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Simulation of the growth pattern of the central retinal artery using a fractal modeling technique.

Assisted by computer, we simulated the typical growth pattern of the central retinal artery by a fractal modeling technique. When processing was started with two fertile sides, an iterative processing function generated a ramiform fractal. The branching angle was chosen at random within a certain range. In addition, a feedback restriction forbade any intersection of branches. We demonstrated that the curved shape of the temporal vessels is caused by simultaneous growth of the vessels and the retina. When processing was begun with only one fertile side, the typical vascular pattern of a retinal coloboma was designed.

Computer Simulation↗

Relationship between incoherent excitation energy migration processes and molecular structures in zinc(II) porphyrin dendrimers.

Multiporphyrin dendrimers are among the most promising architectures to mimic the oxygenic light-harvesting complex because of their structural similarities and synthetic convenience. The overall geometries of dendrimers are determined by the core structure, the type of dendron, and the number of generations of interior repeating units. The rigid core and bulky volume of exterior porphyrin units in multiporphyrin dendrimers give rise to well-ordered three-dimensional structures. As the number of generations of interior repeating units increases, however, the overall structures of dendrimers become disordered and randomized due to the flexibility of the repeating units. To reveal the relationship between molecular structure and processes of excitation-energy migration in multiporphyrin dendrimers, we calculated the molecular structure and measured the time-resolved transient absorption and fluorescence anisotropy decays for various hexaarylbenzene-anchored polyester zinc(II) porphyrin dendrimers along with three types of porphyrin dendrons as references. We found that the congested two-branched type dendrimers exhibit more efficient energy migration processes than one- or three-branched type dendrimers because of multiple energy migration pathways, and the three-dimensional packing efficiency of dendrimers strongly depends on the type of dendrons.

Dendrimers↗

Structural analysis of the formation of glomerular microaneurysms in the Habu venom model.

The goal of this study has been to characterize the process of glomerular microaneurysm formation and to separate it from capillary ballooning. In the Habu venom model glomerular capillary ballooning and glomerular microaneurysm formation are seen regularly. The sequence of glomerular lesions leading to a glomerular microaneurysm has been examined and it is clear that the process starts with local mesangiolysis. This may proceed to mesangial expansion and/or ballooning of glomerular capillaries but in contrast to ballooning the formation of a glomerular microaneurysm is based on endothelial defects. The process occurs as follows: once initiated by mesangial failure lesions extend along the mesangial axis. As long as the extension of the lesion encroaches on divergent capillary branches, capillary ballooning by "coalescence" is the result. This process comes to an end when convergent capillary branching is reached and two capillaries join. At this point endothelial disruptions occur, blood and mesangial spaces merge and a glomerular microaneurysm is established. Further growth of the microaneurysm occurs following damage spreading along the lobular axis. The entire process has been reconstructed and is presented in a three-dimensional model.

Aneurysm↗

Branching mechanism of intergranular crack propagation in three dimensions.

We investigate the process of slow intergranular crack propagation by the finite element method model and show that branching is induced by partial arresting of a crack front owing to the geometrical randomness of grain boundaries. A possible scenario for the branching instability of crack propagation in a disordered continuous medium is also discussed.

Journal Article↗

Mechanism for the removal of residual cytoplasm from spermatids during mouse spermiogenesis.

During spermiogenesis, cytoplasmic processes of Sertoli cells invade spermatid cytoplasm to form a canal complex (Sakai et al., 1988). Thin tubules are formed from the canal complex and intertwine with each other to give rise to the "mixed body." In the present study, analysis of the changes undergone by the intertwining thin tubules indicated that they contribute to the removal of cell organelles from spermatid cytoplasm. Intertwining thin tubules were first detected at step 13. By step 15, their number had greatly increased. In the present study, the membranes of the intertwining thin tubules were clearly observed to be continuous with the spermatid plasma membranes. Thus, the mixed body possibly may be formed as a long pit of the spermatid plasma membrane situated close to the invading Sertoli cell process. With the progress of spermiogenesis, the lumens of the intertwining thin tubules gradually became swollen, and the intertwining swollen tubules fused with each other so that the spermatid cytoplasm enclosed by the intertwining swollen tubules isolated into fragments. This fragmented cytoplasm, which contained a large amount of endoplasmic reticulum, became spherical. Small branches of the invading Sertoli cell processes entered into the lumens of the intertwining swollen tubules and occupied their interior to the point that, finally, they completely engulfed the fragmented spermatid cytoplasm. Because the invading Sertoli cell processes were continuous with Sertoli cell bodies surrounding a spermatid at this step, it is possible for the fragmented cytoplasm to be transported into the latter by way of the invading Sertoli cell processes.

Adenosine Triphosphatases↗

Glial cells in the developing and adult olfactory lobe of the moth Manduca sexta.

The antennal lobe of the moth contains several classes of glial cells that are likely to play functional roles in both the developing and mature lobe. In this study, confocal and electron microscopy were used to examine in detail the morphology of two classes of glial cells, those associated with olfactory receptor axons as they course to their targets in the lobe and those that form borders around the synaptic neuropil of the olfactory glomeruli. The former, the nerve-layer glia, have long processes with multiple expansions that enwrap axon fascicles; the latter, the neuropil glia, constitute two subgroups: complex glia with large cell bodies and branching, vellate arbors; and simple glia, with multiple, mostly unbranched processes with many lamellate expansions along their lengths. The processes of complex glia appear to be closely associated with axon fascicles as they enter the glomeruli, while those of the simple glia surround the glomeruli as part of a multi-lamellar glial envelope, their processes rarely invading the synaptic neuropil of the body of the glomerulus. The full morphological development of antennal-lobe glial cells requires more than two-thirds of metamorphic development. During this period, cells that began as cuboidal or spindle-shaped cells that were extensively dye-coupled to one another gradually assume their adult form and, at least under nonstimulated conditions, greatly reduce their coupling. These changes are only weakly dependent on the presence of olfactory receptor axons. Glial processes are somewhat shorter and less branched in the absence of these axons, but basic structure and degree of dye-coupling are unchanged.

Animals↗

Dendritic spines form 'collars' in hippocampal granule cells.

A quantitative study of the distribution of dendritic spines was carried out in three orders of dendritic branches of granule cells from the dentate gyrus of the rat hippocampus. Golgi-stained preparations (7-19 neurones in each of seven rats) were analysed using computerized microscopy. Identification of spines and quantification of stem-spine geometry was performed using a segmentation algorithm and a line skeleton transformation of dendritic images. Analysis of data using the statistics of point processes revealed that, in all three branch orders, the distribution of visible spines along dendrites was not evenly random, but included dense clusters of spines surrounding the dendritic stem (spine 'collars'). Three-dimensional reconstructions from serial ultrathin sections have confirmed the presence of such spine groups. We speculate the spine collars represent a functional element in which associative synaptic plasticity is fostered by the proximity of individual synapses.

Animals↗

Surface processes of olfactory receptors.

The free surface morphology of olfactory receptor cells from the nasal mucosa of Cynomolgus monkeys was studied electron microscopically. The receptor cell, in addition to showing characteristic cilia, possess several branched or unbranched shorter elevations or spiny processes covered by numerous delicate lace-like filaments not previously described. These filaments diminish in length and number toward the base of the microvillous protrusions.

Animals↗

Universal behavior of one-dimensional multispecies branching and annihilating random walks with exclusion.

A directed percolation process with two symmetric particle species exhibiting exclusion in one dimension is investigated numerically. It is shown that if the species are coupled by branching (A-->AB, B-->BA), a continuous phase transition will appear at the zero-branching-rate limit belonging to the same universality class as that of the two component branching and annihilating random-walk model with two symmetric offsprings. This class persists even if the branching is biased towards one of the species. If the two systems are not coupled by branching but a hard-core interaction is allowed only the transition will occur at finite branching rate belonging to the usual (1+1)-dimensional directed percolation class.

Journal Article↗

Unexpected metal ion requirements specific for catalysis of the branching reaction in a group II intron.

The splicing process catalyzed by group II intron ribozymes follows the same two-step pathway as nuclear pre-mRNA splicing. In vivo, the first splicing step of wild-type introns is a transesterification reaction giving rise to a branched lariat intron-3'-exon intermediate characteristic of this splicing mode. In the wild-type introns, the ribozyme core and the substrate intron-exon junctions are carried by the same precursor molecule, making it difficult to distinguish between RNA folding and catalysis under normal splicing reactions. To characterize the catalytic step of the first transesterification reaction, we studied the reversal of this reaction, reverse branching. In this reverse reaction, the excised lariat intron and the substrate 5'-exon can be preincubated and folded separately, allowing the measure of the catalytic rate of the reaction. To measure the catalytic rate of the second splicing step, purified lariat intron-3'-exon intermediate molecules were preincubated and folded prior to the addition of 5'-exon. Conditions could be found where chemistry appeared rate limiting for both catalytic steps. Study of the metal ion requirements under these conditions resulted in the unexpected finding that, for the intron studied, substitution of magnesium ions by manganese ions enhanced the rate of the first transesterification reaction by two orders of magnitude but had virtually no effect on the second transesterification reaction or the 5' splice site cleavage by hydrolysis. Finally, the catalytic rates measured under optimal conditions for both splicing steps were faster by three orders of magnitude in the branching pathway than in the hydrolytic pathway.

Binding Sites↗

Electron microscopic demonstration of a supraependymal cluster of neuronal cells and processes in the hamster third ventricle.

A supraependymal cluster of neuronal cells and processes consistently present on the floor of the hamster third ventricle was identified and characterized by means of correlative scanning (SEM) and transmission (TEM) electron microscopy. SEM revealed each cluster to be ovoid with the majority of its surface covered by dome-shaped protrusions and fine beaded fibers. A number of processes traveling individually or in groups also entered or exited from the cluster at its base. As these processes passed over the ventricular surface, they contributed to an extensive network on the floor and ventral aspect of the ventricular wall. Some processes terminated on the ependymal surface in bulbous endings while others penetrated the ependyma. The neuronal nature of these clusters and their associated processes was confirmed at the TEM level. The dome-shaped protrusions visible on the surface of the cluster in SEM corresponded to apical surfaces of neurons confined to the peripheral aspect of a core of loosely arranged processes. These cells exhibited a prominent nucleolus, stacks of rough endoplasmic reticulum (RER), polyribosomes, Golgi cisternae, mitochondria and microtubules (MT) and gave rise to dendritic processes which extended into the core. These dendrites gave off branches at acute angles and contained polyribosomes, single cisternae of RER and evenly spaced MT. Other profiles of processes within the core shared these characteristics, suggesting that they also were branches of the peripheral cells. Axons present within the core and on the cluster's surface exhibited vesicle-filled varicosities which frequently established synaptic contact with the peripheral cells and their processes. The presence of an intraventricular cluster of neurons which potentially communicates with centers extrinsic to the ventricle may have important implications in the hypothesized role of cerebrospinal fluid and tanycytic ependyma in the neuroendocrine regulation of anterior pituitary function.

Animals↗

The development of neuronal morphology in insects.

Neurons are highly polarized cells with some regions specified for information input--typically the dendrites--and others specialized for information output--the axons. By extending to a specific location and branching in a specific manner, the processes of neurons determine at a fundamental level how the nervous system is wired to produce behavior. Recent studies suggest that relatively small changes in neuronal morphology could conceivably contribute to striking behavioral distinctions between invertebrate species. We review recent data that begin to shed light on how neurons extend dendrites to their targets and acquire their particular branching morphologies, drawing primarily on data from genetic model organisms. We speculate about how and why the actions of these genes might facilitate the diversification of dendritic morphology.

Animals↗

Glial diffusion barriers during aging and pathological states.

In conclusion, glial cells control not only ECS ionic composition, but also ECS size and geometry. Since ECS ionic and volume changes have been shown to play an important role in modulating the complex synaptic and extrasynaptic signal transmission in the CNS, glial cells may thus affect neuronal interaction, synchronization and neuron-glia communication. As shown in Fig. 2, a link between ionic and volume changes and signal transmission has been proposed as a model for the non-specific feedback mechanism suppressing neuronal activity (Syková, 1997; Ransom, 2000). First, neuronal activity results in the accumulation of [K+]e, which in turn depolarizes glial cells, and this depolarization induces an alkaline shift in glial pHi. Second, the glial cells extrude acid and the resulting acid shift causes a decrease in the neuronal excitability. Because ionic transmembrane shifts are always accompanied by water, this feedback mechanism is amplified by activity-related glial swelling compensated for by ECS volume shrinkage and by increased tortuosity, presumably by the crowding of molecules of the ECS matrix and/or by the swelling of fine glial processes. This, in turn, results in a larger accumulation of ions and other neuroactive substances in the brain due to increased diffusion hinderance in the ECS. Astrocyte hypertrophy, proliferation and swelling influence the size of the ECS volume and tortuosity around neurons, slowing diffusion in the ECS. Their organization may also affect diffusion anisotropy, which could be an underlying mechanism for the specificity of extrasynaptic transmission, including 'cross-talk' between distinct synapses (Barbour and Hausser, 1997; Kullmann and Asztely, 1998). An increased concentration of transmitter released into a synapse (e.g. repetitive adequate stimuli or during high frequency electrical stimulation which induces LTP) results in a significant activation of high-affinity receptors at neighboring synapses. The efficacy of such synaptic cross-talk would be dependent on the extracellular space surrounding the synapses, i.e. on intersynaptic geometry and diffusion parameters. Other recent studies have also suggested an important role for proteoglycans, known to participate in multiple cellular processes, such as axonal outgrowth, axonal branching and synaptogenesis (Hardington and Fosang, 1992; Margolis and Margolis, 1993) that are important for the formation of memory traces. Recent observation of a decrease of fibronectin and chondroitin sulfate proteoglycan staining in the hippocampus of behaviorally impaired aged rats (Syková et al., 1998a,b) supports this hypothesis. It is reasonable to assume that besides neuronal and glial processes, macromolecules of the extracellular matrix contribute to diffusion barriers in the ECS. It is therefore apparent that glial cells play an important role in the local architecture of the CNS and they may also be involved in the modulation of signal transmission, in plastic changes, LTP, LTD and in changes of behavior and memory formation.

Aging↗

Intussusceptive arborization contributes to vascular tree formation in the chick chorio-allantoic membrane.

Various reports indicate that the process of intussusceptive microvascular growth (IMG) plays a crucial role in capillary network formation of the chorio-allantoic membrane (CAM). In the present study we demonstrate by methylmethacrylate (Mercox) casting and in vivo time-lapse observations that intussusception, i.e. insertion of transcapillary tissue pillars, is also strongly involved in vascular tree formation, a process we refer to as intussusceptive arborization (IAR). From day 7 to day 14 of incubation, several arterial and venous branching generations arise from the capillary plexus. The process is initiated by pillar formation in rows, which are demarcating future large vessels in the capillary meshwork. In a subsequent step the pillars undergo reshaping to form narrow tissue septa that successively merge, which results in the production of new generations of blood vessels. This is followed by growth and maturation of all vascular components. The process of IAR in the CAM is very active at days 10 and 11 of incubation and takes place in preferentially perfused capillary regions determining "dynamic areas". The process of intussusception may be preceded by endothelial division, but the transcapillary pillar formation itself occurs primarily by rearrangement and attenuation of the endothelial cells without local endothelial cell proliferation. We conclude that after the early sprouting phase, the process of intussusception is the basic mechanism of CAM vascularization. It leads to capillary network growth and expansion (IMG) and, at the same time to feed vessel formation with several branching generations (IAR).

Allantois↗

Activation of branched-chain keto acid dehydrogenase by exercise.

The present study was conducted to investigate the metabolic regulation of leucine oxidation during exercise. Ten rats per group were run at 27 m/min (0% grade) on a treadmill for 30 and 120 min or until exhausted, and the total and basal activity of branched-chain keto acid dehydrogenase was examined in the muscle, liver, and heart. The total activity of the dehydrogenase in the heart, liver, or skeletal muscle was unchanged by exercise. However, exercise increased the basal activity levels of the dehydrogenase about 10-fold in muscle and 5-fold in heart. The basal dehydrogenase activity in the liver was unchanged by exercise. Activation of the dehydrogenase in both muscle and heart was statistically elevated after 30 min exercise and continued to increase during the remainder of the exercise bout. The basal activity of the dehydrogenase returned to resting levels by 10 min postexercise. The activation of the dehydrogenase in muscle and heart during exercise likely is due to dephosphorylation because activity of the enzyme in mitochondria isolated from exercised muscles reverts to control values when the mitochondria are incubated in the presence of ATP. Thus the increased leucine oxidation observed during exercise is due to activation of the branched-chain keto acid dehydrogenase by dephosphorylation. This is the first example of a large increase in branched-chain keto acid dehydrogenase activity caused by a physiological process. This demonstrates that the muscle's latent capacity of oxidize branched-chain amino acids is much larger than previously thought and that this capacity is used in exercising muscle.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

Micromorphology of the third ventricular ependyma and infundibular recess in the mud-eel Amphipnous cuchia (Ham.).

Ependymal cells of varied size and shape including tanycytes with long basal processes are encountered both in the third ventricle and infundibular recess of A. cuchia. The basal processes of the tanycytes may be branched or unbranched and often lie in contact with each other or even from loose bundles. They also terminate on, or maintain close contact with the neurons of the nucleus lateralis tuberis, blood vessels, axonal tract of the nucleus preopticus and other neuronal elements. Golgi-Cox technique is apparently selective in its impregnation of both neurons and tanycytes. Prominent apical processes of some cells project into the ventricle. Some supraependymal cellular elements have their entire cell body projecting into the ventricle. The ependymal cells react positively to alkaline phosphatase and ascorbic acid tests.

Animals↗

Regulation of ureteric bud branching morphogenesis by sulfated proteoglycans in the developing kidney.

Glycosaminoglycans in the form of heparan sulfate proteoglycans (HSPG) and chondroitin sulfate proteoglycans (CSPG) are required for normal kidney organogenesis. The specific roles of HSPGs and CSPGs on ureteric bud (UB) branching morphogenesis are unclear, and past reports have obtained differing results. Here we employ in vitro systems, including isolated UB culture, to clarify the roles of HSPGs and CSPGs on this process. Microarray analysis revealed that many proteoglycan core proteins change during kidney development (syndecan-1,2,4, glypican-1,2,3, versican, decorin, biglycan). Moreover, syndecan-1, syndecan-4, glypican-3, and versican are differentially expressed during isolated UB culture, while decorin is dynamically regulated in cultured isolated metanephric mesenchyme (MM). Biochemical analysis indicated that while both heparan sulfate (HS) and chondroitin sulfate (CS) are present, CS accounts for approximately 75% of the glycosaminoglycans (GAG) in the embryonic kidney. Selective perturbation of HS in whole kidney rudiments and in the isolated UB resulted in a significant reduction in the number of UB branch tips, while CS perturbation has much less impressive effects on branching morphogenesis. Disruption of endogenous HS sulfation with chlorate resulted in diminished FGF2 binding and proliferation, which markedly altered kidney area but did not have a statistically significant effect on patterning of the ureteric tree. Furthermore, perturbation of GAGs did not have a detectable effect on FGFR2 expression or epithelial marker localization, suggesting the expression of these molecules is largely independent of HS function. Taken together, the data suggests that nonselective perturbation of HSPG function results in a general proliferation defect; selective perturbation of specific core proteins and/or GAG microstructure may result in branching pattern defects. Despite CS being the major GAG synthesized in the whole developing kidney, it appears to play a lesser role in UB branching; however, CS is likely to be integral to other developmental processes during nephrogenesis, possibly involving the MM. A model is presented of how, together with growth factors, heterogeneity of proteoglycan core proteins and glycosaminoglycan sulfation act as a switching mechanism to regulate different stages of the branching process. In this model, specific growth factor-HSPG combinations play key roles in the transitioning between stages and their maintenance.

Animals↗

Distribution of myomodulin-like immunoreactivity in the brain and retrocerebral complex of the locust, Schistocerca gregaria.

The distribution of myomodulin-like immunoreactivity is described for the brain and retrocerebral complex of an insect, the locust, Schistocerca gregaria. The locust brain contains 70-100 neuronal cell bodies and numerous neuropilar processes exhibiting myomodulin-like immunoreactivity. The most marked feature of the staining is a group of lateral tritocerebral neurones that form a highly immunoreactive tract that gives rise to a complex neuropile of stained processes in the dorsal tritocerebrum. This tract continues dorsally and bifurcates into a major branch that exists the brain via nervi corpora cardiaca 1 (NCC1) to innervate the corpora cardiaca and the corpora allata. A minor branch, consisting of several individual axons, combines with immunoreactive processes from the ventral nerve cord and generates a complex immunoreactive neuropile in the anterior and posterior regions of the protocerebrum. Immunoreactive processes are also found in the structured neuropile of the central body complex. Immunoreactive cell bodies are also found in the antennal lobes, in the lateral margins of the protocerebrum, in the optic lobes, and in a few cells in the pars intercerebralis. The results suggest that myomodulin-like neuropeptides may play roles as central neurotransmitters or neuromodulators in insects as well as being released into the circulation as neurohormones or acting as releasing agents for neurohormones in neurohaemal areas. They also further strengthen the idea that myomodulins, which were first identified in molluscs, may represent another interphyletic family of neuropeptides.

Animals↗