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L V Beloussov

Publications and source records attributed to L V Beloussov.

16 recordsLinked to original sources

Tension-dependent collective cell movements in the early gastrula ectoderm of Xenopus laevis embryos.

Ventral ectodermal explants taken from early gastrula embryos of Xenopus laevis were artificially stretched either by two opposite concentrated forces or by a distributed force applied to the internal explant's layer. These modes of stretching reflect different mechanical situations taking place in the normal development. Two main types of kinematic response to the applied tensions were detected. First, by 15 min after the onset of concentrated stretching a substantial proportion of the explant's cells exhibited a concerted movement towards the closest point of the applied stretching force. We define this movement as tensotaxis. Later, under both concentrated and distributed stretching, most of the cell's trajectories became reoriented perpendicular to the stretching force, and the cells started to intercalate between each other, both horizontally and vertically. This was accompanied by extensive elongation of the outer ectodermal cells and reconstruction of cell-cell contacts. The intercalation movements led first to a considerable reduction in the stretch-induced tensions and then to the formation of peculiar bipolar "embryoid" shapes. The type and intensity of the morphomechanical responses did not depend upon the orientation of a stretching force in relation to the embryonic axes. We discuss the interactions of the passive and active components in tension-dependent cell movements and their relations to normal morphogenetic events.

Animals↗

Studies in developmental cytomechanic.

One of the most promising trends in modern developmental and cell biology, recently defined as < >, or < >, is directed towards revealing the role of mechanical stresses, chemomechanical transduction and active stress responses of cells antissues of developing embryos. We review here the results obtained in this field by our research group and compare them with those from other labs. Our studies relate to the buds of hydroid polypes and to amphibian embryos. We describe the space-temporal patterns of mechanical stresses in these species, analyze their morphogenetical role and the tissue responses to the experimental modulations of stress patterns. In hydroid polypes we explore also the molecular events involved in mechanochemical coupling. A model, linking the passive mechanical stresses with the active stress-responses of embryonic tissues is suggested. We consider these investigations as a first approach to a developing embryo as to an < >.

Animals↗

Generative rules for the morphogenesis of epithelial tubes.

A finite elements model imitating the morphogenesis of smoothly curved tubular epithelial rudiments is suggested. It is based upon the experimentally proved assumption of the lateral (tangential) pressure between adjacent epithelial cells. The main idea of the model is that under a non-zero local curvature the lateral cell-cell pressure acquires the radial components which are absent under zero curvature. In the framework of the model we investigate the roles of initial geometry, the different coefficients relating the local curvatures and radial cell shifts, and of visco-elastical cell-cell linkages in the shaping process. We also employ the different temporal regimes (both periodical and constant) of the lateral pressure exerted and the different overall durations of the modelling. As a result, we get a set of biologically realistical shapes, almost all of them belonging to the same basical "trefoiled" archetype. Among the variables explored, shaping was most affected by the changes in visco-elastical coefficients, in the temporal regimes and in the overall duration of the modelling. The model shows that rather complicated and realistical shapes of epithelial rudiments can be obtained without assuming any initial regional differences inside cell layers. The model may be useful for understanding the principles underlying both genetical and epigenetical regulation of the morphogenesis.

Animals↗

Mechanics of animal development.

Morphogenetic movements are active processes created by forces located within the moving cells themselves. As a rule, these forces are generated by cytoskeleton structures (contraction of actin microfilaments organized as subcortical bundles or actine gel) and by the membranes and vacuole mediated processes of osmotic water transport. The intercellular forces lead to contraction and thus give rise to long-range mechanical stresses, mostly tensile ones. Tensile stresses create the regularly space/time arranged fields which remain topologically invariable within certain developmental periods, which then change drastically. The model of epithelial morphogenesis is discussed which postulates the segregation of an initially homogeneous cell sheet to the proportional domains of polarized and tangentially stretched cells as a result of self-organization. Some other models which tend to explain the different kinds of fold formation are also suggested. One of the models implies a simple "curvature increasing rule" which derives a common trefoiled archetype as a fundamental trend of development of an epithelial rudiment.

Amphibians↗

Effects of relaxation of mechanical tensions upon the early morphogenesis of Xenopus laevis embryos.

In Xenopus laevis embryos at the early gastrula stage, circumferential tensions of embryonic ectoderm were relaxed by making sagittal or transversal slits in the ventral parts of embryos and inserting into surgical cuts the sectors of homologous tissue from same-stage embryos. Changes in tensile patterns were controlled by measuring cell surface angles. Immediate decreases in surface cell wall tension as related to transversal wall tension were registered. Within minutes of the operation, the lobopodial activity of the inner ectodermal surface increased. The subsequent gastrulation movements were disturbed, germ layers partially mixed and archenteron reduced. The areas of extensive cell columnarization in the ectoderm of operated embryos were less regularly arranged and were extended much more ventrally than in intact embryos. Ventro-dorsal migration and latero-medial intercalation of mesodermal cells also were suppressed. As the operated embryos developed, we observed increases in the total amount of neural tissue, associated sometimes with duplication and even triplication of neural tubes, duplication of otic vesicles, partial fusion of axial rudiments, suppression of mesodermal segmentation and branching or bending of notochord. In the gravest cases the antero-posterior embryo polarity was disturbed. In some cases we observed the formation of axial rudiments in ventral implants. The role of tensions in determining the patterns of morphogenetic cell movements and in establishing the morphological order of normal development is discussed.

Age Factors↗

The role of external tensions in differentiation of Xenopus laevis embryonic tissues.

Explants extirpated from Xenopus laevis embryos at the early gastrula stage were placed on pieces of hydrophilized latex film which were then either stretched or remained intact. In explants cultivated on the intact films most cells emigrated out of the explants and remained undifferentiated, whereas the explants on the films stretched for 10 min or more developed a normal set of rudiments. In the explants of suprablastoporal zone stretched perpendicularly to the cranio-caudal direction, the axial organs were oriented in the direction of stretching. In the stretched explants, unlike the intact ones, a system of microfilament-associated intercellular contacts was formed within a few minutes.

Animals↗

Model of pattern formation in epithelial morphogenesis.

One of the most universal events in morphogenesis is the formation of domains of morphologically polarized cells in the initially homogeneous epithelial sheets. We investigate the possibility of considering this process as a phenomenon of self-organization which is based upon the following experimentally proven mechanochemical cell properties: (1) a capacity of individual cells for morphological polarization considered as a bistable "all-or-none" transition of a cell from a non-polarized to a polarized state; (2) transmission of this capacity from one cell to another on their contacts; (3) feedback relations between co-operative cell polarization and tangential elastic tensions in a cell sheet: cell polarization increases tangential tensions whereas the latter inhibit further cell polarization. We have constructed a phenomenological model which formally expresses the above properties. Its mathematical description includes but few macroscopic parameters available to experimental investigation and controlled changes. The analysis of the collective dynamic regimes of cell polarization demonstrates that variations of some non-specific parameters leads to spontaneous transition in the morphology of cell layers accompanied by symmetry breaking (Turing's instability). Under these conditions either long-range ordered patterns of cell polarization (including hexagonal cell nets) or non-regular spotted structures can emerge. In the particular case of a sheet having fixed complete dimensions and lacking any external elastic bonds a stable macrostate is created; it corresponds to the sheet's binary subdivision into polarized and non-polarized cell domains of size-invariant proportions. The model conclusions are compared with the morphogenetical processes in sea-urchin development, the morphogenesis of skin derivates and artificially induced budding in hydrozoa.

Animals↗

The action of low-intensity pulsed ultrasound on amphibian embryonic tissues.

Amphibian embryos at different stages of development, and ectomesodermal explants (small pieces of embryonic tissue extirpated from lateral parts of the embryos just after neurulation), have been irradiated with continuous and pulsed ultrasound at a frequency of 0.88 MHz, spatial and temporal average intensities being within the range 0.025-0.1 W cm2. Ultrasound pulse repetition frequencies were varied within 10-1000 Hz, duty factor being constant and equal to 0.5. Destruction of embryonic tissue as an immediate effect, and interruption or distortion of development as delayed (24 hr) effects, were registered. Significant damaging effects of non-cavitational and non-thermal origin and dependent on pulse repetition frequencies, were observed. Highly frequency-dependent resonance-type effects were obtained for irradiated explants. The maximum effect on Rana temporaria explants was obtained at pulse repetition frequencies of 10-20 Hz and on Xenopus laevis explants at 110-130 Hz.

Animals↗

Mechanical stresses and morphological patterns in amphibian embryos.

1. Embryos of Rana temporaria have been dissected and shape alterations of different parts of the embryo, taking place within 1 h of separation, have been studied. Two categories of deformation have been revealed. 2. The first category comprises those deformations which take place immediately after separation. They are insensitive to cooling, cyanide and Cytochalasin B treatment. These deformations, which consist of a shortening of initially elongated cells, are considered to be the passive relaxations of previously established elastic tensil stresses. 3. Deformations of the second category proceed more slowly. They are inhibited by cooling, cyanide and Cytochalasin B treatment, are accompanied by elongation and migration of cells and occasionally lead to rather complex morphodifferentiations of isolated fragments. These processes are considered to be the result of the active work of intracellular contractile systems, either pre-existing or induced de novo. 4. By analysing the arrangement of the passive deformations we have constructed maps of mechanical stresses in embryos from late blastula up to the early tail-bud stage. At several embryonic stages drastic transformations of the stress pattern occur, these transformations being separated by periods during which the pattern of stress distribution remains topologically constant. 5. A correlation between the arrangement of stress lines and the presumptive morphological pattern of the embryo is pointed out. 6. Some possible relations between tensile tissue stresses and active mechanochemical processes are discussed.

Animals↗

Biomechanical feedback in morphogenesis, as exemplified by stretch responses of amphibian embryonic tissues.

We explore the idea that morphogenetical processes may be self-regulated by the biomechanical feedback established between the active stress-generating devices and the passive stresses of stretching and (or) compression, these feedback directed towards hyperrestoration (restoration with overlapping) of the initial stress values. As an example, a stretch-induced behaviour of the pieces of ventral ectoderm of Xenopus laevis early gastrulae is considered. By stretching the explants in 1.3-1.7 times, we induced several active poststretching cell responses, including further autonomous elongation of an explant in the stretch direction and contraction in the perpendicular direction, as well as more complicated shape changes. At the cellular level, these responses were associated with the return of the stretched cells to isodiametrical shapes and with the production of extensive cell protrusions along the stretch direction. As shown by dissections, the stretch-induced tissue tensions were considerably diminished in the poststretching period. The results obtained are discussed within the framework of the hyperrestoration hypothesis.

Animals↗