Density and morphology of dendritic spines in mouse neocortex.

Dendritic spines of pyramidal cells are the main postsynaptic targets of cortical excitatory synapses and as such, they are fundamental both in neuronal plasticity and for the integration of excitatory inputs to pyramidal neurons. There is significant variation in the number and density of dendritic spines among pyramidal cells located in different cortical areas and species, especially in primates. This variation is believed to contribute to functional differences reported among cortical areas. In this study, we analyzed the density of dendritic spines in the motor, somatosensory and visuo-temporal regions of the mouse cerebral cortex. Over 17,000 individual spines on the basal dendrites of layer III pyramidal neurons were drawn and their morphologies compared among these cortical regions. In contrast to previous observations in primates, there was no significant difference in the density of spines along the dendrites of neurons in the mouse. However, systematic differences in spine dimensions (spine head size and spine neck length) were detected, whereby the largest spines were found in the motor region, followed by those in the somatosensory region and those in visuo-temporal region.

Pyramidal cells, patches, and cortical columns: a comparative study of infragranular neurons in TEO, TE, and the superior temporal polysensory area of the macaque monkey.

The basal dendritic arbors of layer III pyramidal neurons are known to vary systematically among primate visual areas. Generally, those in areas associated with "higher" level cortical processing have larger and more spinous dendritic arbors, which may be an important factor for determining function within these areas. Moreover, the tangential area of their arbors are proportional to those of the periodic supragranular patches of intrinsic connections in many different areas. The morphological parameters of both dendritic and axon arbors may be important for the sampling strategies of cells in different cortical areas. However, in visual cortex, intrinsic patches are a feature of supragranular cortex, and are weaker or nonexistent in infragranular cortex. Thus, the systematic variation in the dendritic arbors of pyramidal cells in supragranular cortex may reflect intrinsic axon projections, rather than differences in columnar organization. The present study was aimed at establishing whether cells in the infragranular layers also vary in terms of dendritic morphology among different cortical areas, and whether these variations mirror the ones demonstrated in supragranular cortex. Layer V pyramidal neurons were injected with Lucifer yellow in flat-mounted cortical slices taken from cytoarchitectonic areas TEO and TE and the superior polysensory area (STP) of the macaque monkey. The results demonstrate that cells in STP were larger, had more bifurcations, and were more spinous than those in TE, which in turn were larger, had more bifurcations and were more spinous than those in TEO. These results parallel morphological variation seen in layer III pyramidal neurons, suggesting that increasing complexity of basal dendritic arbors of cells, with progression through higher areas of the temporal lobe, is a general organizational principle. It is proposed that the differences in microcircuitry may contribute to the determination of the functional signatures of neurons in different cortical areas. Furthermore, these results provide evidence that intrinsic circuitry differs across cortical areas, which may be important for theories of columnar processing.

The pyramidal cell in cognition: a comparative study in human and monkey.

Here we present evidence that the pyramidal cell phenotype varies markedly in the cortex of different anthropoid species. Regional and species differences in the size of, number of bifurcations in, and spine density of the basal dendritic arbors cannot be explained by brain size. Instead, pyramidal cell morphology appears to accord with the specialized cortical function these cells perform. Cells in the prefrontal cortex of humans are more branched and more spinous than those in the temporal and occipital lobes. Moreover, cells in the prefrontal cortex of humans are more branched and more spinous than those in the prefrontal cortex of macaque and marmoset monkeys. These results suggest that highly spinous, compartmentalized, pyramidal cells (and the circuits they form) are required to perform complex cortical functions such as comprehension, perception, and planning.

Specialization in pyramidal cell structure in the sensory-motor cortex of the vervet monkey (Cercopethicus pygerythrus).

Recent studies have revealed systematic differences in the pyramidal cell structure between functionally related cortical areas of primates. Trends for a parallel in pyramidal cell structure and functional complexity have been reported in visual, somatosensory, motor, cingulate and prefrontal cortex in the macaque monkey cortex. These specializations in structure have been interpreted as being fundamental in determining cellular and systems function, endowing circuits in these different cortical areas with different computational power. In the present study we extend our initial finding of systematic specialization of pyramidal cell structure in sensory-motor cortex in the macaque monkey [Cereb Cortex 12 (2002) 1071] to the vervet monkey. More specifically, we investigated pyramidal cell structure in somatosensory and motor areas 1/2, 5, 7, 4 and 6. Neurones in fixed, flat-mounted, cortical slices were injected intracellularly with Lucifer Yellow and processed for a light-stable 3,3'-diaminobenzidine reaction product. The size of, number of branches in, and spine density of the basal dendritic arbors varied systematically such that there was a trend for increasing complexity in arbor structure with progression through 1/2, 5 and 7. In addition, cells in area 6 were larger, more branched, and more spinous than those in area 4.

Visuotopic organisation and neuronal response selectivity for direction of motion in visual areas of the caudal temporal lobe of the marmoset monkey (Callithrix jacchus): middle temporal area, middle temporal crescent, and surrounding cortex.

On the basis of extracellular recordings in marmoset monkeys, we report on the organisation of the middle temporal area (MT) and the surrounding middle temporal crescent (MTc). Area MT is approximately 5-mm long and 2-mm wide, whereas the MTc forms a crescent-shaped band of cortex 1-mm wide. Neurones in area MT form a first-order representation of the contralateral hemifield, whereas those in the MTc form a second-order representation with a field discontinuity near the horizontal meridian. The representation of the vertical meridian forms the border between area MT and the MTc. In both areas, the fovea is represented ventrocaudally, and the visual field periphery is represented dorsorostrally. Analysis of single units revealed that 86% of cells in area MT show a strong selectivity for the direction of motion of visual stimuli. The proportion of direction-selective cells in the MTc (53%), whereas lower than that in area MT, is much higher than that observed in most other visual areas. Neurones in the cortex immediately rostral to area MT and the MTc are direction selective, with receptive fields predominantly located in the visual field periphery. In contrast, only a minority of the cells in the cortex ventral to the MTc are direction selective, and their receptive fields emphasise central vision. The results suggest that the MTc is functionally closely related to area MT, and distinct from the areas forming the dorsolateral complex. The MTc may have a role in combining information about motion in the visual field, processed by area MT, with information about stimulus shape, processed by the dorsolateral complex.

Cellular heterogeneity in cerebral cortex: a study of the morphology of pyramidal neurones in visual areas of the marmoset monkey.

The morphological characteristics of the basal dendritic fields of layer III pyramidal neurones were determined in visual areas in the occipital, parietal, and temporal lobes of adult marmoset monkeys by means of intracellular iontophoretic injection of Lucifer yellow. Neurones in the primary visual area (V1) had the least extensive and least complex (as determined by Sholl analysis) dendritic trees, followed by those in the second visual area (V2). There was a progressive increase in size and complexity of dendritic trees with rostral progression from V1 and V2, through the "ventral stream," including the dorsolateral area (DL) and the caudal and rostral subdivisions of inferotemporal cortex (ITc and ITr, respectively). Neurones in areas of the dorsal stream, including the dorsomedial (DM), dorsoanterior (DA), middle temporal (MT), and posterior parietal (PP) areas, were similar in size and complexity but were larger and more complex than those in V1 and V2. Neurones in V1 had the lowest spine density, whereas neurones in V2, DM, DA, and PP had similar spine densities. Neurones in MT and inferotemporal cortex had relatively high spine densities, with those in ITr having the highest spine density of all neurones studied. Calculations based on the size, number of branches, and spine densities revealed that layer III pyramidal neurones in ITr have 7.4 times more spines on their basal dendritic fields than those in V1. The differences in the extent of, and the number of spines in, the basal dendritic fields of layer III pyramidal neurones in the different visual areas suggest differences in the ability of neurones to integrate excitatory and inhibitory inputs. The differences in neuronal morphology between visual areas, and the consistency in these differences across New World and Old World monkey species, suggest that they reflect fundamental organisational principles in primate visual cortical structure.

The second visual area in the marmoset monkey: visuotopic organisation, magnification factors, architectonical boundaries, and modularity.

The organisation of the second visual area (V2) in marmoset monkeys was studied by means of extracellular recordings of responses to visual stimulation and examination of myelin- and cytochrome oxidase-stained sections. Area V2 forms a continuous cortical belt of variable width (1-2 mm adjacent to the foveal representation of V1, and 3-3.5 mm near the midline and on the tentorial surface) bordering V1 on the lateral, dorsal, medial, and tentorial surfaces of the occipital lobe. The total surface area of V2 is approximately 100 mm2, or about 50% of the surface area of V1 in the same individuals. In each hemisphere, the receptive fields of V2 neurones cover the entire contralateral visual hemifield, forming an ordered visuotopic representation. As in other simians, the dorsal and ventral halves of V2 represent the lower and upper contralateral quadrants, respectively, with little invasion of the ipsilateral hemifield. The representation of the vertical meridian forms the caudal border of V2, with V1, whereas a field discontinuity approximately coincident with the horizontal meridian forms the rostral border of V2, with other visually responsive areas. The bridge of cortex connecting dorsal and ventral V2 contains neurones with receptive fields centred within 1 degree of the centre of the fovea. The visuotopy, size, shape and location of V2 show little variation among individuals. Analysis of cortical magnification factor (CMF) revealed that the V2 map of the visual field is highly anisotropic: for any given eccentricity, the CMF is approximately twice as large in the dimension parallel to the V1/V2 border as it is perpendicular to this border. Moreover, comparison of V2 and V1 in the same individuals demonstrated that the representation of the central visual field is emphasised in V2, relative to V1. Approximately half of the surface area of V2 is dedicated to the representation of the central 5 degrees of the visual field. Calculations based on the CMF, receptive field scatter, and receptive field size revealed that the point-image size measured parallel to the V1/V2 border (2-3 mm) equals the width of a full cycle of cytochrome oxidase stripes in V2, suggesting a close correspondence between physiological and anatomical estimates of the dimensions of modular components in this area.

Distribution and patterns of connectivity of interneurons containing calbindin, calretinin, and parvalbumin in visual areas of the occipital and temporal lobes of the macaque monkey.

Immunocytochemical techniques were used to examine the distribution of double-bouquet cells and chandelier cells that were immunoreactive (-ir) for the calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV) in the primary visual area (V1), the second visual area (V2), and cytoarchitectonic area TE in the macaque monkey. Furthermore, the connections between CB-, CR-, and PV-ir neurons in these visual areas were investigated at the light microscope level by using a dual-immunocytochemical staining procedure. The most significant findings were three-fold. First, the number and distribution of CB-ir and CR-ir double-bouquet cells and PV-ir chandelier cells differed considerably between different visual areas. In particular, the different distribution of double-bouquet cells was illustrated dramatically at the V1/V2 border, where CB-ir double-bouquet axons were very few or lacking in V1 but were very numerous in V2. Furthermore, PV-ir chandelier cell terminals were relatively sparse in V1, more frequent in V2, and most frequent in area TE. Second, the percentage of CB-, CR-, and PV-ir neurons receiving multiple contacts on their somata and proximal dendrites from other calcium-binding protein neurons varied between 22% and 85%. The highest percentage of contacts found between immunolabelled cells and multiterminals were for the combinations CR/CB (76-85%; percent of cells immunoreactive for CB that were innervated by multiterminals immunoreactive for CR), followed by the combination PV/CR (42-48%), and then by the other combinations that had similar percentages (22-32% for CR/PV; 26-37% for CB/CR; 29-42% for CR/PV). Third, differences in the relative proportions of CB, CR, and PV terminals in contact with CB-, CR-, and PV-ir neurons were consistent between the different cortical areas studied. Thus, certain characteristics of intraareal circuits differ, whereas others remain similar, in different areas of the occipitotemporal visual pathway. The differences may represent regional specializations related to the different processing of visual stimuli, whereas the similarities may be attributed to general functional requisites for interneuronal circuitry.

Pyramidal cell heterogeneity in the visual cortex of the nocturnal New World owl monkey (Aotus trivirgatus).

Recent studies have revealed marked variation in pyramidal cell structure in the visual cortex of macaque and marmoset monkeys. In particular, there is a systematic increase in the size of, and number of spines in, the arbours of pyramidal cells with progression through occipitotemporal (OT) visual areas. In the present study we extend the basis for comparison by investigating pyramidal cell structure in OT visual areas of the nocturnal owl monkey. As in the diurnal macaque and marmoset monkeys, pyramidal cells became progressively larger and more spinous with anterior progression through OT visual areas. These data suggest that: 1. the trend for more complex pyramidal cells with anterior progression through OT visual areas is a fundamental organizational principle in primate cortex; 2. areal specialization of the pyramidal cell phenotype provides an anatomical substrate for the reconstruction of the visual scene in OT areas; 3. evolutionary specialization of different aspects of visual processing may determine the extent of interareal variation in the pyramidal cell phenotype in different species; and 4. pyramidal cell structure is not necessarily related to brain size.

Cortical integration in the visual system of the macaque monkey: large-scale morphological differences in the pyramidal neurons in the occipital, parietal and temporal lobes.

Layer III pyramidal neurons were injected with Lucifer yellow in tangential cortical slices taken from the inferior temporal cortex (area TE) and the superior temporal polysensory (STP) area of the macaque monkey. Basal dendritic field areas of layer III pyramidal neurons in area STP are significantly larger, and their dendritic arborizations more complex, than those of cells in area TE. Moreover, the dendritic fields of layer III pyramidal neurons in both STP and TE are many times larger and more complex than those in areas forming 'lower' stages in cortical visual processing, such as the first (V1), second (V2), fourth (V4) and middle temporal (MT) visual areas. By combining data on spine density with those of Sholl analyses, we were able to estimate the average number of spines in the basal dendritic field of layer III pyramidal neurons in each area. These calculations revealed a 13-fold difference in the number of spines in the basal dendritic field between areas STP and V1 in animals of similar age. The large differences in complexity of the same kind of neuron in different visual areas go against arguments for isopotentiality of different cortical regions and provide a basis that allows pyramidal neurons in temporal areas TE and STP to integrate more inputs than neurons in more caudal visual areas.

Complex dendritic fields of pyramidal cells in the frontal eye field of the macaque monkey: comparison with parietal areas 7a and LIP.

Layer III pyramidal neurones were injected with Lucifer Yellow in cortical slices taken from the medial subdivision of the frontal eye field (FEF) of the macaque monkey. The average area covered by basal dendritic fields, in the dimension parallel to the cortical layers, was 115.1 +/- 2.9 x 10(3) microm2, significantly larger than that observed among layer III cells in eye movement-related visual areas of the parietal lobe. Furthermore, the dendritic fields of pyramidal cells in the FEF were considerably more complex than those of their counterparts in the parietal lobe, as evaluated by Sholl analysis. Spine density varied along the basal dendritic tree, reaching a maximum of 8.5 +/- 0.8 spines/10 microm at a distance of 70-90 microm from the centre of the cell body. Such highly complex basal dendritic fields of layer III pyramidal neurones in the FEF may enable the integration of a diverse set of inputs from visual, motor, polysensory and memory-related periprincipal cortical areas.

Supragranular pyramidal neurones in the medial posterior parietal cortex of the macaque monkey: morphological heterogeneity in subdivisions of area 7.

Pyramidal neurones were injected with Lucifer Yellow in cortical slices taken from layer III of the medial subdivision of cytoarchitectonic area 7 (7m) of the macaque monkey. Cross-sectional area, branching complexity and spine density of the basal dendritic fields were determined and compared with those of neurones in other areas of the dorsal processing stream. Layer III pyramidal neurones in area 7m have an average basal dendritic field area of 109.57 +/- 13.03 x 10(3) microm2, which is significantly greater than that obtained for neurones in the lateral intraparietal area (LIP) and area 7a. Moreover, Sholl analyses revealed that neurones in area 7m are significantly more complex in their branching patterns than those in LIP and area 7a. These results reinforce the view that, behind the apparent architectural uniformity of Brodmann's area 7, there is a significant diversity of neuronal structure and function.

Variation in the spatial relationship between parvalbumin immunoreactive interneurones and pyramidal neurones in rat somatosensory cortex.

The morphology of pyramidal neurones was revealed by intracellular injection of Lucifer Yellow (LY) in fixed tangential cortical slices taken from rat primary somatosensory cortex. Slices were processed with a combination of antibodies to allow visualization of both the LY-injected neurones and parvalbumin immunoreactive (PV-ir) cell bodies, by confocal microscopy. Basal dendritic fields of pyramidal neurones in layer V were larger and more complex than those of layer III. Furthermore, the number of PV-ir cell bodies contained within the basal dendritic territories of pyramidal neurones in layer V was significantly greater than in layer III (mean +/- s.e.m., 36.3 +/- 3.0 and 20.9 +/- 1.6, respectively). These findings have functional implications both in terms of physiological characteristics, and inhibitory modulation of receptive field properties, of cortical neurones.

Dendritic structure varies as a function of eccentricity in V1: a quantitative study of NADPH diaphorase neurons in the diurnal South American rodent agouti, Dasyprocta prymnolopha.

The cerebral cortex is often described as a composite of repeated units or columns, integrating the same basic circuit. The 'ice-cube' model of cortical organization, and 'canonical' circuit, born from insights into functional architecture, still require systematic comparative data. Here we probed the anatomy of an individual neuronal type within V1 to determine whether or not its dendritic trees are consistent with the 'ice-cube' model and theories of canonical circuits. In a previous report we studied the morphometric variability of NADPH-diaphorase (NADPH-d) neurons in the rat auditory, visual and somatosensory primary cortical areas. Our results suggested that the nitrergic cortical circuitry of primary sensory areas are differentially specialized, probably reflecting peculiarities of both habit and behavior of the species. In the present report we specifically quantified the dendritic trees of NADPH-d type I neurons as a function of eccentricity within V1. Individual neurons were reconstructed in 3D, and the size, branching and space-filling of their dendritic trees were correlated with their location within the visuotopic map. We found that NADPH-d neurons became progressively smaller and less branched with progression from the central visual representation to the intermediate and peripheral visual representation. This finding suggests that aspects of cortical circuitry may vary across the cortical mantle to a greater extent that envisaged as natural variation among columns in the 'ice-cube' model. The systematic variation in neuronal structure as a function of eccentricity warrants further investigation to probe the general applicability of columnar models of cortical organization and canonical circuits.

Alterations in the phenotype of neocortical pyramidal cells in the Dyrk1A+/- mouse.

The gene encoding the dual-specificity tyrosine-regulated kinase DYRK1A maps to the chromosomal segment HSA21q22.2, which lies within the Down syndrome critical region. The reduction in brain size and behavioral defects observed in mice lacking one copy of the murine homologue Dyrk1A (Dyrk1A+/-) support the idea that this kinase may be involved in monosomy 21 associated mental retardation. However, the structural basis of these behavioral defects remains unclear. In the present work, we have analyzed the microstructure of cortical circuitry in the Dyrk1A+/- mouse and control littermates by intracellular injection of Lucifer Yellow in fixed cortical tissue. We found that labeled pyramidal cells were considerably smaller, less branched and less spinous in the cortex of Dyrk1A+/- mice than in control littermates. These results suggest that Dyrk1A influences the size and complexity of pyramidal cells, and thus their capability to integrate information.

Interlaminar differences in the pyramidal cell phenotype in cortical areas 7 m and STP (the superior temporal polysensory area) of the macaque monkey.

Pyramidal neurones were injected with Lucifer Yellow in slices cut tangential to the surface of area 7 m and the superior temporal polysensory area (STP) of the macaque monkey. Comparison of the basal dendritic arbors of supra- and infragranular pyramidal neurones (n = 139) that were injected in the same putative modules in the different cortical areas revealed variation in their structure. Moreover, there were relative differences in dendritic morphology of supra- and infragranular pyramidal neurones in the two cortical areas. Sholl analyses revealed that layer III pyramidal neurones in area STP had considerably higher peak complexity (maximum number of dendritic intersections per Sholl circle) than those in layer V, whereas peak complexities were similar for supra- and infragranular pyramidal neurones in area 7 m. In both cortical areas, the basal dendritic trees of layer III pyramidal neurones were characterized by a higher spine density than those in layer V. Calculations of the total number of dendritic spines in the "average" basal dendritic arbor revealed that layer V pyramidal neurones in area 7 m had twice as many spines as cells in layer III (4535 and 2294, respectively). A similar calculation for neurones in area STP revealed that layer III pyramidal neurones had approximately the same number of spines as cells in layer V (3585 and 3850 spines, respectively). Relative differences in the branching patterns of, and the number of spines in, the basal dendritic arbors of supra- and infragranular pyramidal neurones in the different cortical areas may allow for integration of different numbers of inputs, and different degrees of dendritic processing. These results support the thesis that intra-areal circuitry differs in different cortical areas.

Pyramidal cells of the frontal lobe: all the more spinous to think with.

The basal dendritic arbors of pyramidal cells in prefrontal areas 10, 11, and 12 of the macaque monkey were revealed by intracellular injection in fixed, flat-mounted, cortical slices. The size, number of branches, and spine density of the basal dendrites were quantified and compared with those of pyramidal cells in the occipital, parietal, and temporal lobes. These analyses revealed that cells in the frontal lobe were significantly more spinous than those in the other lobes, having as many as 16 times more spines than cells in the primary visual area (V1), four times more those in area 7a, and 45% more than those in area TE. As each dendritic spine receives at least one excitatory input, the large number of spines reported for layer III cells in prefrontal cortex suggests that they are capable of integrating a greater number of excitatory inputs than layer III pyramidal cells so far studied in the occipital, parietal, and temporal lobes. The ability to integrate a large number of excitatory inputs may be important for the sustained tonic activity characteristic of neurons in prefrontal cortex and their role in memory and cognition.

Morphological variation of layer III pyramidal neurones in the occipitotemporal pathway of the macaque monkey visual cortex.

We compared the morphological characteristics of layer III pyramidal neurones in different visual areas of the occipitotemporal cortical 'stream', which processes information related to object recognition in the visual field (including shape, colour and texture). Pyramidal cells were intracellularly injected with Lucifer Yellow in cortical slices cut tangential to the cortical layers, allowing quantitative comparisons of dendritic field morphology, spine density and cell body size between the blobs and interblobs of the primary visual area (V1), the interstripe compartments of the second visual area (V2), the fourth visual area (V4) and cytoarchitectonic area TEO. We found that the tangential dimension of basal dendritic fields of layer III pyramidal neurones increases from caudal to rostral visual areas in the occipitotemporal pathway, such that TEO cells have, on average, dendritic fields spanning an area 5-6 times larger than V1 cells. In addition, the data indicate that V1 cells located within blobs have significantly larger dendritic fields than those of interblob cells. Sholl analysis of dendritic fields demonstrated that pyramidal cells in V4 and TEO are more complex (i.e. exhibit a larger number of branches at comparable distances from the cell body) than cells in V1 or V2. Moreover, this analysis demonstrated that the dendrites of many cells in V1 cluster along specific axes, while this tendency is less marked in extrastriate areas. Most notably, there is a relatively large proportion of neurones with 'morphologically orientation-biased' dendritic fields (i.e. branches tend to cluster along two diametrically opposed directions from the cell body) in the interblobs in V1, as compared with the blobs in V1 and extrastriate areas. Finally, counts of dendritic spines along the length of basal dendrites revealed similar peak spine densities in the blobs and the interblobs of V1 and in the V2 interstripes, but markedly higher spine densities in V4 and TEO. Estimates of the number of dendritic spines on the basal dendritic fields of layer III pyramidal cells indicate that cells in V2 have on average twice as many spines as V1 cells, that V4 cells have 3.8 times as many spines as V1 cells, and that TEO cells have 7.5 times as many spines as V1 cells. These findings suggest the possibility that the complex response properties of neurones in rostral stations in the occipitotemporal pathway may, in part, be attributed to their larger and more complex basal dendritic fields, and to the increase in both number and density of spines on their basal dendrites.

The occipitoparietal pathway of the macaque monkey: comparison of pyramidal cell morphology in layer III of functionally related cortical visual areas.

The dendritic morphology of pyramidal cells located at the base of layer III in the primary visual area (V1), the second visual area (V2), the middle temporal area (MT), the ventral portion of the lateral intraparietal area (LIPv) and in the portion of cytoarchitectonic area 7a within the anterior bank of the superior temporal sulcus was revealed by injecting neurons with Lucifer Yellow in fixed, flattened slices of macaque monkey visual cortex. These areas correspond to different levels of the occipitoparietal cortical 'stream', which processes information related to motion and spatial relationships in the visual field. The tissue was immunocytochemically processed to obtain a light-stable diaminobenzidine reaction product, revealing the dendritic morphology in fine detail. Retrogradely labelled MT-projecting neurons in supragranular V1 (layer IIIc of Hassler's nomenclature, corresponding to Brodmann's layer IVb) were predominantly pyramidal, although many spiny multipolar (stellate) cells were also found. The average basal dendritic field area of pyramidal neurons in sublamina IIIc of V1 was significantly smaller than that in the homologous layer of V2, within the cytochrome oxidase-rich thick stripes. Furthermore, the average basal dendritic field areas of V1 and V2 pyramidal neurons were significantly smaller than those of neurons in MT, LIPv and area 7a. There was no difference in basal dendritic field area between layer III pyramidal neurons in areas MT, LIPv and 7a. While the shape of most basal dendritic fields was circularly symmetrical in the dimension tangential to the cortical layers, there were significant biases in complexity, with dendritic branches tending to cluster along particular axes. Sholl analysis revealed that the dendritic fields of neurons in areas MT, LIPv and 7a were significantly more complex (i.e. had a larger number of branches) than those of V1 or V2 neurons. Analysis of basal dendritic spine densities revealed regional variations along the dendrites, with peak densities being observed 40-130 microns from the cell body, depending on the visual area. The peak spine density of layer III pyramidal neurons in V1 was lower than that observed in V2, MT or LIPv, which were all similar. Pyramidal neurons in area 7a had the greatest peak spine density, which was on average 1.7 times that found in V1. Calculations based on the average spine density and number of dendritic branches at different distances from the cell body demonstrated a serial increase in the total number of basal dendritic spines per neuron at successive stations of the occipitoparietal pathway. Our observations, comparing dendritic fields of neurons in the homologous cortical layer at different levels of a physiologically defined 'stream', indicate changes in pyramidal cell morphology between functionally related areas. The relatively large, complex, spine-dense dendritic fields of layer III pyramidal cells in rostral areas of the occipitoparietal pathway allow these cells to sample a greater number of more diverse inputs in comparison with cells in 'lower' areas of the proposed hierarchy.

The organization and connections of somatosensory cortex in the brush-tailed possum (Trichosurus vulpecula): evidence for multiple, topographically organized and interconnected representations in an Australian marsupial.

Microelectrode mapping techniques were used to determine the organization of somatosensory cortex in the Australian brush-tailed possum (Trichosurus vulpecula). The results of electrophysiological mapping were combined with data on the cyto- and myeloarchitecture, and patterns of corticocortical connections, using sections cut tangential to the pial surface. We found evidence for three topographically organized representations of the body surface that were coextensive with architectonic subdivisions. A large, discontinuous cutaneous representation in anterior parietal cortex was termed the primary somatosensory area (SI). Lateral to SI we found evidence for two further areas, the second somatosensory area (SII) and the parietal ventral area (PV). While neurones in all of these areas were responsive to cutaneous stimulation, those of SI were non-habituating, whereas those in SII and PV often habituated to the stimuli. Moreover, neuronal receptive fields in SII and PV were, in general, larger than those in SI. Neurones in cortex adjacent to the rostral and caudal boundaries of SI, including cortex that interdigitated between the discontinuous SI head and body representations, required stimulation of deep receptors in the periphery to elicit responses. Within the region of cortex containing neurones responsive to stimulation of deep receptors, body parts were represented in a mediolateral progression. Injections of anatomical tracers placed in electrophysiologically identified locations in SI revealed ipsilateral connections with other parts of SI, as well as cortex rostral to, caudal to, and interdigitating between, SI. Injections in SI also resulted in labelling in PV, SII, motor cortex, posterior parietal cortex and perirhinal cortex. The patterns of contralateral projections reflected those of ipsilateral projections, although they were relatively less dense. The present findings support recent observations in other marsupials in which multiple representations of the body surface were described, and suggest that multiple interconnected sensory representations may be a common feature of cortical organization and function in marsupials.