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J DeFelipe

Publications and source records attributed to J DeFelipe.

At least 55 records · Page 3Linked to original sources
PubMed1995

Immunocytochemical localization of non-NMDA ionotropic excitatory amino acid receptor subunits in human neocortex.

The distribution of immunocytochemically localized subunits that comprise ionotropic non-NMDA excitatory amino acid receptors was examined in human frontal, parietal and temporal association neocortex. AMPA/kainate receptor subunits were identified using a monoclonal antibody (3A11) that recognizes an epitope common to GluR2 and GluR4 [GluR2(4)], as well as polyclonal antisera that recognize GluR2 and GluR3 (GluR2/3). Kainate receptor subunits were identified using a monoclonal antibody (4F5) that recognizes an epitope common to GluR5/6/7. For all three antibodies used, labeling was observed in a large number of neurons throughout the human association neocortex with the highest immunoreactivity present in pyramidal-like neurons, a cellular pattern largely similar to that observed in the monkey neocortex. These data demonstrate the cellular localization patterns for some non-NMDA receptor subunits in human neocortex, details upon which further studies on the roles of these subunits in human neurological diseases can be based.

Adolescent↗
PubMed1994

Microzonal decreases in the immunostaining for non-NMDA ionotropic excitatory amino acid receptor subunits GluR 2/3 and GluR 5/6/7 in the human epileptogenic neocortex.

Potential alterations in glutamate-utilizing excitatory circuits in resected human epileptogenic frontal and temporal neocortex were investigated by using immunocytochemical methods to visualize receptor subunits which comprise the AMPA/kainate (GluR2/3) and kainate (GluR5/6/7) receptor subtypes. Examination of the patterns of immunostaining in regions of neocortex that were identified as spiking and non-spiking based on intraoperative electrocorticography revealed dramatic, microzonal decreases in immunoreactivity for the receptor subunits examined. The patches of decreased immunostaining for GluR2/3 and for GluR5/6/7 were often coincident with respect to each other. However, such abnormal regions were not necessarily correlated with any particular electrocorticographically defined regions nor any overtly abnormal cytoarchitectural features in adjacent Nissl-stained sections. Moreover in many but not all cases, the focal regions of decreased receptor subunit immunoreactivity coincided with small patches of decreased parvalbumin immunoreactivity a calcium-binding protein which labels a subpopulation of powerful inhibitory GABAergic interneurons. These results indicate that in the human epileptogenic neocortex there may be alterations in particular excitatory and/or inhibitory synaptic systems at small, multiple neocortical foci, and that these alterations are found mostly in the same regions. We suggest that these alterations may contribute to the initiation and/or propagation of seizure activity.

Adolescent↗
PubMed1994

A study of SMI 32-stained pyramidal cells, parvalbumin-immunoreactive chandelier cells, and presumptive thalamocortical axons in the human temporal neocortex.

Immunocytochemical studies in the primate neocortex have shown that particular populations of pyramidal cells can be identified by antibody SMI 32 that recognizes a nonphosphorylated epitope of neurofilament protein, while chandelier cells (a powerful type of cortical inhibitory interneuron) and presumptive thalamocortical axons can be identified by antibodies directed against the calcium-binding protein parvalbumin (PV). We used these antibodies in correlative light and electron microscopic immunocytochemical studies to analyze certain aspects of the synaptic circuitry of human temporal neocortex. In sections cut in the tangential plane, many PV-immunoreactive chandelier cell axon terminals and apical dendrites of SMI 32-stained pyramidal cells were distributed in small clusters. Combination of immunocytochemistry for PV and SMI 32 revealed four subpopulations of pyramidal cells with regard to the immunocytochemical staining by SMI 32 and the innervation of their axon initial segments by PV-positive or -negative chandelier cell axon terminals, but there were differences in the concentration and proportion of these subpopulations by layers. Furthermore, we present electron microscopic evidence suggesting that the characteristic layer III dense band of PV-immunoreactive puncta is made up mainly of presumptive thalamocortical axon terminals. Besides, coincidence was found between the dense PV-immunoreactive band and the dendritic plexus formed by the SMI 32-stained pyramidal cells in the lower half of layer III, which leads us to think that they are probably a major target of PV-immunoreactive thalamic terminations.

Antibodies, Monoclonal↗
PubMed1993

A study of NADPH diaphorase-positive axonal plexuses in the human temporal cortex.

Nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry was used to study the morphology of labeled axonal plexuses in the human lateral temporal cortex. Strongly stained non-pyramidal neurons and a dense NADPH diaphorase-positive network of fibers were observed in all cortical layers. Certain stained fibers are found to give rise to basket-like formations. Notably other fibers seem to innervate small blood vessels. In addition, numerous blood vessels show a punctate labeling over their surfaces. These findings provide new morphological and chemical details of the axonal innervation of the human neocortex.

Axons↗
PubMed1993

A simple and reliable method for correlative light and electron microscopic studies.

We describe in detail a simple method for flat-embedding that can be subsequently used in correlative light and electron microscopic studies. The method can be applied to any material suitable for electron microscopy and is especially useful for study of the synaptology and ultrastructural characteristics of immunocytochemically or morphologically identified neurons or their processes. We present here an example to show how accurately one can delineate the fine details of a complex axonal arborization impregnated with the Golgi method in the mouse cerebral cortex. Golgi-impregnated sections to be studied at the electron microscopic level are osmicated, dehydrated, infiltrated with Araldite resin, flat-embedded, and identified cells or processes photographed. Serial semi-thin sections (1-2 microns thick) are then cut with an ultramicrotome, examined with the light microscope, and the elements rephotographed. Selected semi-thin sections are then resectioned on the ultramicrotome at 60-70 nm and examined electron microscopically. This method allows the systematic and accurate localization of stained cells and processes throughout the successive steps of the procedure.

Animals↗
PubMed1992

Santiago Ramón y Cajal and methods in neurohistology.

Controversy, misunderstanding or uninformed opinion abound over the extent to which the great Spanish neurohistologist, Santiago Ramón y Cajal, specified his staining methods in his analytical papers, the methods by which he analysed and presented his data, and the microscopes available to him. In this paper, we have attempted to outline the information on these points that we have been able to obtain from a detailed examination of his writings and a study of the evidence remaining in the Cajal Museum in Madrid.

Animals↗
PubMed1991

Parvalbumin immunoreactivity reveals layer IV of monkey cerebral cortex as a mosaic of microzones of thalamic afferent terminations.

Correlative light and electron microscopic immunocytochemical methods were used to study the pattern of staining for the calcium-binding protein parvalbumin (PV) in the primary visual area (area 17) and area 3b of the first somatic sensory area of the monkey cerebral cortex. A conspicuous feature of the light microscopic staining pattern is the presence of focal aggregations of immunoreactive terminal-like puncta within the major thalamic recipient layers (IV and VI). At the electron microscopic level these aggregations of puncta are found to be immunoreactive terminals most of which form asymmetric synapses, principally on dendritic spines and, to a lesser extent, on dendritic shafts. Outside the aggregations, most PV-immunoreactive terminals form symmetric synapses. Correlative observations in the present and other studies indicate that the aggregations of PV-immunoreactive terminals forming asymmetric synapses arise from thalamic afferent fibers while those forming symmetric synapses arise from intrinsic gamma-aminobutyric acid neurons. The aggregations of PV immunoreactivity in layers IV and VI form microzones of preferred thalamic afferent terminations which may contribute to the formation of functional columns based upon focussed thalamic inputs.

Afferent Pathways↗
PubMed1991

Patterns of synaptic input on corticocortical and corticothalamic cells in the cat visual cortex. I. The cell body.

Immunocytochemical and electron microscopic methods were used to examine the ultrastructure and synaptology of callosal and corticothalamic pyramidal cell somata in the cat visual cortex (area 17). Callosal and corticothalamic cells were labeled after injection of horseradish peroxidase (HRP) in the contralateral visual cortex or in the ipsilateral lateral geniculate nucleus. The synaptic relationship between each of the two populations of pyramidal cells and cells containing the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) was examined at the light and electron microscope level using the combined techniques of retrograde transport of HRP and GABA immunocytochemistry. We found that callosal and corticothalamic cell somata have an ultrastructure and synaptology that distinguishes them from each other. Reconstructions from electron micrographs of serial sections revealed that the vast majority of synapses (89-96%) on the cell body of pyramidal cells were formed by GABAergic axon terminals, and that within each population of pyramidal cells there was variability in the number and density of axosomatic synapses. Callosal pyramidal cells received a greater number and higher density of axosomatic synapses than corticothalamic cells. These data suggest that callosal cells receive more inhibition than corticothalamic cells at the level of their somata.

Animals↗
PubMed1991

Patterns of synaptic input on corticocortical and corticothalamic cells in the cat visual cortex. II. The axon initial segment.

In the present study we examined the synaptology of the axon initial segments (AISs) of three populations of retrogradely labeled pyramidal cells: callosal, corticothalamic, and ipsilateral corticocortical projecting cells. The cells were labeled by horseradish peroxidase injected into the contralateral visual cortex, the ipsilateral lateral geniculate nucleus, or area 19 of the ipsilateral hemisphere. The AISs of these cells were completely reconstructed from tracings of serial electron micrographs and the number and type of synapses ending on them determined. These data were compared with that described in the companion paper (Fariñas and DeFelipe, J Comp Neurol 1991; 304:53-69), in which the ultrastructure and synaptology of the somata of callosal and corticothalamic cells was investigated. Together, these data permit comparisons to be made between the somatic and axonic innervation of the same cells. Our results show that each population of efferent cells examined received a characteristic and rather homogeneous number (in comparison with the number of axosomatic synapses) of axoaxonic synapses: 16 to 23 for AISs of callosal cells, 22 to 28 for ipsilateral corticocortical cells, and 1 to 5 for corticothalamic cells. Since most axoaxonic synapses are likely to be GABAergic, it is concluded that each population of pyramidal cells receives a distinct GABAergic inhibitory synaptic profile.

Animals↗
PubMed1990

A microcolumnar structure of monkey cerebral cortex revealed by immunocytochemical studies of double bouquet cell axons.

Immunocytochemical methods were used to study 28,000 mol. wt calbindin and tachykinin immunoreactivity in the monkey cerebral cortex. Calbindin and tachykinin immunoreactivity give rise to a generally different pattern of staining of cell bodies and terminal-like puncta. However, the staining of long, vertically-oriented bundles of processes--identical to classical double bouquet cell axonal arborizations--is the most prominent feature of the pattern of both calbindin- and tachykinin-immunoreactive staining. These bundles form a widespread and regular columnar system descending from layer II to layers III-V. The bundles are most evident in layer III where, in tangential sections, they have a density of 7-15 bundles/10,000 microns 2 with a center-to-center spacing of 15-30 microns. The distribution of immunoreactive bundles through the cortex is not homogeneous; somatic sensory, auditory, and visual areas display a large number of calbindin-immunoreactive bundles while tachykinin-immunoreactive bundles are only numerous in the auditory areas and in area 18 of the visual cortex. In the motor cortex (area 4) few or no immunoreactive bundles are visualized with either antibody. Correlative light and electron microscope analysis of tachykinin immunoreactive bundles in the primary auditory cortex shows that the tachykinin-positive axons of the bundles form symmetrical synaptic contacts with dendritic shafts (57%) and spines (43%). Frequently, several immunoreactive boutons that arise from the same fiber are seen climbing along the surfaces of vertically-oriented, non-immunoreactive processes which include myelinated and unmyelinated axons and probably glial processes. The same ultrastructural features and a similar synaptic distribution were found in a previous study [DeFelipe et al. (1989) Brain Res. 503, 49-54] of calbindin-positive bundles in the somatic sensory cortex (areas 3a and 1). Despite the virtually identical morphological features of tachykinin- and calbindin-immunoreactive bundles, colocalization studies demonstrate little coexistence of the two antigens in somata and none in the axonal bundles of double bouquet cells. These data suggest that the double bouquet cell is a chemically heterogeneous, but ubiquitous morphological type of cortical interneuron, whose uniquely bundled axonal system, which is probably GABAergic, imposes a fundamental microcolumnar organization upon the cerebral cortex.

Animals↗
PubMed1989

Glutamate-positive neurons and axon terminals in cat sensory cortex: a correlative light and electron microscopic study.

Immunocytochemical methods were used to perform a correlative light and electron microscopic study of neurons and axon terminals immunoreactive to the antiglutamate (Glu) serum of Hepler et al. ('88) in the visual and somatic sensory areas of cats. At the light microscopic level, numerous Glu-positive neurons were found in all layers except layer I of both cortical areas. On the basis of the dendritic staining of Glu-positive cells, two major morphological categories were found: pyramidal cells, which were the most frequent type of immunostained neuron, and multipolar neurons, which were more numerous in layer IV of area 17 than in any other layer. A large number of Glu-positive neurons, however, did not display dendritic labelling and were considered unidentified neurons. Counts of labelled neurons were performed in the striate cortex; approximately 40% were Glu-positive. Numerous lightly stained punctate structures were observed in all cortical layers: the majority of these Glu-positive puncta were in the neuropil. After resectioning the plastic sections for electron microscopy it was observed that: 1) the majority of neurons unidentifiable at light microscopic level were indeed pyramidal neurons except in layer IV of area 17, where many stained cells were probably spiny stellate neurons. Some Glu-positive neurons, however, exhibited clear ultrastructural features of nonspiny nonpyramidal cells; 2) all synaptic contacts made by Glu-positive axon terminals were of the asymmetric type, but not all asymmetric synaptic contacts were labelled. The vast majority of postsynaptic targets of Glu-positive axons were unlabelled dendritic spines and shafts. The present results provide further evidence that Glu (or a closely related compound) is probably the neurotransmitter of numerous excitatory neurons in the neocortex.

Animals↗
PubMed1989

Synapses of double bouquet cells in monkey cerebral cortex visualized by calbindin immunoreactivity.

In the monkey neocortex, immunoreactivity for the 28-kDa vitamin D-dependent calcium binding protein (Calbindin) is contained in a set of GABAergic intrinsic neurons whose small size and laminar locations render them very distinct from a second set of GABAergic intrinsic neurons that show immunoreactivity for another calcium-binding protein, parvalbumin. A conspicuous feature of many calbindin-immunoreactive cells is their possession of long, vertically oriented bundles of immunoreactive processes that descend or ascend vertically through several cortical layers. These are components of the radial fasciculi of the cortex and are here shown by correlative electron microscopic immunocytochemistry to consist of both immunoreactive dendrites and unmyelinated axons. The morphology of the bundles and the parent cells indicates that the cells are classical double bouquet cells. The calbindin-positive axons in the radial fasciculi in the present study formed symmetric synapses on unlabeled dendritic shafts (62%) and spines (38%). Despite the close-packed nature of the immunoreactive axons, relatively few terminals of the same axon converged on a single postsynaptic profile. The postsynaptic profiles were identified in certain cases as side branches of pyramidal cell apical and basal dendrites. Mainstem apical dendrites generally did not receive synapses derived from the calbindin-positive axons. These results indicate that double bouquet cells can be distinguished both by their GABAergic character and by their possession of calbindin immunoreactivity. They are probably major contributors to the vertical flow of inhibitory influences across laminae of the cerebral cortex.

Animals↗
PubMed1989

Visualization of chandelier cell axons by parvalbumin immunoreactivity in monkey cerebral cortex.

Antibodies directed against the calcium-binding protein parvalbumin label a subpopulation of gamma-aminobutyric acid-releasing neurons in the cerebral cortex that is thought to have particular metabolic and physiological properties. The chandelier cell is a well-characterized morphological type of gamma-aminobutyric acid-releasing cortical interneuron, the axon of which possesses very distinctive terminal portions located around the initial axon segments of pyramidal cells. In the pre- and postcentral gyri of the monkey, we found that these distinctive terminal portions of chandelier cell axons were immunocytochemically stained for parvalbumin in a manner that reveals their complete structure. The chandelier cell axons were identified light-microscopically as short, vertically oriented rows of parvalbumin-positive puncta (PV-Rs). The PV-Rs varied in both length and complexity and were located beneath unstained pyramidal cells. PV-Rs were very numerous in layers II-III, where most pyramidal cells appeared to have a PV-R beneath them. Fewer PV-Rs were found in deeper layers, and in layer VI PV-Rs were rare. With EM all PV-Rs could be seen to form multiple synaptic contacts of the symmetrical type on the initial segments of pyramidal cell axons. Parvalbumin immunoreactivity can therefore be used as a reliable marker for chandelier cell axons.

Animals↗
PubMed1988

Demonstration of glutamate-positive axon terminals forming asymmetric synapses in cat neocortex.

Electron microscopic examination of sections immunocytochemically processed with an anti-glutamate serum reveals that many asymmetric synapses in the cat neocortex contain elevated levels of immunodetectable glutamate. These labelled axon terminals are likely to use glutamate as neurotransmitter. Axon terminals forming symmetric contacts were never labelled. Since glutamate is known to exert potent excitatory effects on neocortical neurons, the present finding gives immunocytochemical evidence that asymmetric synapses are excitatory.

Animals↗
PubMed1988

A light and electron microscopic study of serotonin-immunoreactive fibers and terminals in the monkey sensory-motor cortex.

Immunocytochemical methods were used to study the distribution and ultrastructure of serotonin (5-hydroxytryptamine; 5-HT) immunoreactive fibers innervating the monkey sensory-motor cortex. Beaded 5-HT positive fibers were found in all cortical layers of both areas but with relatively fewer in middle cortical layers. Examination of 2 micron-thick plastic sections at the light microscope level, revealed that the vast majority of the bouton-like structures on the fibers lay in the neuropil and not adjacent to neuronal somata. A few beaded immunoreactive fibers were seen around certain pyramidal and non-pyramidal cell somata, very occasionally forming modest pericellular ramifications. Serial reconstructions made from electron micrographs after resectioning the 2 micron-thick sections, revealed that the dilatations of the fibers are 5-HT positive boutons but the boutons examined rarely formed morphologically identifiable synaptic contacts. Of 191 reconstructed boutons only 5 made contacts with obvious membrane specializations, all of which were of the asymmetrical type. No immunoreactive synaptic contacts were seen on pyramidal cell somata in the cortex, nor on dendrites or somata in the white matter underlying the cortex, although 5-HT positive boutons commonly lay closely adjacent to neuronal profiles in both sites. 5-HT fibers in the cortex and white matter have a similar morphological appearance and both myelinated and unmyelinated types are seen.

Animals↗
PubMed1988

Local connections in transplanted and normal cerebral cortex of rats.

Injections of the fluorescent tracer Fluoro-Gold were made in transplanted and normal cerebral cortex of rats in order to investigate and compare the local connectivities of both. In the normal somatosensory cortex, small injections in superficial layers (I to III) produced retrograde cell labeling below the injection site in two bands: in layer V and in the deep part of layer VI. Pieces of embryonic rat neocortical tissue were transplanted into a cavity made in the somatosensory cortex of young adult rats. After a survival period of 2-3 months, small injections of Fluoro-Gold were made in the superficial part of the grafts. These injections revealed multiple clusters of intratransplant-projecting cells. No callosal or thalamic neurons were labeled in these experiments. On occasion, a bilaminated pattern of retrograde cell labeling was observed inside the transplants. In both transplanted and normal cortices, pyramidal and non-pyramidal cells were retrograde-labeled. We conclude that in the neocortical transplants there is a pattern of local connectivity that is reminiscent of the pattern of intracortical connectivity in the normal neocortex in at least two aspects: first, the retrograde-labeled cells tended to form clusters or bands; second, both pyramidal and non-pyramidal cells were labeled.

Animals↗
PubMed1988

Synaptic connections of an interneuron with axonal arcades in the cat visual cortex.

A single, isolated interneuron with axonal arcades in the cat visual cortex was analysed in detail by both light and electron microscopy. The neuron was impregnated by the Golgi-Kopsch method, gold-toned, and processed for electron microscopy using the ethanolic phosphotungstic acid (PTA) staining method of Bloom & Aghajanian (1968). These methods, in combination, resulted in the successful identification of a large number of synaptic boutons arising from the axon of the cell under study. We examined serially at the electron microscope level 210 boutons of the axonal arborization of the cell. Of these, 152 formed identifiable symmetrical synaptic contacts with a variety of postsynaptic elements. The vast majority of the postsynaptic targets were dendritic profiles, which represented 95.7% of all the synaptic contacts identified. Only one example was observed of two labelled boutons making contacts with the same postsynaptic element; the rest were apparently on different elements. This distribution of synapses, characterized by the lack of convergence, is very similar to that reported by other authors for a certain kind of double bouquet cell which, in turn, shares some morphological features with the neurons with axonal arcades. It is suggested that fine details of the geometry of the axonal arborization of a given cell are an important reflection of the distribution of its synapses.

Animals↗