemi-sphere of six cats. The number of neurons/mm³ of tissue was obtained for each lamina by using the method of size-frequency distribution applied to neuronal nuclei. The number of neurons per unit of cortical surface could then be calculated from measurements of layer, thickness. The number of neurons/mm³ of tissue for trie total cortical thickness is on the order of 48,000 to 50,000 neurons, with no statistically significant differences be-tween binocular and monocular regions. There are no significant differences for any of the layers except layer IV, in which the numerical density is 20% higher in the monocular region. The thickness of the cortex and of many of its layers, however, do vary between the two regions. Consequently there are significant differences in the number of neurons under 1 mm² the total cortical thickness there are significantly more (27%) neurons in the binocular (78,440) than in the monocular region (61,900). This overall difference is due to significant changes in layers II, IIIA, IVA, and especially in layers V and VIA where neurons are 40% more numerous in the binocular region. These findings could signify either that the binocular region contains additional interneurons specifically related to binocular interactions or that it has a greater number of neurons projecting to other cortical and subcortical areas, or both.     

Single retinal ganglion cells projecting in bilaterally to the lateral geniculate nuclei or superior colliculi by way of axon collaterals in the cat

In most mammals with frontalized eyes, retinal ganglion cells in the nasal or temporal retina send their axons to the contralateral or ipsilateral half, respectively, of the brain. Previous studies in the cat, however, have indicated a retinal region of “nasotemporal overlap” from which arise the retinal projections to both the contralateral and ipsilateral halves of the brain. The present study thus examined in the cat whether any retinal ganglion cells give rise to bifurcating axons that innervate both halves of the brain. By employing fluorescent retrograde double labeling, we investigated whether or not single retinal ganglion cells project bilaterally to the lateral geniculate nuclei or superior colliculi by way of axon collaterals. After Fast Blue was injected into the lateral geniculate nucleus on one side and Diamidino Yellow was injected contralaterally into the lateral geniculate nucleus, 100–200 ganglion cells in each retina were double labeled with both tracers. These double-labeled cells were distributed primarily in the temporal retina, including the region around the vertical meridian and, additionally, in the nasal retina. About 60–80% of the double-labeled cells had large cell bodies (more than 25 μm in diameter), and the others had medium-sized ones (15–25 μm in diameter). The pattern of distribution of double-labeled cells, which was observed after the combined injection into both superior colliculi, was similar to that seen after the combined injection into both lateral geniculate nuclei; more than 9% of double-labeled cells, however, were large. The results indicate that a certain population of ganglion cells in the cat retina send their axons bilaterally to the lateral geniculate nuclei or superior colliculi by way of axon collaterals. The bilaterally projecting ganglion cells are mostly large, corresponding probably to α cells (the morphological counterparts of Y cells). In comparison with the patterns of bilateral projections of single retinal ganglion cells in the rat and monkey, the pattern of the bilateral retinofugal projections in the cat could represent an intermediate between those in the rat and monkey. © 1994 Wiley-Liss, Inc.     

Divergence of single axons in afferent projections to the cat's visual cortical areas 17, 18, and 19: a parametric study

The proportions of neurons projecting via axon collaterals to two areas in the cat's occipital cortex (diverging neurons) were determined quantitatively in subcortical and cortical afferents by making use of the retrograde axonal transport of two different tracers. The proportions of diverging neurons were determined for that part of the afferent sites in which neurons filled with tracers from both injected areas occurred (overlap zone). A number of experimental variables were tested for their role in possibly influencing the results of quantitative double-label experiments, among them the types and the combinations of retrograde tracers, the position of the injections, the survival time, and the histological procedure. The most important variable was the position of the cortical injection, which had to be restricted clearly to the cortical grey matter and to one cortical area in order to avoid false-positive double labeling. Other experimental variables affected the total number of retrogradely labeled neurons and/or the ratio between neurons labeled with the two different tracers rather than the proportions of double-labeled neurons. In particular DL proportions were largely independent of the number and density of labeled neurons. They only deviated significantly from mean values in those sections in which the number of labeled neurons amounted to less than 20% of the maximal number of labeled neurons found in one section throughout the overlap zone. Our results show that divergence is common in afferents to the cat visual cortex. The amount of divergence, however, varies considerably according to the origin of the afferent projection. The proportion of diverging neurons expressed as the percentage of the total number of neurons projecting to areas 17 and 18 was 3% in the A-laminae of the dorsal part of the lateral geniculate nucleus, about 8% in the posteromedial lateral suprasylvian area, and about 15% in the C-laminae of the dorsal part of the lateral geniculate nucleus, in the medial interlaminar nucleus, in the lateral part of the lateral posterior nucleus, and in the claustrum. The proportions of diverging neurons in the afferent projections to areas 17 and 19, and to areas 18 and 19 were about 10%. Diverging neurons were also found in the projections of the intralaminar thalamic nuclei to the visual cortex.(ABSTRACT TRUNCATED AT 400 WORDS)     

Ultrastructural analysis of projections to the pulvinar nucleus of the cat. I: Middle suprasylvian gyrus (areas 5 and 7)

The mammalian pulvinar nucleus (PUL) establishes heavy interconnections with the parietal lobe, but the precise nature of these connections is only partially understood. To examine the distribution of corticopulvinar cells in the cat, we injected the PUL with retrograde tracers. Corticopulvinar cells were located in layers V and VI of a wide variety of cortical areas, with a major concentration of cells in area 7. To examine the morphology and distribution of corticopulvinar terminals, we injected cortical areas 5 or 7 with anterograde tracers. The majority of corticopulvinar axons were thin fibers (type I) with numerous diffuse small boutons. Thicker (type II) axons with fewer, larger boutons were also present. Boutons of type II axons formed clusters within restricted regions of the PUL. We examined corticopulvinar terminals labeled from area 7 at the ultrastructural level in tissue stained for gamma-aminobutyric acid (GABA). By correlating the size of the presynaptic and postsynaptic profiles, we were able to quantitatively divide the labeled terminals into two categories: small and large (RS and RL, respectively). The RS terminals predominantly innervated small-caliber non-GABAergic (thalamocortical cell) dendrites, whereas the RL terminals established complex synaptic arrangements with dendrites of both GABAergic interneurons and non-GABAergic cells. Interpretation of these results using Sherman and Guillery's recent theories of thalamic organization (Sherman and Guillery [1998] Proc Natl Acad Sci U S A 95:7121-7126) suggests that area 7 may both drive and modulate PUL activity.     

Laminar and columnar patterns of geniculocortical projections in the cat: Relationship to cytochrome oxidase

We examined the laminar and columnar arrangement of projections from different layers of the lateral geniculate nucleus (LGN) to the visual cortex in the cat. In light of recent reports that cytochrome oxidase blobs (which in primates receive specific geniculate inputs) are also found in the visual cortex of cats, the relationship between cytochrome oxidase staining and geniculate inputs in this species was studied. Injections of wheat germ agglutinin-conjugated horseradish peroxidase were made into the anterior “genu” of the LGN, where isoelevation contours of the geniculate layers are distorted due to the curvature of the nucleus. Consequently, anterograde labeling from the various LGN layers was topographically separated across the surface of the cortex, and labeling in a particular isoelevation representation of the cortex could be associated with a specific layer of the LGN. Labeling from the A layers, which contain X and Y cells, was coextensive with layers 4 and 6 in both area 17 and area 18, as previously reported. Labeling from the C layers, which contain Y and W cells, occupied a zone extending from the 4a/4b border to part way into layer 3 in area 17. The labeling extended throughout layer 4 in area 18. There was also labeling in layer 5a and layer 1 in both area 17 and area 18. Except in layer 1, labeling from the C layers was patchy. In the tangential plane, adjacent sections stained for cytochrome oxidase showed that the patches of labeling from the C laminae aligned with the cytochrome oxidase blobs. The cytochrome blobs were visible in layers 3 and 4a, but not in layer 4b in both areas 17 and 18. These results suggest that W cells project specifically to the layer 3 portion of the blobs, while Y cells, at least those of the C layers, project specifically to the layer 4a portion of the blobs in area 17. The heavy synaptic drive of the Y cells is probably the cause of the elevated metabolism, and thus, higher cytochrome oxidase activity, of the blobs. © 1996 Wiley-Liss, Inc.     

Coincidence of patchy inputs from the lateral geniculate complex and area 17 to the cat's Clare-Bishop area

Pathways from a variety of structures to the largest of the cat's suprasylvian visual areas, the Clare-Bishop area, were found to patchy. These inputs arose from the lateral geniculate complex, from area 18, from area 19, and, as noted by Montero (Brain Behav. Evol. 18:194-218, '81), from area 17. The Clare-Bishop area was previously delineated on the basis of its uniform pattern of connections with cortex and thalamus (Sherk: J. Comp. Neurol. 247:1-31, '86) and found to incorporate pieces of several retinotopically defined areas (Tusa, Palmer, Rosenquist: Cortical Sensory Organization. Vol 2. Multiple Visual Areas. Clifton, NJ: Humana Press, pp. 1-31, '81). However, since individual patches did not correspond to particular retinotopically defined areas, other explanations of afferent patchiness were sought. An obvious question is whether the patches originating from different sources are systematically related to each other. Two hypotheses were considered. First, different inputs--for example, from the lateral geniculate nucleus (LGN) and from area 17--might terminate in intermingled but mutually exclusive zones in the Clare-Bishop area. Second, multiple patches of input might reflect duplicate representations of the corresponding visual field segment in the Clare-Bishop area. Both hypotheses were tested by injecting the lateral geniculate complex and either area 17 or area 19 with different anterograde tracers. In each case the two injections involved regions of the visual field that coincided to some degree, ranging from near-total overlap to almost complete exclusion. The first hypothesis predicted that the different labels in the Clare-Bishop area would never be found to overlap, while the second hypothesis predicted that when injections were closely matched retinotopically, there would be extensive overlap between patches. The results supported the second hypothesis: the better the retinotopic match between injections, the greater the overlap found between labeled geniculate and cortical input in the Clare-Bishop area. However, the multiplicity of patches seen in some experiments, and the close spacing between some patches, suggested that an additional, nonretinotopic mechanism also contributes to patchiness in the projections to the Clare-Bishop area.     

Four types of neuron in layer IVab of cat cortical area 17 accumulate 3H-GABA

Roughly 10% of the neurons in layer, IVah.of cat area 17 accumulate exogenous ³H-gamma-aminobutyric acid (GABA) but how many types of neuron comprise this population was unknown. We characterized these neurons by partial reconstruction of their somas from serial electron microscope autoradiograms and distinguished four types. GABA 1 was large (> 16.5 μm) and dark with a dense distribution of synaptic terminals, substantial geniculate input to the soma, and a moderate accumulation of GABA. GABA 2 was small (< 13 μm) and pale, also with a dense distribution of terminals but without evidence of somatic geniculate input, and a moderate accumulation of GABA. GABA 3 was radially fusiform (20μm × 8 μm) with varicose dendrites, a sparse distribution of synaptic terminals, and a heavy accumulation of GAB A. GAB A 4 was medium in size (15 μm) with a moderate distribution of synaptic terminals and a heavy accumulation of GABA. Reasons are presented for believing that each of these four categories of GABA-accumulating neuron represents a fundamental cell type.     

Branched projections from the nucleus prepositus hypoglossi to the oculomotor nucleus and the cerebellum. A retrograde fluorescent double-labeling study in the cat

The retrograde transport of fluorescent substances was used in order to investigate divergent axon collaterals of neurons in the nucleus prepositus hypoglossi (Ph). Fast blue (FB) was injected into the flocculus, paraflocculus and/or the vermis, while nuclear yellow (NY) was injected into the oculomotor nucleus alone or combined with injections in the nucleus of Darkschewitsch, the interstitial nucleus of Cajal and the medial longitudinal fascicle. Within optimal survival time, separate populations of single-labeled neurons of both dyes were found in Ph in all cases. Double-labeled neurons were seen in the rostral Ph following FB injections into the flocculus and the paraflocculus and NY injections restricted to the oculomotor nucleus. The present findings demonstrate that many neurons in the rostral Ph give collateral branches to the cerebellum and to the oculomotor nucleus.     

Characterization of transient cortical projections from auditory, somatosensory, and motor cortices to visual areas 17, 18, and 19 in the kitten

We have examined the anatomical features of ipsilateral transient cortical projections to areas 17, 18, and 19 in the kitten with the use of axonal tracers Fast Blue and WGA-HRP. Injections of tracers in any of the three primary visual areas led to retrograde labeling in frontal, parietal, and temporal cortices. Retrogradely labeled cells were not randomly distributed, but instead occurred preferentially at certain loci. The pattern of retrograde labeling was not influenced by the area injected. The main locus of transiently projecting neurons was an isolated region in the ectosylvian gyrus, probably corresponding to auditory area A1. Other groups of transiently projecting neurons had more variable locations in the frontoparietal cortex. The laminar distribution of neurons sending a transient projection to the visual cortex is characteristic and different from that of parent neurons of other cortical pathways at the same age. In the frontoparietal cortex, transiently projecting neurons were located mainly in layer 1 and the upper part of layers 2 and 3. In the ectosylvian gyrus, nearly all the neurons are located in layers 2 and 3. In addition, a few transiently projecting neurons are found in layer 6 and in the white matter. Transiently projecting neurons have a pyramidal morphology except for the occasional spindle-shaped cell of layer 1 and multipolar cells observed in the white matter. Anterograde studies were used to investigate the location of transient fibers in the visual cortex. Injections of WGA-HRP at the site of origin of transient projections gave rise to few retrogradely labeled cells in areas 17, 18, and 19, demonstrating that transient projections to these areas are not reciprocal. Although labeled axons were found over a wide area of the posterior cortex, they were more numerous over certain regions, including areas 17, 18, and 19, and absent from other more lateral cortical regions. Transient projecting fibers were present in all cortical layers at birth. Plotting the location of transient fibers in numerous sections and at all ages showed that these fibers are not more plentiful in the white matter than they are in the gray matter. We found no evidence that the white/gray matter border constituted a physical barrier to the growth of transient axons. Comparison of the organization of this transient pathway to that of other transient connections is discussed with respect to the development of the cortex.     

Morphological and immunocytochemical observations on the visual callosal projections in the cat

The connections between the left and right 17-18 border regions of the cat's visual cortex were labeled by axonal transport of peroxidase-conjugated wheat-germ agglutinin (WGA-HRP) and examined by light and electron microscopy. The cells of origin of the pathway were further characterized by transport of fluorescent microspheres ("beads") followed by in vitro injection of cells with Lucifer Yellow, and by beads transport followed by immunocytochemistry with antibodies to gamma-aminobutyric acid (GABA). The cells of origin of the callosal pathway were located in the lower part of layer 2/3, the upper part of layer 4, and layer 6. In layers 2/3 and 6, they were pyramidal cells; in layer 4 they were star pyramids or spiny stellate cells. None of them were spinefree or sparsely spinous cells, and none were GABA-positive. The axon terminals of the callosal pathway formed type 1 (asymmetric) synapses, and most of them contacted dendritic spines. Both the cells of origin and the terminals were arranged in patches. The findings suggest that the direct action of the callosal pathway is excitatory. The callosal system appears to represent only a subset of the cell types that have intrinsic horizontal projections within areas 17 or 18.