Biomedical subjects

Kathleen S Rockland

Publications and source records attributed to Kathleen S Rockland.

At least 19 recordsLinked to original sources

PubMed2006

Comparative analysis of layer-specific genes in Mammalian neocortex.

We examined the expression patterns of 4 layer-specific genes in monkey and mouse cortices by fluorescence double in situ hybridization. Based on their coexpression profiles, we were able to distinguish several subpopulations of deep layer neurons. One group was characterized by the expression of ER81 and the lack of Nurr1 mRNAs and mainly localized to layer 5. In monkeys, this neuronal group was further subdivided by 5-HT2C receptor mRNA expression. The 5-HT2C(+)/ER81(+) neurons were located in layer 5B in most cortical areas, but they intruded layer 6 in the primary visual area (V1). Another group of neurons, in monkey layer 6, was characterized by Nurr1 mRNA expression and was further subdivided as Nurr1(+)/connective tissue growth factor (CTGF)(-) and Nurr1(+)/CTGF(+) neurons in layers 6A and 6B, respectively. The Nurr1(+)/CTGF(+) neurons coexpressed ER81 mRNA in monkeys but not in mice. On the basis of tracer injections in 3 monkeys, we found that the Nurr1(+) neurons in layer 6A send some corticocortical, but not corticopulvinar, projections. Although the Nurr1(+)/CTGF(-) neurons were restricted to lateral regions in the mouse cortex, they were present throughout the monkey cortex. Thus, an architectonic heterogeneity across areas and species was revealed for the neuronal subpopulations with distinct gene expression profiles.

Animals↗

PubMed2006

Long-range interneurons within the medial pulvinar nucleus of macaque monkeys.

Like other thalamic nuclei, the primate pulvinar is considered not to have long-range intrinsic connections, either excitatory or inhibitory. Injections of biotinylated dextran amine (BDA) in the medial pulvinar, however, reveal retrogradely filled neurons up to 2.0 mm from the injection edge. Serial section reconstruction (n = 18) confirmed that retrogradely filled neurons projected to the injection site and showed that they had additional long-range collaterals within the posterior pulvinar. Arrays of small, beaded terminations occurred in multiple foci along the collaterals. Terminal arrays were up to 1.0 mm in length; foci were separated by about 0.7 mm. Somata were large (average area = 220 microm2), and dendritic arbors were radiate and also large (about 1.0 mm in diameter), but without either the appendages of classical interneurons or the hairlike spines characteristic of radiate pulvinocortical projection neurons. Double labeling for BDA and parvalbumin (PV) or BDA and gamma-aminobutyric acid (GABA) indicated that these large neurons were positive for both PV and GABA. Double labeling for PV and GABA, or PV and glutamic acid decarboxylase 67 (GAD67) revealed a small number of similarly large neurons in the posterior pulvinar that were positive for both substances. Thus, we propose that these neurons are a novel class of inhibitory interneuron, longer range than the classic thalamic local circuit interneurons. Future questions include how these neurons relate to other inhibitory systems and specific postsynaptic populations and whether they are located preferentially within the posterior pulvinar, possibly related to the multimodal character of this thalamic region.

Animals↗

PubMed2006

Neocortical areas, layers, connections, and gene expression.

Cortical patterns of gene expression provide a new approach to long standing issues of lamination, and area identity and formation. In this review, we summarize recent findings where molecular biological techniques have revealed a small number of area-specific genes in the nonhuman primate cortex. One of these (occ1) is strongly expressed in primary visual cortex and is associated with thalamocortical connections. Another gene, RBP, is more strongly expressed in association areas. It is not clear whether RBP might be linked with any particular connectional system, but several possibilities are raised. We also discuss possible roles of area-specific genes in postnatal development, and conclude with a brief sketch of future directions.

Animals↗

PubMed2006

Transient synaptic zinc-positive thalamocortical terminals in the developing barrel cortex.

In rat barrel cortex, layer 4 has a transiently high density of zinc-positive terminations from postnatal day (P)9 to P12 [P.W. Land & L. Shamalla-Hannah (2002)J. Comp. Neurol., 447, 43-56]. These terminations have been proposed to originate from cortico-cortical connections, but their exact origin is unknown. To determine their sources, we injected sodium selenite into the barrel cortex of two adult rats and 32 pups, from P5 to P28. As predicted, abundant zinc-positive cortically projecting neurons were visible around the injection sites and in distant cortical areas. From P9 to P13, however, neurons retrogradely labeled by zinc selenite occurred in the thalamus, in topographically appropriate regions of the ventroposterior medial (VPM) and posterior nuclei (Po). Because there are no previous reports of zinc-positive sensory thalamocortical connections, we sought corroboration of this unexpected finding by electron microscopy. This revealed a subset of boutons in layers 4 and 1, positive for both zinc and vesicular glutamate transporter 2, a protein used by thalamocortical terminations. Finally, in an additional nine rats, we carried out in situ hybridization for zinc transporter 3 mRNA. Moderate signal was detected in VPM and Po at P10, but this disappeared by P28. In contrast, a strong signal was apparent in the anterodorsal nucleus, which projects to limbic areas, and this persisted at P28. The timing of the transient zinc-positive terminations in the sensory thalamus roughly coincides with the onset of exploratory and whisking behavior in the middle of the second postnatal week; and this suggests zinc is important for activity-related refinement of circuitry.

Animals↗

PubMed2005

Improved Golgi-like visualization in retrogradely projecting neurons after EGFP-adenovirus infection in adult rat and monkey.

An adenovirus vector was generated using a neuron-specific promoter synapsin I and enhanced green fluorescent protein (EGFP) reporter (AdSynEGFP). In addition, two modifications were identified that resulted in robust and reliable retrograde transport and EGFP expression after injection of the virus into three different brain regions in adult rats (medial prefrontal cortex, posterior thalamic nuclear group, and CA1). These are postinjection survival times of 14 days and addition of high concentrations of NaCl (>or=600 mM) to the injection buffer. These modifications resulted in obvious improvement in the intensity of the EGFP signal and in the number of labeled cells. Use of anti-EGFP in immunofluorescence or immunoperoxidase processing further enhanced the signal so that Golgi-like filling of dendritic spines and axon collaterals was routinely achieved. Effectiveness of the AdSynEGFP for Golgi-like filling was confirmed in one rhesus monkey with injections in visual area V4. Because of the long-term viability of the infected neurons (at least up to 28 days in rats and 22 days in monkey), this AdSynEGFP is suitable for use in microcircuitry studies in combination with other fluorescently tagged elements, including anterogradely labeled extrinsic projections. The native EGFP signal (without antibody enhancement) may be sufficient for studies involving cultured cells or slices.

Adenoviridae↗

PubMed2005

Distinctive morphology of hippocampal CA1 terminations in orbital and medial frontal cortex in macaque monkeys.

The fine morphology of hippocampal connections to the orbital and medial frontal cortex (OMFC) was investigated by placing injections of anterograde tracers in the CA1 in two monkeys. The axons terminated mainly in layers 2 and 3, of areas 11, 13, 14c, 25, and 32, and were widely divergent in these layers, traversing 2-4 mm. Boutons were scattered along the main axon, but also occurred as distinctive small, spherical clusters of terminations ("mini-clusters"; diameter <50 microm). Occasional larger terminal arbors were observed in layer 3 and these were often unusually tortuous or convoluted. These features may imply a specialized microcircuitry of hippocampal-OMFC connections, including an origin from a particular subpopulation of CA1 projection neurons.

Animals↗

PubMed2005

Distribution of synaptic zinc in the macaque monkey amygdala.

We have mapped the macaque amygdala for the distribution of synaptic zinc (Zn), a co-factor of glutamate implicated in plasticity, as well as in several excitotoxic and other pathophysiological conditions. In brief, we found that the amygdala is Zn enriched in all nuclear groups (i.e., basolateral and cortical groups, as well as central and medial nuclei) but with marked differences in density. By comparing parallel tissue series histologically reacted for Zn and parvalbumin (PV), we further found that regions high in Zn are typically low in PV neuropil. In the basolateral group, there is a particularly distinct dorsoventral gradation such that Zn levels are most dense ventrally, i.e., in the paralaminar nucleus, the ventral division of the lateral nucleus, and the parvicellular divisions of both the basal nucleus and the accessory basal nucleus. PV levels are least dense in these same regions. For the central and medial nuclei, there is a slight mediolateral gradient, with Zn levels being higher medially. PV is low overall in these nuclei. Electron microscopic results confirmed that Zn is contained in synaptic boutons. These form asymmetrical, presumably excitatory, synapses, and the postsynaptic targets are mainly spines of projection neurons. The inhomogeneous distribution of Zn in the monkey amygdala may be related to different types or degrees of plasticity among the amygdaloid subnuclei. The complementary distribution with PV parallels that of several other substances and is interesting in the context of subnuclear vulnerability for human neuronal disease, such as seizure and Alzheimer's disease.

Amygdala↗

PubMed2005

Direct projections from CA1 to the superior temporal sulcus in the monkey, revealed by single axon analysis.

Anterograde tracer injections in the middle sector of CA1 in macaque monkeys demonstrate a direct projection to the fundus of the anterior superior temporal sulcus, in area IPa. Terminations are predominantly in layer 3. With regard to both terminal and arbor configuration, these hippocampal-cortical connections are morphologically similar to corticocortical connections to temporal association cortex. This report provides additional evidence of direct CA1 connections to particular multimodal cortical areas.

Animals↗

PubMed2005

Zinc-enriched amygdalo- and hippocampo-cortical connections to the inferotemporal cortices in macaque monkey.

Synaptic zinc (Zn), a co-factor in some glutamatergic synapses, has been implicated in plasticity effects, as well as in several excitotoxic and other pathophysiological conditions. In this study, we provide information about the distribution of Zn in inferotemporal cortex, a region at the interface of the visual and hippocampal networks. In brief, we found a lateral to medial increase in Zn, where TEad, a unimodal visual area, showed low levels of Zn; TEav, intermediate levels; and perirhinal cortex, a multimodal limbic area, high levels. The distribution of parvalbumin, a calcium binding protein, showed a reverse gradient to that of Zn. The neurons of origin of the Zn+ termination were identified by making intracortical injections sodium selenite (Na2SeO3). This substance interacts with Zn to form precipitates of ZnSe and in this form is transported retrogradely to the soma. A mixed population of labeled neurons was visualized, which included Zn+ neurons in CA1 of the hippocampus and in several amygdala subnuclei. In CA1, Zn+ neurons were restricted to the upper part of stratum pyramidale. Zn is thought to contribute to activity-dependent synaptic plasticity. The specifically high level in perirhinal cortex, and its origin from neurons in CA1 and the amygdala, may relate to cellular events involved in visual long-term memory formation.

Amygdala↗

PubMed2004

Transgenic mice expressing a fluorescent in vivo label in a distinct subpopulation of neocortical layer 5 pyramidal cells.

The neuronal components of cortical circuits have been characterized on the basis of their morphological and functional properties, and further refined by correlation of marker proteins with particular cell types. This latter approach has been very fruitful for GABA-containing neurons, but comparable diagnostic markers for subpopulations of excitatory pyramidal cells have been more elusive. An emerging new approach consists of transgenic mice that express fluorescent proteins under the control of promoters that are active in specific cell types. Here, we analyzed a line of transgenic mice that carries a transgene consisting of regulatory sequences of the potassium channel Kv3.1 and enhanced yellow fluorescent protein (EYFP). In these mice, a set of neurons in neocortical layer 5 expresses high levels of the transgenic marker protein. EYFP-expressing, and nonexpressing layer 5 cells were easily identified in living tissue under conditions suitable for patch-clamp electrophysiology. By using immunolabeling, retrograde Fast Blue labeling and electrophysiological recordings with biocytin injections, we identified the fluorescent neurons as a population of pyramidal cells with distinct morphological and electrophysiological properties when compared with nonfluorescent neighboring layer 5 pyramidal cells. The most prominent morphological difference between these two populations was a much smaller number of apical oblique dendrites in EYFP-positive as compared with the EYFP-negative cells. The most prominent electrophysiological feature was a steady spike frequency adaptation in EYFP-positive cells, whereas EYFP-negative cells responded to a depolarizing current injection with a closely spaced spike doublet followed by constant frequency firing. The in vivo labeled transgenic mice provide an experimental tool for further functional differentiation of these populations of layer 5 pyramidal cells.

Action Potentials↗

PubMed2004

Some thoughts on cortical minicolumns.

Although a columnar geometry is one of the defining features of cortical organization, major issues regarding its basic nature, key features, and functional significance remain unclear and often controversial. This review is intended to survey some of the basic anatomical features of columnar organization, and in particular the smaller scale dendritic minicolumns. One motive was simply to clarify what seem to be differences in terminology, where "minicolumn" can be used to refer to vertical rows of cells, pyramidal cell modules, or apical dendritic bundles. A second aim was to review anatomical details which over the years have tended increasingly to be overlooked. A third aim was to expand on recent results concerning the border of layers 1 and 2 as a specialized zone with its own micromodular organization. Views on columnar organization have arguably been heavily influenced by a desire for general principles; but re-examination of the complex underlying features may be both timely and worthwhile. We point out that what are defined as dendritic bundles do not extend through the full cortical thickness and are not strictly repetitive, but rather display significant inter- and intra-areal variation.

Animals↗

PubMed2004

Region specific micromodularity in the uppermost layers in primate cerebral cortex.

A micromodularity specific to the uppermost cortical layers, layers 1 and 2, is demonstrated in primates. This is most pronounced as patches of zinc-positive (Zn+) terminations, preferentially in the pre-Rolandic and limbic areas. The upper layer modularity can frequently be demonstrated by parvalbumin-immunoreactive (PV-ir) GABAergic terminations and by bundles of apical dendrites. Double-labeling or alternate section analysis shows that PV-ir and Zn+ terminations co-mingle at the layer 1,2 border and appear to coincide with dendritic bundles, proposed to originate from layer 2 pyramidal neurons. This model is basically similar to the prominent wall-and-hollow honeycomb organization in rat visual cortex. The organization of the PV-ir and dendritic components, however, is more difficult to define in primate than in rat. Moreover, the micromodularity is not uniform across areas. In some areas (motor and limbic), the modularity can be visualized by both zinc and PV. In other areas (i.e. primary sensory, sensory associational and prefrontal areas 46 and 8), although PV immunohistochemisty shows a periodic distribution, there is no detectable Zn+ modularity. These results add to the evidence for the complexity of layers 1 and 2 and raise the possibility that patches of Zn+ terminations correspond to zones of area-specific zinc-related plasticity. This might figure in the context of top-down or feedback influences, as often associated with layer 1.

Animals↗

PubMed2004

Connectional neuroanatomy: the changing scene.

This article gives a brief overview of cortical connectivity, as this has been investigated by anatomic tracer studies over the past 25 years. Open questions are discussed, from the perspective that connectivity studies are in a transition period when both techniques and conceptual frameworks are rapidly changing.

Animals↗

PubMed2003

Connections between the anterior inferotemporal cortex (area TE) and CA1 of the hippocampus in monkey.

In addition to the trisynaptic perforant pathway from entorhinal cortex to CA1, there are multiple direct parallel pathways between several cortical regions and CA1. These may be supposed to function cooperatively, in conjunction with the perforant pathway; but neither the functional nor anatomical organization of the extended network is well understood. In this report, we further investigate the connections between anterior inferotemporal cortex (area TE) and CA1. Injections of tracer substances demonstrate that part of the dorsal subdivision of TE sends projections to CA1, but does not receive reciprocating projections back. This contrasts with the bi-directional connections between the more ventral subdivision, TEav, and CA1, as reported by previous studies (and corroborated by tracer injections in this report). The corticohippocampal projections from dorsal TE are likely to be unimodal visual. They partially converge in the posterior portion of CA1 with connections from posterior TE and from the inferior parietal lobule, perhaps constituting a network related to visual or visuospatial processes.

Animals↗

PubMed2003

Neuron-specific distribution of P2X7 purinergic receptors in the monkey retina.

Extracellular ATP is a signaling molecule, working through P2X purinoceptors in the nervous system. P2X7 is a major subtype of the purinoceptors in the brain, where it is expressed mostly in glia cells and considered to work as a trigger of cytolysis. In the rodent retina, however, P2X7 is expressed in several classes of neurons including ganglion cells. In the present study we identified cells immunopositive for P2X7 by double immunolabeling. Immunoreactivity for P2X7 was observed in the inner nuclear layer (INL), the inner plexiform layer (IPL), and the ganglion cell layer (GCL). In the INL, strongly immunopositive cells corresponded to the subpopulation of horizontal cells. In the IPL, fine processes were immunopositive. In the GCL, most of the ganglion cells showed P2X7 immunoreactivity. At the ultrastructural level, immunoreactivity was confirmed in the cytoplasm of ganglion cells. No P2X7 immunoreactivity was found in non-neural cells, i.e., Müller cells or microglia. The immunohistochemical distribution of other purinoceptor subtypes (P2X1, P2X2, and P2X4) was also examined in the monkey retina. Immunoreactivity for P2X1 was strongly detected in a band, in sublamina a of the IPL. The band existed at almost the same level as tyrosine hydroxylase immunoreactivity, but did not seem to actually overlap. P2X2 was not expressed in the retina, and P2X4 was only faintly expressed at the scleral margin of the INL. Because P2X7 in the primate retina is expressed exclusively in neurons, it may in this location be involved in neural transmission rather than in cytolysis, as found for glia cells.

Animals↗

PubMed2003

Honeycomb-like mosaic at the border of layers 1 and 2 in the cerebral cortex.

In this report, we present evidence of a small-scale modularity (<100 microm) at the border of layers 1 and 2 in neocortical areas. The modularity is best seen in tangential sections, with double-labeling immunohistochemistry to reveal overlapping or complementary relationships of different markers. The pattern is overall like a reticulum or mosaic but is described as a "honeycomb," in which the walls and hollows are composed of distinct afferent and dendritic systems. We demonstrate the main components of the honeycomb in rat visual cortex. These are as follows: (1) zinc-enriched, corticocortical terminations in the walls, and in the hollows, thalamocortical terminations (labeled by antibody against vesicular glutamate transporter 2 and by cytochrome oxidase); (2) parvalbumin-dense neuropil in the walls that partly colocalizes with elevated levels of glutamate receptors 2/3, NMDAR receptor 1, and calbindin; and (3) dendritic subpopulations preferentially situated within the walls (dendrites of layer 2 neurons) or hollows (dendrites of deeper neurons in layers 3 and 5). Because the micromodularity is restricted to layers 2 and 1b, without extending into layer 3, this may be another indication of a laminar-specific substructure at different spatial scales within cortical columns. The suggestion is that corticocortical and thalamocortical terminations constitute parallel circuits at the level of layer 2, where they are segregated in association with distinct dendritic systems. Results from parvalbumin staining show that the honeycomb mosaic is not limited to rat visual cortex but can be recognized at the layer 1-2 border in other areas and species.

Animals↗

PubMed2003

Multisensory convergence in calcarine visual areas in macaque monkey.

The neural substrates of multisensory perception and integration are still obscure, especially at the cortical level. Alternative viewpoints emphasize (1) 'bottom-up' processes, where different modalities converge in higher order multisensory areas, or (2) 'top-down' projections from multimodal to unimodal areas. In this anatomic study, we use anterograde tracer injections in parietal (8 monkeys) and auditory (3 monkeys) association areas, and demonstrate direct projections to areas V1 and V2 in the calcarine fissure (i.e. the peripheral visual field representation). The laminar signature, with terminations in layers 1 and/or 6, could be consistent with feedback-type connections. A subset of connections from parietal areas, however, branch to both V1 and a ventral extrastriate area (TEO or TEp). Thus, the direct connections to early visual areas V1 and V2 may well operate in conjunction with polysynaptic pathways in a densely parallel network.

Animals↗

PubMed2003

Developmental study of dendritic bundles in layer 1 of the rat granular retrosplenial cortex with special reference to a cell adhesion molecule, OCAM.

In the granular retrosplenial cortex (GRS) of adult rats, callosally projecting pyramidal neurons in layer 2 form dendritic bundles, 30-100 micro m wide, in layer 1. The distinctness of these bundles makes the GRS an attractive model system for investigating the developmental, microcircuitry, and basic organizational features related to dendritic modularity. In this report, we investigate the developmental time course of the dendritic bundles, visualized by immunohistochemistry for microtubule-associated protein 2 (MAP2) and glutamate receptor subunits 2/3 (GluR2/3). Bundles in layer 1 are apparent as early as postnatal day 5, first with GluR2/3 and then, from postnatal day 14, with MAP2. As a step toward understanding the mechanisms of dendritic aggregation, we further investigated the ontogeny of expression of the cell adhesion molecule OCAM. OCAM exhibits a patchy distribution in layer 1 from postnatal day 3 to adult, and the regions of weak OCAM immunoreactivity selectively correspond to the dendritic bundles (in both GluR2/3 and MAP2). The periodic geometry of OCAM-immunoreactive regions, the time course of their appearance and the distinct localization complementary to the bundles support the possibility that this molecule is one contributor to the establishment and maintenance of dendritic modules. The interdigitating relationship between regions of high OCAM immunoreactivity and the dendritic bundles in layer 1 suggests that OCAM may have a repellent influence on the formation of these bundles.

Aging↗