Cells Swim Research Articles

Evaluation of women's aging influence on sperm passage inside the fallopian tube using 3D dynamic mechanical modeling.

The fallopian tubes play an important role in human fertility by facilitating the spermatozoa passage to the oocyte as well as later actively facilitating the fertilized oocyte transportation to the uterus cavity. The fallopian tubes undergo changes involving biological, physical, and morphological processes due to women aging, which may impair fertility. Here, we have modelled fallopian tubes of women at different ages and evaluated the chances of normal and pathological sperm cells reaching the fertilization site, the ampulla. By utilizing a unique combination of simulative tools, we implemented dynamic three-dimensional (3D) detailed geometrical models of many normal and pathological sperm cells swimming together in 3D geometrical models of three fallopian tubes associated with different women's age groups. By tracking the sperm cell swim, we found that for all age groups, the number of normal sperm cells in the ampulla is the largest, compared with the pathological sperm cells. On the other hand, the number of normal sperm cells in the fertilization site decreases due to the morphological and mechanical changes that occur in the fallopian tube with age. Moreover, in older ages, the normal sperm cells swim with lower velocities and for shorter distances inside the ampulla toward the ovary. Thus, the changes that the human fallopian tube undergoes due to women's aging have a significant influence on the human sperm cell motility. Our model of sperm cell motility through the fallopian tube in relation to the woman's age morphological changes provides a new scope for the investigation and treatment of diseases and infertility cases associated with aging, as well as a potential personalized medicine tool for evaluating the chances of a natural fertilization per specific features of a man's sperm and a woman's reproductive system.

Reduced surface accumulation of swimming bacteria in viscoelastic polymer fluids

Surface-associated bacterial communities flourish in nature and in the body of animal hosts with abundant macromolecular polymers. It is unclear how the endowed viscoelasticity of polymeric fluids influences bacterial motile behavior in such environments. Here, we combined experiment and theory to study near-surface swimming of flagellated bacteria in viscoelastic polymer fluids. In contrast to the swimming behavior in Newtonian fluids, we discovered that cells swim in less curved trajectories and display reduced near-surface accumulation. Using a theoretical analysis of the non-Newtonian hydrodynamic forces, we demonstrated the existence of a generic lift force acting on a rotating filament near a rigid surface, which arises from the elastic tension generated along curved flow streamlines. This viscoelastic lift force weakens the hydrodynamic interaction between flagellated swimmers and solid surfaces and contributes to a decrease in surface accumulation. Our findings reveal previously unrecognized facets of bacterial transport and surface exploration in polymer-rich environments that are pertinent to diverse microbial processes and may inform the design of artificial microswimmers capable of navigating through complex geometries.

Prediction of Sperm Progression in Three Dimensions Using Rapid Optical Imaging and Dynamic Mechanical Modeling.

We present a multidisciplinary approach for predicting how sperm cells with various morphologies swim in three-dimensions (3D), from milliseconds to much longer time scales at spatial resolutions of less than half a micron. We created the sperm 3D geometry and built a numerical mechanical model using the experimentally acquired dynamic 3D refractive-index profiles of sperm cells swimming in vitro as imaged by high-resolution optical diffraction tomography. By controlling parameters in the model, such as the size and shape of the sperm head and tail, we can then predict how different sperm cells, normal or abnormal, would swim in 3D, in the short or long term. We quantified various 3D structural factor effects on the sperm long-term motility. We found that some abnormal sperm cells swim faster than normal sperm cells, in contrast to the commonly used sperm selection assumption during in vitro fertilization (IVF), according to which sperm cells should mainly be chosen based on their progressive motion. We thus establish a new tool for sperm analysis and male-infertility diagnosis, as well as sperm selection criteria for fertility treatments.

Controlling Cell Motion and Microscale Flow with Polarized Light Fields

We investigate how light polarization affects the motion of photoresponsive algae, Euglena gracilis. In a uniformly polarized field, cells swim approximately perpendicular to the polarization direction and form a nematic state with zero mean velocity. When light polarization varies spatially, cell motion is modulated by local polarization. In such light fields, cells exhibit complex spatial distribution and motion patterns which are controlled by topological properties of the underlying fields; we further show that ordered cell swimming can generate directed transporting fluid flow. Experimental results are quantitatively reproduced by an active Brownian particle model in which particle motion direction is nematically coupled to local light polarization.

Elasto-hydrodynamic interaction of two swimming spermatozoa

Due to its biological importance in the fertilization process, the swimming of a solitary spermatozoon has been investigated intensively. However, the elasto-hydrodynamic interactions between spermatozoa remain unclear, and the collective swimming of cells has not been fully clarified. In this study, we numerically investigated pairwise interactions of sperm cells in terms of fluid and solid mechanics. To describe fluid–structure interactions between sperm cells, we developed a boundary element–finite element coupling method. When two sperm cells swim side-by-side, their swimming speed may increase compared to solitary swimming. On the other hand, when two sperm cells swim in line, the front sperm swims faster, while the rear sperm swims slower. To reproduce the experimentally observed flagellar synchronization, we employed a geometric clutch hypothesis and proposed a curvature-associated wave-propagation model. The elasto-hydrodynamic synchronization of flagella resulted in an increase in the swimming speeds of side-by-side sperm cells of up to 16%, indicating that elasto-hydrodynamic synchronization is beneficial for cells in terms of swimming speed. The results clarify the fluid–structure interactions of flagellar mechanics and are important in understanding the collective swimming of spermatozoa.

Random walk patterns of a soil bacterium in open and confined environments

We used microfluidic tools and high-speed time-lapse microscopy to record trajectories of the soil bacterium Pseudomonas putida in a confined environment with cells swimming in close proximity to a glass-liquid interface. While the general swimming pattern is preserved, when compared to swimming in the bulk fluid, our results show that cells in the presence of two solid boundaries display more frequent reversals in swimming direction and swim faster. Additionally, we observe that run segments are no longer straight and that cells swim on circular trajectories, which can be attributed to the hydrodynamic wall effect. Using the experimentally observed parameters together with a recently presented analytic model for a run-reverse random walker, we obtained additional insight on how the spreading behavior of a cell population is affected under confinement. While on short time scales, the mean square displacement of confined swimmers grows faster as compared to the bulk fluid case, our model predicts that for large times the situation reverses due to the strong increase in effective rotational diffusion.

Paramecium swimming and ciliary beating patterns: a study on four RNA interference mutations.

Paramecium cells swim and feed by beating their thousands of cilia in coordinated patterns. The organization of these patterns and its relationship with cell motility has been the subject of a large body of work, particularly as a model for ciliary beating in human organs where similar organization is seen. However the rapid motion of the cells makes quantitative measurements very challenging. Here we provide detailed measurements of the swimming of Paramecium cells from high-speed video at high magnification, as they move in microfluidic channels. An image analysis protocol allows us to decouple the cell movement from the motion of the cilia, thus allowing us to measure the ciliary beat frequency (CBF) and the spatio-temporal organization into metachronal waves along the cell periphery. Two distinct values of the CBF appear at different regions of the cell: most of the cilia beat in the range of 15 to 45 Hz, while the cilia in the peristomal region beat at almost double the frequency. The body and peristomal CBF display a nearly linear relation with the swimming velocity. Moreover the measurements do not display a measurable correlation between the swimming velocity and the metachronal wave velocity on the cell periphery. These measurements are repeated for four RNAi silenced mutants, where proteins specific to the cilia or to their connection to the cell base are depleted. We find that the mutants whose ciliary structure is affected display similar swimming to the control cells albeit with a reduced efficiency, while the mutations that affect the cilia's anchoring to the cell lead to strongly reduced ability to swim. This reduction in motility can be related to a loss of coordination between the ciliary beating in different parts of the cell.

PAR and UV Effects on Vertical Migration and Photosynthesis in Euglena gracilis†

Recently it was shown that the unicellular flagellate Euglena gracilis changes the sign of gravitaxis from negative to positive upon excessive radiation. This sign change persists in a cell culture for hours even if subsequently transferred to dim light. To test the ecological relevance of this behavior, a vertical column experiment was performed (max. depth 65 cm) to test distribution, photosynthetic efficiency and motility in different horizons of the column (surface, 20, 40 and 65 cm). One column was covered with a UV cut-off filter, which transmits photosynthetically active radiation (PAR) only, the other with a filter which transmits PAR and UV. The columns were irradiated with a solar simulator (PAR 162 W m(-2), UV-A 32.6 W m(-2), UV-B 1.9 W m(-2)). The experiment was conducted for 10 days, normally with a light/dim light cycle of 12 h:12 h, but in some cases the light regime was changed (dim light instead of full radiation). Under irradiation the largest fraction of cells was found at the bottom of the column. The cell density decreased toward the surface. Photosynthetic efficiency, determined with a pulse amplitude modulated fluorometer, was negligible at the surface and increased toward the bottom. While the cell suspension showed a positive gravitaxis at the bottom, the cells in the 40 cm horizon were bimodally oriented (about the same percentage of cells swimming upward and downward, respectively). At 20 cm and at the surface the cells showed negative gravitaxis. Positive gravitaxis was more pronounced in the UV + PAR samples. At the surface and in the 20 and 40 cm horizons photosynthetic efficiency was better in the PAR-only samples than in the PAR + UV samples. At the bottom photosynthetic efficiency was similar in both light treatments. The data suggest that high light reverses gravitaxis of the cells, so that they move downward in the water column. At the bottom the light intensity is lower (attenuation of the water column and self shading of the cells) and the cells recover. After recovery the cells swim upward again until the negative gravitaxis is reversed again.

Microrobotic visual control of motile cells using high-speed tracking system

We propose a visual control system for motile cells. Our goal is to control microorganisms as microscale smart robots for various applications. As a first step, we have developed a visual feedback control system for Paramecium caudatum cells. In order to ensure both detailed measurements and a large working space, lock-on tracking of a free-swimming cell with a high frame rate is essential. In our system, high-speed (1-kHz frame rate) tracking hardware and software are used for the continuous observation of moving cells with high magnification. Cells swim in a chamber, and their positions and other properties are measured and computed in real time. The chamber position is visually controlled automatically to track a specific cell. The cell motion is controlled electrically by utilizing the galvanotaxis (intrinsic reaction to electrical stimulus) of microorganisms. Experimental results for open-loop control (periodic zigzag motion) and closed-loop control (trapping within a small region that is 1 mm wide) demonstrate the possibility of using microorganisms as micromachines.

Sensory transduction of gravitaxis in Euglena gracilis

Euglena gracilis is a single-celled freshwater flagellate which shows, among other movement behaviors, a negative gravitaxis (cells swim upward in the water column) and a pronounced phototaxis. Gravitaxis is most likely an active physiological mechanism, and the influence of various inhibitors on the gravitaxis in Euglena gracilis was tested using the fully automatic image analysis system ECOTOX to elucidate the possible cellular mechanism of gravity perception and signal transduction. Substances such as the phosphodiesterase inhibitors theophylline, IBM-X and forskolin, which increase the intracellular cAMP concentration, enhance the precision of graviorientation of the cells measured shortly after incubation. Indomethacin, which decreases the intracellular cAMP concentration by inhibition of the adenylate cyclase, inhibited graviorientation. All inhibitors of calmodulin metabolism tested reduced the precision of gravitaxis in Euglena gracilis. Heparin and LiCl, which inhibit the IP3-pathway (block IP3 channels or formation of IP3, respectively) had no effect. The potassium channel blockers BaCl2 and CsCl showed an effect on the orientation behavior, while tetraethylammonium (TEA) had no influence. The protein phosphatase inhibitor, ocadaic acid, inhibited graviorientation at low concentrations. The data are discussed with respect to the current working model for gravitaxis in Euglena gracilis

MORPHOLOGICAL AND GENETIC CHARACTERIZATION OF PHAEOCYSTIS CORDATA AND P. JAHNII (PRYMNESIOPHYCEAE), TWO NEW SPECIES FROM THE MEDITERRANEAN SEA

Two new Phaeocystis species recently discovered in the Mediterranean Sea are described using light and electron microscopy, and their systematic position is discussed on the basis of an analysis of their nuclear‐encoded small‐subunit ribosomal RNA gene (SSU rRNA) sequences. Phaeocystis cordata Zingone et Chrétiennot‐Dinet was observed only as flagellated unicells. Cells are heart shaped, with two flagella of slightly unequal length and a short haptonema. The cell body is covered with two layers of thin scales. The outermost layer scales are oval, with a faint radiating pattern, a raised rim, and a modest central knob. The inner‐layer scales are smaller and have a faint radiate pattern and an inflexed rim. Cells swim with their flagella close together, obscuring the haptonema, pushing the cell, and causing it to rotate about its longitudinal axis while moving forward. Phaeocystis jahnii Zingone was isolated as a nonmotile colony. It forms loose aggregates of cells embedded in a mucilaginous, presumably polysaccharide matrix without a definite shape or visible external envelope. The flagellated stage has the features typical of other Phaeocystis species. Cells are rounded in shape and slightly larger than P. cordata. The cell body is covered with extremely thin scales of two different sizes with a very faint radiating pattern toward their margin. Swimming behavior is similar to that of P. cordata, with the flagella in a posterior position as the cells swim. The SSU rRNA sequence analysis indicated that both species are distinct from other cultivated Phaeocystis species sequenced to date. Regions previously identified as specific for the genus Phaeocystis are not found in P. jahnii, and new genus‐specific regions have been identified. P. cordata is more closely related to the colonial species P. globosa, P. antarctica, and P. pouchetii and has branched prior to the divergence of the warm‐water P. globosa species complex from the cold‐water species P. antarctica and P. pouchetii. These results are discussed within a framework ofthe available data on the evolution of the world’s oceans.

Metabolic Oxidative Stress-induced HSP70 Gene Expression Is Mediated through SAPK Pathway: ROLE OF Bcl-2 AND c-Jun NH2-TERMINAL KINASE

In previous reports we demonstrated that glucose deprivation induces metabolic oxidative stress in drug-resistant human breast carcinoma MCF-7/ADR cells (Lee, Y. J., Galoforo, S. S., Berns, c. M., Chen, J. C., Davis, B. H., Swim, J. E., Corry, P. M., and Spitz, D. R. (1998) J. Biol. Chem. 273, 5294-5299). In the study described here, we investigated intracellular responses to metabolic oxidative stress. Northern blots show an increase in the level of HSP70 and HSP28 mRNA in cells exposed to glucose-free medium for 1 h. One- and two-dimensional polyacrylamide gel analyses confirmed that glucose deprivation induced a family of HSPs, particularly an inducible HSP70. Overexpression of bcl-2 suppressed glucose deprivation-induced HSP70 gene expression, heat shock transcription factor-heat shock element binding activity, as well as c-Jun NH2-terminal kinase (JNK1) activation. Expression of a dominant-negative mutant of JNK1 also suppressed glucose deprivation-induced JNK1 activation as well as HSP70 gene expression. Taken together, the stress-activated protein kinase signal transduction pathway is involved in glucose deprivation-induced heat shock gene expression.

Touring machines

Touring machines

Chapter 40 Assay of Chlamydomonas Phototaxis

Publisher Summary Phototaxis depends on slight light-dependent changes in the waveform of the cis and trans flagella, as revealed by step-up and step-down illumination of immobilized cells. Chlamydomonas responds phototactically to blue illumination with a peak response at 503 nm. The response can be either positive (cells swim toward light source) or negative (cells swim away from light source) depending on actinic beam intensity, history of cells' exposure to light, genetic background of cells, and other factors. The assay of phototaxis offers the most stringent test of Chlamydomonas flagellar function currently available, as it measures not only motility but also the ability of the flagella to respond to intracellular signals. This chapter describes a method for analysis of the swimming direction of a population of individual cells in response to light. The method has been particularly useful for the characterization of mutants with defects in their ability to phototax. The chapter also discusses culture and handling of Chlamydomonas cells and phototaxis room setup. Dark-adapted Chlamydomonas are quite light-sensitive. Therefore, the room must be lighttight when the door is closed to prevent biasing of light-dependent swimming. During the phototaxis assay, it is critical that the microscope objective be aligned with an imaginary line running between the outlets of the two fiberoptic cables so that the actinic light enters the field of view strictly from the left or right. There is description of microscopy and video equipment. Dark-field microscopy is preferred because the cells are then brightly illuminated against a dark background. Virtually any moderately sensitive video camera is acceptable except for cameras with noisy signals that lead to thresholding difficulties in the video processor; any monochrome monitor is also acceptable. The general procedure for data acquisition is examined.

Theory and experimental results on gravitational effects on monocellular algae.

The orientation of a body which has an anisotropic distribution of mass and which is suspended in water is biased by gravitational torque, so that the center of gravity lies below the center of buoyancy. Many species of unicellular swimming algae are gravitationally oriented in this manner. Their axis of propulsion is essentially fixed within their bodies, so that when the cells swim, they swim upwards. Gravitaxis is an exotaxis, which requires no sensory processing. Nevertheless, gravity affects the lives of these cells both individually and collectively. For single cells, gravity intervenes in the execution and mechanism of sense-dependent taxes, such as phototaxis, it provides for fail-safe locomotion toward the upper interface of their habitat, the source of light and air, and it may cause up-accumulation. Populations of single cells, swimming in the presence of gravity, are coupled through fluid-mechanical interactions which cause spatial and temporal patterns of fluid convection and cell concentration. These patterns modify the cell's environmental interactions, by facilitating downward migrations of cell populations, by mixing the embedding fluid and its contents, and by providing a collective mechanism for controlling light intensity at the individual cell level. Summarizing, gravity modulates the interaction of algal cells with each other and with their environment.

Hydrodynamic Phenomena in Suspensions of Swimming Microorganisms

Vast numbers of microorganisms are suspended in temperate aqueous environments. Oceans and rivers, puddles and droplets, the fluid interiors of animals, all host an array of splendidly varied creatures. Although their presence is usually not casua1\y obvious, they constitute the major part of the world's biomass. Their population dynamics-replication and decay, accumulation and dispersal-modulates and regulates their own life, the life of the larger creatures that feed on them, and even the climate (Charlson et al 1987). Microorganisms interact with each other and with the world, at length scales that vary upward from the size of an individual, say 10-4 cm, to the dimensions of the entire body of fluid in which they live. Fluid mechanics, in concert with the organisms' behavior, governs the dynamics of many of these interactions. However, we are concerned here with relatively localized, small-scale phenomena. We consider only single­ celled microorganisms which are motile, i .e. self-propelled, and so small that inertial effects can be ignored in describing their locomotion. Examples to be discussed include species of algae, bacteria, and protozoa, not to mention spermatozoa. The trajectories along which individual cells swim are determined by thc