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BDCA-2, BDCA-3, and BDCA-4: three markers for distinct subsets of dendritic cells in human peripheral blood.

We have generated a panel of mAbs that identify three presumably novel human dendritic cell Ags: BDCA-2, BDCA-3, and BDCA-4. In blood, BDCA-2 and BDCA-4 are expressed on CD11c(-) CD123(bright) plasmacytoid dendritic cells, whereas BDCA-3 is expressed on small population of CD11c(+) CD123(-) dendritic cells. All three Ags are not detectable on a third blood dendritic cell population, which is CD1c(+) CD11c(bright) CD123(dim), or on any other cells in blood. BDCA-4 is also expressed on monocyte-derived and CD34(+) cell-derived dendritic cells. Expression of all three Ags dramatically changes once blood dendritic cells undergo in vitro maturation. BDCA-2 is completely down-regulated on plasmacytoid CD11c(-) CD123(bright) dendritic cells, expression of BDCA-3 is up-regulated on both plasmacytoid CD11c(-) CD123(bright) dendritic cells and CD1c(+) CD11c(bright) CD123(dim) dendritic cells, and expression of BDCA-4 is up-regulated on CD1c(+) CD11c(bright) CD123(dim) dendritic cells. BDCA-2 is rapidly internalized at 37 degrees C after mAb labeling. The three presumably novel Ags serve as specific markers for the respective subpopulations of blood dendritic cells in fresh blood and will be of great value for their further analysis and to evaluate their therapeutic potential.

Animals↗

Production of IL-12 by human monocyte-derived dendritic cells is optimal when the stimulus is given at the onset of maturation, and is further enhanced by IL-4.

Dendritic cells produce IL-12 both in response to microbial stimuli and to T cells, and can thus skew T cell reactivity toward a Th1 pattern. We investigated the capacity of dendritic cells to elaborate IL-12 with special regard to their state of maturation, different maturation stimuli, and its regulation by Th1/Th2-influencing cytokines. Monocyte-derived dendritic cells were generated with GM-CSF and IL-4 for 7 days, followed by another 3 days +/- monocyte-conditioned media, yielding mature (CD83(+)/dendritic cell-lysosome-associated membrane glycoprotein(+)) and immature (CD83(-)/dendritic cell-lysosome-associated membrane glycoprotein(-)) dendritic cells. These dendritic cells were stimulated for another 48 h, and IL-12 p70 was measured by ELISA. We found the following: 1) Immature dendritic cells stimulated with CD154/CD40 ligand or bacteria (both of which concurrently also induced maturation) secreted always more IL-12 than already mature dendritic cells. Mature CD154-stimulated dendritic cells still made significant levels (up to 4 ng/ml). 2) Terminally mature skin-derived dendritic cells did not make any IL-12 in response to these stimuli. 3) Appropriate maturation stimuli are required for IL-12 production: CD40 ligation and bacteria are sufficient; monocyte-conditioned media are not. 4) Unexpectedly, IL-4 markedly increased the amount of IL-12 produced by both immature and mature dendritic cells, when present during stimulation. 5) IL-10 inhibited the production of IL-12. Our results, employing a cell culture system that is now being widely used in immunotherapy, extend prior data that IL-12 is produced most abundantly by dendritic cells that are beginning to respond to maturation stimuli. Surprisingly, IL-12 is only elicited by select maturation stimuli, but can be markedly enhanced by the addition of the Th2 cytokine, IL-4.

Adjuvants, Immunologic↗

CD1a dendritic cells predominate in transitional cell carcinoma of bladder and kidney but are minimally activated.

PURPOSE: In this study, we employed similar techniques to detail dendritic cell subsets within bladder transitional cell carcinoma and kidney transitional cell carcinoma. MATERIALS AND METHODS: To identify both the CD1a+ and CD1a- antigen-expressing dendritic cell populations we employed a double labeling technique to identify non-lineage-expressing leukocytes similar to that employed to isolate blood dendritic cells. RESULTS: Dendritic cells were found in significant numbers within both bladder and kidney derived transitional cell carcinoma. Almost all the dendritic cells among the tumor cells belonged to the CD1a+ subset of epithelial dendritic cells. Similar numbers of dendritic cells were observed in the lamina propria adjacent to the tumor. These dendritic cells belonged predominantly to the CD1a- subset. These differences appear to reflect the different dendritic cell phenotypes reported for the epidermis and dermis. CONCLUSIONS: The number of dendritic cells increased as the grade of the tumor increased, reflecting an overall higher leukocyte density in higher grade tumors. However, a possible trend for less dendritic cell activation in higher grade cancers was noted, raising the intriguing possibility that this might be a relevant prognostic factor, to be confirmed in a larger study.

Antigens, CD1↗

Dendritic spines lost during glutamate receptor activation reemerge at original sites of synaptic contact.

During cerebral ischemia, neurons undergo rapid alterations in dendritic structure consisting of focal swelling and spine loss. We used time-lapse microscopy to determine the fate of dendritic spines that disappeared after brief, sublethal hypoxic or excitotoxic exposures. Dendrite and spine morphology were assessed in cultured cortical neurons expressing yellow fluorescent protein or labeled with the fluorescent membrane tracer, DiI. Neurons exposed to NMDA, kainate, or oxygen-glucose deprivation underwent segmental dendritic beading and loss of approximately one-half of dendritic spines. Most spine loss was observed in regions of local dendritic swelling. Despite widespread loss, spines recovered within 2 hr after termination of agonist exposure or oxygen-glucose deprivation and remained stable over the subsequent 24 hr. Recovery was slower after NMDA than AMPA/kainate receptor activation. Time-lapse fluorescence imaging showed that the vast majority of spines reemerged in the same location from which they disappeared. In addition to spine recovery, elaboration of dendritic filopodia was observed in new locations along the dendritic shaft after dendrite recovery. Spine recovery did not depend on actin polymerization because it was not blocked by application of latrunculin-A, which eliminated filamentous actin staining in spines and blocked spine motility. Throughout spine loss and recovery, presynaptic and postsynaptic elements remained in physical proximity. These results suggest that elimination of dendritic spines is not necessarily associated with loss of synaptic contacts. Rapid reestablishment of dendritic spine synapses in surviving neurons may be a substrate for functional recovery after transient cerebral ischemia.

Animals↗

Postnatal differentiation of "presynaptic dendrites" in the lateral geniculate nucleus of the rhesus monkey.

The most characteristic synaptic arrangement in the LGN is the triadic unit, in which a retinal terminal is presynaptic to a principal cell (P cell) and to a Golgi interneuron (I cell) dendrite, which contains synaptic vesicles and is in turn presynaptic to the same P-cell element. The ontogenetic differentiation of these "presynaptic dendrites" was studied in monkey LGN by standard and quantitative electron microscopy. The dendrites and axonal arborization of I cells are well developed in the newborn monkey. Scattered synaptic vesicles are present in the dendrites, but these profiles exhibit only postsynaptic sites. The dendrodendritic synapse of the triadic arrangement is missing, although contacts between P-cell and I-cell dendrites can be observed. Conversely, the I-cell axons in the newborn establish numerous synapses with dendrites and perikarya of P cells. At about 2 weeks of age, presynaptic sites appear in the I-cell dendrites, resulting in the formation of synaptic triads. Parallel to the development of "axonal" properties in the dendrites, the number of true I-cell axonal profiles decreases sharply. These transformations become progressively more frequent with age, and beyond 8 weeks the LGN ultraarchitectonics approaches that of the mature animal. The percentage of the surface of interneuron processes occupied by presynaptic sites is similar at all developmental stages at about the 2.6% level. The relative contribution of presynaptic dendrites and of axons changes, however, so that the actual length of contacts in the mature monkey is only one-half that of the newborn for the axonal sites, and over 50 times longer for the dendritic sites. The correlation of these findings with electrophysiologic and behavioral references suggests that some but not all axonal functions may be taken over by the dendrites, that the possible inhibitory phasing of P-cell discharge would appear only after the second postnatal week, and that the triadic arrangement may not be indispensable for brightness or total luminous flux discrimination but could be required for more complex forms of visually guided behavior.

Animals↗

Local and target-derived brain-derived neurotrophic factor exert opposing effects on the dendritic arborization of retinal ganglion cells in vivo.

The dendritic and axonal arbors of developing retinal ganglion cells (RGCs) are exposed to two sources of BDNF: RGC dendrites are exposed to BDNF locally within the retina, and RGC axons are exposed to BDNF at the target, the optic tectum. Our previous studies demonstrated that increasing tectal BDNF levels promotes RGC axon terminal arborization, whereas increasing retinal BDNF levels inhibits RGC dendritic arborization. These results suggested that differential neurotrophic action at the axon versus dendrite might be responsible for the opposing effects of BDNF on RGC axonal versus dendritic arborization. To explore this possibility, we examined the effects of altering BDNF levels at the optic tectum on the elaboration of RGC dendritic arbors in the retina. Increasing tectal BDNF levels resulted in a significant increase in dendritic branching, whereas neutralizing endogenous tectal BDNF with function-blocking antibodies significantly decreased dendritic arbor complexity. Thus, RGC dendritic arbors react in opposing manners to retinal- versus tectal-derived BDNF. Alterations in retinal BDNF levels, however, did not affect axon terminal arborization. Thus, RGC dendritic arborization is controlled in a complementary manner by both local and target-derived sources of BDNF, whereas axon arborization is modulated solely by neurotrophic interactions at the target. Together, our results indicate that developing RGCs modulate dendritic arborization by integrating signals from discrete sources of BDNF in the eye and brain. Differential integration of spatially discrete neurotrophin signals within a single neuron may therefore finely tune afferent and efferent neuronal connectivity.

Animals↗

Human monocyte-derived dendritic cells pulsed with wild-type p53 protein efficiently induce CTLs against p53 overexpressing human cancer cells.

PURPOSE: Dendritic cells are the most potent antigen-presenting cells for initiating cellular immune responses. Dendritic cells are attractive immunoregulatory cells for cancer immunotherapy, and their efficacy has been investigated in clinical trials. The tumor suppressor gene p53 is pivotal in the regulation of apoptosis, and p53-based immunization is an attractive approach to cancer immunotherapy because of the accumulation of p53 protein in malignant but not in normal cells. It has been shown that dendritic cells transduced with an adenoviral wild-type p53 (wt-p53) construct mediate the antitumor immune responses against p53-overexpressing tumor cells. We examined whether monocyte-derived human dendritic cells pulsed with the purified full-length wt-p53 protein were also capable of inducing the specific antitumor responses against p53-overexpressing tumors in vitro. EXPERIMENTAL DESIGN: Immature dendritic cells generated in the presence of interleukin-4 and granulocyte/macrophage colony-stimulating factor from monocytes of HLA-A2- or HLA-A24-positive healthy individuals were pulsed with the purified p53 protein. Uptake of p53 protein by human dendritic cells was assessed by Western blotting and immunohistochemical staining using anti-p53 antibody. Induction of p53-specific CTL response was also evaluated by the cytotoxic assay against p53-overexpressing human tumor cells. RESULTS: Both Western blot and immunohistochemical analysis showed the accumulation of p53 protein in human immature dendritic cells. T cells obtained from HLA-A2- or HLA-A24-positive healthy donors were stimulated twice with p53 protein-pulsed dendritic cells and then applied to the cytotoxicity assay against p53-overexpressing target cells. The CTL activity was specific for p53-overexpressing tumor cells and MHC class I restricted. Moreover, the CTL activity generated by p53 protein-pulsed dendritic cells was nearly identical with that induced by adenoviral wt-p53-transduced dendritic cells. CONCLUSIONS: Our results indicate that monocyte-derived human dendritic cells pulsed with the wt-p53 protein could induce the specific antitumor effect against p53-overexpressing tumors and that this in vitro model offers a new and more simple approach to the development of p53-based immunotherapy.

Antigens, CD↗

Altered pyramidal cell dendritic development in the motor cortex of lead intoxicated neonatal rats. A Golgi study.

Neonatal Long-Evans rat pups were given the standard dose, 600 mg of lead acetate per kg of body weight every 24 hours beginning one day after birth until a cumulative dose of 2400 mg/kg (4 doses) had been administered via stomach intubation. Blood lead levels in lead treated rats averaged 526.35 micrograms/dl at 10 days of age. Blood lead values in age-matched controls averaged 0.079 microgram/dl. The body weights of the lead treated rats were not significantly different than control rats at 30 days of age. The brain weights were significantly greater than those of control rats at 30 days. Camera lucida drawings of pyramidal cells from motor cortex of control rats contained significantly (54%) more secondary and tertiary branches extending laterally from the primary apical dendrite than pyramidal cells of lead treated rats. The dendritic branches were numbered according to their branching point away from the soma. There was a significant reduction in the number of 4, 5, 6, and 7th order branches extending from the apical dendrite and 3 and 4th order branches extending from the basal dendrites in the leaded-rats. The mean dendritic length was reduced by 19% in basal dendrites and by 28% in apical dendrites. Measurements of dendritic material by the Scholl method revealed 17% reduction in the basal dendrites and a 36% reduction in the apical dendritic material in treated rats. These results suggest that neonatal lead exposure alters the dendritic development of pyramidal cells of rat motor cortex.

Animals↗

Preferential localization of polyribosomes under the base of dendritic spines in granule cells of the dentate gyrus.

Electron microscopic studies of the dentate gyrus of the rat have revealed an apparent association between polyribosomes and dendritic spines. The present study was designed to elucidate the nature of this association. Our qualitative observations revealed that polyribosomes appeared primarily in two locations within the dendrite: (1) beneath the base of identified spines just subjacent to the intersection of the spine neck with the main dendritic shaft and (2) beneath mounds in the dendritic membrane which had the appearance of the base of a spine which extended out of the plane of section. To quantitatively define the nature of the apparent association, we attempted to determine (1) the proportion of spines with associated polyribosomes and (2) the proportion of the polyribosomes within dendrites which are associated with spine bases. Evaluation of profiles which were identifiable as spine neck-dendritic shaft intersections in a single section revealed that an average of 12.2% had associated polyribosomes. A serial section analysis revealed a somewhat higher incidence, however. Of a collection of 34 through-sectioned spines, 29% had polyribosomes which were revealed in one or more of the sections comprising the series. To evaluate what proportion of polyribosomes within the dendrite was associated with spines, we evaluated a series of photographs covering approximately 1250 micrometer2 of the dentate molecular layer from five animals, identifying all polyribosomes within dendrites and scoring their location as being (1) under spines, (2) under mounds, or (3) other. An average of 9.6% of the polyribosomes were found under processes identifiable as spine neck-dendritic shaft intersections, while 71.4% of the polyribosomes were found under mounds. Only 19% were not obviously associated with spines or mounds. Spine bases and mounds comprise only 3 of 35%, respectively, of the outline of dendritic profiles, however, indicating that the high incidence of polyribosomes under these elements cannot be accounted for by chance. To attempt to determine whether the mounds represent the base of dendritic spines, 68 mounds in 21 dentritic profiles were selected from the middle of the series of 20 serial sections. Ninetine of these mounds (28%) were continuous with an identified spine, and an additional 31% were continuous with thin processes of the size and appearance of spine necks. Thus, most of the mounds probably do represent the base of spines which extend out of the plane of a single section.

Animals↗

[Metric examinations on the development of apical dendrites of cortical pyramidal cells during the postnatal time in the rat].

1. It is explained the postnatal evolution of some selected features of Golgi-impregnated layer V pyramidal neurons in the rat sensorimotor cortex from the 1st to the 90th day after birth (13 stages) quantitatively. Especially it is reported about the development of the length and the quantity of apical main dendrites and of the apical lateral branches from the 1st to the 3rd order as well as the branching behaviour of the dendrites (free ends of dendrites, branching points, ramification index). 2. The evolution of mean length of apical dendrites is represented by a sigmoidal polygon trace - like the classic growing curve. 3. During the 3rd month of the postnatal life the lateral branches of the apical dendrites show a distinct growing increase relating to the mean values of the total length of dendrites, their quantity and the branching behaviour. This intensification of growth is in contrast to the growing stagnation of the apical main dendrites in this time. 4. From this point of view it is deduced that relating to the maturation of pyramidal cells - contrary to the papers of other writers and contrary to a previous private conception - albino rats in the age of 3 months do not are differentiated cortical completely. 5. The several marks of pyramidal dendrites do not form themselves in the same time but successively. The maturing process of new dendrites or dendrite branches is the base of significant increase of the length. 6. It seems, that the formation of dendritic "tree" takes place in growing batches. In this context there are discussed the correlations between the morphometric conceivable changes in the cortical pyramids and the accession of functions in the animal.

Animals↗

Expansion of immunostimulatory dendritic cells among the myeloid progeny of human CD34+ bone marrow precursors cultured with c-kit ligand, granulocyte-macrophage colony-stimulating factor, and TNF-alpha.

Human CD34+ bone marrow progenitors cultured in the presence of granulocyte-macrophage CSF (GM-CSF) develop along a myeloid pathway, and the addition of exogenous TNF-alpha leads to the differentiation of dendritic cells among the myeloid progeny. These bone marrow CD34+ -derived dendritic cell that develop during 2-wk culture have the same morphologic, phenotypic, and functional properties that distinguish mature dendritic cells in blood. c-kit ligand does not directly influence dendritic cell differentiation per se, but rather increases the total cell number in synergistic combination with GM-CSF and TNF-alpha. This degree of expansion translates into an effective yield of approximately 1.7 x 10(6) mature dendritic cells per single ml of normal adult human bone marrow, compared with approximately 10(6) dendritic cells usually obtained from 450 to 500 ml of peripheral blood. In addition to dendritic cells that constitute approximately 10 to 15% of the total myeloid progeny, the cultures contain monocytes/macrophages and intermediate granulocytic precursors. Monocytes/macrophages and dendritic cells together comprise all of the class II MHC-positive progeny. Sorted cells bearing the CD14+ HLA-DR+ phenotype of mature monocytes are at least 1.5 to 2 logs less active than CD14- HLA-DR+ dendritic cells as stimulators in the allogeneic MLR, even though both CD14+ and CD14- subpopulations share expression of several costimulatory ligands. The synergistic combination of c-kit ligand, GM-CSF, and TNF-alpha therefore expands substantial numbers of immunostimulatory CD14- HLA-DR+ dendritic cells from defined CD34+ progenitors in human bone marrow. This should facilitate the use of dendritic cells in the manipulation of T cell-mediated immune responses.

Antigens, CD↗

Receptive fields and dendritic structure of directionally selective retinal ganglion cells.

We studied the relationship between the receptive fields of directionally selective retinal ganglion cells and the dendritic arbors of the same cells. The cells were recorded from extracellularly under visual control and then injected with Lucifer yellow. The arbor of Lucifer-filled dendrites could then be directly compared with the properties of the receptive field. A large population of on-off directionally selective cells was injected and drawn. The directionally selective ganglion cells had bistratified receptive fields similar to those previously described by others in the central retina. In the periphery, the dendritic fields became larger, rounder, and sparser than centrally. The diameters of the dendrites were measured in living or lightly fixed retinas; they were found to be somewhat larger than previously estimated by electron microscopy. The local structure of the dendritic arbor bore no obvious relation to the directional properties of the cell. The receptive fields of most cells were centered symmetrically around their dendritic fields. For about 10% of the cells, however, the receptive field was displaced. The displacement was always toward the preferred direction, relative to the dendritic field. The meaning of these shifts is not clear. In both cases, the diameter of the receptive field exceeded the diameter of the dendritic field only slightly; in our sample, the diameters of the receptive fields averaged 6% larger than the dendritic fields. This means that the neurons afferent to the directionally selective ganglion cells must either have narrow dendritic fields or, if they are wide spreading, have dendrites that do not conduct effectively along their length. It also means that the observed spread of neurobiotin between DS ganglion cells (Vaney, 1991) must be due to a very few gap junctions, or to some mechanism other than a gap junction.

Animals↗

Contributions of voltage-gated Ca2+ channels in the proximal versus distal dendrites to synaptic integration in prefrontal cortical neurons.

The electrogenesis of synaptically activated dendritic Ca2+-mediated potentials, which may contribute to synaptic signal integration in pyramidal cells, was examined in rat layers V-VI prefrontal cortical (PFC) neurons in vitro. Intrasomatically recorded suprathreshold synaptic responses evoked by stimulation of the distal dendrites were attenuated by focal Cd2+ application to the proximal apical dendritic stem (100-200 micron from soma), but not to the apical dendritic tuft (>500 micron from soma). With use of intracellular QX-314 and Cs+ to block Na+ and K+ currents, intrasomatic recordings revealed that the Cd2+-induced attenuation of synaptic responses was attributable to the blockade of a dendritic Ca2+-mediated "hump" potential and high-threshold Ca2+ spike activated by NMDA EPSPs. The hump potential was not blocked by bath application of Ni2+ (100 microM) but was blocked by focal application of Cd2+ to the proximal but not distal apical dendrites, suggesting that it was generated by Ca2+ channels located in the proximal dendrites. Direct patch-clamp recordings made from the distal apical tuft of layers V-VI PFC neurons revealed that layers I-II synaptic stimulation or intradendritic depolarizing current pulses evoked tetrodotoxin- and QX-314-sensitive Na+ spikes. Unlike in the stem of the apical dendrite, Ca2+ spikes were not easily evoked in the distal apical tuft when Na+ channels were blocked. When triggered, the Cd2+-sensitive Ca2+ spikes in the dendritic tuft were nonregenerative and had very high activation thresholds (approximately +10 mV). These results suggested that the high voltage-activated Ca2+ potentials that amplify distal EPSPs are primarily generated in the proximal stem of the apical dendrite and not within the fine dendritic branches of the apical tuft of layers V-VI PFC neurons.

Animals↗

Gap junctions between non-pyramidal cell dendrites in the rat hippocampus (CA1 and CA3 regions): a combined Golgi-electron microscopy study.

Non-pyramidal cells in the rat hippocampus were examined with a combined Golgi-electron microscopic method. The somata of non-pyramidal cells were ovoid, about 15 X 30 micron, and several smooth and/or varicose dendrites extended from them. With electron microscopy, Golgi-impregnated gold-toned non-pyramidal cells showed distinctive fine structural features. The somata displayed large nuclei and an extensive perikaryal cytoplasm. The nuclei showed extensive cytoplasmic invaginations, little heterochromatin, conspicuous nucleoli, and intranuclear rods composed of filamentous bundles. The perikaryal cytoplasm was rich in cell organelles such as well-developed cisternae of rough endoplasmic reticulum and Golgi apparatus, and numerous clusters of free ribosomes and mitochondria. Many synaptic boutons, most of which formed asymmetrical synapses, impinged upon the somata and dendrites. Gap junctions were seen on varicose dendrites of Golgi-impregnated non-pyramidal cells. These gap junctions were patch-like, about 0.1-0.6 micron in diameter, and situated in the stratum radiatum or stratum oriens of the CA1 and CA3 regions 70-230 micron from the soma. They displayed a characteristic cytoplasmic semidense material undercoating the junctional membranes. The gap junctions were usually formed between impregnated and unimpregnated varicose dendrites. Thirteen of a total of 22 gap junctions involving the impregnated dendrites were situated singly, whereas the remaining nine were on four impregnated dendrites in clusters of two or three side by side. In the latter cases, two pairs of junctions were formed between pairs of dendrites running parallel to each other, and each of the other two pairs was formed among three dendrites, appearing to make a dendritic network bridged by gap junctions. One gap junction was seen between two impregnated dendrites originating from two identified Golgi-impregnated non-pyramidal cells. These observations revealed unequivocally that non-pyramidal cells in the hippocampus form gap junctions with one another on their dendrites.

Animals↗

The dendritic architecture of the visual pretectal nuclei of the rat: a study with the Golgi-Cox method.

The dendritic architecture of the neurons of the visual pretectal nuclei in the rat was studied with the Golgi-Cox method. The olivary pretectal nucleus (PO) is characterized by distinctive neurons with a gnarled, tufted, richly branched dendritic arbor forming a dense neuropil within the nucleus. The distinct dendritic morphology of the olivary pretectal neurons enables this nucleus to be identified at all levels of the pretectum in Golgi-impregnated preparations. Rostromedially, the PO is surrounded by peripheral neurons whose dendrites wrap around the surface of the PO. The nucleus of the optic tract (NTO) contains three types of cells: (1) superficial horizontal cells whose dendrites extend out transversely; (2) large multipolar neurons whose dendrites spread out predominantly in a transverse plane, and (3) small to medium multipolar neurons with varying dendritic architecture. The posterior pretectal nucleus (PP) is composed predominantly of (1) multipolar cells with horizontally and vertically oriented dendrites extending out transverse to the optic axons; (2) piriform cells with dendrites extending dorsally toward the brachium; and (3) small multipolar neurons. The presence of superficial horizontal and large multipolar neurons in the NTO distinguishes the NTO from the PP in Golgi preparations. The horizontally oriented dendrites of many of the multipolar neurons in the PP give this nucleus an appearance distinct from that of the NTO. The differences in dendritic morphology between the visual pretectal nuclei in the rat permit identification of these nuclei at all levels within the pretectum. The boundaries of these nuclei, as determined in the Golgi-Cox preparations, correlate quite well with the boundaries defined by studying retinal projections (Scalia and Arango, '79).

Animals↗

Magnopyramidal neurons in the anterior motor speech region. Dendritic features and interhemispheric comparisons.

OBJECTIVE: To test the hypothesis that the larger somal size of layer III magnopyramidal neurons in left than in right Brodmann area 45 is associated with greater measures of dendritic arborization in the left hemisphere neurons. DESIGN: A case series involving postmortem human brain specimens was used to compare dendritic parameters of Golgi-impregnated layer III pyramidal neurons in Brodmann area 45 in the left and right hemispheres. SUBJECTS: A convenience sample consisting of 9 subjects with no known neurologic or psychiatric disorders was obtained at autopsy. MAIN OUTCOME MEASURES: Dendritic parameters of the 10 largest Golgi-impregnated layer III pyramidal neurons were measured in each hemisphere of each brain using a eutectic neuron tracing system. The measures examined were somal size, neuron depth, mean segment length, number of branch points, maximal branch order, combined dendritic diameter, total dendritic length, horizontal extent of the dendritic field, and spine density. The Golgi-impregnated slides were also compared with Nissl-stained slides from adjacent blocks to determine the laminar location of the sampled neurons. RESULTS: Although the mean somal size was greater in the left than in the right hemisphere, none of the dendritic parameters examined was larger for the left hemisphere neurons. As expected, total dendritic length was positively correlated to somal size (r = 0.43, P < 001) in the left hemisphere. However, there was no correlation between these parameters in the right hemisphere (r = 0.02, P = .76). CONCLUSIONS: The difference between left and right Brodmann area 45 magnopyramidal neurons in the correlation of somal size and dendritic length provides additional evidence of anatomical differences between these 2 populations of neurons. However, the lack of interhemispheric differences in measures of dendritic arborization suggests that additional factors contribute significantly to the marked difference in somal size between the large magnopyramidal neurons in the left and right hemispheres.

Adult↗

Synaptic input to cochlear nucleus dendrites that receive medial olivocochlear synapses.

Axons of olivocochlear neurons originate in the superior olivary complex and project to the cochlea. Along their course, medial olivocochlear axons give off branches to the cochlear nucleus. We labeled these branches with horseradish peroxidase and used electron microscopy to determine their target dendrites. Target dendrites were of two classes: "large" dendrites and "varicose" dendrites. Using serial sections, we reconstructed the dendrites and, in addition to the labeled olivocochlear input, we determined the synaptic profile of unlabeled inputs onto the dendrites. We classified the terminals on the basis of the shape and size of their synaptic vesicles. On large dendrites, the predominant type of unlabeled terminal had small round (SmRnd) vesicles. These terminals are likely to be excitatory, and some of them may originate from unlabeled medial olivocochlear branches. On varicose dendrites, the predominant type of terminal had pleomorphic vesicles. These terminals are likely to be inhibitory. They may be from descending inputs that arise in higher centers. A final type of terminal onto large dendrites exhibited signs of neuronal degeneration, possibly because the cell body of origin was damaged during the injection procedure. These terminals often had long, perforated synaptic densities and may originate from type II primary afferents. Thus, medial olivocochlear efferents and type II afferents, which both contact outer hair cells in the periphery, appear to synapse onto the same targets in the cochlear nucleus. In contrast, where examined, the target dendrites did not receive terminals with large vesicles from afferents that contact inner hair cells. Thus, target neurons appear to function in a neural circuit associated more closely with outer than with inner hair cells.

Animals↗

Expansion of the dendritic tree of motoneurons innervating neck muscles of the adult cat after permanent axotomy.

The morphologic characteristics of neck motoneurons with intact axons were compared with those of neck motoneurons that had been permanently axotomized for 11 to 17 weeks. Motoneurons were identified antidromically, intracellularly stained with horseradish peroxidase (HRP) and examined after reconstructions of their entire dendritic tree. Axotomized motoneurons differed qualitatively and quantitatively from motoneurons with intact axons. The distal branches of axotomized motoneurons exhibited two novel features: some gave rise to tangled appendages that exhibited growth cone-like specializations resembling lamellipodia and filopodia; others followed a meandering path and had unusually large diameters. These branches showed a discontinuous pattern of staining that was similar to the appearance of myelinated axons stained intra-axonally with HRP. A quantitative analysis of the dendritic trees of 13 completely reconstructed dendritic trees (five axotomized motoneurons and eight motoneurons with intact axons) showed that total dendritic surface area, total dendritic length, and total number of branches increased 38, 34, and 215%, respectively, after axotomy. These measurements were confirmed by comparing the sizes of a larger number of motoneurons (16 axotomized and 21 intact), calculated on the basis of correlations between dendritic tree size and proximal dendritic diameter. We conclude, therefore, that neck motoneurons, in contrast to other types of motoneurons, expand their dendritic trees after axotomy. It is suggested that this expansion is a consequence of two mechanisms: one involves dendritic growth, possibly leading to new synaptic connections; the other causes a conversion of some dendrites into axons.

Animals↗