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Migrating Cells (migrating + cell)
Selected AbstractsIn vivo imaging of microglial cell traffickingACTA OPHTHALMOLOGICA, Issue 2009M PAQUES Purpose Microglial cells (MCs) are active sensors of neural tissues that are rapidly mobilized upon disruption of homeostasis. OUr goal was to observe in vivo the migration of MCs, which has not been done yet. Methods Following acute laser damage, the behavior of MCs in the retina of adult Cx3cr1gfp/+ and gfp/gfp mice was observed noninvasively using time-lapse confocal scanning laser ophthalmoscopy. Observation were done at various time-points up to 8 days after laser damage. Results Focal damage elicite prompt migratory response of MCs within 200 to 400 µm around laser burns. This migratory response was preceded in all case by dendritic reorientation. Convergent and nonconvergent migration were observed. Such migratory activity persisted several days after laser damage. At day 8, the microglia network was restored and microglial locomotion had ceased. Conclusion To our knowledge, this is the first observation of microglial locomotion in vivo. A Morphological evidence of microglial activation starts with dendritic reorganization. Migrating cells were only of the dendritic type (i.e. not ameboid). There appears to be a notable heterogeneity in the locomotor response of MCs. MCs within and around scars remain highly motile and mobile several days after laser damage. [source] Neutrophils display biphasic relationship between migration and substrate stiffnessCYTOSKELETON, Issue 6 2009Kimberly M. Stroka Abstract Neutrophils are one type of migrating cell in the body's innate immune system and are the first line of defense against inflammation or infection. While extensive work exists on the effect of adhesive proteins on neutrophil motility, little is known about how neutrophil motility is affected by the mechanical properties of their physical environment. This study investigated the effects of substrate stiffness on the morphology, random motility coefficient, track speed (v), spreading area, and distribution of turning angles of neutrophils during chemokinesis. Human neutrophils were plated onto polyacrylamide gels of varying stiffness, ranging from 3 to 13 kPa, and coated with the extracellular matrix protein fibronectin, and timelapse images were taken with phase contrast microscopy. Our results show a biphasic behavior between neutrophil motility and substrate stiffness, with the optimum stiffness for motility depending on the concentration of fibronectin on the surface of the gel. On 100 ,g/mL fibronectin, the optimum stiffness is 4 kPa (v = 6.9 ± 0.6 ,m/min) while on 10 ,g/mL fibronectin, the optimum stiffness increases to 7 kPa (v = 4.5 ± 2.0 ,m/min). This biphasic behavior most likely arises because neutrophils on soft gels are less adherent, preventing production of traction forces, while neutrophils on stiff gels adhere strongly, resulting in decreased migration. At intermediate stiffness, however, neutrophils can attain optimal motility as a function of extracellular matrix coating. Cell Motil. Cytoskeleton 2009. © 2009 Wiley-Liss, Inc. [source] Regulation of the Neurofibromatosis 2 gene promoter expression during embryonic developmentDEVELOPMENTAL DYNAMICS, Issue 10 2006Elena M. Akhmametyeva Abstract Mutations in the Neurofibromatosis 2 (NF2) gene are associated with predisposition to vestibular schwannomas, spinal schwannomas, meningiomas, and ependymomas. Presently, how NF2 is expressed during embryonic development and in the tissues affected by neurofibromatosis type 2 (NF2) has not been well defined. To examine NF2 expression in vivo, we generated transgenic mice carrying a 2.4-kb NF2 promoter driving ,-galactosidase (,-gal) with a nuclear localization signal. Whole-mount embryo staining revealed that the NF2 promoter directed ,-gal expression as early as embryonic day E5.5. Strong expression was detected at E6.5 in the embryonic ectoderm containing many mitotic cells. ,-gal staining was also found in parts of embryonic endoderm and mesoderm. The ,-gal staining pattern in the embryonic tissues was corroborated by in situ hybridization analysis of endogenous Nf2 RNA expression. Importantly, we observed strong NF2 promoter activity in the developing brain and in sites containing migrating cells including the neural tube closure, branchial arches, dorsal aorta, and paraaortic splanchnopleura. Furthermore, we noted a transient change of NF2 promoter activity during neural crest cell migration. While little ,-gal activity was detected in premigratory neural crest cells at the dorsal ridge region of the neural fold, significant activity was seen in the neural crest cells already migrating away from the dorsal neural tube. In addition, we detected considerable NF2 promoter activity in various NF2-affected tissues such as acoustic ganglion, trigeminal ganglion, spinal ganglia, optic chiasma, the ependymal cell-containing tela choroidea, and the pigmented epithelium of the retina. The NF2 promoter expression pattern during embryogenesis suggests a specific regulation of the NF2 gene during neural crest cell migration and further supports the role of merlin in cell adhesion, motility, and proliferation during development. Developmental Dynamics 235:2771,2785, 2006. © 2006 Wiley-Liss, Inc. [source] Reelin is essential for neuronal migration but not for radial glial elongation in neonatal ferret cortex,DEVELOPMENTAL NEUROBIOLOGY, Issue 5 2008Alisa Schaefer Abstract Numerous functions related to neuronal migration are linked to the glycoprotein reelin. Reelin also elongates radial glia, which are disrupted in mutant reeler mice. Our lab developed a model of cortical dysplasia in ferrets that shares features with the reeler mouse, including impaired migration of neurons into the cerebral cortex and disrupted radial glia. Explants of normal ferret cortex in coculture with dysplastic ferret cortex restore the deficits in this model. To determine if reelin is integral to the repair, we used explants of P0 mouse cortex either of the wild type (WT) or heterozygous (het) for the reelin gene, as well as P0 reeler cortex (not containing reelin), in coculture with organotypic cultures of dysplastic ferret cortex. This arrangement revealed that all types of mouse cortical explants (WT, het, reeler) elongated radial glia in ferret cortical dysplasia, indicating that reelin is not required for proper radial glial morphology. Migration of cells into ferret neocortex, however, did not improve with explants of reeler cortex, but was almost normal after pairing with WT or het explants. We also placed an exogenous source of reelin in ferret cultures at the pial surface to reveal that migrating cells move toward the reelin source in dysplastic cortex; radial glia in these cultures were also improved toward normal. Our results demonstrate that the normotopic position of reelin is important for proper neuronal positioning, and that reelin is capable of elongating radial glial cells but is not the only radialization factor. © 2008 Wiley Periodicals, Inc. Develop Neurobiol, 2008. [source] Self-produced extracellular stimuli modulate the Pseudomonas aeruginosa swarming motility behaviourENVIRONMENTAL MICROBIOLOGY, Issue 10 2007Julien Tremblay Summary Pseudomonas aeruginosa presents three types of motilities: swimming, twitching and swarming. The latter is characterized by rapid and coordinated group movement over a semisolid surface resulting from morphological differentiation and intercellular interactions. A striking feature of P. aeruginosa swarming motility is the formation of migrating tendrils producing colonies with complex fractal-like patterns. Previous studies have shown that normal swarming motility is intimately related to the production of extracellular surface-active molecules: rhamnolipids (RLs), composed of monorhamnolipids (mono-RLs) and dirhamnolipids (di-RLs), and 3-(3-hydroxyalkanoyloxy) alkanoic acids (HAAs). Here, we report that (i) di-RLs attract active swarming cells while HAAs behave as strong repellents, (ii) di-RLs promote and HAAs inhibit tendril formation and migration, (iii) di-RLs and HAAs display different diffusion kinetics on a surface as di-RLs spread faster than HAAs in agar, (iv) di-RLs and HAAs have no effect on swimming cells, suggesting that swarming cells are different from swimming cells not only in morphology but also at the regulatory level and (v) mono-RLs act as wetting agents. We propose a model explaining how HAAs and di-RLs together modulate the behaviour of swarming migrating cells by acting as self-produced negative and positive chemotactic-like stimuli. [source] Ultrastructural characteristics and lectin-binding properties of M cells in the follicle-associated epithelium of chicken caecal tonsilsJOURNAL OF ANATOMY, Issue 4 2000HIROSHI KITAGAWA To clarify the nature of M cells, the detailed ultrastructural characteristics and lectin-binding properties of M cells were investigated in follicle-associated epithelium (FAE) of chicken caecal tonsils. M cells presented various outlines from columnar to dome shaped. Their polymorphism was dependent on the number of harboured intraepithelial migrating cells. The lighter and larger nuclei of M cells were situated at more apical levels in the epithelial lining compared with those of neighbouring microvillous epithelial cells. The microvilli, which were significantly shorter and thicker than those of adjacent microvillous epithelial cells, were sparsely distributed or completely absent on the apical surfaces of M cells. In general, the apical cytoplasm of M cells without microvilli protruded slightly into the intestinal lumen. Numerous small vesicles were often contained in the apical cytoplasm. The numerous small invaginations of the apical and lateral cell surfaces suggested active transportation of luminal substances. No canaliculi existed in the apical cytoplasm of M cells whereas they were often detected in the neighbouring microvillous epithelial cells. A noteworthy finding was the frequent detection of multivesicular bodies in the apical cytoplasm of M cells. These multivesicular bodies suggest some degradation of ingested luminal substances during transcytoplasmic transportation. WGA and 4 other lectins strongly reacted with all epithelial cells except for M cells, this negativity suggesting a means of detecting M cells in chicken caecal tonsils. Three lectins, DSL, ConA and Jacalin, reacted weakly with the glycocalyx on M cells. The positive reactivity might allow chicken M cells to be utilised for specific antigen delivery into the mucosal immune system in some parenteral vaccinations. [source] Glial-guided neuronal migration in P19 embryonal carcinoma stem cell aggregatesJOURNAL OF NEUROSCIENCE RESEARCH, Issue 1 2005Marcelo F. Santiago Abstract During development of the nervous system, neuronal precursors that originated in proliferative regions migrate along radial glial fibers to reach their final destination. P19 embryonal carcinoma (EC) stem cells exposed to retinoic acid (RA) differentiate into neurons, glia, and fibroblast-like cells. In this work, we induced P19 aggregates for 4 days with RA and plated them onto tissue culture dishes coated with poly-L-lysine. Several cells migrated out of and/or extended processes from the aggregates after 24 hr. Some cell processes were morphologically similar to radial glial fibers and stained for glial fibrillar acidic protein (GFAP) and nestin. Large numbers of migrating cells showed characteristics similar to those of bipolar migrating neurons and expressed the neuronal marker microtubule-associated protein 2. Furthermore, scanning electron microscopy analysis revealed an intimate association between the radial fibers and the migrating cells. Therefore, the migration of neuron-like cells on radial glia fibers in differentiated P19 aggregates resembled some of the migration models used thus far to study gliophilic neuronal migration. In addition, HPTLC analysis in this system showed the expression of 9-O-acetyl GD3, a ganglioside that has been associated with neuronal migration. Antibody perturbation assays showed that immunoblockage of 9-O-acetyl GD3 arrested neuronal migration in a reversible manner. In summary, we have characterized a new cell culture model for investigation of glial-guided neuronal migration and have shown that 9-O-acetyl GD3 ganglioside has an important role in this phenomenon. © 2005 Wiley-Liss, Inc. [source] Seeding density modulates migration and morphology of rabbit chondrocytes cultured in collagen gelsBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2009Ali Baradar Khoshfetrat Abstract The cultures of rabbit chondrocytes embedded in collagen gels were conducted to investigate the cell behaviors and consequent architectures of cell aggregation in an early culture phase. The chondrocyte cells seeded at 1.0,×,105 cells/cm3 underwent a transition to spindle-shaped morphology, and formed the loose aggregates with a starburst shape by means of possible migration and gathering. These aggregates accompanied the poor production of collagen type II, while the cells seeded at 1.6,×,106 cells/cm3 exhibited active proliferation to form the dense aggregates rich in collagen type II. Stereoscopic observation was performed at 5 days to define the migrating cells in terms of a morphology-relating parameter of sphericity determined for individual cells in the gels. The frequency of migrating cells decreased with increasing seeding density, while the frequency of dividing cells showed the counter trend. The culture seeded at 1.0,×,105 cells/cm3 gave the migrating cell frequency of 0.25, the value of which was 25 times higher than that at 1.6,×,106 cells/cm3. In addition, the analysis of mRNA expression revealed that the chondrocyte cells seeded at 1.0,×,105 cells/cm3 showed appreciable down-regulation in collagen type II relating to differentiation and up-regulation in matrix metalloproteinases relating to migration, as compared to the cells seeded at 1.6,×,106 cells/cm3. These data supports the morphological analyses concerning the cell migration and aggregate formation in the cultures with varied seeding densities. It is concluded that the seeding density is an important factor to affect the cell behaviors and architecture of aggregates and thereby to modulate the quality of cultured cartilage. © 2008 Wiley Periodicals, Inc. Biotechnol. Bioeng. 2009;102: 294,302. © 2008 Wiley Periodicals, Inc. [source] | ||||||||||