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András Czirók

Publications and source records attributed to András Czirók.

13 recordsLinked to original sources

Mesodermal cell displacements during avian gastrulation are due to both individual cell-autonomous and convective tissue movements.

Gastrulation is a fundamental process in early development that results in the formation of three primary germ layers. During avian gastrulation, presumptive mesodermal cells in the dorsal epiblast ingress through a furrow called the primitive streak (PS), and subsequently move away from the PS and form adult tissues. The biophysical mechanisms driving mesodermal cell movements during gastrulation in amniotes, notably warm-blooded embryos, are not understood. Until now, a major challenge has been distinguishing local individual cell-autonomous (active) displacements from convective displacements caused by large-scale (bulk) morphogenetic tissue movements. To address this problem, we used multiscale, time-lapse microscopy and a particle image velocimetry method for computing tissue displacement fields. Immunolabeled fibronectin was used as an in situ marker for quantifying tissue displacements. By imaging fluorescently labeled mesodermal cells and surrounding extracellular matrix simultaneously, we were able to separate directly the active and passive components of cell displacement during gastrulation. Our results reveal the following: (i) Convective tissue motion contributes significantly to total cell displacement and must be subtracted to measure true cell-autonomous displacement; (ii) Cell-autonomous displacement decreases gradually after regression from the PS; and (iii) There is an increasing cranial-to-caudal (head-to-tail) cell-autonomous motility gradient, with caudal cells actively moving away from the PS faster than cranial cells. These studies show that, in some regions of the embryo, total mesodermal cell displacements are mostly due to convective tissue movements; thus, the data have profound implications for understanding cell guidance mechanisms and tissue morphogenesis in warm-blooded embryos.

Animals↗

Humoral and contact interactions in astroglia/stem cell co-cultures in the course of glia-induced neurogenesis.

Astroglial cells support or restrict the migration and differentiation of neural stem cells depending on the developmental stage of the progenitors and the physiological state of the astrocytes. In the present study, we show that astroglial cells instruct noncommitted, immortalized neuroectodermal stem cells to adopt a neuronal fate, while they fail to induce neuronal differentiation of embryonic stem cells under similar culture conditions. Astrocytes induce neuron formation by neuroectodermal progenitors both through direct cell-to-cell contacts and via short-range acting humoral factors. Neuron formation takes place inside compact stem cell assemblies formed 30- 60 h after the onset of glial induction. Statistical analyses of time-lapse microscopic recordings show that direct contacts with astrocytes hinder the migration of neuroectodermal progenitors, while astroglia-derived humoral factors increase their motility. In non-contact co-cultures with astrocytes, altered adhesiveness prevents the separation of frequently colliding neural stem cells. By contrast, in contact co-cultures with astrocytes, the restricted migration on glial surfaces keeps the cell progenies together, resulting in the formation of clonally proliferating stem cell aggregates. The data indicate that in vitro maintained parenchymal astrocytes (1) secrete factors, which initiate neuronal differentiation of neuroectodermal stem cells; and (2) provide a cellular microenvironment where stem cell/stem cell interactions can develop and the sorting out of the future neurons can proceed. In contrast to noncommitted progenitors, postmitotic neuronal precursors leave the stem cell clusters, indicating that astroglial cells selectively support the migration of maturing neurons as well as the elongation of neurites.

Animals↗

Dystroglycan is involved in laminin-1-stimulated motility of Müller glial cells: combined velocity and directionality analysis.

We investigate the role of dystroglycan, a major laminin-1 receptor and central member of the dystrophin-glycoprotein complex, in the laminin-1 induced motility of cultured Muller glial cells. Binding of laminin-1 to dystroglycan was prevented by IIH6, a function-blocking monoclonal antibody against alpha-dystroglycan. As an alternative means of inhibition, we used heparin to mask the dystroglycan binding site of the laminin-1, known to overlap with heparin binding sites. Cell motility was characterized in a two-dimensional motility assay based on computer-controlled videomicroscopy and statistical analysis of cellular trajectories. We obtained data on both the cell velocity and the diffusion index, a measure of direction-changing frequency. Both means of inhibition of dystroglycan function led to a significant decrease in the ability of laminin-1 to stimulate cell migration. At the same time, dystroglycan function does not appear to be involved in laminin-1-dependent increase in process dynamism and direction-changing activity.

Animals↗

Cerebellar granule cells show age-dependent migratory differences in vitro.

Developmental differences between cerebellar granule cells during their migratory period were revealed using dissociated granule cell cultures isolated from 4, 7, or 10 days old (P4, P7, P10) mice. Under all culture conditions, the great majority of cultivated cell populations consisted of those granule cells that had not reach their final destination in the internal granule cell layer (IGL) by the age of isolation. In vitro morphological development and the expression of migratory markers (TAG-1, astrotactin, or EphB2) showed similar characteristics between the cultures. The migration of 1008 granule cells isolated from P4, P7, and P10 cerebella and cultivated under identical conditions were analyzed using statistical methods. In vitro time-lapse videomicroscopy revealed that P4 cells possessed the fastest migratory speed while P10 granule cells retained their migratory activity for the longest time in culture. Cultures obtained from younger postnatal ages showed more random migratory trajectories than P10 cultures. Our observations indicate that despite similar morphological and molecular properties, migratory differences exist in granule cell cultures isolated from different postnatal ages. Therefore, the age of investigation can substantially influence experimental results on the regulation of cell migration.

Age Factors↗

A digital image-based method for computational tissue fate mapping during early avian morphogenesis.

The early stages of vertebrate development, encompassing gastrulation, segmentation, and caudal axis formation, presumably involve large (finite) morphogenetic deformations; however, there are few quantitative biomechanical data available for describing such large-scale or tissue-level deformations in the embryo. In this study, we present a new method for automated computational "tissue fate mapping," by combining a recently developed high-resolution time-lapse digital microscopy system for whole-avian embryo imaging with particle image velocimetry (PIV), a well-established digital image correlation technique for measuring continuum deformations. Tissue fate mapping, as opposed to classical cell fate mapping or other cell tracking methods, is used to track the spatiotemporal trajectories of arbitrary (virtual) tissue material points in various layers of the embryo, which can then be used to calculate finite morphogenetic deformation or strain maps. To illustrate the method, we present representative tissue fate and strain mapping data for normal early-stage quail embryos. These data demonstrate, to our knowledge, for the first time, large tissue-level deformations that are shared between different germ layers in the embryo, suggesting a more global morphogenetic patterning mechanism than had been previously appreciated.

Animals↗

alphavbeta3 integrin-dependent endothelial cell dynamics in vivo.

A major challenge confronting developmental cell biologists is to understand how individual cell behaviors lead to global tissue organization. Taking advantage of an endothelial cell-specific marker and scanning time-lapse microscopy, we have examined the formation of the primary vascular pattern during avian vasculogenesis. Five types of distinguishable endothelial cell motion are observed during formation of a vascular plexus: (1) global tissue deformations that passively convect endothelial cells; (2) vascular drift, a sheet-like medial translocation of the entire vascular plexus; (3) structural rearrangements, such as vascular fusion; (4) individual cell migration along existing endothelial structures; and (5) cell process extension into avascular areas, resulting in new links within the plexus. The last four types of motion are quantified and found to be reduced in the presence of an alphavbeta3 integrin inhibitor. These dynamic cell motility data result in new hypotheses regarding primordial endothelial cell behavior during embryonic vasculogenesis.

Animals↗

Extracellular matrix dynamics during vertebrate axis formation.

The first evidence for the dynamics of in vivo extracellular matrix (ECM) pattern formation during embryogenesis is presented below. Fibrillin 2 filaments were tracked for 12 h throughout the avian intraembryonic mesoderm using automated light microscopy and algorithms of our design. The data show that these ECM filaments have a reproducible morphogenic destiny that is characterized by directed transport. Fibrillin 2 particles initially deposited in the segmental plate mesoderm are translocated along an unexpected trajectory where they eventually polymerize into an intricate scaffold of cables parallel to the anterior-posterior axis. The cables coalesce near the midline before the appearance of the next-formed somite. Moreover, the ECM filaments define global tissue movements with high precision because the filaments act as passive motion tracers. Quantification of individual and collective filament "behaviors" establish fate maps, trajectories, and velocities. These data reveal a caudally propagating traveling wave pattern in the morphogenetic movements of early axis formation. We conjecture that within vertebrate embryos, long-range mechanical tension fields are coupled to both large-scale patterning and local organization of the ECM. Thus, physical forces or stress fields are essential requirements for executing an emergent developmental pattern-in this case, paraxial fibrillin cable assembly.

Animals↗

Dynamic imaging of cell, extracellular matrix, and tissue movements during avian vertebral axis patterning.

Vertebrate axis patterning depends on cell and extracellular matrix (ECM) repositioning and proper cell-ECM interactions. However, there are few in vivo data addressing how large-scale tissue deformations are coordinated with the motion of local cell ensembles or the displacement of ECM constituents. Combining the methods of dynamic imaging and experimental biology allows both cell and ECM fate-mapping to be correlated with ongoing tissue deformations. These fate-mapping studies suggest that the axial ECM components "move" both as a composite meshwork and as autonomous particles, depending on the length scale being examined. Cells are also part of this composite, and subject to passive displacements resulting from tissue deformations. However, in contrast to the ECM, cells are self-propelled. The net result of cell and ECM displacements, along with proper ECM-cell adhesion, is the assembly of new tissue architecture. Data herein show that disruption of normal cell-ECM interactions during axis formation results in developmental abnormalities and a disorganization of the ECM. Our goal in characterizing the global displacement patterns of axial cells and ECM is to provide critical information regarding existing strain fields in the segmental plate and paraxial mesoderm. Deducing the mechanical influences on cell behavior is critical, if we are to understand vertebral axis patterning. Supplementary material for this article is available online at http://www.mrw.interscience.wiley.com/suppmat/1542-975X/suppmat/72/v72.266.html.

Animals↗

Patterning of embryonic blood vessels.

Morphometric methods were developed to characterize the geometry of vascular patterns in avian and murine embryos. By using these methods, we found that networks of blood vessels formed during vasculogenesis share similar geometric properties (i.e., mean blood vessel diameters and avascular space diameters) regardless of developmental stage, location, or species in which they form. We also found that endothelial cell density within a unit area of an embryonic vasculature could be used to accurately distinguish between a small diameter, capillary-like vascular network (low endothelial cell density) and a large diameter, presinusoidal network (high endothelial cell density). Furthermore, we show that endothelial cell size remains constant in small and large diameter vessels, indicating that increased endothelial cell size is not the basis for diversity in vessel diameter. These observations serve as a foundation for future studies seeking to evaluate the effects of agents or genetic mutations on aspects of vasculogenesis.

Allantois↗

Novel approaches for the study of vascular assembly and morphogenesis in avian embryos.

Dynamic imaging of primary capillary bed formation in a warm-blooded embryo now is readily accomplished with the use of modern digital cameras, software, and instrumentation. The precise dynamic behavior of endothelial cells and their emergent vascular patterns are easily recorded and quantified in exquisite detail. As an example, we present data regarding vasculogenesis and vascular remodeling under normal and vascular endothelial growth factor-stimulated conditions, including corresponding computational analyses of endothelial cell behavior. The results show that the quail embryo provides an excellent experimental system to test reagents hypothesized to play a role in vascularization of tumors, engineered tissues, or wound sites.

Animals↗

Laminin-1 increases motility, path-searching, and process dynamism of rat and mouse Muller glial cells in vitro: implication of relationship between cell behavior and formation of retinal morphology.

Spatial correlation was observed between the localization of laminin-1 at the inner limiting membrane (ILM) and extensive Muller glial process arborization in the same area, as demonstrated by immunolabeling of Muller glial processes and laminin-1 in rat retinae in situ. To test if this spatial correlation is due to a functional relationship, we investigated the impact of laminin-1 on the motility of cultured primary rat and mouse retinal Muller glial cells by statistical analysis of computer-controlled videomicroscopic time-lapse images. We demonstrate that laminin-1 increases motility and path-searching activity of Muller cells in vitro and it also enhances the cells' process formation/withdrawal dynamism. The increase in path-searching activity and cell process dynamism indicates that there is a functional relationship between laminin-1 and Muller glial cells presumably involving signaling towards the cytoskeleton. We hypothesize that laminin-1 is involved in process arborization of Muller cells at the vitread border of the retina resulting in the formation of the functional barrier made up of Muller glial endfeet.

Animals↗

Is bioconvection enhancing bacterial growth in quiescent environments?

Bioconvection is an intriguing pattern-forming phenomenon driven by the swimming activity of various aquatic microorganisms. It is generally assumed that bioconvection has a positive effect on the entire microbial population by carrying oxygen into deep layers of non-aerated suspensions. In order to examine the presence of such a biological benefit, we analysed the correlation between bioconvective pattern formation and population growth of several Bacillus subtilis and Bacillus licheniformis strains under non-aerated conditions. Bioconvection is a robust phenomenon, we observed its development in numerous cultures of various strains and growth phases. Nevertheless, evaluation of the data has not revealed detectable positive effects on population growth, questioning the potential biological relevance of bioconvection in natural habitats.

Bacillus subtilis↗

Irradiation and Taxol treatment result in non-monotonous, dose-dependent changes in the motility of glioblastoma cells.

OBJECTIVE: Three-dimensional conformal radiotherapy, stereotactic radiosurgery and concurrent chemoradiotherapy are among the most important postoperative therapeutic measures in the treatment of malignant gliomas. We investigated in vitro how these modalities affect cell motility, a key factor in tumor invasiveness and malignancy. METHODS: A highly motile glioblastoma cell line was exposed to clinically relevant (2-20 Gy) radiation doses. Some cultures were also subjected to radiosensitizing treatment, in which 5 and 10 nM Taxol was added to the medium for 2 h before the irradiation. The surviving cell fraction was continuously monitored during a 3 day-long time period using an automatized scanning videomicroscope system. Cell motility on a two-dimensional substrate was analyzed by following a large population of cells in each culture. Average velocities, their distribution within the population and persistence of migration were calculated from the cell trajectories. RESULTS: Irradiation increases both the persistence of migration and the heterogeneity of the cell population. Moreover, it results in a non-monotonous alteration of cell motility: While > 10 Gy doses impair motion, exposure to 2 Gy increases velocities by 20%. Taxol treatment reduced the motility of irradiated cells, while slightly increased the velocities of non-irradiated cells. We thus show that - at least for certain glioblastoma cells - both irradiation and Taxol treatment can substantially and synergistically influence cell motility. CONCLUSIONS: High grade gliomas are characterized by bad prognosis and poor response to therapy. The unexpected motogenic effect of low-dose radiation and paclitaxel treatments highlight the importance of similar investigations to develop more effective clinical treatments.

Actins↗