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A kinetic Ising model for dynamical behavior of anisotropic system--its significance in streaming phenomena.

A time-delayed kinetic Ising model is constructed for a one-dimensional case in order to study dynamical behaviors of systems including anisotropic interactions between elements. Spin-spin interactions are mediated by a unidirectional signal decaying exponentially with space and time. Non-equilibrium order-disorder transition occurs when the frictional coefficient for signal transmission is varied. Then the model is extended by including a macroscopic field which is produced by local fields caused by spins. The macroscopic field also shows a transition at a certain value of the frictional coefficient. This model well reproduces the features of the transition observed in streaming in the stream cell and Nitella cells.

Actins↗

Progress in plasmodial differentiation improves regularity of oscillating contractions in Physarum polycephalum.

Based on the knowledge about subcellular morphogenetic processes in the acellular slime mold Physarum polycephalum, we hypothesized that during differentiation of undifferentiated endoplasm to the highly differentiated complex structure of the contractile apparatus of this organism, the regularity of oscillating contractions must improve. We measured the endogenous contraction automaticity starting from the de novo generation within minutes after sampling small portions of undifferentiated endoplasm. The standard deviation of the normalized period duration of these samples was compared to the respective values of radial contractions of differentiated protoplasmic plasmodial strands. The mean normalized standard deviation in endoplasmic drops was 28.3+/-12.2%. Respective values in protoplasmic strands were 10.0+/-3.7%. The difference between the experimental groups was highly significant (p<<0.0001). We interpret the verification of our hypothesis as an indication that the very regular oscillating contractions in fully differentiated stages of Physarum require the complex structure of the sophisticated contractile apparatus, represented by the circular plasmalemma invagination system of protoplasmic strands, while the regularity is lower in stages, where the differentiation is still in progress. We believe that this is due to deficits in coordination capabilities, which need a directional and spatially oriented protoplasmic streaming as a precondition.

Animals↗

Cytoskeleton-dependent transport of cytoplasmic particles in previtellogenic to mid-vitellogenic ovarian follicles of Drosophila: time-lapse analysis using video-enhanced contrast microscopy.

In Drosophila oogenesis, several morphogenetic determinants and other developmental factors synthesized in the nurse cells have been shown to accumulate in the oocyte during pre- to mid-vitellogenic stages. However, the mechanisms of the involved intercellular transport processes that seem to be rather selective have not been revealed so far. We have investigated in vitro, by means of video-enhanced contrast time-lapse microscopy, the transport of cytoplasmic particles from the nurse cells through ring canals into the oocyte during oogenesis stages 6-10A. At stage 7, we first observed single particles moving into the previtellogenic oocyte. The particle transfer was strictly unidirectional and seemed to be selective, since only some individual particles moved whereas other particles lying in the vicinity of the ring canals were not transported. The observed transport processes were inhibitable with 2,4-dinitrophenol, cytochalasin B or N-ethylmaleimide, but not with microtubule inhibitors. At the beginning of vitellogenesis (stage 8), the selective translocation of particles through the ring canals became faster (up to 130 nm/second) and more frequent (about 1 particle/minute), whereas during mid-vitellogenesis (stages 9-10A) the velocity and the frequency of particle transport decreased again. Following their more or less rectilinear passage through the ring canals, the particles joined a circular stream of cytoplasmic particles in the oocyte. This ooplasmic particle streaming started at stage 6/7 with velocities of about 80 nm/second and some reversals of direction at the beginning. The particle stream in the oocyte was sensitive to colchicine and vinblastine, but not to cytochalasin B, and we presume that it reflects the rearrangement of ooplasmic microtubules described recently by other authors. We propose that during stages 7-10A, a selective transport of particles into the oocyte occurs through the ring canal along a polarized scaffold of cytoskeletal elements in which microfilaments are involved. This transport might be driven by a myosin-like motor molecule. Either attached to, or organized into, such larger particles or organelles, specific mRNAs and proteins might become selectively transported into the oocyte.

2,4-Dinitrophenol↗

Actomyosin-based motility of endoplasmic reticulum and chloroplasts in Vallisneria mesophyll cells.

Intracellular localization and motile behaviour of the endoplasmic reticulum (ER), plastids and mitochondria were studied in living mesophyll cells of Vallisneria using the vital fluorochrome 3,3'-dihexyloxacarbocyanine iodide (DIOC6(3)). In quiescent cells, the ER was composed of a three-dimensional network of tubular and lamellar elements. Chloroplasts were distributed evenly throughout the cell periphery and appeared embedded within the ER network. The ER network was relatively stationary, with the exception of rare motile episodes occurring as movement of tubular ER strands and adjacent areas of the polygonal network in localized areas of the cell. During experimental induction of streaming, most of the lamellar ER elements transformed into tubules and together with the chloroplasts they began to translocate to the anticlinal walls to establish the circular streaming around the circumference of the cell. Microwave-accelerated fixation followed by immunofluorescence revealed an hitherto unknown phase of actin reorganization occurring within the cells and most interestingly at the surface of the chloroplasts during streaming induction. Myosin was localized in an ER-like pattern in quiescent as well as in streaming cells, with bright fluorescent label localized on mitochondria and proplastids. In addition, myosin label appeared on the surface of the chloroplasts, preferentially in streaming mesophyll cells. Motile activities were impeded by the actin-depolymerizing drug cytochalasin D (CD), the thioreagent N-ethylmaleimide (NEM), and thapsigargin, an inhibitor of the ER-Ca(2+)-ATPase. These inhibitors also interfered with the integrity of actin filaments, the intracellular distribution of myosin and calcium-homeostasis, respectively. These effects suggested an obligate association of at least one type of myosin with the membranes of ER and smaller organelles and are consistent with the appearance of another type of myosin on the chloroplast surface upon streaming induction.

Actins↗

Protein phosphorylation regulates actomyosin-driven vesicle movement in cell extracts isolated from the green algae, Chara corallina.

In Characean cells endoplasmic streaming stops upon membrane depolarization accompanied by Ca(2+) entry. We investigated the mechanism of this cessation of endoplasmic streaming by reconstituting the vesicle movement in vitro. In a living cell of Chara corallina, there are a number of vesicles moving along actin cables. Vesicles in the endoplasm squeezed out of the cell into a medium containing Mg-ATP showed directional movements under a dark field microscope. When the extracted endoplasm was treated with 20 nM okadaic acid, vesicles showed only movements like the Brownian motion. When it was treated with 50 nM staurosporine, directional movements of vesicles were activated. These movements were analyzed by image processing of videomicroscopic records. Vesicle movements along F-actin filaments were also observed by merging both images of the same field by dark field microscopy and fluorescence microscopy, indicating that myosin on the vesicle surface was responsible for vesicle movements. We also examined the effects of okadaic acid and staurosporine on in vitro sliding of F-actin on Chara myosin. When Chara myosin was treated with 20 nM okadaic acid in the cell extract, the number of sliding F-actin filaments was greatly reduced. In contrast, it increased when Chara myosin was treated with 50 nM staurosporine. In addition, Chara myosin treated with protein kinase C greatly diminished its motility. These results suggest that inactivation of Chara myosin via its phosphorylation is responsible for cessation of endoplasmic streaming.

Actin Cytoskeleton↗

Cytochalasin B stabilises the sub-cortical actin bundles of Chara against a solution of low ionic strength.

The properties of the sub-cortical actin bundles in the perfused Chara cell model are altered by concentrations of cytochalasin B (CB) which inhibit streaming. This is demonstrated by treating the bundles with intracellularly introduced CB after first using ATP to strip away their associated motile organelles. Such CB-treated bundles are resistant to a solution of low ionic strength which, as judged by light and electron microscopy, extracts untreated bundles. It is therefore concluded that a site of action for CB lies within the actin bundles and that, while the alteration detected may not itself be of direct physiological significance, it may nonetheless indicate the site at which CB inhibits streaming in both the cell model and the intact cell.

Actins↗

A continuum model of contraction waves and protoplasm streaming in strands of Physarum plasmodium.

We present a mathematical model for continuously distributed mechanochemical autooscillations (autowaves) in a protoplasmic strand of Physarum polycephalum. The model is based on a hypothesis of local positive feedback between deformation and contraction of the contractile apparatus. This feedback is mediated through a cell regulatory system whose kinetics involves coupling to mechanical strain. Mathematical analysis and computer simulations have demonstrated that the solutions of the model agree quantitatively with the available experimental data. In particular, hydrodynamic interaction alone, between different sections of the strand via the streaming endoplasm, is capable of inducing the characteristic contractile behavior.

Actins↗

[Heat resistance of 2 types of protoplasmic movement in Physarum polycephalum plasmodia].

The thermostabilities of the "unordered" and shuttle protoplasmic streamings in myxomycete Physarum polycephalum plasmodia was studied. A comparison of these thermostabilities has revealed that the cessation of the former streaming occurs at temperatures higher than those required for arresting the shuttle streaming. The difference between the two types of protoplasmic streamings is better seen in the rate of repair of protoplasmic streaming halted by a 10 minutes heating at 38-41 degrees C. For example, the unordered streaming is restored 2 minutes after heating plasmodia at 39 degrees for 10 min., while the shuttle streaming is resumed in 24 minutes. It is supposed that the two protoplasmic streamings are independent to an appreciable extent, and that the shuttle streaming, being more complex and coordinated, has appeared in the evolution at later stages than the unordered one. The higher heat sensitivity of the shuttle streaming substantiates a view of the lower stability to injury in regulatory mechanisms if compared to the stability of motile mechanisms.

Acclimatization↗

Mutations in the par genes of Caenorhabditis elegans affect cytoplasmic reorganization during the first cell cycle.

A dramatic reorganization of cytoplasm occurs during the first cell cycle in embryos of the nematode, Caenorhabditis elegans. We present here the results of a quantitative study of some of the events during this reorganization in wild-type embryos and in par mutant embryos. The par mutations define a set of genes required for cytoplasmic localization in early embryos. We show that par mutations lead to defects in several events of the reorganization. Mutations in all four of the par genes we studied lead to defects in pseudocleavage and asymmetric redistribution of cortical microfilaments. In addition, some of the par mutations affect streaming of cytoplasm, migration of the pronuclei, and asymmetric shortening of the embryo. We propose that the major function of the par genes might be to orchestrate this initial reorganization of cytoplasm.

Actin Cytoskeleton↗

A mathematically modelled cytogel cortex exhibits periodic Ca++-modulated contraction cycles seen in Physarum shuttle streaming.

If each of many cells of an embryo (or different zones in a single cell) possess identical active cytogel machinery, having the 'right' mechanochemical response properties, then the collective interaction among those identical participants leads automatically to the globally coherent tissue deformations seen in embryogenesis, and to shuttle streaming in the plasmodial slime mould Physarum polycephalum. Biologically plausible, and experimentally verifiable hypotheses are proposed concerning how the tension generated by a strand of cytogel is determined by the deformation it suffers and by the concentration of a contraction trigger chemical, Ca2+, whose kinetics involve coupling to mechanical strain. The consequences of these hypotheses, deduced by solving the appropriate differential equation systems numerically, and displayed in computer-animated films, closely imitate diverse tissue deformation events seen in developing embryos. The same hypotheses on cytogel behaviour are used to model a thick-walled Physarum vein segment, and two such segments are set up to be able to pump endoplasm back and forth between them. Under certain conditions, this model exhibits spontaneous rhythmic mechanochemical oscillations, many features of which correlate well with shuttle streaming in Physarum. Small gradual variations of parameters, presumably under genetic control, are shown to cause abrupt and biologically interesting bifurcations of the qualitative behaviour of the model.

Calcium↗

Ca++ regulation in caffeine-derived microplasmodia of Physarum polycephalum.

Caffeine-derived microplasmodia possess a Ca++-sequestering system which can initiate motility. The experiments presented here suggest that this system is membranous and nonmitochondrial in nature. Therefore, it is proposed that the shuttle streaming in the plasmodium is controlled by the localized release and uptake of free Ca++ from an intracellular storage system analogous to the sarcoplasmic reticulum.

Caffeine↗

Protoplasmic streaming of an internodal cell of Nitella flexilis; its correlation with electric stimulus.

The sudden cessation or sudden decrease in velocity of the protoplasmic streaming of Nitella flexilis is observed whenever an action potential is elicited. The action potential can be generated by an electric stimulus after its refractory period, whether the flow is at a complete standstill or on the way to recovery. The membrane potential is generally decreased more or less when the rate of flow is decreased on application of salts or other agents. There is, however, no parallelism between these two. The membrane potential decreases proportionally with applied voltage of subthreshold intensity, while the rate of flow does not change appreciably. Only on application of a superthreshold voltage does the flow stop suddenly. In one case the rate of flow decreased to half without appreciable decrease in membrane potential. In another case it continued flowing at about one-half rate, although the membrane potential was almost zero. The Q(10) of the rate of flow is about 2, while it is 1.1 to 1.5 for the membrane potential. The sudden cessation of the protoplasmic streaming is supposed to be caused by the temporary formation of certain interlinkages among contractile protein networks in the endoplasm during excitation at the cathodal half of Nitella.

Action Potentials↗