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51.
Structural analysis of human neutrophil migration: Centriole, microtubule, and microfilament orientation and function during chemotaxis 总被引:41,自引:21,他引:20 下载免费PDF全文
Orientation of nucleus, centriole, microtubules, and microfilaments within human neutrophils in a gradient of chemoattractant (5 percent Escherichia coli endotoxin-activated serum) was evaluated by electron microscopy. Purified neutropils (hypaque-Ficoll) were placed in the upper compartment of chemotactic chambers. Use of small pore (0.45 μm) micropore filters permitted pseudopod penetration, but impeded migration. Under conditions of chemotaxis with activated serum beneath the filter, the neutrophil population oriented at the filter surface with nuclei located away from the stimulus, centrioles and associated radial array of microtubules beneath the nuclei, and microfilament-rich pseudopods penetrating the filter pores. Reversal of the direction of the gradient of the stimulus (activated serum above cells) resulted in a reorientation of internal structure which preceded pseudopod formation toward the activated serum and migration off the filter. Coordinated orientation of the entire neutrophil population did not occur in buffer (random migration) or in a uniform concentration of activated serum (activated random migration). Conditions of activated random migration resulted in increased numbers of cells with locomotory morphology, i.e. cellular asymmetry with linear alignment of nucleus, centriole, microtubule array, and pseudopods. Thus, activated serum increased the number of neutrophils exhibiting locomotory morphology, and a gradient of activated serum induced the alignment of neutrophils such that this locomotory morphology was uniform in the observed neutrophil populayion. In related studies, cytochalasin B and colchicines were used to explore the role of microfilaments and microtubules in the neutrophil orientation and migration response to activated serum. Cytochalasin B (3.0 μg/ml) prevented migration and decreased the microfilaments seen, but allowed normal orientation of neutrophil structures. In an activated serum gradient, colchicines, but not lumicolchicine, decreased the orientation of nuclei and centrioles, and caused a decrease in centriole-associated microtubules in concentrations as low as 10(-8) to 10(-7) M. These colchicines effects were associated with the rounding of cells and impairment of pseudopod formation. The impaired pseudopod formation was characterized by an inability to form pseudopods in the absence of a solid substrate, a formation of narrow pseudopods within a substrate, and a defect in pseudopod orientation in an activated serum gradient. Functional studies of migration showed that colchicines, but not lumicolchicine, minimally decreased activated random migration and markedly inhibited directed migration, but had not effect on random migration. These studies show that, although functioning microfilaments are probably necessary for neutrophil migration, intact microtubules are essential for normal pseudopod formation and orientation, and maximal unidirectional migration during chemotaxis. 相似文献
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Worldwide patterns of mitochondrial DNA differentiation in the harbor seal (Phoca vitulina) 总被引:3,自引:0,他引:3
Stanley HF; Casey S; Carnahan JM; Goodman S; Harwood J; Wayne RK 《Molecular biology and evolution》1996,13(2):368-382
The harbor seal (Phoca vitulina) has one of the broadest geographic
distributions of any pinniped, stretching from the east Baltic, west across
the Atlantic and Pacific Oceans to southern Japan. Although individuals may
travel several hundred kilometers on annual feeding migrations, harbor
seals are generally believed to be philopatric, returning to the same areas
each year to breed. Consequently, seals from different areas are likely to
be genetically differentiated, with levels of genetic divergence increasing
with distance. Differentiation may also be caused by long-standing
topographic barriers such as the polar sea ice. We analyzed samples of 227
harbor seals from 24 localities and defined 34 genotypes based on 435 bp of
control region sequence. Phylogenetic analysis and analysis of molecular
variance showed that populations in the Atlantic and Pacific Oceans and
east and west coast populations of these oceans are significantly
differentiated. Within these four regions, populations that are
geographically farthest apart generally are the most differentiated and
often do not share genotypes or differ in genotype frequency. The average
corrected sequence divergence between populations in the Atlantic and
Pacific Oceans is 3.28% +/- 0.38% and those among populations within each
of these oceans are 0.75% +/- 0.69% and 1.19% +/- 0.65%, respectively. Our
results suggest that harbor seals are regionally philopatric, on the scale
of several hundred kilometers. However, genetic discontinuities may exist,
even between neighboring populations such as those on the Scottish and east
English coasts or the east and west Baltic. The mitochondrial data are
consistent with an ancient isolation of populations in both oceans, due to
the development of polar sea ice. In the Atlantic and Pacific, populations
appear to have been colonized from west to east with the European
populations showing the most recent common ancestry. We suggest the recent
ancestry of European seal populations may reflect recolonization from Ice
Age refugia after the last glaciation.
相似文献
53.
Christian P. Fetzer Janelle Sauvageau Gerd Kock Carsten Berghaus Jan-Amadé Bangert Markus Dicks Rolf Heumann Kai S. Erdmann Raphael Stoll 《Biomolecular NMR assignments》2007,1(2):151-153
Protein tyrosine phosphatase-basophil like (PTP-BL) represents a large multi domain non-transmembrane scaffolding protein
that contains five PDZ domains. Here we report the backbone assignments of the PDZ2/PDZ3 tandem domain of PTP-BL. These assignments
now provide a basis for the detailed structural investigation of the interaction between the PDZ domains 2 and 3 of PTP-BL.
It will lead to a better understanding of the proposed scaffolding function of this tandem domain in multi-protein complexes
assembled by PTB-BL.
Christian P. Fetzer, Janelle Sauvageau and Gerd Kock contributed equally to this work. 相似文献
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David Heinzmann Anna Bangert Anna-Maria Müller Saskia N. I. von Ungern-Sternberg Frederic Emschermann Tanja Sch?nberger Madhumita Chatterjee Andreas F. Mack Karin Klingel Reinhard Kandolf Miroslav Malesevic Oliver Borst Meinrad Gawaz Harald F. Langer Hugo Katus Gunter Fischer Andreas E. May Ziya Kaya Peter Seizer 《PloS one》2015,10(4)
Cyclophilins are a group of highly conserved cytosolic enzymes that have a peptidylprolyl cis/trans isomerase activity. Cyclophilin A (CyPA) can be secreted in the extracellular space by inflammatory cells and upon cell death. The presence of CyPA in patients with non-ischemic cardiomyopathy is associated with poor clinical prognosis. Here, we investigated the inhibition of extracellular CyPA in a mouse model of troponin I-induced autoimmune myocarditis using the strictly extracellular CyPA-inhibitor MM284. Since A/J mice develop severe inflammation and fibrosis after immunization with murine cardiac troponin I (mcTn I), we used this model to analyze the effects of an extracellular CyPA inhibition. As extracellular CyPA-inhibitor we used the recently described CsA-derivate MM284. In vitro studies confirmed that MM284 inhibits CyPA-induced monocytic migration and adhesion. A/J mice immunized with mcTnI were treated with MM284 or vehicle every second day. After 28 days, we found a considerable reduction of myocardial injury and fibrosis. Further analysis revealed a reduced myocardial presence of T-cells and macrophages compared to control treated animals. Whereas MMP-9 expression was reduced significantly by MM284, we observed no significant reduction of inflammatory cytokines such as IL-6 or TNFα. Extracellular CyPA plays an important role in autoimmune myocarditis for myocardial damage and fibrosis. Our data suggest a new pharmacological approach for the treatment of myocardial inflammation and reduction of cardiac fibrosis by inhibition of extracellular CyPA. 相似文献
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Kadioglu A De Filippo K Bangert M Fernandes VE Richards L Jones K Andrew PW Hogg N 《Journal of immunology (Baltimore, Md. : 1950)》2011,186(10):5907-5915
Neutrophils and T cells play an important role in host protection against pulmonary infection caused by Streptococcus pneumoniae. However, the role of the integrins in recruitment of these cells to infected lungs is not well understood. In this study we used the twin approaches of mAb blockade and gene-deficient mice to investigate the relative impact of specific integrins on cellular recruitment and bacterial loads following pneumococcal infection. We find that both Mac-1 (CD11b/CD18) and α(4)β(1) (CD49d/CD29) integrins, but surprisingly not LFA-1 (CD11a/CD18), contribute to two aspects of the response. In terms of recruitment from the circulation into lungs, neutrophils depend on Mac-1 and α(4)β(1), whereas the T cells are entirely dependent on α(4)β(1). Second, immunohistochemistry results indicate that adhesion also plays a role within infected lung tissue itself. There is widespread expression of ICAM-1 within lung tissue. Use of ICAM-1(-/-) mice revealed that neutrophils make use of this Mac-1 ligand, not for lung entry or for migration within lung tissue, but for combating the pneumococcal infection. In contrast to ICAM-1, there is restricted and constitutive expression of the α(4)β(1) ligand, VCAM-1, on the bronchioles, allowing direct access of the leukocytes to the airways via this integrin at an early stage of pneumococcal infection. Therefore, integrins Mac-1 and α(4)β(1) have a pivotal role in prevention of pneumococcal outgrowth during disease both in regulating neutrophil and T cell recruitment into infected lungs and by influencing their behavior within the lung tissue itself. 相似文献
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Lucy J. Hathaway Silvio D. Brugger Brigitte Morand Mathieu Bangert Jeannine U. Rotzetter Christoph Hauser Werner A. Graber Suzanna Gore Aras Kadioglu Kathrin M��hlemann 《PLoS pathogens》2012,8(3)
The polysaccharide capsule of Streptococcus pneumoniae defines over ninety serotypes, which differ in their carriage prevalence and invasiveness for poorly understood reasons. Recently, an inverse correlation between carriage prevalence and oligosaccharide structure of a given capsule has been described. Our previous work suggested a link between serotype and growth in vitro. Here we investigate whether capsule production interferes with growth in vitro and whether this predicts carriage prevalence in vivo. Eighty-one capsule switch mutants were constructed representing nine different serotypes, five of low (4, 7F, 14, 15, 18C) and four of high carriage prevalence (6B, 9V, 19F, 23F). Growth (length of lag phase, maximum optical density) of wildtype strains, nontypeable mutants and capsule switch mutants was studied in nutrient-restricted Lacks medium (MLM) and in rich undefined brain heart infusion broth supplemented with 5% foetal calf serum (BHI+FCS). In MLM growth phenotype depended on, and was transferred with, capsule operon type. Colonization efficiency of mouse nasopharynx also depended on, and was transferred with, capsule operon type. Capsule production interfered with growth, which correlated inversely with serotype-specific carriage prevalence. Serotypes with better growth and higher carriage prevalence produced thicker capsules (by electron microscopy, FITC-dextran exclusion assays and HPLC) than serotypes with delayed growth and low carriage prevalence. However, expression of cpsA, the first capsule gene, (by quantitative RT-PCR) correlated inversely with capsule thickness. Energy spent for capsule production (incorporation of H3-glucose) relative to amount of capsule produced was higher for serotypes with low carriage prevalence. Experiments in BHI+FCS showed overall better bacterial growth and more capsule production than growth in MLM and differences between serotypes were no longer apparent. Production of polysaccharide capsule in S. pneumoniae interferes with growth in nutrient-limiting conditions probably by competition for energy against the central metabolism. Serotype-specific nasopharyngeal carriage prevalence in vivo is predicted by the growth phenotype. 相似文献
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Human subjects were exposed to a concurrent-chain procedure in which amount of reinforcement in the terminal links was varied. The experimental procedure was designed to resemble as closely as possible animal operant procedures: verbal instructions were eliminated, the key-press operant response was shaped, and a “consummatory” response was required to receive reward. In addition to varying amount of reward, three different pairs of initial-link values in the concurrent chain were studied. The human subjects showed undermatching to amount of reinforcement (as do animal subjects). Moreover, the degree of undermatching tended to increase as the values of the initial links increased, consistent with Fantino's delay reduction hypothesis (1977) that choice for a larger reward decreases as the length of the initial link increases. 相似文献