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Dendritic morphology and development in the ferret lateral superior olivary nucleus.

The dendritic morphology of cells in the lateral superior olivary nucleus was studied with the Golgi method in adult and postnatal ferrets. The lateral superior olivary nucleus in the adult ferret is a convoluted structure with an M-shape in frontal sections. The major cell type appears to have disk-shaped dendritic trees. Most dendritic trees appear to be approximately orthogonal to the curved medial-lateral axis of the nucleus. Depending on their position in the limb and on the plane of section with respect to the dendritic tree, the disk-shaped cells are either bipolar or radiate in orientation. One subclass of disk-shaped cells has secondary dendritic branches that end as tufts of tendril-like processes. In a second subclass of cells, the dendrites exhibit several orders of dichotomous branching and lack obvious tufts of terminal processes. Marginal cells are observed at the border of the nucleus and have dendrites restricted to the margins of the cell plate. The bipolar orientation of disk-shaped cells orthogonal to the axis of the limbs is already apparent by the time of birth. Transient spines and other appendages are abundant on somata and dendrites during the first postnatal week. By the end of the first postnatal month only distal appendages are found. Tufts of fine tendril-like processes appear at the ends of dendrites between postnatal days 28 and 56.

Animals↗

Self-organization of tree form: a model for complex social systems.

The branches of a tree can be viewed as redundant organs: they have the same functional roles and the same unlimited developmental potential. The most prominent aspect of tree form, here considered as an example of dynamic self-organization, is the relations among these branches. A branch is a source of information about its size, state and environment. A response to this information is the oriented vascular differentiation which connects the branch with the rest of the plant. A known phytohormone, auxin, is formed by shoot tissues and replaces this orienting influence of a shoot. This accounts for neighboring branches having specific, even competing, effects without any other branch specificity. The same phytohormone also elicits different processes throughout the plant, integrating their occurrence without recourse to a central control. Self-organization requires a positive feedback that enhances the development of a successful branch, and an inhibitory influence that reduces similar branch growth. Both roles could depend on the oriented vascular differentiation that is specified by auxin. Actual tree form at any given time is a transient dynamic state. One reason for this is that competition between branches is a long-term, gradual process. A second reason is that a periodic surfeit of root supplies enhances a seasonal formation of many branches which 'search' environmental possibilities, and most of these branches decline only gradually. The various aspects of tree form could thus depend on remarkably simple and robust principles, requiring only limited genetic information. Much of the necessary information about the details of tree form is generated by a Darwinian-like process of developmental selection, reaching a predictable overall outcome without following any strict plan. It is a case of self-organization in which the component structures, the tree branches, do not move, but do undergo long term developmental changes.

Animals↗

Type 1 nitrergic (ND1) cells of the rabbit retina: comparison with other axon-bearing amacrine cells.

NADPH diaphorase (NADPHd) histochemistry labels two types of nitrergic amacrine cells in the rabbit retina. Both the large ND1 cells and the small ND2 cells stratify in the middle of the inner plexiform layer, and their overlapping processes produce a dense plexus, which makes it difficult to trace the morphology of single cells. The complete morphology of the ND1 amacrine cells has been revealed by injecting Neurobiotin into large round somata in the inner nuclear layer, which resulted in the labelling of amacrine cells whose proximal morphology and stratification matched those of the ND1 cells stained by NADPHd histochemistry. The Neurobiotin-injected ND1 cells showed strong homologous tracer coupling to surrounding ND1 cells, and double-labelling experiments confirmed that these coupled cells showed NADPHd reactivity. The ND1 amacrine cells branch in stratum 3 of the inner plexiform layer, where they produce a sparsely branched dendritic tree of 400-600 microm diameter in ventral peripheral retina. In addition, each cell gives rise to several fine beaded processes, which arise either from a side branch of the dendritic tree or from the tapering of a distal dendrite. These axon-like processes branch successively within the vicinity of the dendritic field before extending, with little or no further branching, for 3-5 mm from the soma in ventral peripheral retina. Consequently, these cells may span one-third of the visual field of each eye, and their spatial extent appears to be greater than that of most other types of axon-bearing amacrine cells injected with Neurobiotin in this study. The morphology and tracer-coupling pattern of the ND1 cells are compared with those of confirmed type 1 catecholaminergic cells, a presumptive type 2 catecholaminergic cell, the type 1 polyaxonal cells, the long-range amacrine cells, a novel type of axon-bearing cell that also branches in stratum 3, and a type of displaced amacrine cell that may correspond to the type 2 polyaxonal cell.

Amacrine Cells↗

Comparative analysis of response to phenotypic and marker-assisted selection for multiple lateral branching in cucumber ( Cucumis sativus L.).

Yield increase in processing cucumber ( Cucumis sativus L.) is positively correlated with an increase in number of fruit-bearing branches. Multiple lateral branching (MLB) is a metric trait controlled by at least five effective factors. Breeding efficacy might be improved through marker-assisted selection (MAS) for MLB. Experiments were designed to independently confirm previously determined linkage of molecular markers (L18-2-H19A SNP, CSWTAAA01 SSR, CSWCT13 SSR, W7-2 RAPD and BC-551 RAPD) to MLB, and to determine their utility in MAS. These markers were present in significantly higher frequency than expected (1, presence:3, absence; p < 0.001) in BC(2) plants selected based on a high MLB phenotype (BC(2)PHE). However, markers that were considered selectively neutral fit the expected segregation of donor parent DNA in BC(2) progeny. Markers linked to MLB were used in MAS of BC(1) and BC(2) plants to produce BC(2)MAS, and BC(3)MAS progeny. Means for MLB in MAS populations were compared with backcross populations developed through phenotypic selection (BC(2)PHE, BC(3)PHE) and by random mating where no selection had been applied (BC(2)RND, BC(3)RND). Statistical analysis showed no significant differences ( p < 0.001) between means of phenotypic (BC(2)PHE = 3.02, BC(3)PHE = 3.29) and marker-aided selection (BC(2)MAS = 3.12, BC(3)MAS = 3.11) for MLB. However, both phenotypic and MAS population means were significantly higher than the random control (BC(2)RND = 2.27, BC(3)RND = 2.41) for MLB. Thus, given the observed response to selection and the rapid life-cycle of cucumber (4 months), markers linked to MLB when used in MAS will most likely be effective tools in cucumber improvement.

Cucumis sativus↗

Jet-directed catheter for interventional radiology.

A new catheter of dual lumen construction, suitable for clinical use, is capable of navigating acute vessel branches by selective retrojet fluid thrusts issuing from cowlings built into its distal sidewalls. Conventional radiopaque liquids can be used both to drive the system and to fluoroscopically locate its position relative to the vascular network. In vitro studies have shown the catheter to traverse straight vessels and negotiate difficult lateral turns over short radii in the process of entering progressively smaller branches. In vivo studies have verified these capabilities and suggest a means for embolizing tumors and arteriovenous malformations located at otherwise inaccessible regions. For arterial infusion of drugs to treat tumors, the turbulent jet action is utilized to enhance mixing within the blood stream to overcome the streaming phenomenon common to other catheters. The result is better distribution of the agent and increased opportunity for the drug to reach more of the tumor. The catheter system is undergoing clinical trials.

Angiography↗

Three-dimensional morphology of astrocytes and oligodendrocytes in the intact mouse optic nerve.

The three-dimensional morphology of astrocytes and oligodendrocytes was analysed in the isolated intact mature mouse optic nerve, by correlating laser scanning confocal microscopy and camera lucida drawings of single cells, dye-filled with lysinated rhodamine dextran or horseradish peroxidase, respectively. These techniques enabled the entire process field of single dye-filled cells to be visualized in all planes and resolved the fine details of glial morphology. Morphometric analysis showed that the processes of all astrocytes had branches ending at the pial surface, on blood vessels, and freely in the nerve; branches ending in the nerve were described to end at nodes of Ranvier in the accompanying paper. Astrocytes were classified into a single morphological population in which each cell subserved multiple functions. The results of this study do not support the contention that astrocytes can be subdivided into two morphological and functional subtypes, namely type-1 and type-2, which have process ending either at the glia limitans or at nodes, respectively. Three-dimensional analysis of oligodendrocyte units, defined as the oligodendrocyte, its processes and the axons it ensheaths, showed the provision of single myelin segments for an average of 19 nearby axons (range 12-35) with a mean internodal length of 138 microns (range 50-350 microns). Mouse optic nerve oligodendrocytes were a homogeneous population and were markedly similar to those in the rat optic nerve. The results of our analysis of oligodendrocyte morphology are consistent with the view that the number and internodal length of myelin sheaths supported by a single oligodendrocyte are related to the diameter of the ensheathed axons.

Animals↗

A demonstration of glial filament distribution in astrocytes isolated from rat cerebral cortex.

Astrocytes have been classically described based on morphological characteristics as either protoplasmic or fibrous. However, the appearance of astrocytes at the light microscopic level following immunoreaction with glial filament acidic protein antisera or Cajal's gold-chloride method is markedly different from the morphology of Golgi-impregnated or horseradish peroxidase-filled astrocytes. The present study combines immunohistochemistry using glial fibrillary acidic protein antisera and a cellular suspension of structurally intact astrocytes isolated from the rat cerebral cortex to demonstrate that the distribution of glial filaments is limited to major astrocytic processes. Because the sheet-like processes which decorate the major branches of protoplasmic astrocytes do not contain glial filaments, the entire protoplasmic astrocyte is not visible when viewed in situ in the gray matter of the cerebral cortex. Thus staining techniques that have a propensity for the glial filaments such as antisera to glial fibrillary acidic protein and the gold-chloride method provide only a skeletal outline of the major processes of the protoplasmic astrocyte. On the other hand, fibrous astrocytes which do not have feathery decorations on major processes are visible in the cerebral cortex in their structural entirety. The present investigation also offers additional evidence of the specificity of glial fibrillary acidic protein antisera for astrocytes.

Animals↗

ATP hydrolysis on actin-related protein 2/3 complex causes debranching of dendritic actin arrays.

Extension of lamellipodia, an important dissipative process in cell motility, is driven by the turnover of a polarized dendritic array of actin filaments. Motility is driven by catalytic cycles of filament attachment to Wiskott-Aldrich syndrome protein (WASP)-activated actin-related protein (Arp)2/3 complex at the leading edge, branch formation, and detachment, allowing subsequent growth of branched filaments. The morphology, mechanical strength, and lifetime of the array are determined by the processes of filament branching, debranching, and treadmilling. All three processes are controlled by ATP hydrolysis. ATP hydrolysis on F-actin is known to be at the origin of treadmilling. Here, by using radiolabeled ATP covalently bound to Arp2/3, we show that ATP is hydrolyzed on Arp2, not on Arp3, after a delay following filament branching. Hydrolysis of ATP on Arp2 promotes debranching of filaments and acts as a clock that controls the stability of dendritic actin arrays in lamellipodia. Finally, we propose that hydrolysis of ATP on G-actin in the ternary G-actin-WASP-Arp2/3 complex on branch formation destabilizes the WASP-actin interface and energetically facilitates the detachment step in the branching reaction.

Actin-Related Protein 2↗

Cloning of drebrin A and induction of neurite-like processes in drebrin-transfected cells.

The developmentally-regulated neuron-specific protein, drebrin A, is expressed first at the time of outgrowth and maturation of dendrites, and is localized within dendrites of the adult brain. A cDNA clone of adult rat drebrin A was isolated and sequenced. There is no overall homology with other reported protein sequences except chicken drebrins. We constructed the expression vector MIW-DA containing the drebrin A cDNA. Transfection of nonneuronal cells with MIW-DA induced the formation of highly branched neurite-like cell processes. In these process-bearing transfectants, expressed debrin A is concentrated in submembraneous regions of the cell. Furthermore, actin concentration is higher in these cells than other fibroblasts. These results suggest a possible role of drebrin A in neurite outgrowth.

Amino Acid Sequence↗

Identification of Asp 549 as the catalytic nucleophile of glycogen-debranching enzyme via trapping of the glycosyl-enzyme intermediate.

Glycogen-debranching enzyme catalyzes the removal of branching from glycogen via a two-step process involving first the transfer of a maltotriosyl unit from the branch to the main chain and second the hydrolysis of the residual alpha-(1,6)-linked glucose moiety. Since the transfer occurs with retention of anomeric configuration, a mechanism involving a maltotriosyl-enzyme species is presumed. 4-Deoxy-alpha-maltotriosyl fluoride functions as an incompetent substrate for this transferase activity since a glycosyl-enzyme species in formed, as witnessed by a "burst" of fluoride release, but turned over only very slowly unless a suitable acceptor such as maltotriose is added, at which point 4-deoxymaltohexaose is released. Peptic proteolysis of this trapped enzyme generated a mixture of peptides which was separated by reverse phase high-performance liquid chromatography, and the glycosylated peptide was located by use of tandem mass spectrometry in the neutral loss mode. Subsequent tandem mass spectrometric experiments on this peptide identified it as one surrounding Asp 549. This amino acid is completely conserved in all alpha-glucanotransferases and alpha-glucosidases belonging to this sequence -related family and is hereby identified as the catalytic nucleophile.

Amino Acid Sequence↗

On SIF cell-neuron relationships in autonomic ganglion.

Seldom found SIF cell processes in the lower mesenteric ganglion of the cat, in close proximity to principal ganglionic neurons either their axons or dendrites are described. Contacts between SIF cells and neurons could not always be regarded as unambiguous synapses. But the close proximity of both cells wrapped in the same satellite cells and same basal lamina, suggests the possibility of such synapses. Formations of narrow branches from main SIF cell processes were found in serial sections. In addition, these processes sometimes lost their Schwann cell envelope not only in the vicinity of blood capillaries, but also in the vicinity of the neuron. Some functional similarity between individual SIF cell groups in autonomic ganglia has been supposed. The hypothesis of adrenergic afferent and efferent synapse on SIF cell as a part of an inhibitory feedback loop between a SIF cell group and principal ganglionic neuron has been proposed. The endocrine function of SIF cells is considered to be a main role of these cells in autonomic ganglia.

Animals↗

Three dimensional reconstruction of branched tree structures from serial sections.

A process is described for the automatic registration and reconstruction of anatomical tree structures such as capillary networks in the microstructure of tissue. The technique uses images of, or images representing, a set of closely spaced parallel slices called "serial sections". The source of the serial sections may be MRI, multi-planar X-ray, multi-planar infra-red scans, or simple histological sections. A tree structure is defined as a structure consisting of a network of nested ducts, vessels or solid cores which branch and join with another such that one structure may appear as more than one distinct area in a single section, or in a series of sections. The reconstruction of such an object poses many problems which do not occur when restructuring objects which are so shaped that they can give rise to only one area in each section.

Animals↗

Dendritic growth and remodeling of cat retinal ganglion cells during fetal and postnatal development.

We have studied the development of retinal ganglion cell morphology in the cat's visual system from early fetal to postnatal times. In particular, we have examined the contribution of growth and remodeling to the establishment of mature retinal ganglion cell form. Ganglion cells were identified by retrograde labeling with rhodamine latex microspheres deposited in the superior colliculus and lateral geniculate nucleus between embryonic day 34 (E34; birth = E65) and adulthood. To reveal the fine morphological details of retrogradely labeled ganglion cells, 48 hr later Lucifer yellow was injected intracellularly in living retinae that had been dissected and maintained in vitro. Our results show that at E35-37 the majority of ganglion cells are very simple in morphology, with a few dendritic processes that are generally aligned in a radial direction towards or away from the optic disc. During the ensuing 2 week period, there is a progressive growth and elaboration of dendrites. By E50, some ganglion cells resembling the adult alpha, beta, and gamma classes can be identified based on comparisons of the appearance and dimensions of their dendritic trees and somata with neighboring filled cells. However, ganglion cell dendrites and axons at this age express several transient morphological features. The axons of ganglion cells give rise to delicate processes originating from the intraretinal portion of the axon, including side branches, present in about half of the cells, and occasionally bifurcations that give rise to axon collaterals. These transient axonal features are present throughout development, including the neonatal period; no axon collaterals were observed after postnatal day 15, while axonal side branches persisted even at P31 but were gone by adulthood. Ganglion cell dendrites exhibit excessive branches and exuberant somatic and dendritic spines. Quantitative analysis of these processes shows that after E45 dendritic trees increase dramatically in complexity, reaching the peak number of spines and branch points by the first week of postnatal life. The number of dendritic processes then falls abruptly to reach near-adult levels by the end of the first postnatal month. Even though dendritic morphology closely resembles that seen in the adult at this age, ganglion cell bodies and dendrites must continue to grow to reach their adult size.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Dynamic changes in Sertoli cell processes invading spermatid cytoplasm during mouse spermiogenesis.

Studies using thick sections stained by ATPase cytochemistry and scanning electron microscopy were carried out to determine three-dimensional ultrastructural alterations in Sertoli cell processes invading neighboring spermatids during mouse spermiogenesis. Sertoli cell processes start invading spermatid cytoplasm at the acrosomal phase of development and undergo considerable change at the maturation phase of development. At step 14, these processes elongate and begin to branch in the spermatid cytoplasm, and by step 15, they extend in various directions to form a complex of canals that the authors have designated the canal complex. The present observations also clarify that the complicated canal complex undergoes regional modification. At the late stages of maturation, the endoplasmic reticulum has gathered with other cell organelles to form aggregates of endoplasmic reticulum in the vicinity of which invading Sertoli cell processes extensively ramify further into thin tubules that intertwine with each other to form a region of thin tubules. In thin sections, each such region was a complex, consisting of small vesicles and endoplasmic reticulum, and corresponded to what has been defined as a mixed body by Morales and Clermont (Anat. Rec., 203:233-244, 1982). During the course of the formation of the region, the invading Sertoli cell processes are continuous at all times with the cell body of the surrounding Sertoli cell.

Adenosine Triphosphatases↗

The role of the SAP motif in promoting Holliday junction binding and resolution by SpCCE1.

Holliday junctions are four-way branched DNA structures that are formed during recombination and by replication fork regression. Their processing depends on helicases that catalyze junction branch migration, and endonucleases that resolve the junction into nicked linear DNAs. Here we have investigated the role of a DNA binding motif called SAP in binding and resolving Holliday junctions by the fission yeast mitochondrial resolvase SpCCE1. Mutation or partial/complete deletion of the SAP motif dramatically impairs the ability of SpCCE1 to resolve Holliday junctions in a heterologous in vivo system. These mutant proteins retain the ability to recognize the junction structure and to distort it upon binding. However, once formed the mutant protein-junction complexes are relatively unstable and dissociate much faster than wild-type complexes. We show that binding stability is necessary for efficient junction resolution, and that this may be due in part to a requirement for maintaining the junction in an open conformation so that it can branch migrate to cleavable sites.

Amino Acid Sequence↗

A mathematical model for conduction of action potentials along bifurcating axons.

1. A mathematical model based on the Hodgkin-Huxley equations is derived to describe quantitatively the propagation of action potentials in a branching axon. 2. The model treats the case of a bifurcating axon with branches of different diameters. The solution takes into account the changes in space constant in the different regions. 3. The model allows for investigating parameters leading to preferential conduction of action potentials in one daughter branch as seen experimentally. 4. Assuming that the only difference between the various daughter branches is in their diameters, conduction blocks should occur simultaneously rather than differentially into all daughter branches when the geometrical ratio is greater than 10. 5. In order to obtain differential conduction into the two branches changes in ionic concentrations due to the repetitive action potentials had to be introduced into the equations. 6. We find that conditions which allow differential buildup of K concentration around the two branches, produce differential conduction block. These conditions may be different periaxonal spaces around the branches or different time constant for recovery processes that eliminate K from the periaxonal space. 7. The effects of an inexcitable branch on conduction of action potentials in the second branch are described. 8. We find that the membrane current which is associated with the action potential is much more sensitive than the action potential itself and shows more distinct changes near regions of inhomogeneity such as a branch point, a step increase in diameter or an inexcitable branch.

Action Potentials↗

The pruned gene encodes the Drosophila serum response factor and regulates cytoplasmic outgrowth during terminal branching of the tracheal system.

We identified a Drosophila gene, pruned, that regulates formation of the terminal branches of the tracheal (respiratory) system. These branches arise by extension of long cytoplasmic processes from terminal tracheal cells towards oxygen-starved tissues, followed by formation of a lumen within the processes. The pruned gene is expressed in terminal cells throughout the period of terminal branching. pruned encodes the Drosophila homologue of serum response factor (SRF), which functions with an ETS domain ternary complex factor as a growth-factor-activated transcription complex in mammalian cells. In pruned loss of function mutants, terminal cells fail to extend cytoplasmic projections. A constitutively activated SRF drives formation of extra projections that grow out in an unregulated fashion. An activated ternary complex factor has a similar effect. We propose that the Drosophila SRF functions like mammalian SRF in an inducible transcription complex, and that activation of this complex by signals from target tissues induces expression of genes involved in cytoplasmic outgrowth.

Animals↗

The function and structure of the marsupial enamel.

The aims of this study are to clarify the structure of tubular enamel and the function of enamel tubules on the marsupial of opossum (Monodelphis domestica). Almost all enamel prisms, surrounded by interprismatic enamel, ran obliquely from the dentinoenamel junction (DEJ), and bent near the enamel surface. The enamel tubules are distributed in both enamel prisms and the interprismatic enamel near the DEJ. From the middle to the surface of the enamel, one enamel tubule ran within a single enamel prism. Most of enamel tubules continued from the DEJ to near the enamel surface. It is suggested that each enamel tubule developed in relation to one ameloblast. The fibers of odontoblastic process penetrated the DEJ from the dentinal tubules into the enamel tubules, and some branched across the enamel prisms. The odontoblastic process may be actively cross into the ameloblastic layer and may be involved in the formation of enamel tubules. After in vivo injection of tetracycline, tetracycline labeling showed that the odontoblastic tubules continued to enamel tubules. And strontium was detected in enamel tubules from the DEJ to the enamel surface, as was the dentinal tubules. In conclusion, there was active transport by the odontoblast and it's process through the enamel tubules.

Animals↗