The estimation of the motive force for protoplasmic streaming inNitella

Summary By use of a theoretical model for a section of a cell ofNitella and assumptions regarding the form of the stress/rate of strain relation for the streaming protoplasm, it has been possible to determine possible velocity profiles for the streaming in normal and disturbedNitella cells. A match of velocities from these theoretical studies to those measured in real systems has led to a re-estimation of the motive force and of the viscosity coefficient as well as to a first estimate of the thickness of the layer over which the force must be distributed.These new results show the motive force field to be restricted to a layer of about 0.1m thickness alongside the sol/gel interface (the outside boundary of the streaming layer), the force per unit area of this interface to be about 0.36 Nm^–2 (3.6 dyne cm^–2) and a possible stress/rate of strain relation to be of the form (stress)=(viscosity coefficient) × (rate of strain)1/3.Although this latter relation is similar to that obtained by Kamiya and Kuroda (1965) for isolated protoplasm, their viscosity coefficient is about twelve times the present estimate (0.027 Nm^–2s 1/3) suggesting that the fluidin situ is much less viscous than their isolated material. The estimate for the motive force is about double that of previous workers. (Kamiya andKuroda 1958,Tazawa 1968).     

Measurement of the motive force of the protoplasmic rotation inNitella

Summary The present work is a first attempt at calculating the absolute amount of the motive force responsible for the rotational protoplasmic streaming. The calculation was made on the basis of the conclusion we arrived at previously through the analysis of intracellular velocity distribution, namely, that the active driving mechanism responsible for the rotational streaming is located at the interface between the cortical gel and the outer edge of the endoplasmic layer. The motive force, which is the shifting force generated at this interface, was determined in the internodal cell ofNitella flexilis to be within the range of 1–2 dynes/cm² at room temperature.Supported by a Grant for Fundamental Scientific Research from the Japanese Ministry of Education.     

Measurement of motive force for cytoplasmic streaming in characean internodes

Summary With an attempt to measure the motive force responsible for cytoplasmic streaming in characean internodal cells, the difference between densities of cytoplasm and vacuolar sap was heightened by about 10 times (density of vacuolar sap was made larger than that of cytoplasm) by replacing the natural vacuolar sap ofChara corallina with an artificial one of higher density. Endoplasmic flow contiguous to the peripheral actin cables (peripheral flow of endoplasm) in the centrifugal direction was not influenced at all by the application of centrifugal acceleration up to 1400 g. We thus concluded that the motive force for the peripheral flow should be much larger than 12dyn/cm², a figure more than 10 times larger than that for bulk endop lasmic flow so far reported.Dedicated to Emeritus Professor Noburo Kamiya on the occasion of his 80th birthday     

Endoplasmic filaments generate the motive force for rotational streaming in Nitella

The streaming endoplasm of characean cells has been shown to contain previously unreported endoplasmic filaments along which bending waves are observed under the light microscope using special techniques. The bending waves are similar to those propagated along sperm tails causing propulsion of sperm. In Nitella there is reason to believe that nearly all of the filaments are anchored in the cortex and that their beating propels the endoplasm in which they are suspended. This hypothesis is supported by calculations in which typical and average wave parameters have been inserted into the classical hydrodynamic equations derived for sperm tail bending waves. These calculations come within an order of magnitude of predicting the velocity of streaming and they show that waves of the character described, propagated along an estimated 52 m of endoplasmic filaments per cell, must generate a total motive force per cell within less than an order of magnitude of the forces measured experimentally by others. If we assume that undulating filaments produce the force driving the endoplasm, then the method described for measuring the motive force could lead to a lower than actual value for the motive force, since both centrifugation and vacuolar perfusion would reverse the orientation of some filaments. Observations of the initiation of particle translation in association with the filaments suggest that particle transport and wave propagation, which occur at the same velocity, may both be dependent on the same process. The possibility that some form of contractility provides the motive force for filament flection and particle transport is discussed.     

Videomicroscopy in the study of protoplasmic streaming and cell movement

Summary Various types of cell motility have been observed and analyzed with techniques of increasing sensitivity and sophistication. Photokymography, cinemicrography and laser-Doppler spectroscopy have all made important contributions to our knowledge of cytoplasmic streaming and cell movement.Now videomicroscopy is finding applications in recording and analyzing two different kinds of images. Video intensification microscopy by image intensifiers and silicon intensified target (SIT) video cameras is used to intensify images too dim to be viewed by eye or photographed. On the other hand, video enhanced microscopy uses a less sensitive chalnicon or other vidicon camera with adjustable amplification and offset to enhance the contrast and improve the resolution of microscopes that employ instrumental compensators.Both of these videotechniques have greatly extended the usefulness of the optical microscope: image intensification to brighten dim images and video enhancement to improve the contrast and resolution so that even submicroscopic structures and events can be recorded. These video techniques can both be further extended by a frame memory, with which images can be further enhanced by computer processing. Still to be developed, however, are appropriate methods for automatic tracking of particle motions.     

Fluid-dynamical study of protoplasmic streaming in a plant cell

A mathematical model of protoplasmic streaming in a plant cell such as Nitella and Chara is studied. General rheological equations for the non-Newtonian fluid is derived theoretically, and the boundary value problem for the model is solved. The pattern of motion of cytoplasm in a living cell is obtained, and the rheological property of protoplasm is evaluated in vivo.     

A study of protoplasmic streaming inPhysarum by laser Doppler spectroscopy

Summary Protoplasmic streaming in the slime moldPhysarum polycephalum has been characterized using laser Doppler spectroscopy. Measurement of the spectrum of scattered laser light permits simultaneous determination of the velocities of all particles in the laser beam, with the relative intensity from each particle proportional to its light scattering cross-section. Simple experimental modifications allow the tracking of the oscillations of the streaming velocities. Rhythmic wall contractions can be monitored simultaneously with the flow velocities. Interpretation of the Doppler spectra shows that a small fraction of the particles in the flowing protoplasm are moving with velocities two to four times greater than the characteristic velocities reported by optical microscopy. Transverse velocities in the tubes are nearly as great as the longitudinal velocities. The shape of the Doppler spectrum at the maximum of the oscillation cycle is consistent with a spatial velocity profile which is sharper than parabolic, presumably because of a viscosity gradient from the center to the walls of the plasmodial tubes. The shape of the Doppler spectrum of depolarized scattered light is of approximately the same form. The response of the plasmodium to increased temperature is an increase in the frequency of the velocity oscillations with little change in the magnitude of the velocities. The response of the plasmodium to very high intensities of laser light is to gel at the point of incidence.     

Studies on the velocity distribution of the protoplasmic streaming in the myxomycete plasmodium

Summary It was shown that the velocity distribution of the intracapillary streaming of protoplasm in a plasmodium ofPhysarum polycephalum is the same no matter whether the flow is spontaneous or whether it is induced artificially by external local air pressure applied to the plasmodium. Thus we conclude that the protoplasmic flow in the plasmodium is caused by local difference in endoplasm pressure. The view that the seat of the motive force responsible for the flow is located in the streaming protoplasm itself is untenable for this type of streaming.     

Laser light-scattering analysis of protoplasmic streaming in the slime mold Physarum polycephalum

Laser light scattering is shown to be an effective means of obtaining a rapid, objective assessment of dynamic changes in the intact plasmodium of the myxomycete Physarum polycephalum during bidirectional (shuttle) streaming. The motion of material in a 100 mum diameter region of a plasmodial vein was studied by following changes in the autocorrelation function of the fluctuations in the scattered light intensity. The autocorrelation function was recorded at 10 s intervals and analyzed to follow changes in the flow velocity of protoplasm associated with shuttle streaming. Rhythmic velocity changes and a "beating" pattern of velocity maxima were readily observed. In an attempt to locate the site of underlying structural changes in the vein responsible for the changing pattern of flow, the average scattered intensity was separated into components derived from moving and stationary scatterers. Periodic variations in the light intensity due to stationary scatterers are related to the streaming cycle and indicate the occurrence of important structural changes in the vein walls. Two possible interpretations of the data are offered; one involving gross dynamic changes in vein structure, the other involving the formation, contraction, or breakdown of fibrillar material in the vein wall during the streaming cycle.     

Steady and transient behaviors of protoplasmic streaming in Nitella internodal cell

Steady and transient behaviors of protoplasmic streaming in Nitella internodal cell have been investigated for various temperatures from 30°C to near 0°C. It has been found that steady velocity of the streaming linearly decreases with increasing inverse temperature but its proportionality coefficient changes at ~ 10°C. Velocity distribution, which reflects temporal fluctuations of the protoplasmic streaming, is nonGaussian and its half width becomes larger at higher temperatures. On the other hand, recovery of the protoplasmic streaming, which is observed after stopping the streaming with a current stimulus to the internodal cell, has been found to show more clear sigmoidal time courses at higher temperatures.     

Locomotive mechanism of Physarum plasmodia based on spatiotemporal analysis of protoplasmic streaming

We investigate how an amoeba mechanically moves its own center of gravity using the model organism Physarum plasmodium. Time-dependent velocity fields of protoplasmic streaming over the whole plasmodia were measured with a particle image velocimetry program developed for this work. Combining these data with measurements of the simultaneous movements of the plasmodia revealed a simple physical mechanism of locomotion. The shuttle streaming of the protoplasm was not truly symmetric due to the peristalsis-like movements of the plasmodium. This asymmetry meant that the transport capacity of the stream was not equal in both directions, and a net forward displacement of the center of gravity resulted. The generality of this as a mechanism for amoeboid locomotion is discussed.     

Ca2+ effect on protoplasmic streaming in Nitella internodal cell

Ca²⁺ ion effect on protoplasmic streaming in an internodal cell of Nitella has been investigated for various temperatures. We have found that the protoplasmic streaming at low temperature is remarkably affected by the Ca²⁺ ions in the internodal cell but larger concentrations of the Ca²⁺ ions are needed to suppress the streaming velocity at higher temperatures. These streaming behaviors of the protoplasm, furthermore, have been elucidated on the basis of the reaction equations which take into account ATP hydrolysis due to actin-myosin molecules and inactivity of the molecules due to the Ca²⁺ ions.     

A study of protoplasmic streaming in Nitella by laser Doppler spectroscopy.

Laser light scattered from particles in the streaming protoplasm of a living cell is shifted in frequency by the Doppler effect. The spectrum of the scattered light can be measured and interpreted to infer details of the velocity distribution in the protoplasm. We have developed this approach to study the protoplasmic streaming in the fresh-water alga Nitella. Our results indicate a characteristic flow pattern to which diffusion makes a negligible contribution. No difference in the velocity of particles of different size is indicated. The streaming velocity linearly with temperature with a supraoptimal temperature of 34 degrees C, and the velocity distribution becomes narrower at high temperatures. The protoplasmic streaming can be inhibited by laser light, and this effect has been used to study the photoresponse of the algae. Using beam diameters of about 50 mum, we have shown that the inhibition is very local, becoming minimal at a displacement of about 200 mum in the upstream direction and 400 mum in the downstream direction. Prolonged exposure produces a bleached area free of chloroplasts, which is three orders of magnitude less sensitive to photoinhibition.