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Electrical characteristics of dendrites and dendritic spines in intracellularly stained CA3 and dentate hippocampal neurons.

Theoretical parameters of synaptic efficacy were studied in a detailed cable model of in vitro hippocampal neurons. CA3 pyramidal cells (n = 9) and dentate granule neurons (n = 6) were injected with horseradish peroxidase (HRP) after brief physiological analysis. The dendrites of these HRP-stained neurons were measured and approximated by a series of cylindrical segments. Specific electrical values of the neurons were calculated on a steady-state basis, using a cable analysis of the dendritic segments. The evaluation was expanded to include idealized dendritic spines. Average spine dimensions were determined by electron microscopic measurements. The density distribution of spines was patterned after reported Golgi measurements of similar neurons. The average electrotonic length of the CA3 apical dendrites was determined to be 0.69 length constant. These CA3 neurons were not well approximated by a single equivalent cylinder coupled to a soma (E-C model). The dentate granule cells exhibited an average electrotonic length of 1.12 length constants and could be adequately represented by the E-C model. Synaptic efficacy was estimated by the transfer of either charge or steady-state voltage from a spine input to the soma. Charge transfer varied as a function of the electrotonic distance from the soma to the input site. Voltage transfer, however, did not vary as a simple function of electrotonic distance. Voltage and charge transfer averaged less than 10% loss across the dendritic spine neck. These calculations, based on specific neuronal anatomy, predict that dendritic spines do not significantly attenuate steady-state electrical signals.

Animals↗

High-dose UV-B radiation alters human dendritic cell costimulatory activity but does not allow dendritic cells to tolerize T lymphocytes to alloantigen in vitro.

Human blood dendritic cells require UV-B radiation (290 to 320 nm) in excess of 1,000 J/m2 to inhibit their stimulation of primary T-cell responses to alloantigen by 60% to 70% or more. The effect is twofold to threefold greater in the allogeneic mixed leukocyte reaction (MLR) than in polyclonal mitogenesis using comparable numbers of dendritic cells and doses of UV-B radiation. UV-B radiation does not significantly alter dendritic cell viability at the doses administered. Dendritic cell expression of important costimulatory ligands, eg, B7/BB1 and ICAM-1/CD54, is reduced in proportion to the dose of UV-B light administered. UV-B irradiated dendritic cells nevertheless initiate stable contacts with primary alloreactive T lymphocytes that are sufficient to prime T-cell responsiveness to interleukin-2 (IL-2). Subsequent proliferation is severely abrogated without supplemental lymphokine, while T-cell alloreactivity is preserved in a secondary response, irrespective of primary exposure to UV-B irradiated dendritic cells.

Antigen-Presenting Cells↗

Two populations of splenic dendritic cells detected with M342, a new monoclonal to an intracellular antigen of interdigitating dendritic cells and some B lymphocytes.

A monoclonal has been isolated that labels an intracellular antigen in dendritic cells and some B cells. The M342 hamster immunoglobulin was selected because it stained cells in the periarterial sheaths of spleen, the deep cortex of lymph node, and the thymic medulla--the same regions in which one finds interdigitating cells, the presumptive in situ counterparts of isolated lymphoid dendritic cells. M342 labeled an antigen within granules of isolated dendritic cells, but only in cells that had been cultured for a day and not in fresh isolates. This extends recent findings that most freshly isolated spleen dendritic cells are located in the periphery of the white pulp nodule and may serve as precursors for the periarterial pool of interdigitating cells, the site for M342 staining in situ. By electron microscopic immunolabeling, the M342 antigen was found exclusively in a type of multivesicular body. M342 staining was not found in mononuclear phagocytes from blood and peritoneal cavity. Peritoneal B cells expressed M342+ granules, and upon appropriate stimulation splenic B cells developed reactive granules as well. We conclude that M342 is a strong marker for interdigitating cells. Its existence reveals intracellular specializations in the vacuolar system of antigen-presenting cells including subsets of dendritic cells.

Animals↗

Calcium accumulation by dendritic mitochondria declines along the apical dendrites of pyramidal neurons in area CA1 of guinea pig hippocampal slices.

An electron microscopic histochemical study was performed in stratum radiatum of area CA1 of guinea pig hippocampal slices in order to determine the spatial distribution of a dendritic mitochondrial subpopulation which accumulated calcium during in vitro incubation. A distribution gradient was found along the course of apical dendrites exhibiting the highest density values at the base of the dendrites and decaying to baseline values at about 50 microns distal from the cell body layer. The pronounced calcium accumulation by mitochondria in the proximal apical dendrites was markedly but not completely reduced by blocking L-type Ca-channels. These results (i) support the observation of a clustered distribution of L-type Ca-channels at the base of apical dendrites, (ii) designate these voltage dependent Ca2+ channels as one of the possible routes for calcium influx caused by hypoxia/ischemia induced during slice preparation, and (iii) emphasize the role of mitochondrial calcium sequestering under ischemic/hypoxic conditions.

Animals↗

Dendritic growth cones and recurrent basal dendrites are typical features of newly generated dentate granule cells in the adult hippocampus.

Granule cells in the hippocampal dentate gyrus are generated throughout adulthood of mammals, and recent studies indicate that they are incorporated into neural circuitry and mature into functional neurons. To determine whether newly generated granule cells form dendritic growth cones during this process of synaptogenesis, we used the immunocytochemical method to localize doublecortin, a protein associated with microtubules in newborn neurons. Here we show that both dendritic growth cones and recurrent basal dendrites are common features of newly generated granule cells. This study is the first to show dendritic growth cones in the dentate gyrus of the adult nervous system and suggests that dendrites in adult brains grow in a similar way as those found in immature brains.

Animals↗

Activation of N-methyl-D-aspartate receptor induces a shift of drebrin distribution: disappearance from dendritic spines and appearance in dendritic shafts.

Drebrin is a major actin-filament-binding protein localized in mature dendritic spines. A recent in vivo immunoelectron microscopic study suggests that drebrin content at each dendritic spine is regulated by some unknown mechanisms. In the present in vitro study, we examined whether glutamate stimulation alters drebrin content in dendritic spines. Glutamate stimulation induced disappearance of drebrin immunostaining from dendritic spines but led to appearance of drebrin immunostaining in dendritic shafts and somata. The glutamate-induced shift of drebrin immunostaining was blocked by an NMDA receptor antagonist. Immunoblot analyses showed that both the total and the cytosolic drebrin remained unchanged and revealed that the drebrin shift was not due to drebrin degradation. These findings indicate that NMDA receptor activation induces a shift in subcellular distribution of drebrin associated with actin filaments, and that the shift might be a molecular basis for actin reorganization accompanied with synaptic plasticity.

Actin Cytoskeleton↗

Dendritic Ca2+ channels characterized by recordings from isolated hippocampal dendritic segments.

Dendritic arbors are critical for the information processing capability of central neurons, but quantitative analysis of their membrane properties has been hampered by their geometrical complexity. Here, we have focused on an important source of Ca2+ entry in dendrites, the voltage-gated Ca2+ channels, by applying the whole-cell voltage-clamp technique to isolated dendritic segments ("dendrosomes") from rat hippocampal neurons. We found that low voltage-activated T-type Ca2+ channels provide a significantly larger fraction of the Ca2+ influx in dendrites than their counterparts in cell bodies. Surprisingly, 60%-70% of the high voltage-activated Ca2+ current in dendrosomes was N and P/Q type, and these channels were susceptible to neurotransmitter inhibition, suggesting a novel physiological role for G protein-regulated Ca2+ channel modulation in controlling dendritic excitability and Ca2+ signaling.

Animals↗

Alterations in the morphology of dendrites and dendritic spines in the nucleus accumbens and prefrontal cortex following repeated treatment with amphetamine or cocaine.

Repeated treatment with psychostimulant drugs produces changes in brain and behaviour that far outlast their initial neuropharmacological actions. The nature of persistent drug-induced neurobehavioural adaptations is of interest because they are thought to contribute to the development of dependence and addiction, and other forms of psychopathology, e.g. amphetamine psychosis. There are many reports that psychostimulants produce biochemical adaptations in brain monoamine systems, especially dopamine systems. The purpose of the present study was to determine if they might also alter the morphology of neurons in brain regions that receive monoaminergic innervation. Rats were given repeated injections of either amphetamine or cocaine, or, to control for general motor activity, allowed access to a running wheel. They were then left undisturbed for 24-25 days before their brains were processed for Golgi-Cox staining. Treatment with either amphetamine or cocaine (but not wheel running experience) increased the number of dendritic branches and the density of dendritic spines on medium spiny neurons in the shell of the nucleus accumbens, and on apical dendrites of layer V pyramidal cells in the prefrontal cortex. Cocaine also increased dendritic branching and spine density on the basilar dendrites of pyramidal cells. In addition, both drugs doubled the incidence of branched spines on medium spiny neurons. It is suggested that some of the persistent neurobehavioural consequences of repeated exposure to psychostimulant drugs may be due to their ability to reorganize patterns of synaptic connectivity in the nucleus accumbens and prefrontal cortex.

Amphetamine↗

iC3b arrests monocytic cell differentiation into CD1c-expressing dendritic cell precursors: a mechanism for transiently decreased dendritic cells in vivo after human skin injury by ultraviolet B.

Our previous data indicated that C3, its bioactive product iC3b, and the iC3b ligand CD11b are critical for ultraviolet-induced immunosuppression. We thus hypothesized that iC3b is an important skin-based factor regulating CD11b+ monocytic cell function in the acute post-ultraviolet period. Although monocytic cell migration peaked at 1-3 d after ultraviolet exposure of skin, dermal CD1c dendritic cells underwent a rapid and prolonged depletion that did not recover until day 7. Because ultraviolet-induced iC3b deposits are reciprocally maximal on day 3, but fade by day 7, we next hypothesized that iC3b can be responsible for the delay in differentiation into dendritic cells of monocytic cells migrating into ultraviolet-exposed skin. Analysis of dermal cells derived from keratome biopsies suggested that iC3b exposure could inhibit the development of CD1c+ dermal cells. To model newly immigrating blood monocytes entering ultraviolet-exposed, iC3b-containing dermis, purified monocytes from human blood were induced with granulocyte-macrophage colony stimulating factor to generate a population of dendritic cell precursors expressing CD1c. Incubation with iC3b markedly inhibited the appearance of CD1c+ cells (p<0.05) and induced CD1c-CD14+ cells. This inhibition was reversed by coincubation with an anti-CD11b antibody that blocks the iC3b binding site. Other functions associated with dendritic cell maturation were also inhibited by iC3b, such as interleukin-12p70 production as well as CD80 and CD40 expression. Restimulation of monocytes for DC maturation revealed that iC3b induced a temporary inhibition of DC differentiation. Thus, a human skin response in which iC3b is transiently (3-7 d) generated in dermis, such as ultraviolet, can arrest monocytic skin-infiltrating cells from undergoing dendritic cell precursor differentiation.

Animals↗

Altered expression and endocytic function of CD205 in human dendritic cells, and detection of a CD205-DCL-1 fusion protein upon dendritic cell maturation.

CD205 (DEC-205) is a member of the macrophage mannose receptor family of C-type lectins. These molecules are known to mediate a wide variety of biological functions including the capture and internalization of ligands for subsequent processing and presentation by dendritic cells. Although its ligands await identification, the endocytic properties of CD205 make it an ideal target for those wishing to design vaccines and targeted immunotherapies. We present a detailed analysis of CD205 expression, distribution and endocytosis in human monocyte-derived dendritic cells undergoing lipopolysaccharide-induced maturation. Unlike other members of the macrophage mannose receptor family, CD205 was up-regulated upon dendritic cell maturation. This increase was a result of de novo synthesis as well as a redistribution of molecules from endocytic compartments to the cell surface. Furthermore, the endocytic capacity of CD205 was abrogated and small amounts of the recently identified CD205-DCL-1 fusion protein were detected in mature DC. Our results suggest that CD205 has two distinct functions -- one as an endocytic receptor on immature dendritic cells and a second as a non-endocytic molecule on mature dendritic cells -- and further highlight its potential as an immuno-modulatory target for vaccine and immunotherapy development.

Antigens, CD↗

Dendritic subunits determined by dendritic morphology.

A theoretical framework is presented in which arbitrarily branched dendritic structures with nonhomogeneous membrane properties and nonuniform geometry can be transformed into an equivalent unbranched structure (equivalent cable). Rall's equivalent cylinder is seen to be one part of the equivalent cable in the special case of dendrites satisfying the Rall criteria. The relation between the branched dendrite and its equivalent unbranched representation is uniquely defined by an invertible mapping that connects configurations of inputs on the branched structure with those on the unbranched structure, and conversely. This mapping provides a new definition of dendritic subunit and provides a mechanism for characterizing local and nonlocal signal processing within dendritic structures.

Animals↗

Cellular mechanisms of dendrite pruning in Drosophila: insights from in vivo time-lapse of remodeling dendritic arborizing sensory neurons.

Regressive events that refine exuberant or inaccurate connections are critical in neuronal development. We used multi-photon, time-lapse imaging to examine how dendrites of Drosophila dendritic arborizing (da) sensory neurons are eliminated during early metamorphosis, and how intrinsic and extrinsic cellular mechanisms control this deconstruction. Removal of the larval dendritic arbor involves two mechanisms: local degeneration and branch retraction. In local degeneration, major branch severing events entail focal disruption of the microtubule cytoskeleton, followed by thinning of the disrupted region, severing and fragmentation. Retraction was observed at distal tips of branches and in proximal stumps after severing events. The pruning program of da neuron dendrites is steroid induced; cell-autonomous dominant-negative inhibition of steroid action blocks local degeneration, although retraction events still occur. Our data suggest that steroid-induced changes in the epidermis may contribute to dendritic retraction. Finally, we find that phagocytic blood cells not only engulf neuronal debris but also attack and sever intact branches that show signs of destabilization.

Abdomen↗

Bottlebrush dendritic endings and large dendritic fields: motion-detecting neurons in the mammalian tectum.

The widefield vertical neurons of the lower stratum griseum superficiale (SGS3) and upper stratum opticum (SO) of the superior colliculus provide an extrageniculate route for visual information to reach the pulvinar. Previous physiological studies indicate that SGS3/SO neurons have large receptive fields and respond to small moving stimuli. We sought to better characterize the dendritic morphology of SGS3/SO neurons with intracellular filling in slice preparations of the ground squirrel superior colliculus. We found that dendrites of widefield vertical cells end in monostratified arrays of spiny terminal specializations called "bottlebrush" dendritic endings. Two major subtypes of neurons are described. Type I neurons have somata restricted to the SGS3 and bottlebrush endings in the most superficial sublayer of the SGS. Type II neurons are found at the base of the SGS and in the upper SO, and have bottlebrush endings arrayed within the middle sublayers of the SGS. Bottlebrush endings may sample and integrate laminated afferents to the superior colliculus, and cellular subtypes may underlie multiple information streams within the tectopulvinar pathway. A similar dendritic morphology and projection pattern can be found in cells of the avian optic tectum that project upon the nucleus rotundus, a thalamic nucleus homologous to the mammalian caudal/inferior pulvinar. Because motion processing is a dominant feature of the avian tectorotundal pathway, the current results suggest that both dendritic morphology and motion processing are conserved features of widefield vertical cells in the tectopulvinar pathway of vertebrates.

Animals↗

Maturational steps of bone marrow-derived dendritic murine epidermal cells. Phenotypic and functional studies on Langerhans cells and Thy-1+ dendritic epidermal cells in the perinatal period.

The adult murine epidermis harbors two separate CD45+ bone marrow (BM)-derived dendritic cell systems, i.e., Ia+, ADPase+, Thy-1-, CD3- Langerhans cells (LC) and Ia-, ADPase-, Thy-1+, CD3+ dendritic epidermal T cells (DETC). To clarify whether the maturation of these cells from their ill-defined precursors is already accomplished before their entry into the epidermis or, alternatively, whether a specific epidermal milieu is required for the expression of their antigenic determinants, we studied the ontogeny of CD45+ epidermal cells (EC). In the fetal life, there exists a considerable number of CD45+, Ia-, ADPase+ dendritic epidermal cells. When cultured, these cells become Ia+ and, in parallel, acquire the potential of stimulating allogeneic T cell proliferation. These results imply that CD45+, Ia-, ADPase+ fetal dendritic epidermal cells are immature LC precursors and suggest that the epidermis plays a decisive role in LC maturation. The day 17 fetal epidermis also contains a small population of CD45+, Thy-1+, ADPase-, CD3- round cells. Over the course of 2 to 3 wk, they are slowly replaced by an ever increasing number of round and, finally, dendritic CD45+, Thy-1+, CD3+ EC. Thus, CD45+, Thy-1+, ADPase-, CD3- fetal EC may either be DETC precursors or, alternatively, may represent a distinctive cell system of unknown maturation potential. According to this latter theory, these cells would be eventually outnumbered by newly immigrating CD45+, Thy-1+, CD3+ T cells--the actual DETC.

Animals↗

Dendro axonic neurotransmission. II. Morphological sites for the synthesis, binding and release of neurotransmitters in dopaminergic dendrites in the substantia nigra and cholinergic dendrites in the neostriatum.

Morphological evidence is presented indicating sites of synthesis, storage, and release of neurotransmitters in dendrites of dopaminergic cells of the substantia nigra and cholinergic cells of the neostriatum. Smooth endoplasmic reticulum can be identified in dopaminergic neurons touching the dendritic surface. The false transmitter for dopamine, 5-hydroxydopamine (5-OHDA), is localized to smooth endoplasmic reticulum or large vesicular structures which approach the dendritic surface. The dopamine synthesizing enzyme, tyrosine hydroxylase (TH), is localized to microtubules and smooth endoplasmic reticulum which approach the postsynaptic membrane. In the neostriatum, dopaminergic nerve endings make asymmetrical axospinous contacts. The postsynaptic spines often contain a few 'vesicles' near the postsynaptic thickenings. The surface and subsurface structures stain preferentially for choline acetyltransferase (CAT), the synthesizing enzyme for acetylcholine. It is hypothesized that neurotransmitters are released from dendrites as a general phenomenon in the CNS and that they can act upon axonal endings.

Acetylcholine↗

The dendritic varicosity: a mechanism for electrically isolating the dendrites of cat retinal amacrine cells?

Amacrine dendritic varicosities from cat retina were reconstructed using serial electron micrographs. Each varicosity contained a synaptic input and a synaptic output, suggesting that they may function as isolated local circuits. A passive steady state electrical model demonstrated that for a given conductance change the varicose dendrite maximizes the local membrane potential and minimizes th distance membrane potential change as compared to other possible dendritic shapes. We, therefore, suggest that the function of the varicosities on amacrine cell dendrites might be to electrically isolate these local input-output circuits.

Animals↗

Dendritic cells freshly isolated from human blood express CD4 and mature into typical immunostimulatory dendritic cells after culture in monocyte-conditioned medium.

A procedure has been developed to isolate dendritic cells to a high degree of purity from fresh blood. Prior enrichment methods have relied upon an initial 1-2-d culture period. Purified fresh isolates lack the characteristic morphology, phenotype, and immunostimulatory function of dendritic cells. The purified cells have the appearance of medium sized lymphocytes and express substantial levels of CD4, but lack the T cell molecules CD3, CD8, and T cell receptor. When placed in culture, the cells mature in a manner resembling the previously described, cytokine-dependent maturation of epidermal dendritic cells (Langerhans cells). The cells enlarge and exhibit many cell processes, express much higher levels of major histocompatibility complex class II and a panel of accessory molecules for T cell activation, and become potent stimulators of the mixed leukocyte reaction. Among the many changes during this maturation process are a fall in CD4 and the appearance of high levels of B7/BB1, the costimulator for enhanced interleukin 2 production in T cells. These changes are not associated with cell proliferation, but are dependent upon the addition of monocyte-conditioned medium. We suggest that the freshly isolated CD4-positive blood dendritic cells are recent migrants from the bone marrow, and that subsequent maturation of the lineage occurs in tissues in situ upon appropriate exposure to cytokines.

Antigen-Presenting Cells↗

The role of graft-derived dendritic leukocytes in the rejection of vascularized organ allografts. Recent findings on the migration and function of dendritic leukocytes after transplantation.

Dendritic cells isolated from lymphoid tissues are potent stimulators of primary allogeneic T-cell responses in vitro and in vivo. Similar major histocompatibility complex class II-bearing dendritic-shaped leukocytes are contained within transplanted organs and these are thought to be important passenger leukocytes that trigger rejection. Recent findings on the migration, phenotype, and function of cardiac dendritic leukocytes (DLs) are reviewed. After transplantation donor DLs migrate rapidly from mouse cardiac allografts into the recipients's spleens. Within the spleens donor DLs associate with recipient CD4+ T cells. Isolated cardiac DLs, like lymphoid dendritic cells, are potent stimulators of T-cell proliferation in vitro. This suggests that DLs function as passenger leukocytes by migrating from grafts into the lymphoid tissues of the recipient and that sensitization to vascularized organ allografts may occur centrally within lymphoid tissues rather than peripherally in the graft itself.

Animals↗