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Biomedical subjects

D A Steindler

Publications and source records attributed to D A Steindler.

53 records · Page 3Linked to original sources
PubMed1986

Monoclonal antibody to glial fibrillary acidic protein reveals a parcellation of individual barrels in the early postnatal mouse somatosensory cortex.

The relative dispositions of cells in immature and mature mouse barrel field cortices that bind antibody to glial fibrillary acidic protein (GFAP) were examined and photographed under the light microscope. Light micrographs demonstrate that radially oriented glial cells are present in the barrel field of postnatal day 6 cortices and that they are located predominantly within the presumptive barrel sides and/or septae, thus sharply delineating individual barrels from each other. The relative dispositions of radial glial fibers observed at this time implicate glia in development of topographic order during early postnatal development of the somatosensory cortex. In contrast, no such delineation could be detected in the cortices of more mature mice, because GFAP-positive astrocytes are present throughout the barrel field and are not confined to barrel sides. This ephemeral nature of the GFAP-delineated barrel field is of interest with respect to the recently reported ephemeral lectin-delineated barrel field.

Aging↗
PubMed1986

Lectins demarcate the barrel subfield in the somatosensory cortex of the early postnatal mouse.

Plant lectins were used to examine the disposition of glycosylated molecules in vibratome sections through the barrel subfield of mouse somatosensory cortex at selected times during postnatal development. The peroxidase conjugates of peanut agglutinin (PNA, specific for N-acetylgalactosamine), concanavalin A (specific for mannose), and wheat germ agglutinin (specific for N-acetylglucosamine and N-acetylneuraminic acid) were used to study lectin binding in aldehyde-fixed tissue sections of cortex. Following peroxidase cytochemistry and light microscopy, it was found that all three lectins bound in the region of the barrel subfield as early as postnatal day 3 (day of birth = postnatal day 1). The lectins bound to the prospective sides and/or septae of individual barrels in preference to the prospective hollows. This lectin demarcation of the barrel field occurred prior to the detection of this region with cresyl violet staining and was still demonstrable on postnatal day 6, when the individual barrels became discernible with cresyl violet. This suggests that the lectin binding material is present before the barrel field becomes a fully formed and organized region. A decrease in lectin affinity for binding sites in these tissue sections occurs during postnatal development (Cooper and Steindler: Soc. Neurosci. (Abstr.) 10: 43a, '84) and this study demonstrates that lectins do not delineate the barrel field of more mature animals (2-3 months old), whereas barrels can be detected with cresyl violet at this time. A preliminary electron microscope analysis of the postnatal day 6 somatosensory cortex demonstrates that the lectin PNA binds to elements of the forming neuropil and also to Golgi apparatus intermediate saccules in neuronal cells. The prospective barrel field can be detected with lectins during a critical period in development in which alterations can occur in the barrel field in response to peripheral deprivation (Jeanmonod et al: Neuroscience 6:1503-35, '81) and therefore we suggest that the glycans visualized with lectin-peroxidase conjugates denote possible candidates for molecules involved in shaping barrel structure.

Animals↗
PubMed1986

Wheat germ agglutinin binding sites in the adult mouse cerebellum: light and electron microscopic studies.

The binding properties of derivatized wheat germ agglutinin (WGA) have been examined in fixed tissue sections from the adult mouse cerebellum and also in axonal tracing paradigms following cerebellar injections. The aim of these studies is to begin to distinguish the roles different binding sites may play in generating diverse biological activities which lead to neuronal uptake and axonal transport of lectins or glycoconjugates. Vibratome sections from aldehyde-fixed cerebellum were incubated in N-[acetyl-3H] WGA or WGA conjugated to horseradish peroxidase (WGA-HRP). Sections from this in vitro binding paradigm and those from cerebellar pressure injection cases using those tracers (in vivo binding paradigm) were processed for light microscopic autoradiography, histochemistry, and electron microscopy. Blocking experiments were also performed with various sugar haptens to confirm the binding specificity of these lectin preparations. Light microscopy of lectin binding patterns within the cerebellar cortex has revealed that both derivatized WGA preparations bind most intensely to the molecular layer. Within the deep cerebellar nuclei, binding is unique and produces a punctate delineation of cell bodies and dendrites. Electron microscopy revealed that these binding sites are associated with glial processes which abut the plasma membrane of deep nuclei cells. Cerebellar WGA-HRP injection sites contain labeled profiles involved in uptake and axonal transport of the labeled lectin (e.g., multivesicular and dense bodies) in addition to label associated with synapses, glia, undetermined components of the extracellular space, and neuronal plasma membranes. These sites are therefore presumed to possess a high affinity or capacity for binding derivatized WGA. Binding studies performed here thus reveal, for the first time, the existence of discrete glial sites that display an extraordinary attraction for lectins such as WGA. The roles such glial glycoconjugates play in diverse biological activities including neuronal uptake and transport of macromolecules need further study.

Animals↗
PubMed1986

The organization of divergent axonal projections from the midbrain raphe nuclei in the rat.

The intranuclear organization of divergently projecting neurons of the midbrain raphe in the rat was studied by using double retrograde axonal tracing. Paired injections of the tracers N-[acetyl-3H] WGA and horseradish peroxidase were made within known projection targets of the midbrain raphe (caudate-putamen, amygdala, hippocampus, substantia nigra, and locus coeruleus). After injections of either tracer in the aforementioned targets, retrograde labeled neurons were found mainly ipsilaterally and within midline portions of the dorsal raphe nucleus, its caudal B6 portion, and within the linear and superior central nuclei of the median raphe complex. There are discrete intranuclear distributions of raphe neurons that project to these forebrain and brainstem sites, and there is an overall rostrocaudal topographic order within the raphe with neurons projecting to the neostriatum, amygdala, and substantia nigra residing most rostrally and neurons projecting to the hippocampus and/or locus coeruleus occupying caudal portions of the B6 and superior central nuclei. Such distributions of projection neurons suggest the existence of an "encephalotopic" intranuclear organization within the raphe; that is, each central nervous system structure that receives midbrain raphe projections has its own unique representation within a topographically distinct portion of one or more of the raphe subgroups. These findings suggest an overall functional organization within the midbrain raphe nuclear complex whereby rostral portions are associated with the basal ganglia and related nuclei, and caudal portions relate to the limbic system. An intermediate representation of amygdala-projecting raphe neurons functionally conjoins the two. Collateralized neurons are found within complex zones of overlap in the topographically organized distributions of raphe neurons projecting to functionally related structures.

Amygdala↗
PubMed1986

The morphology and divergent axonal organization of midbrain raphe projection neurons in the rat.

The morphology of dorsal raphe neurons was examined using intracellular injections of horseradish peroxidase (HRP) and the Golgi technique. Light microscopic examination of HRP-labeled projection neurons revealed a neuron type with radiating, poorly branched and sparsely spined dendrites and terminal dendritic thickets. The stem axon of these neurons left the nucleus ventrally but gave off a beaded collateral while still within the parent cell's dendritic domain. Somatodendritic morphology from Golgi-Kopsch stained material coincided with intracellular HRP findings and the dorsal raphe may consist of varieties of one basic morphological type of neuron. Intracellular recordings made during the HRP injection experiments confirmed that stimulation of the ventral medial tegmentum elicited an antidromic action potential and an inhibitory postsynaptic potential in dorsal raphe projection neurons. The order of axonal projections arising from the midbrain raphe nuclei was examined using a double retrograde axonal tracing technique. After paired HRP and [3H] wheat germ agglutinin injections within certain projection targets of the dorsal and median raphe neurons (caudate-putamen, amygdala, hippocampus, substantia nigra and locus coeruleus), each target structure was found to have its own unique representation within a topographically distinct portion of one or more of the raphe subgroups. Neurons projecting to the caudate-putamen and substantia nigra occupied rather rostral portions. Neurons projecting to the hippocampus and locus coeruleus resided more caudally. Neurons projecting to the amygdala were situated intermediately. Overall, rostrocaudal topography in the intranuclear distributions of raphe projection neurons resulted in the formation of complex overlap zones where collateralized neurons always resided.

Action Potentials↗
PubMed1985

Trigeminocerebellar, trigeminotectal, and trigeminothalamic projections: a double retrograde axonal tracing study in the mouse.

Double retrograde axonal tracing experiments were carried out in order to reveal potential patterns of divergence in axonal projections from the two major sensory nuclei of the mouse brainstem trigeminal complex: the principal sensory and spinal trigeminal nuclei (oralis, interpolaris, and caudalis divisions). The tracers wheat germ agglutinin, N-[acetyl-3H] and horseradish peroxidase were used in paired injection strategies within portions of the cerebellum, superior colliculus, and thalamic ventrobasal complex and/or posterior group of adult ICR white mice. Trigeminal neurons with projections to tactile areas of the cerebellar cortex or underlying deep cerebellar nuclei were found scattered throughout the principal sensory nucleus and interpolaris division, and mainly in dorsal regions of the oralis division of the spinal trigeminal nucleus. Injections of either tracer which involved lateral portions of the rostral half of the superior colliculus labeled trigeminotectal neurons mainly in the contralateral interpolaris division, ventral half of the oralis division, and a ventral region of the principal sensory nucleus near the oralis border. Fewer trigeminotectal neurons were found scattered throughout the principal sensory nucleus and the magnocellular layer of the caudalis divisions, although an occasional labeled neuron wa also found in the marginal layer. Contralaterally projecting trigeminothalamic neurons were observed throughout the principal sensory nucleus, interpolaris division, and within the marginal and magnocellular layers of caudalis. Double-labeled neurons were observed only after paired injections of the tracers in the thalamus and ipsilateral superior colliculus, and they were found within the caudoventral portion of the principal sensory nucleus near the oralis border, throughout the interpolaris division, within the magnocellular layer of caudalis, and only a few double-labeled neurons were also found within the marginal layer. After such injections, 50% of the labeled tectum-projecting neurons in the principal sensory nucleus, 64% in the interpolaris division, and 57% in the caudalis division are branched neurons which have collateralized projections to both the superior colliculus and thalamus. These projections, which have not been described before, appear to arise from more than one class of projection neuron which is differentially distributed within different regions of the trigeminus.

Animals↗
PubMed1985

Corticocortical and collateral thalamocortical connections of postcentral somatosensory cortical areas in squirrel monkeys: a double-labeling study with radiolabeled wheatgerm agglutinin and wheatgerm agglutinin conjugated to horseradish peroxidase.

Experiments were performed to determine whether thalamic neurons send collaterals to more than one subdivision of postcentral somatosensory cortex, and to evaluate the fidelity of somatotopic connections between cortical representations of the body surface. After microelectrode recordings identified the locations of the representations of the fingertips along the border of area 3b with area 3a and along the border of area 1 with area 2, one tracer, tritiated N-acetyl wheatgerm agglutinin ([3H]WGA), was injected into the rostral representation at the 3a-3b border, and another, wheatgerm agglutinin conjugated to horseradish peroxidase (WGA:HRP), was injected into the caudal representation at the 1-2 border. The results indicate that three thalamic regions are interconnected with both border zones. Regions of overlapping retrograde and anterograde label included the ventromedial portion of the lateral division of the ventroposterior nucleus (VPL), which is known to represent the digits; the medial extent of a dorsal capping zone of VPL that we term the ventroposterior superior nucleus (VPS); and the anterior pulvinar (Pa). Cells labeled with one or the other tracer, as well as cells labeled with both tracers, were found in VPL, VPS, and Pa. Our procedures necessarily underestimated the numbers of cells projecting to both rostral and caudal injection zones, but at least 15-20% of VPL and 20-40% of VPS neurons that were autoradiographically labeled appeared to have such collaterals. While only a few cells in Pa were autoradiographically labeled from injections at the 3a-3b border, the presence of several double-labeled cells suggests that at least some Pa neurons project to both injected zones. By relating the present results to those from studies where connections were determined for single architectonic fields, we conclude that many neurons in VP project to both areas 3b and 1, and that many neurons in VPS project to both areas 3a and 2. In addition, both areas 3a and 2 appear to have interconnections with Pa. Connections of Pa with area 2 have only recently been reported (Pons and Kaas, in press), and connections of Pa with area 3a have not been described. The results imply that, in part, the same information from cutaneous receptors is relayed to areas 3b and 1, and the same information from deep receptors is relayed to areas 3a and 2.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1983

Rapid and simple determination of delivery after iontophoretic and pressure injections of radiolabeled tracer substances.

A fluorographic method is described using X-ray film analysis for the determination of delivery of radiolabeled tracer substances both in Agar plates and in tissue sections. This method is most useful in neuroanatomical autoradiographic studies for providing rapid identification of delivery, placement, and extent of an injection site after iontophoresis or pressure injections of radiolabeled axonal tracer substances.

Animals↗
PubMed1983

Combined immunocytochemistry and autoradiographic retrograde axonal tracing for identification of transmitters of projection neurons.

A double labeling method is described which combines immunocytochemistry for identification of the neurotransmitter serotonin with autoradiographic retrograde axonal tracing using wheat germ agglutinin (WGA), N-[acetyl-3H]. The permanence, sensitivity, and distinctness of the two labels provide a valuable means for analyzing transmitter-identified projection neurons in the central nervous system. Combined immunohistochemical/autoradiographic preparations, following injections of WGA, N-[acetyl-3H] in the caudate-putamen of mice, revealed large numbers of serotonergic and fewer non-serotonergic raphe-striatal projection neurons.

Animals↗
PubMed1983

N-[acetyl-3H] wheat germ agglutinin: anatomical and biochemical studies of a sensitive bidirectionally transported axonal tracer.

Both retrograde and anterograde autoradiographic axonal labeling were associated with the appropriate pathways after injections of N-[acetyl-3H] wheat germ agglutinin within numerous structures in the central nervous system of mice. Sections processed for light microscopic autoradiography following the placement of injections mainly within the neocortex, neostriatum, or cerebellum have revealed patterns of bidirectional axonal labeling in various thalamic, monoaminergic, deep cerebellar and precerebellar nuclei that were similar to that seen after similarly placed horseradish peroxidase injections. Small injections of N-[acetyl-3H] wheat germ agglutinin, by way of an extremely limited extracellular spread of the tracer, yield large amounts of autoradiographic retrograde and anterograde axonal labeling. High specific activities, in addition to the sensitivity displayed by this tracer, allow the use of relatively short autoradiographic exposure times that still lead to an extensive signal over labeled neurons. Biochemical analysis of this radiolabeled derivative of wheat germ agglutinin was carried out using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results from Coomassie blue staining and fluorography of the gels have revealed that affinity-purified N-[acetyl-3H] wheat germ agglutinin migrates predominantly as a monomer (molecular weight 18,000 +/- 1000 daltons) as well as a dimer (35,100 +/- 1000 daltons) when compared with native wheat germ agglutinin and molecular weight standards. Gels run on the native preparation, prior to derivatization, radiolabeling and affinity purification, in addition to containing species that co-migrate with the monomeric and dimeric forms of the tagged lectin, also reveal several other bands with molecular weights above and below those values for the monomer and dimer. When scintillation counting was performed on fresh gels of N-[acetyl-3H] wheat germ agglutinin, the predominant species was again found to be the monomeric and dimeric forms; however, small peaks were also present around 12,000 daltons as well as above 43,000 daltons. Coomassie blue staining and fluorography never revealed banding below 18,000 daltons, though some trace of labeling was present, but not in the form of discrete bands, above 36,000 daltons. The presence, albeit small in comparison to the monomer and dimer, of these other higher and lower molecular weight species might in part result from fragments produced by proteolytic digestion of intact subunits of N-[acetyl-3H] wheat germ agglutinin that may or may not possess intact binding sites.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1982

Differences in the labeling of axons of passage by wheat germ agglutinin after uptake by cut peripheral nerve versus injections within the central nervous system.

Injections of a radiolabeled derivative of wheat germ agglutinin (WGA) in the subcortical white matter of the cerebral cortex in mice do not give rise to autoradiographic labeling of axon systems coursing through this fiber mass. Exposing the cut-end of mouse tibial nerve to WGA does, however, produce labeling within dorsal root ganglia and the spinal cord. These findings are discussed with consideration for dissimilarity in mode of central versus peripheral administration of the tracer, as well as in the light of potential relative differences in the uptake and transport of WGA and HRP.

Afferent Pathways↗
PubMed1981

Locus coeruleus neurons have axons that branch to the forebrain and cerebellum.

Utilizing a recently introduced double retrograde axonal tracing technique, the existence of collateralized axonal projections of locus coeruleus neurons to both forebrain and cerebellar cortical structures has been examined in adult mice. Following paired injection strategies using horseradish peroxidase and radiolabeled wheat germ agglutinin within portions of the forebrain and cerebellar cortices, substantial numbers of double labeled 'branched' neurons have been found that reside within all regions of the nucleus.

Afferent Pathways↗
PubMedDate not supplied

Neuron-glial interactions during the in vivo and in vitro development of the nigrostriatal circuit.

This paper examines a particular aspect of glial-neuronal interactions during central nervous system development: the possible influence of growing neurites on the expression of glial-associated extracellular matrix (ECM) molecules. In particular, using in vivo manipulations of the dopaminergic projections from the midbrain substantia nigra, as well as an in vitro model of the developing nigrostriatal circuit, we look at the reciprocal interactions between growing dopaminergic axons and astrocyte-derived ECM molecules in the striatum. Glial-derived glycoconjugates, including tenascin and a proteoglycan designated DSD-1, are developmentally expressed ECM molecules which have been shown to have different effects on immature neurons and their growing processes. Here we show that the glial expression of these ECM constituents in a target region (the caudate-putamen or neostriatum) may be affected by the presence or absence of an appropriate, maturing afferent projection (in this case, dopaminergic nigrostriatal axons). In general, our results reveal complex glial-neuronal interactions during the normal development of central nervous system circuits, and the ability to create in vivo and in vitro models which may be useful toward understanding these complex cellular and molecular interactions in degeneration and plasticity of the nigrostriatal circuit in diseases including Parkinson's.

Animals↗
PubMedDate not supplied

A unique mosaic in the visual cortex of the reeler mutant mouse.

Numerous studies have revealed abnormal cytoarchitectonics in the reeler mouse brain. In the present study, acetylcholinesterase (AChE) histochemistry has revealed a distinctive mosaic within the occipital cortex of the reeler mouse. The mosaic does not appear until after the second postnatal week, perhaps in association with eye opening. AChE staining in the visual cortex of normal littermates does not exhibit a mosaic pattern, but rather, is present within bands or laminae. The AChE mosaic in reeler persists into adulthood. Immunocytochemical staining of the tenascin glycoprotein, an astrocyte-derived extracellular matrix molecule that is concentrated in boundaries around emerging functional patterns in the CNS, reveals a boundary-mosaic pattern in the first postnatal week. Dil axonal tracing in normal versus reeler mice indicates that the thalamocortical projections may also be associated with the AChE mosaic. The observation that a mosaic is unique to the occipital cortex of reeler mice suggests that it may evolve through abnormal cell and molecular interactions in the mutant cortex that normally lead to the development of functional visual representations.

Acetylcholinesterase↗