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Morphology and frequency of axon terminals on the somata, proximal dendrites, and distal dendrites of dorsal neck motoneurons in the cat.

The purpose of the present study was to compare the frequency of different classes of axon terminals on selected regions of the somatodendritic surface of dorsal neck motoneurons. Single motoneurons supplying neck extensor muscles were antidromically identified and intracellularly stained with horseradish peroxidase. By using light microscopic reconstructions as a guide, axon terminals on the somata, proximal dendrites (within 250 microns of the soma), and distal dendrites (more than 540 microns from the soma) were examined at the electron microscopic level. Axon terminals were divided into several classes based on the shape, density, and distribution of their synaptic vesicles. The proportion of axon terminals belonging to each axon terminal class was similar on the somata and proximal dendrites. However, there were major shifts in the relative frequency of most classes of axon terminals on the distal dendrites. The most common classes of axon terminals on the somata and proximal dendrites contained clumps of either spherical or pleomorphic vesicles. These types of axon terminals accounted for more than 60% of the axon terminals on these regions. In contrast, only 11% of the axon terminals found on distal dendrites belonged to these types of axon terminals. The most commonly encountered axon terminal on distal dendrites contained a dense collection of uniformly distributed spherical vesicles. These types of axon terminals accounted for 40% of all terminals on the distal dendrites, but only 5-7% of the axon terminals on the somata and proximal dendrites. Total synaptic density on each of the three regions examined was similar. However, the percentage of membrane in contract with axon terminals was approximately four times smaller on distal dendrites than somata or proximal dendrites. Axon terminals (regardless of type) were usually larger on somata and proximal dendrites than distal dendrites. These results indicate that there are major differences in the types and arrangement of axon terminals on the proximal and distal regions of dorsal neck motoneurons and suggest that afferents from different sources may preferentially contact proximal or distal regions of the dendritic trees of these cells.

Animals

Serotonin-containing structures in the nucleus raphe dorsalis of the cat: an ultrastructural analysis of dendrites, presynaptic dendrites, and axon terminals.

In the nucleus raphe dorsalis of the cat, an electron microscopic immunocytochemistry method was used to identify the fine structure of serotoninergic dendritic profiles and axon terminals analyzed in serial sections. Two classes of serotoninergic dendrites were distinguished in the nucleus. The first class was constituted by conventional serotonin (5-HT) dendrites that were contacted by unlabeled axon terminals containing differing populations of synaptic vesicles. The second class consisted of serotoninergic dendrites that contained vesicles in their dendritic shafts. Such 5-HT dendrites were further subdivided into two groups according to their synaptic contacts. In some 5-HT vesicle-containing dendrites, the vesicles were densely packed in small clusters and were associated with a well-defined synaptic specialization. These dendrites were classified as serotoninergic presynaptic dendrites and established synaptic contacts with unlabeled and labeled dendrites and were contacted by unlabeled axon terminals. In other 5-HT vesicle-containing dendrites, extensive serial section examination showed that the vesicles could be observed near the membrane but were never found to be associated with any synaptic membrane specialization. Serotoninergic axon terminals that were presumed to be recurrent collaterals of 5-HT neurons were present in the nucleus. Some of them were observed in synaptic contact with dendrites or dendritic protrusions whereas others did not exhibit synaptic specializations. The existence of serotoninergic dendrodendritic synaptic contacts and axon terminals suggests direct local interactions between serotoninergic neurons within the nucleus raphe dorsalis.

Animals

Postnatal development of cat hind limb motoneurons. II: In vivo morphology of dendritic growth cones and the maturation of dendrite morphology.

The maturation of dendrite morphology was studied by light and electron microscopy in cat spinal alpha-motoneurons intracellularly labeled with horseradish peroxidase. Alpha-motoneurons supplying the triceps surae (TS) and the intrinsic foot sole (SP) muscles were investigated in kittens from birth to 44-46 days of postnatal (d.p.n.) age. At birth, a large number of dendritic branches displayed growth cones, filopodia, and fusiform processes. The growth cones were of lamellipodial and filopodial types, but intermediate forms also occurred. The growth cones shared several morphological features with the neuritic growth cones studied in vitro. It was suggested that the occurrence of different types of growth cones--even in the same dendrite--may reflect their transformation from one type to the other and the level of growth activity could be inferred from the number and form of the growth cones. About 50-70% of the terminal branches in the dendrites of newborn kittens possessed growth cones, filopodia, and/or fusiform processes. The corresponding figure for preterminal branches was 20-30%, with a clear decrease in incidence when approaching the soma. During the period under study, most of these growth-associated processes disappeared from the dendrites so that at 44-46 d.p.n. of age only about 10% of the terminal and less than 1% of the preterminal branches had growth-associated processes. Analysis of the three-dimensional distribution of dendritic branches with such processes disclosed that they were relatively more frequent in the medial, rostral, and caudal dendritic territories. It was concluded that the pattern of distribution and disappearance of growth cones, filopodia, and fusiform processes coincided with postnatal longitudinal dendritic growth and the development of the adult dendritic territories described in a preceding paper (Ulfhake et al., '88). Dendritic growth, with respect to length and caliber, also occurred in the absence of growth cones and filopodia. It is suggested that the important role of these processes may be to act as a steering device in establishing the adult distribution and synaptology of the dendrites. Comparison of TS and SP alpha-motoneuron dendrite morphology at birth and at 22-24 d.p.n. age showed that the SP neurons lagged in the maturation process. Light and electron microscopic observations indicated that postnatally direct contacts might exist between dendrites and fine blood vessels in the neuropil without any interposing glial sheath. The number of such suspected contacts diminished during the period under study, indicating that the glial ensheathment of the blood vessel takes place, in part, postnatally.

Animals

Anatomy of dendrites in motoneurons supplying the intrinsic muscles of the foot sole in the aged cat: evidence for dendritic growth and neo-synaptogenesis.

Motoneurons (MNs) supplying the intrinsic muscles of the foot sole (IFS) were studied in the aged cat (greater than 15y). Axon conduction velocity of IFS MNs was 30-40% slower in the aged than in young adult cats. IFS MNs that appeared intact during intracellular recordings and labeling with horseradish peroxidase (HRP) were subjected to anatomical investigation of their dendrites. The results were compared with corresponding data from young adult (less than 3y) cats. The average number of dendrites per IFS MN was twelve in both the aged and young adults. However, the branching was significantly more extensive in the aged cat, thus indicating that proliferation of dendritic branches may occur during the later part of life. Topological analysis revealed a significant difference in the frequency distributions of nodal vertices between young adult and aged cats. In the young adult, the dendritic branching pattern was compatible with trees generated by outgrowth from terminal segments, while in the aged there was a clear indication of collateral outgrowth of branches. The dendritic path distance and the length of terminal branches were similar in young adults and aged. The length of preterminal branches was shorter in the aged, while the combined dendritic length of a dendrite was larger compared to young adults. These data are consistent with the topological data, and add further evidence that the proliferation of branches in the aged cat may also take place from preterminal branches. Light microscopic analysis revealed the presence of "growth cone-like" extensions in the dendrites of the aged cats. Such profiles were not encountered in dendrites from young adults. Electron microscopic observations showed that these "growth cone-like" formations were not artifacts and that they were apposed by numerous axonal boutons, of which a number made synaptic contact. A distinct feature of the extensions was their rich content of mitochondria and membranous elements. It was suggested that these "growth cone-like" formations were sites at which novel synaptic connections are established, and that they may represent the initial stage of an outgrowth of new dendritic branches in the aged cat. Local dendritic branch diameter related closely to the amount of dendritic membrane area located distally in both young adults and aged. Curve fitting disclosed that this relationship was quite similar for both age groups, despite concurrent differences in combined dendritic length and branching degree.

Aging

The function of Ia+ dendritic cells and Ia- dendritic cell precursors in thymocyte mitogenesis to lectin and lectin plus interleukin 1.

The response of thymocytes to lectin is a standard tissue culture model for identifying cytokines such as IL-1 that are required for thymocyte mitogenesis. To study accessory cell requirements for these responses, it was necessary to deplete endogenous accessory cells with two techniques: anti-Ia and complement, and passage over nylon wool. Proliferation to Con A was then restored with 0.1-0.3% exogenous splenic dendritic cells, or 30-fold higher levels of peritoneal macrophages. The "costimulatory" action of IL-1, whereby responses to lectin were enhanced 3-10-fold, required the presence of dendritic cells. This effect of IL-1 could be reproduced by culturing the dendritic cells for 12 h in 1 U/ml human or murine rIL-1 alpha before addition to the thymocyte proliferation assay. The function of IL-1-treated dendritic cells was not blocked by a neutralizing anti-IL-1 antibody. The endogenous population of thymic accessory cells was partially characterized. A trace (0.1-0.3%) fraction of Ia+, Ig-, plastic nonadherent dendritic cells was visualized and enriched to a level of 1-10% by depleting CD4+,CD8+, and Ig+ lymphocytes. When this double-negative population was cultured with IL-1 and washed, the treated thymic dendritic cells were 10-fold more active as accessory cells. When the CD4-,CD8-, Ig- populations were depleted of dendritic cells with anti-Ia and complement, the subsequent addition of IL-1 had a second effect. Ia+ dendritic cells redeveloped over a 2-d interval, and they exhibited the same properties as resident dendritic cells in thymus and spleen. The majority were lysed by 33D1 anti-dendritic cell mAb and complement, lacked Fc receptors, and acted as powerful stimulators of the MLR and Con A mitogenesis. The development of dendritic cells did not occur with IL-2, -3, -4 or granulocyte/macrophage colony-stimulating factor or in nylon-nonadherent populations. The IL-1-dependent, Ia- precursor was not detectable in bone marrow. These results begin to analyze the endogenous accessory function of the thymus in culture. Dendritic cells actively stimulate thymocyte mitogenesis. The mitogenic action of IL-1 involves effects on resident Ia+ dendritic cells as well as a new population of thymic, Ia- precursors.

Animals

Demonstration of local protein synthesis within dendrites using a new cell culture system that permits the isolation of living axons and dendrites from their cell bodies.

The presence of polyribosomes within dendrites suggests a capability for local dendritic protein synthesis. However, local synthesis is difficult to evaluate because of rapid somatodendritic protein transport. The present study describes a two-surfaced culture system that allowed the separation of living axons and dendrites from their cell bodies of origin. Because this system eliminates the transport of proteins produced in the cell body, it was possible to study the extent of dendritic protein synthesis directly. Hippocampal neurons were plated on a Nucleopore polycarbonate membrane that was mounted on a thick matrix of proteins (Matrigel) fixed on a coverslip. As the neurons grew, axons and dendrites grew through the membrane into the Matrigel. To evaluate local protein synthesis within dendrites, the membrane with the cell bodies was removed, leaving a dense array of transected dendrites and axons on the coverslip with few contaminant cell bodies. Absence of cell bodies was confirmed by staining with the nuclear stain Hoechst 33258. Coverslips with isolated neurites were pulse labeled with 3H-leucine for 30 min, and fixed for autoradiography to identify sites of protein synthesis. Autoradiographic analyses revealed that isolated dendrites (immunochemically identified using antibodies against MAP2) became heavily labeled, whereas axons exhibited little if any labeling. The labeling was essentially eliminated when the neurites were pulse labeled with 3H-leucine in the presence of puromycin, whereas labeling was affected only minimally by chloramphenicol. The puromycin-sensitive incorporation of 3H-leucine in dendrites demonstrates that the polyribosomes previously described are active in protein synthesis. This system will allow a characterization of synthetic activity within isolated neurites and provide a new approach to identifying proteins that are produced within dendrites.

Animals

Dendritic arbors and dendritic excrescences of abnormally positioned neurons in area CA3c of mice carrying the mutation "hippocampal lamination defect".

BALB/cJ and BALB/cByJ mice are homozygous for the autosomal gene "hippocampal lamination defect" (provisional gene symbol: Hld) which produces an abnormality in the lamination of the pyramidal cell layer of area CA3c of the hippocampus such that early-generated neurons are superficial and late-generated neurons are deep. Other inbred strains of mice are wild-type (+/+) at the Hld locus and do not have this inversion in cell position in area CA3c. The Golgi method was used to analyze the dendritic arbors of the abnormally positioned pyramidal cells and to compare the distribution of dendritic excrescences (i.e., the termination sites of the mossy fibers) in +/+ and Hld/Hld mice. It was found that in +/+ mice the late-generated pyramidal cells (whose cell bodies are positioned just below the suprapyramidal mossy fiber layer) have one set of dendritic excrescences on their apical dendrites as they extend through the suprapyramidal mossy fiber layer and a second set on their basal dendrites as they pass through the infrapyramidal mossy fiber layer. In contrast, in Hld/Hld mice the late-generated pyramidal cells (whose cell bodies are abnormally positioned just below the intrapyramidal mossy fiber layer) have two sets of dendritic excrescences on their apical dendrites, as they pass through the intrapyramidal and suprapyramidal mossy fiber layers, and none on their basal dendrites. In addition, in the vicinity of the apparent point of contact of the intrapyramidal mossy fibers, the apical dendrites of some of the abnormally positioned pyramidal cells have several fine-caliber branches.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

The cell surface of mouse dendritic cells: FACS analyses of dendritic cells from different tissues including thymus.

The surface of dendritic cells from mouse spleen, thymus, and epidermis has been compared with a panel of monoclonal antibodies and the FACS. A method was first developed to isolate populations of large, adherent, thymic dendritic cells that were greater than 90% pure. These were released by collagenase digestion and separated from adherent macrophages after overnight culture. Enrichment was based on the facts that most macrophages remained plastic adherent and rosetted strongly with antibody-coated erythrocytes. As in spleen, thymic dendritic cells were stellate in shape, had abundant class I and II MHC products, lacked many standard macrophage and lymphocyte markers, and actively stimulated the mixed leukocyte reaction. Most spleen and thymic dendritic cells could be lysed by the 7D4 mAb, to the low-affinity IL-2 receptor, and complement but the levels of 7D4 by FACS were low and sometimes not above background. Differences among dendritic cells from different tissues were noted with other mAb. Adherent dendritic cells from thymus all expressed the J11d "B cell" antigen and the NL145 interdigitating cell marker, but lacked the 33D1 spleen dendritic cell antigen. Eighty to ninety percent of spleen dendritic cells were J11d-, NL145-, 33D1+ but the remainder expressed the J11d+, NL145+, 33D1- thymic phenotype. The latter phenotype also was identical to that of epidermal Langerhans' cells. We postulate that the major 33D1+ cell in spleen represents a migratory stage in which dendritic cells are moving from tissues to lymphoid organs.

Animals

Dendritic morphology of interstitial amacrine cells with monostratified dendrites in different-sized carp retinas.

The dendritic morphology of a class of interstitial (IS) amacrine cells in retinas of different-sized carp (body length, 9.1-32.3 cm) was investigated by identifying their fluorescent nuclei pre-loaded with 4,6-diamidino-2-phenylindole (DAPI), followed by iontophoretic injection of Lucifer yellow (LY) in isolated and formaldehyde-fixed flat-mounts under microscopic control. The LY-injected fusiform or pyriform cell bodies were found to locate at the middle of the inner plexiform layer (IPL) or immediately beneath the amacrine cell layer, and their dendrites monostratified in sublamina b of the IPL. The pyriform cells had a short stem from which extended 4-5 stout dendrites, while the fusiform cells extended similar dendrites from the soma. The dendrites of both types of cell were decorated with spines and a few long axon-like processes. The pyriform cells were found more frequently in smaller retinas than in larger retinas, suggesting that the former may migrate proximally during retinal growth. The dendritic field sizes of these IS amacrine cells were wider as the fish became larger, while the dendritic morphology, analyzed by the Sholl's branching model, was very similar in smaller and larger retinas. The results indicate that the IS amacrine cells do not add dendrites, but that their dendritic trees simply expand during retinal growth.

Aging

The dendritic extent of motoneurons in frog brachial spinal cord: a computer reconstruction of HRP-filled cells. With comments on dendritic reconstruction methodologies.

A lateral and a medial motoneuron in the brachial spinal cord of the leopard frog, Rana pipiens, were labeled by horseradish peroxidase applied to the ventral root. Their dendritic trees were traced, analyzed, and plotted using a computer-microscope system. Some dendrites of the medial motoneuron crossed the midline of the spinal cord, but no dendrites of the lateral motoneuron crossed the midline. Nevertheless, the total dendritic length of the lateral motoneuron exceeded that of the medial motoneuron. The peak number of dendritic branch segments of the medial motoneuron was located at a greater distance from its soma than that of the lateral motoneuron. Three-dimensional reconstruction and rotation of the dendritic trees revealed that the dendrites of the medial motoneuron had a greater rostrocaudal extent than those of the lateral motoneuron. When compared to reports of Golgi-impregnated motoneurons, our results suggest that the HRP technique labels dendrites more completely. However, use of the HRP technique may introduce greater errors in the subsequent measurement of dendritic segments due to nonuniform tissue shrinkage.

Animals

Architecture of apical dendrites in the murine neocortex: dual apical dendritic systems.

A monoclonal antibody (5F9) against microtubule-associated protein 2 is a selective and sensitive marker for neocortical dendrites in the mouse. The marker stains all dendrites. It affords a particularly comprehensive picture of the patterns of arrangements of apical dendrites which are most intensely stained with this antibody. Dual systems of apical dendrites arise from the polymorphic neurons of layer VI, on the one hand, and the pyramidal neurons of layers II-V, on the other. Terminal arborization of the former is concentrated principally at the interface of layers V and IV, while that of the latter is in the molecular layer. Apical dendrites of both systems are grouped into fascicles. In supragranular layers and in upper layer VI-lower layer V, where apical dendrites are most abundant, the fascicles coalesce into septa. These generate a honeycomb-like pattern, subdividing these cortical levels into columnar spaces of approximately 20-40 micron diameter. At the level of layer IV, where the number of apical dendrites is greatly reduced, the fascicles are isolated bundles. These bundles have the form of circular, elliptical or rectangular columns in the primary somatosensory, temporal and frontal regions, respectively. Those in the barrel field are preferentially concentrated in the sides of barrels and the interbarrel septa. The configurations of the dendritic fascicles, particularly the midcortical bundles, may conform to the spatial configuration of investing axons of interneurons.

Animals

CD45 epitope mapping of human CD1a+ dendritic cells and peripheral blood dendritic cells.

The authors studied the pattern of leukocyte common antigen (CD45) epitope expression on dendritic cells in sections of human epidermis, tonsillar epithelium, dermatopathic lymph nodes, and in isolates from blood. The monoclonal antibodies (MAb) used were specific for all known CD45 epitopes, including the seven different CD45 common epitopes as well as the four known CD45R epitopes (two CD45RA, one CD45RB, and one CD45RO). Dendritic cells in all sites were uniformly reactive for the CD45 common epitopes tested except 2B11, which may recognize a CD45R rather than CD45 epitope. By single-label immunoperoxidase and double-label immunofluorescence epitope mapping of CD1a+ dendritic cells in tissue sections, it was generally difficult or impossible to detect expression of CD45RA, CD45RB, CD45RO, or 2B11. In blood dendritic cells, however, low levels of these CD45R epitopes were detected consistently using single-label immunoperoxidase staining of cytocentrifuge preparations. Monocytes were similar to blood dendritic cells except that the staining with MAb to CD45RO and 2B11 was slightly stronger. The authors conclude that dendritic cells differ from most subpopulations of lymphocytes in that CD45 common epitopes are readily detectable but the existing RA, RB, and RO epitopes are either undetectable or expressed at relatively low levels. These studies raise the possibility that CD1a+ dendritic cells may express a novel dominant CD45 isoform.

Antigens, CD

Deprived somatosensory-motor experience in stumptailed monkey neocortex: dendritic spine density and dendritic branching of layer IIIB pyramidal cells.

Infant macaque monkeys (Macaca arctoides) were individually raised to age 6 months in large clear cubes built into one wall of a control colony that allowed them visual access to it but not tactile contact. The two deprivation conditions (Cond 2 and Cond 3) were equal both in physical size and with respect to partial social isolation. They differed in the degree of somatosensory-motor opportunity available during development in that the Cond 2 chamber was empty, whereas Cond 3 contained ladders, a trapeze, and play objects. Four monkeys from each of these conditions were compared with four colony-reared (Cond CR) monkeys. Neuroanatomical changes were evaluated by using light microscopy in Golgi-Cox-stained neocortex. Dendritic spines on the apical shafts of layer IIIB pyramidal cells were counted in primary motor (MI), somatosensory (SI), and visual (area 17, V1) cortical regions. Layer IIIB pyramidal neurons with somas of medium size were selected from each cortical region and the density of apical dendritic spines determined. The basilar dendritic branches of these same neurons were traced, and the dendritic branching complexity was assessed in order to compare the sensitivity of the dendritic spine and branching measures consequent to deprived rearing. The number of apical dendritic spines was significantly reduced in Cond 2 when compared with either Cond 3 or Cond CR (which did not differ from each other). This occurred in both MI and SI cortex, but not in the visual cortex, the region used as a control for a global brain effect. Branching complexity measured on the same pyramidal neurons was reduced only in MI cortex of Cond 2. These results show spine density, a more direct measure of neuronal connectivity, to be the more sensitive measure of early environmental deprivation. Also, the enriched environment provided by Cond 3 relative to Cond 2 offset the effect of partial social isolation such that both morphometric measures had values comparable to Cond CR monkeys.

Animals

Quantitative analysis of dendritic branching. I. Simple formulae for the quantitative analysis of dendritic branching.

Dendritic branching of neurons may be quantitatively studied using applications of graph theory. Dendritic ramifications may be considered as 'forests' of planted, stemmed, binary 'trees' with simple mathematical properties. Two fundamental numbers, that of dendritic tips and that of dendritic stems, usually suffice to quantify the dendritic branching of one neuron as, in most cases, they allow the computation of all other numbers of dendritic points and segments.

Animals

Signal enhancement in distal cortical dendrites by means of interactions between active dendritic spines.

Pyramidal neurons in the cerebral cortex characteristically give rise to an apical dendrite, whose distal dendritic branches in layer I are covered with spines. These spines are known to be sites of synaptic connections, but the physiological properties of the spines and the functional significance of their responses are still largely unknown. The main function attributed thus far to these synaptic responses, situated at a great distance from the neuronal cell body, is slow background modulation of impulse output in the axon. In pursuing computer simulation analysis of electrical properties of dendrites, we have obtained results suggesting interactions between distal dendritic spines. If the heads of dendritic spines have excitable membrane properties, the spread of current from one or several spines could bring adjacent spines to their thresholds for impulse generation. This could give rise to a sequence of spine head action potentials, representing a saltatory propagation, from one or more excitable spine heads to nearby excitable spine heads, in the distal dendritic branches. Both the amplification due to several spine action potentials and the possibility of propagation into more proximal branches would increase the efficacy of distal synaptic inputs. Because of nonlinear dependence upon several modifiable parameters (such as spine stem resistance and membrane excitability) and upon the spatio-temporal pattern of synaptic input, such contingent synaptic enhancement would be particularly relevant to cortical functions underlying information processing and to plasticity underlying learning and memory.

Dendrites

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

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