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Imaging the spatial distribution of membrane receptors during neutrophil phagocytosis.

Optical microscopy and image processing have been employed to study the distribution of several cell surface receptors on living human neutrophils during opsonin-dependent and opsonin-independent phagocytosis. Receptors were labeled using fluorescein-, rhodamine-, or AMCA-conjugated F(ab')2 fragments of anti-Fc gamma RIIIB (CD16), anti-CR3 (CD11b/CD18), and anti-uPAR (urokinase-type plasminogen activator receptor) antibodies, intact phycoerythrin-labeled interleukin 8, and fluorescein- or rhodamine-labeled Con A (concanavalin A), Boc-PLPLP (tert-butyl-oxycarbonyl-Phe(D)-Leu-Phe(D)-Leu-Phe-OH), and N-formyl-Nle-Leu-Phe-Nle-Tyr-Lys. Labeled neutrophils were observed during the phagocytosis of IgG-opsonized erythrocytes and nonopsonized latex beads, Escherichia coli, and Staphylococcus aureus. To quantitate receptor distribution, cells were divided into four quadrants with the first being the point of attachment and the fourth being opposite the point of attachment. Ligated formyl peptide receptors, and to a lesser extent CR3, accumulated at the sites of target internalization for all forms of phagocytosis examined. However, Fc gamma RIIIB, uPAR, IL-8, Con A, and the FPR antagonist FBoc-PLPLP were not polarized on cells during phagocytosis. These data suggest that agonist-labeled formyl peptide receptors may play a broader role in leukocyte function than previously suggested, including possible participation in phagocytosis.

Amino Acid Sequence↗

Modeling the influence of ectodomain affinities on the spatial distribution of membrane receptors.

We have previously shown that the glycosylphosphatidyl-inositol (GPI)-linked urokinase-type plasminogen activator receptor (uPAR) reversibly associates with the integrins complement receptor type 3 (CR3; alphaMbeta2) and CR4 (alphaxbeta2) during leukocyte motility. These receptor-to-receptor interactions could potentially be accounted for by diffusion-controlled reactions or by directed transport phenomena. To address these alternatives, we have used computer simulation techniques. Our results show that a diffusion-controlled interaction between uPAR and CR4 during accumulation at lamellipodia is not physically reasonable. This suggests that a directed transport mechanism participates in establishing uPAR-integrin association.

Animals↗

Aggregation of individual trees and patches in forest succession models: capturing variability with height structured, random, spatial distributions.

Individual based, stochastic forest patch models have the potential to realistically describe forest dynamics. However, they are mathematically intransparent and need long computing times. We simplified such a forest patch model by aggregating the individual trees on many patches to height-structured tree populations with theoretical random dispersions over the whole simulated forest area. The resulting distribution-based model produced results similar to those of the patch model under a wide range of conditions. We concluded that the height- structured tree dispersion is an adequate population descriptor to capture the stochastic variability in a forest and that the new approach is generally applicable to any patch model. The simplified model required only 4.1% of the computing time needed by the patch model. Hence, this new model type is well-suited for applications where a large number of dynamic forest simulations is required.

Bias↗

The spatial distribution of Borrelia burgdorferi-infected Ixodes ricinus in the Connemara region of County Galway, Ireland.

Studies were carried out in the Connemara area of County Galway in the west of Ireland in order to determine the abundance and distribution of the tick, Ixodes ricinus and the prevalence of its infection with Borrelia burgdorferi. The tick was very abundant locally, in particular when associated with cattle, sheep and enclosed red deer. Large numbers of ticks not only occurred on the pastures, but also on adjacent roadside verges. No infections with B. burgdorferi could be demonstrated when nymphal ticks were sampled from central areas of the pastures, suggesting that livestock and red deer are probably not significant reservoirs of the spirochaete. Small numbers of infected nymphal and adult ticks were associated with hedges, dry stone walls, the margins of woodland adjoining infested pastures and in woodland from which livestock were excluded. Woodmice (Apodemus sylvaticus) were most numerous in such habitats and the majority were infected with B. burgdorferi.

Animals↗

Spatial distribution of cortical proteins in cells of epithelial sheets.

In the differentiated pigmented epithelial cells of the retina (RPE) of chick embryos cytoskeletal proteins are found in polygonal rings located in the cell cortex. Within the cortical rings of the RPE cells vinculin and spectrin occupy a characteristic position closest to the plasma membrane; actin is found farther away, while tropomyosin and myosin are located farthest from the plasma membrane. The differences in the distribution of these proteins might reflect the functional specialization of different parts of the cortical ring required to develop and transmit tension from individual cells throughout the entire epithelial sheet.

Actins↗

Spatial distribution of synapses onto thoracic motor neurones in locusts.

Aggregates of synaptic vesicles, stained black by the zinc iodideosmium procedure, can be visualised with the light microscope in 1 mum plastic sections. This allows the main branches of a neurone to be reconstructed relatively rapidly and the associated vesicle aggregates to be plotted. By resectioning, the identity of the vesicle aggregates has been confirmed with the electron microscope. Two flight motor neurones in the mesothoracic ganglion of the locust have been examined. One is identified as a dorsal longitudinal muscle motor neurone (muscle 112) and the other is probably a subalar neurone (muscle 99). Both have a large density of vesicle aggregates on the neuropilar segment, the widest part of the main neuronal axis, but few on the neurite within 250 mum of the cell body. The larger branches arising from the neuropilar segment tend to have a lower density of aggregates than fine branches, which suggests that synapses to the branches may occur mainly on the distal twigs. These results are an important preliminary step in determining the integrative functions of such neurons and have immediate implications in the interpretation of microelectrode recordings.

Animals↗

Spatial distribution of various types of bulbar respiratory neurons in the rabbit.

In anesthetized rabbits, the burst activity of 277 single respiratory neurons was recorded extracellularly. The neurons were classified according to their spike incidence distribution within the respiratory cycle and to their response to lung distension or collapse (alpha or beta type). About one third of the neurons found in all animals were located at the level of the caudal end of the promontorium gliosum, widely scattered over the bulbar cross-section. More rostrally the units were located within a dorsal area neighbouring the tractus solitarius, more caudally within a ventral field surrounding the nucleus ambiguus. Most of the inspiratory neurons (the most frequently encountered type) were found within an antero-medial region of the medulla, extending down to about two thirds of its depth. Most of the inspiratory-expiratory units were found within an intermediate zone extending from antero-dorsal to postero-ventral. Most of the expiratory cells were found postero-ventrally. Most of the expiratory-inspiratory neurons (the rarest type) were again found within an antero-medial region at an intermediate depth. The alpha neurons were located in the rostral and the beta units in the caudal part of the medulla. The average proportion of respiratory neurons to non-respiratory units was estimated as about 1:36 and the total number of bulbar respiratory neurons as 500 on each side.

Animals↗

Spatial distribution of afferent information to the anterior lobe of the cat's cerebellum.

In the decerebrate cat discharges of Purkinje cells with simple and complex spikes as well as granule cell discharges in the c1-zone of lobules Va, b and c of the cerebellar anterior lobe were analyzed during a passive movement of the cat's forepaw. Penetrations were made 50 micrometers apart along the mediolateral and parasagittal directions, the depth never exceeding 500 micrometers. The response of the Purkinje cells to the climbing fiber input was surprisingly constant, while simple spike responses of the same cells showed a great variability to the same input. The variability between granule cell discharges recorded at a 50 micrometer distance from each other was similar to that of the simple spikes of the Purkinje cell. It is assumed that because of a patchy distribution of excited granule cells, two neighbouring Purkinje cells may receive a different information via their parallel fiber inputs. This difference is considered to be responsible for the great variability of their responses to mossy fiber inputs.

Afferent Pathways↗

Spatial distribution of excitatory innervation on a muscle fiber of the lobster (Homarus americanus).

The distribution of nerve terminals from a single excitatory motor axon has been followed along a fiber from the proximal accessory flexor muscle of the lobster by thin serial sectioning at periodic intervals. The excitatory motor axon provides small axonal branches (diameter 5--12 micron) of varying lengths that travel along the surface of the muscle fiber. Each of these branches gives rise to discrete synaptic terminals which are not uniformly distributed. Individual terminals vary in length from 10--60 micron and the majority possess neuromuscular synapses. The greatest distance between synapses is slightly over 1 mm which is well within the length constant (2.6 mm) of this muscle fiber. Thus the spatail distribution of synapses is such as to ensure adequate depolarization along the entire length of the muscle fiber.

Animals↗

Spatial distribution of individual medial lemniscal axons in the thalamic ventrobasal complex of the cat.

Neurons responding to deep or cutaneous stimuli are situated in different parts of the ventrobasal complex. Within the cutaneous region elongated clusters of cells with common place and modality properties project to single columns in the somatic sensory cortex. The present study sought to determine to what extent single lemniscal axons contribute terminals to different regions and to different cell clusters. Lemniscal axons, anterogradely labelled by horseradish peroxidase injected into the medial lemniscus of cats were examined light and electron microscopically. Labelled axons bore one or two, mainly anteroposteriorly oriented, terminal ramifications. These ramifications were relatively small when compared to the length of the complex. Some of the axons bore one or two collaterals that ascended towards the dorsal edge of the complex and formed an additional small ramification there. Electron microscopic analyses of labelled lemniscal axons provided further evidence to that already available that most of their boutons synapse on proximal dendrites of relay neurons and on presynaptic dendrites, presumably belonging to interneurons. A concurrent study of Golgi-impregnated ventrobasal neurons showed three morphological types all with dendritic fields of similar extent. From measurements of the lemniscal terminal ramifications and the counting of counterstained cells it was calculated that 50-120 neurons may receive input from a single terminal ramification. However, because of the restricted extent of the ramifications, the elongated clusters of cells projecting to a single cortical column probably receive input from multiple lemniscal axons and not all members of the cluster receive inputs from the same axons.

Afferent Pathways↗

Control of DNA replication and spatial distribution of defined DNA sequences in salivary gland cells of Drosophila melanogaster.

In dividing cells, each sequence replicates exactly once in each S-phase, but in cells with polytene chromosomes, some sequences may replicate more than once or fail to replicate during S-phase. Because of this differential replication, the control of replication in polytene cells must have some unusual features. Dennhöfer (1982a) has recently concluded that the total DNA content of the polytene cells of Drosophila salivary glands exactly doubles in each S-phase. This observation, along with previous studies demonstrating satellite underreplication in salivary gland cells, led us to consider the hypothesis that there is a "doubling of DNA" mechanism for the control of DNA replication in polytene cells. With this mechanism, a doubling of DNA content, rather than the replication of each sequence, would signal the end of a cycle of DNA replication. To test this hypothesis, we have reinvestigated the replication of several sequences (satellite, ribosomal, histone and telomere) in salivary gland cells using quantitative in situ hybridization. We find that underreplication of some sequences does occur. In addition we have repeated Dennhöfer's cytophotometric and labeling studies. In contrast to Dennhöfer, we find that the total DNA contents of nonreplicating nuclei do reflect this partial replication, in accord with Rudkin's (1969) result. We conclude that DNA replication in polytene cells is controlled by modifications of the mechanism operating in dividing cells, where control is sequence autonomous, and not by a "doubling of DNA" mechanism. In situ hybridization to unbroken salivary gland nuclei reveals the distribution of specific sequences. As expected, satellite, histone and 5S sequences are usually in a single cluster.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Evidence for non-random spatial distribution of meiotic exchanges in Podospora anserina: comparison between linkage groups 1 and 6.

In Podospora anserina, positive and very efficient chiasma interference is observed. However, its modalities are different for the two linkage groups 1 (LG1) and 6 (LG6) studied here. In the right arm of LG1, two zones exist in which always occurs only one crossing-over. They are formed independently each other. Moreover, the genetic map consists of clusters of genes located near the centromere and at the limit between the two interference zones. It is postulated that this structure of the map results from the localization of crossing-over in the middle of each zone. We suppose that the type of chiasma interference, in Podospora, is a typical one as it is in Drosophila. It seems that both these phenomena are under common genetical control. In the LG6, we observe a weaker positive chiasma interference without crossing-over localization.

Ascomycota↗

Chemical induction of quadruple and octuple chromosomes in Chinese hamster CHO-K1 cells and relationship between their three-dimensional structure and spatial distribution of BrdU-labeled chromatids.

Double endoreduplication of Chinese hamster CHO-K1 cells that exhibited quadruple chromosomes at metaphase was induced by a combination of rotenone and ammonium vanadate treatments. Analysis of sister chromatid differential staining patterns (using 5-bromo-2'-deoxyuridine) revealed that approximately 50% of the quadruple chromosomes did not keep the scheme of "outside replication" of DNA. Based on the ratio of the staining patterns observed, we suggest that the two diplochromosomes forming a quadruple chromosome are held together by a physical link connecting the two original chromatids. Metaphases with octuple chromosomes were also produced by the same treatment. Each chromosome constituting an octuple chromosome was longer and thinner than ordinary metaphase chromosomes. This suggests incomplete chromosome condensation at metaphase. The majority of octuple chromosomes showed the eight constituent chromosomes to be so enmeshed that a planar alignment could not be observed in air-dried preparations.

Aneuploidy↗