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D M Landis

Publications and source records attributed to D M Landis.

At least 37 records · Page 2Linked to original sources
PubMed1991

A novel epitope expressed on the surface of developing and mature astrocytes.

Plasmalemmal fractions from cultured astrocytes have been used as the immunogen in generating a monoclonal antibody, termed 8C10, which binds to the surface of cultured astrocytes of the rat. 8C10 immunoreactivity is present on the membrane surface of cultured type 1 astrocytes, type 2 astrocytes, oligodendrocytes, meningeal cells, and 02A progenitor cells, and it persists after aldehyde fixation. The antibody also stains aldehyde-fixed central nervous system, in a pattern which suggests that the plasma membranes of fine astrocytic processes in adult neuropil express the epitope. Astrocytic perikarya and processes in white matter are also stained, but there is no immunoreactivity present in neuronal processes or perikarya. Astrocytic processes in developing cerebellar cortex are stained at postnatal ages when some of these processes are guiding the migration of neuronal perikarya.

Animals↗
PubMed1990

Serum influences the differentiation of membrane structure in cultured astrocytes.

The membranes of mammalian astrocytic processes apposed to blood vessels or forming the surface of the brain contain high concentrations of a characteristic intramembrane particle aggregate, termed "assemblies." In order to identify developmental processes which contribute to this remarkable regional specialization of membrane structure, we have devised culture conditions which support the differentiation of assemblies in secondary cultures of astrocytes derived from neonatal rat forebrain. We report here that different lots of fetal calf serum vary dramatically in their capacity to support the differentiation of assemblies. Fetal calf serum thus appears to exert two distinct influences on astrocyte development: it promotes the differentiation of type 2 astrocytes from bipotential precursor cells, as shown by others, and it influences the density of assemblies in type 1, flat, GFAP-immunoreactive astrocytes in our secondary cultures. Horse serum and defined media also support the appearance of assemblies in flat, GFAP-immunoreactive astrocytes. The separate effects of serum supplementation upon cell lineage and membrane differentiation have to be carefully considered in studies designed to examine factors influencing astrocytic development in vitro.

Animals↗
PubMed1989

Changes in the structure of synaptic junctions during climbing fiber synaptogenesis.

Synaptogenesis between climbing fiber axons and Purkinje cells involves both an orderly translocation of synaptic junctions over the Purkinje cell surface and an elimination of all but one innervating axon. We used thin-section and freeze-fracture electron microscopic techniques to study structural changes in synaptic junctions during this interval of synapse translocation and elimination. In freeze-fractured preparations, virtually all climbing fiber synaptic junctions with the perisomatic processes and somatic spines lacked the particle aggregates that characterized the extracellular half of the postsynaptic membrane of mature synaptic junctions with dendritic spines. Some climbing fiber junctions with the dendritic shaft in the second postnatal week were associated with such aggregates, despite the fact that these junctions are transient. Thus, during the interval when Purkinje cells initially were innervated by multiple climbing fibers, and subsequently denervated of all but one climbing fiber afferent per cell, only a few of the transient synaptic junctions on the cell body and proximal dendrites have associated particles. The presence of a particle aggregate at a synaptic junction does not appear to be correlated with the permanence of that junction and probably is not correlated with the capacity to support synaptic transmission. The particle aggregates might be indicative of relatively long-lived junctions, or might occur only at junctions formed by the climbing fiber that will persist in synaptic contact with the mature Purkinje cell.

Aging↗
PubMed1989

Substructure in the assemblies of intramembrane particles in astrocytic membranes.

Assemblies are a specialization of intramembrane structure in astrocytes which is concentrated in astrocytic processes at the interface with vascular and cerebrospinal fluid compartments. When astrocytic processes are rapidly frozen and then freeze-fractured at very low stage temperature, the constituent particles of assemblies fracture in at least two planes in the lipid bilayer. The true outer surface of the astrocytic membrane can be exposed by etching rapidly frozen tissue and in such preparations the assemblies are seen to extend through the extracellular half of the membrane to be exposed on the surface to the extracellular fluid. The dimensions of the particles, their tendency to fracture at several levels and their exposure to the extracellular space all indicate that they are not composed solely of lipid. We conclude that assemblies represent a protein which partially or entirely bridges the membrane and which may serve a transport function.

Animals↗
PubMed1988

Membrane and cytoplasmic structure at synaptic junctions in the mammalian central nervous system.

Application of rapid freezing, freeze substitution fixation, and freeze fracture techniques to the study of synaptic junctions in the mammalian central nervous system has revealed new aspects of synaptic structure that are consistent with and partially explicate advances in synaptic biochemistry and physiology. In the axoplasm adjacent to the presynaptic active zone, synaptic vesicles are linked to large spectrin-like filamentous proteins by shorter proteins that resemble synapsin I in morphology. This mesh of presynaptic filamentous proteins serves to concentrate synaptic vesicles in the vicinity of the active zone. The affinity with which the vesicles are bound by the mesh is probably modulated by the extent of phosphorylation at specific sites on the constituent filamentous proteins, and changes in the binding affinity result in changes in transmitter release. The structural organization of the postsynaptic density in Purkinje cell dendritic spines consists of very fine strands with adherent, heterogeneous globular proteins. Some of these globular proteins probably correspond to protein kinases and their substrates. The postsynaptic density, positioned at the site of the maximal depolarization caused by synaptic currents, apparently serves as a supporting framework for a variety of proteins, which respond to and transduce postsynaptic depolarization. At least two classes of filamentous protein fill the cytoplasm of spines with a complex mesh, which presumably contributes to maintenance of the spine shape. Membrane bound cisterns are a ubiquitous feature of Purkinje cell dendritic spines. Studies of rapidly frozen tissue with electron probe microanalysis and elemental imaging reveal that these cisterns take up and sequester calcium, which is derived from the extracellular space, and which probably enters the spine as part of the synaptic current.

Animals↗
PubMed1988

The organization of cytoplasm at the presynaptic active zone of a central nervous system synapse.

The axoplasm at the presynaptic active zone of excitatory synapses between parallel fibers and Purkinje cell spines contains a meshwork of distinct filaments intermingled with synaptic vesicles, seen most clearly after the rapid freezing, freeze-etch technique of tissue preparation. One set of filaments extends radially from synaptic vesicles and intersects similar filaments associated with vesicles as well as larger filaments arising from the presynaptic membrane. The small, vesicle-associated filaments appear to link synaptic vesicles to one another and to enmesh them in the vicinity of the synaptic junction. The vesicle-associated filaments could be synapsin I because they have the same molecular dimensions and are distributed in the same pattern as synapsin I immunoreactivity.

Animals↗
PubMed1988

Activity-dependent accumulation of calcium in Purkinje cell dendritic spines.

The calcium content of synapses of parallel fibers on Purkinje cell dendritic spines was determined by electron probe x-ray microanalysis of freeze-dried cryosections from directly frozen slices of mouse cerebellar cortex. In fresh slices frozen within 20-30 sec of excision, calcium concentrations ranging from 0.8 to 18.6 mmol/kg of dry weight were measured in cisterns of smooth endoplasmic reticulum within Purkinje cell dendritic spines. The average calcium content of spine cisterns in rapidly excised slices (6.7 +/- 0.6 mmol/kg of dry weight +/- SEM) was higher than the average calcium content of spine cisterns in brain slices incubated without stimulation for 1-2 hr before direct freezing (2.5 +/- 0.4 mmol/kg of dry weight). Depolarization of incubated cerebellar slices by isotonic 55 mM KCl resulted in the accumulation within spine cisterns of very high amounts of calcium or isotonically substituted strontium, both derived from the extracellular fluid. These results suggest that one function of spine cisterns is to sequester free calcium that enters the spine through ligand-gated or voltage-gated channels during synaptic transmission.

Animals↗
PubMed1987

Initial junctions between developing parallel fibers and Purkinje cells are different from mature synaptic junctions.

During postnatal development of cerebellar cortex, junctions are formed between parallel fiber axons and the shafts of Purkinje cell dendrites. These shaft junctions resemble synaptic junctions on spines in thin sections, in that the axon contains a cluster of synaptic vesicles, and the pre- and postjunctional membranes are lined by electron-dense material. The shaft junctions do not have the aggregate of particles arrayed on the extracellular half of the postjunctional membrane that is characteristic of mature spine synaptic junctions in freeze-fractured preparations, however, and so presumably have a different protein composition. Shaft junctions are transient specializations, present only in developing tissue, but do not appear simply to be intermediates in the formation of mature spine synaptic junctions. In normal development, spines are formed by the Purkinje cell dendrite at sites not occupied by shaft junctions. Moreover, in certain neurological mutant mice, shaft junctions form in the absence of spine synapses, and in other systems spines develop in the absence of shaft junctions. We suggest that shaft junctions are a class of synapse formed during development which is distinguished by its capacity to dissociate, and that only some fraction of the parallel fibers forming shaft synapses with a developing Purkinje cell will have established spine synapses in the adult.

Aging↗
PubMed1987

Morphology of striatal neurons containing VIP-like immunoreactivity.

We have used light and electron microscopic immunocytochemical techniques to study the distribution and morphology of neurons that contain vasoactive intestinal polypeptide-like immunoreactivity (VIP-Ir) in the adult rat striatum. VIP-Ir cells were sparsely distributed throughout all rostrocaudal levels of the striatum. Cell bodies were of medium size (12-17 microns) and gave rise to three to five primary dendrites, which branched close to the soma and became varicose. These dendrites appeared aspiny at the light microscope level and could be traced up to 250 microns in length. Dendrites frequently traversed axonal bundles in the striatum, a pattern not exhibited by neurons containing somatostatin-like or substance P-like immunoreactivity. In several instances, very fine varicose processes arborized extensively within 40 microns of the VIP-Ir soma; these may represent axons. In thin-sectioned preparations, examined under the electron microscope, the nucleus of VIP-Ir neurons was eccentrically located and showed several deep invaginations. Immunoreactive dendrites of VIP-Ir cells appeared virtually spine-free. Synapses with asymmetric or symmetric junctional specializations were present on the dendritic surface. Several VIP-Ir varicosities were found to terminate on the VIP-Ir cell body, forming synaptic junctions with symmetric specializations; these synapses may derive from recurrent collaterals. VIP-Ir cells thus resemble other aspiny striatal neurons considered likely to be local circuit neurons.

Animals↗
PubMed1987

Substructure in the postsynaptic density of Purkinje cell dendritic spines revealed by rapid freezing and etching.

In tissue prepared by rapid freezing, freeze fracture, and shallow etching, the postsynaptic density of Purkinje cell dendritic spines has a substructure consisting of fine filaments and irregular, globular adherent proteins. The number and packing density of the globular proteins vary from region to region within a single density and are even more variable when different junctions are compared. Whereas actinlike microfilaments and spectrinlike filaments are juxtaposed to the postsynaptic density, they do not appear to be continuous with the constituent filaments of the density. We suggest that the postsynaptic density at this class of synapse is composed of fine filamentous proteins that insert on the postsynaptic membrane and serve as a supporting framework for a variety of globular proteins. The globular proteins may vary qualitatively and quantitatively from junction to junction, and are positioned in the region of the spine that has the greatest concentration of ionized calcium entering with the synaptic current, and the greatest extent of postsynaptic depolarization.

Animals↗
PubMed1987

Distribution of calcium and potassium in presynaptic nerve terminals from cerebellar cortex.

The elemental composition of the presynaptic nerve terminals in rapidly frozen synapses of the cerebellar molecular layer was determined by electron probe x-ray microanalysis and elemental imaging of characteristic x-rays. Elemental imaging of thin freeze-dried cryosections from fresh cerebellar slices frozen within 20 sec of removal from the brain showed normal concentrations of potassium (95 +/- 6 mmol/liter wet tissue +/- SEM) and calcium (0.8 +/- 0.4 mmol/liter) in whole presynaptic terminals, even though mitochondrial and nonmitochondrial sites containing up to 30 mmol of calcium per liter were present elsewhere in the neuropil. Quantitative electron probe analysis of synaptic vesicle clusters and intraterminal mitochondria indicated that their calcium concentrations were 0.4 +/- 0.1 and 1.2 +/- 0.2 mmol/liter, respectively. The low calcium content of presynaptic organelles was confirmed by the absence of detectable deposits in preparations freeze-substituted so as to stabilize calcium content. Similar experiments were carried out on cerebellar slices rapidly frozen after incubation in vitro. The distribution of potassium and calcium in presynaptic terminals of resting and depolarized (55 mM potassium) slices was qualitatively and quantitatively similar to that in freshly excised cortex, although resting slices lacked the few calcium-rich sites that appeared in other areas of the neuropil after stimulation. The calcium concentrations in whole terminals, synaptic vesicles, and mitochondria of resting slices were 1.4 +/- 0.7, 0.7 +/- 0.2, and 0.9 +/- 0.2 mmol/liter, respectively. Thus, amounts of calcium typical of storage organelles in other tissues are not present within cerebellar synaptic vesicles, suggesting that they have a limited role in calcium storage and release.

Animals↗
PubMed1986

Membrane structure at synaptic junctions in area CA1 of the rat hippocampus.

In tissue from area CA1 of the rat hippocampus prepared for electron microscopic study by thin-sectioning, asymmetric synaptic junctions were found on dendritic spines, spiny dendritic shafts, and non-spiny dendritic shafts. In freeze-fractured preparations, aggregates of large particles were found on the extracellular half of the postsynaptic membrane at each of these synaptic junctions. Particle aggregate areas were measured and particle packing densities were computed at dendritic spine synapses and dendritic shaft synapses in area CA1, and compared to similar measures of particle aggregates on dendritic spines of cerebellar Purkinje cells. All of these CA1 and cerebellar synapses are excitatory and are thought to use glutamate as a neurotransmitter. There was a tendency for the dispersion of particles within individual aggregates to be less uniform in larger aggregates in both area CA1 and cerebellar cortex. Distinct particle-free zones could be distinguished in the center of particle aggregates on large "mushroom-shaped" spines in area CA1. There was no statistically significant difference between the particle densities at CA1 dendritic spines (2848 +/- 863 particles/micron2) and CA1 dendritic shafts (2707 +/- 718 particles/micron2). However, the average density of particles at cerebellar dendritic spine synapses (3614 +/- 1081 particles/micron2) was significantly greater than at dendritic spine or shaft synapses found in area CA1. Symmetric synaptic junctions were observed on the CA1 pyramidal cell somas and dendritic shafts in thin-sectioned preparations. These synapses typically exert an inhibitory action mediated by gamma-aminobutyric acid. In freeze-fracture preparations, large varicosities were found apposed to the pyramidal somal and dendritic membranes, but there were no specializations of particle distribution on either the extracellular or the cytoplasmic half of the fractured postsynaptic membranes. This finding parallels observations from freeze-fracture preparations of other GABAergic synapses in the central nervous system.

Animals↗
PubMed1986

Somatostatin immunocytochemistry in the rabbit retina.

In previous work, radioimmunoassay was used to document the presence of somatostatin-like immunoreactive material in the rabbit retina. The present study was undertaken to determine the cellular localization of that material by light microscopic immunocytochemistry. Rabbit retinas were fixed by immersion in paraformaldehyde-lysine-periodate and reacted, either as whole retinas or as 50 microns Vibratome cross-sections, with an antiserum directed against somatostatin-14. Consistent staining of neuronal perikarya was seen only in the retinas of rabbits that had been pretreated with intravitreal injections of colchicine. Specifically stained cell bodies are present in the ganglion cell layer; the cells give rise to fibers in both the innermost and outermost sublaminas of the inner plexiform layer. In retinal whole mounts, the cells possess two or three primary dendrites with sparse branching. The dendritic fields are up to 1 mm in diameter, and adjacent dendritic fields overlap. Many cells have a thin varicose process arising from the soma or a proximal primary dendrite; these processes branch repeatedly within the retina and resemble intraretinal axons. The somatostatin-reactive cells may be associational ganglion cells or displaced amacrine cells; it is less likely that they are ganglion cells with axons projecting to the brain.

Animals↗
PubMed1985

Excitotoxin lesions do not mimic the alteration of somatostatin in Huntington's disease.

Huntington's disease is accompanied by severe neuronal death in the striatum, but despite this cell loss, there is a marked increase in the striatal concentration of somatostatin-like immunoreactivity (SLI). We attempted to examine the mechanism of this increase by using kainic or ibotenic acid to effect a unilateral lesion in the rat neostriatum. Graded doses of toxin cause a proportional decrease in the concentration of somatostatin-like immunoreactivity to a maximum of 50% of control, which is stable over an interval of 3 months. The increased somatostatin-like immunoreactivity in Huntington's disease is not mimicked by the excitotoxin lesions in rats. In addition we find that unilateral kainic acid lesions in the striatum reduce SLI in the contralateral striatum as well, although histologic evidence and assay of choline acetyltransferase activity indicate that damage is confined to the injected side. Immunocytochemistry demonstrates a loss of somatostatin-containing neurons on the lesioned side but no discernible loss in the contralateral striatum. The bilateral decrease in SLI following unilateral lesions suggests damage to a somatostatin projection to the contralateral striatum or a compensatory interaction between the two striatal nuclei.

Animals↗
PubMed1985

High-resolution 2-deoxyglucose autoradiography in quick-frozen slabs of neonatal rat olfactory bulb.

We have used rapid freezing and freeze-substitution fixation to permit electron microscopic study of [3H]2-deoxyglucose autoradiographs. The techniques minimize diffusion of label into processing fluids and, by inference, migration of label within tissue. Slabs of olfactory bulbs from 12-day-old rats were quick-frozen after one hour of exposure to physiological olfactory stimuli. In light microscopic autoradiographs at low magnification, the neuropil of individual olfactory glomeruli appeared uniformly labeled with different levels of labeling in different glomeruli. At higher magnification, glomerular neuropil labeling consisted of small unlabeled regions surrounded by label clusters, suggesting greater deoxyglucose uptake by olfactory nerve terminals as compared with their postsynaptic dendrites. Periglomerular neurons were labeled differentially. Some microglia and glia precursor cells were heavily labeled in all bulbar laminae. The ultrastructure of cells and neuropil in all bulbar laminae was well-preserved. Cell processes and organelles could be identified in both stained sections and unstained electron microscopic autoradiographs. These experiments demonstrate the feasibility of combining quick-freezing with freeze substitution, in order to extend the resolution of studies using diffusable tracers such as 2-deoxyglucose. The results suggest that this is a promising method for assessing several controversies concerning deoxyglucose incorporation and neuronal and glial metabolism.

Animals↗
PubMed1985

Immunoreactive somatostatin in the rat retina: light microscopic immunocytochemistry and chromatographic characterization.

The retinas of adult, male Long-Evans rats contain somatostatin-like immunoreactive material (SLI) as detected by radioimmunoassay. The SLI co-chromatographs with synthetic somatostatin-14 on both gel permeation chromatography and reversed phase high performance liquid chromatography; no somatostatin-28-like material or higher molecular weight forms have been detected. Immunocytochemical methods detect SLI in at least two cell populations. The more abundant stained cells are at the inner margin of the inner nuclear layer and give off processes which form a dense meshwork of fine, varicose fibers at the outer border of the inner plexiform layer, as well as processes which pass into other sublaminas of the inner plexiform layer. Varicose immunoreactive fibers run vertically or obliquely through the inner nuclear layer and bifurcate at its outer margin, giving rise to horizontally running fibers in the outer plexiform layer. These observations are consistent with rat retinal SLI being contained within amacrine cells, at least some of which are interplexiform cells. With cholchicine pretreatment, a more sparse population of stained cells is detected in the ganglion cell layer. These cells give rise to processes which enter the inner plexiform layer. It is not known if these are ganglion cells or displaced amacrine cells.

Animals↗
PubMed1985

Huntington's disease is accompanied by changes in the distribution of somatostatin-containing neuronal processes.

The distribution of somatostatin-like immunoreactivity in the caudate, putamen, globus pallidus and ventral mesencephalon of the normal human brain has been studied with immunocytochemical techniques, and compared to that seen in Huntington's disease. Within the normal striatum, sparsely distributed varicose fibers and a population of medium-sized neurons were stained. In Huntington's disease, somatostatin immunoreactive striatal neurons appear to degenerate in proportion to the loss of striatal tissue, but there is an increase in the density of immunostained varicose fibers. In contrast, the pattern and amount of fiber staining in the substantia nigra appeared virtually unchanged from that seen in the normal brain. The morphology of striatal perikarya containing somatostatin-like immunoreactivity and the patterns of fiber staining in normal and Huntington's disease pallidum and substantia nigra suggest that striatal neurons containing somatostatin-like immunoreactivity are local circuit neurons.

Adult↗