188 research outputs found
Bend propagation in the flagella of migrating human sperm
A pre-requisite for sexual reproduction is successful unification of the male and female gametes; in externally-fertilising echinoderms the male gamete is brought into close proximity to the female gamete through chemotaxis, the associated signalling and flagellar beat changes being elegantly characterised in several species. In the human, sperm traverse a relatively high-viscosity mucus coating the tract surfaces, there being a tantalising possible role for chemotaxis. To understand human sperm migration and guidance, studies must therefore employ similar viscous in vitro environments. High frame rate digital imaging is used for the first time to characterise the flagellar movement of migrating sperm in low and high viscosities. While qualitative features have been reported previously, we show in precise spatial and temporal detail waveform evolution along the flagellum. In low viscosity the flagellum continuously moves out of the focal plane, compromising the measurement of true curvature, nonetheless the presence of torsion can be inferred. In high viscosities curvature can be accurately determined and we show how waves propagate at approximately constant speed. Progressing waves increase in curvature approximately linearly except for a sharper increase over a distance 20-27 m from the head/midpiece junction. Curvature modulation, likely influenced by the outer dense fibres, creates the characteristic waveforms of high viscosity swimming, with remarkably effective cell progression against greatly increased resistance, even in high viscosity liquids. Assessment of motility in physiological viscosities will be essential in future basic research, studies of chemotaxis and novel diagnostics
Beating patterns of filaments in viscoelastic fluids
Many swimming microorganisms, such as bacteria and sperm, use flexible
flagella to move through viscoelastic media in their natural environments. In
this paper we address the effects a viscoelastic fluid has on the motion and
beating patterns of elastic filaments. We treat both a passive filament which
is actuated at one end, and an active filament with bending forces arising from
internal motors distributed along its length. We describe how viscoelasticity
modifies the hydrodynamic forces exerted on the filaments, and how these
modified forces affect the beating patterns. We show how high viscosity of
purely viscous or viscoelastic solutions can lead to the experimentally
observed beating patterns of sperm flagella, in which motion is concentrated at
the distal end of the flagella
The influence of viscosity on the motility and sensory ability of the dinoflagellate Heterocapsa triquetra
Seawater viscosity is influenced by temperature as well as through excretion of exopolymers by some plankton.
We examined the role of viscosity on the movement patterns and sensory abilities of the dinoflagellate Heterocapsa
triquetra, manipulating the viscosity of seawater to simulate a 10+1.58C temperature change. In a second treatment,
we seeded the water with microbeads to examine swimming behaviours in the presence of a mechanical stimulus.
Increased viscosity reduced distances between conspecifics 4.7-fold and increased distances between protists and
microbeads by 3.4-fold. Increased viscosity also affected other aspects of motility, with an overall reduction in swimming speed of 2.0- and 7.0-fold for treatments with and without mechanical stimuli. Higher viscosities were associated with upward vertical migration, in both the presence and absence of microbeads. Cells were highly sensitive to disturbances to the velocity field, by as little as 1.5%, and different approach distances of H. triquetra to conspecifics over mechanical stimuli suggest sensory capacity to distinguish types of particles. Mediation of motility and migratory
behaviours through viscosity implies ramifications for the distribution of protists and their encounters with resources,
predators and conspecifics triggered by events such as temperature changes and phytoplankton bloom events
Physical Aspects of Axonemal Beating and Swimming
We discuss a two-dimensional model for the dynamics of axonemal deformations
driven by internally generated forces of molecular motors. Our model consists
of an elastic filament pair connected by active elements. We derive the dynamic
equations for this system in presence of internal forces. In the limit of small
deformations, a perturbative approach allows us to calculate filament shapes
and the tension profile. We demonstrate that periodic filament motion can be
generated via a self-organization of elastic filaments and molecular motors.
Oscillatory motion and the propagation of bending waves can occur for an
initially non-moving state via an instability termed Hopf bifurcation. Close to
this instability, the behavior of the system is shown to be independent of
microscopic details of the axoneme and the force-generating mechanism. The
oscillation frequency however does depend on properties of the molecular
motors. We calculate the oscillation frequency at the bifurcation point and
show that a large frequency range is accessible by varying the axonemal length
between 1 and 50m. We calculate the velocity of swimming of a flagellum
and discuss the effects of boundary conditions and externally applied forces on
the axonemal oscillations.Comment: 14 pages, 8 figures, REVTE
Human sperm swimming in a high viscosity mucus analogue
Remarkably, mammalian sperm maintain a substantive proportion of their progressive swimming speed within highly viscous fluids, including those of the female reproductive tract. Here, we analyse the digital microscopy of a human sperm swimming in a highly viscous, weakly elastic mucus analogue. We exploit principal component analysis to simplify its flagellar beat pattern, from which boundary element calculations are used to determine the time-dependent flow field around the sperm cell. The sperm flow field is further approximated in terms of regularized point forces, and estimates of the mechanical power consumption are determined, for comparison with analogous low viscosity media studies. This highlights extensive differences in the structure of the flows surrounding human sperm in different media, indicating how the cell-cell and cell- boundary hydrodynamic interactions significantly differ with the physical microenvironment. The regularized point force decomposition also provides cell-level information that may ulti- mately be incorporated into sperm population models. We further observe indications that the core feature in explaining the effectiveness of sperm swimming in high viscosity media is the loss of cell yawing, which is related with a greater density of regularized point force singularities along the axis of symmetry of the flagellar beat to represent the flow field. In turn this implicates a reduction of the wavelength of the distal beat pattern – and hence dynamical wavelength selection of the flagellar beat – as the dominant feature governing the effectiveness of sperm swimming in highly viscous media
Application of a numerical simulation to improve the separation efficiency of a sperm sorter
This paper describes a study in which numerical simulations were applied to improve the separation efficiency of a microfluidic-based sperm sorter. Initially, the motion of 31 sperm were modeled as a sinusoidal wave. The modeled sperm were expected to move while vibrating in the fluid within the microchannel. In this analysis, the number of sperm extracted at the outlet channel and the rate of movement of the highly motile sperm were obtained for a wide range of flow velocities within the microchannel. By varying the channel height, and the width and the position of the sperm-inlet channel, we confirmed that the separation efficiency was highly dependent on the fluid velocity within the channel. These results will be valuable for improving the device configuration, and might help to realize further improvements in efficiency in the future
Probing human sperm metabolism using 13C-magnetic resonance spectroscopy
STUDY QUESTION: Can 13C-Magnetic Resonance Spectroscopy (MRS) of selected metabolites provide useful information about human sperm metabolism and how glycolysis or oxidative phosphorylation are used by different sperm populations? SUMMARY ANSWER: Sperm populations, prepared by density gradient centrifugation (DGC) and incubated with either 13Cu-glucose, 13Cu-fructose or 13C1-pyruvate, showed consistent evidence of metabolism generating principally lactate and more intermittently bicarbonate, and significantly more lactate was produced from 13Cu-glucose by vital or motile sperm recovered from the 40/80% interface compared to those from the pellet, which could not be accounted for by differences in the non-sperm cells present. WHAT IS KNOWN ALREADY: Previous studies have focused on CO2 or other specific metabolite production by human sperm and there remains considerable debate about whether glycolysis and/or oxidative phosphorylation is the more important pathway for ATP production in sperm. STUDY DESIGN SIZE, DURATION: Sperm populations were prepared by DGC and subjected to 13C-MRS to answer the following questions. (i) Is it possible to detect human sperm metabolism of 13C substrates implicated in energy generation? (ii) What are the kinetics of such reactions? (iii) Do different sperm populations (e.g. '80%' pellet sperm and '40%' interface sperm) utilise substrates in the same way? Semen samples from 97 men were used in these experiments; 52 were used in parallel for aims (i) and (ii) and 45 were used for aim (iii). PARTICIPANTS/MATERIALS, SETTING, METHODS: Sperm populations were prepared from ejaculates of healthy men using a Percoll/Phosphate Buffered Saline (PBS) DGC and then incubated with a range of 13C-labelled substrates (13Cu-glucose, 13Cu-fructose, 13C1-pyruvate, 13C1-butyrate, 13C3-lactate, 13C2,4-D-3-hydroxybutyrate, 13C5-L-glutamate, 13C1,2-glycine or 13Cu-galactose) along with penicillin/streptomycin antibiotic at 37°C for 4 hours, 24 hours or over 48 hours for an estimated rate constant. Sperm concentration, vitality and motility were measured and, for a subset of experiments, non-sperm cell concentration was determined. A 9.4T magnetic resonance spectrometer was used to acquire 1D 13C, inverse gated 1H decoupled, MRS spectra. Spectrum processing was carried out using spectrometer software and Matlab scripts to determine peak integrals for each spectrum. MAIN RESULTS AND THE ROLE OF CHANCE: 13Cu-glucose, 13Cu-fructose and 13C1-pyruvate were consistently converted into lactate and, to a lesser extent, bicarbonate. There was a significant correlation between sperm concentration and lactate peak size for 13Cu-glucose and 13Cu-fructose, which was not observed for 13C1-pyruvate. The lactate peak did not correlate with the non-sperm cell concentration up to 6.9 × 106/ml. The concentration of 13Cu-glucose, 13Cu-fructose or 13C1-pyruvate (1.8, 3.6, 7.2 or 14.4 mM) had no influence on the size of the observed lactate peak over a 4 hour incubation. The rate of conversion of 13C1-pyruvate to lactate was approximately three times faster than for 13Cu-glucose or 13Cu-fructose which were not significantly different from each other. After incubating for 4 hours, the utilisation of 13Cu-glucose, 13Cu-fructose or 13C1-pyruvate by sperm from the '40%' interface of the DGC was no different from those from the pellet when normalised to total sperm concentration. However, after normalising by either the vital or motile sperm concentration, there was a significant increase in conversion of 13Cu-glucose to lactate by '40%' interface sperm compared to pellet sperm (Vital = 3.3 ± 0.30 × 106 vs 2.0 ± 0.21 × 106; p = 0.0049; Motile = 7.0 ± 0.75 × 106 vs 4.8 ± 0.13 × 106; p = 0.0032. Mann-Whitney test p < 0.0055 taken as statistically significant). No significant differences were observed for 13Cu-fructose or 13C1-pyruvate. LARGE SCALE DATA: Not applicable. LIMITATIONS REASONS FOR CAUTION: Only 13C labelled metabolites that accumulate to a sufficiently high concentration can be observed by 13C MRS. For this reason, intermediary molecules in the metabolic chain are difficult to observe without trapping the molecule at a particular step using inhibitors. Non-sperm cell concentration was typical of the general population and no link was found between these cells and the magnitude of the 13C-lactate peak. However, it is possible that higher concentrations than the maximum observed (6.9 × 106/ml) may contribute to exogenous substrate metabolism in other experiments. WIDER IMPLICATIONS OF THE FINDINGS: 13C-MRS can provide information on the underlying metabolism of multiple pathways in live sperm. Dysfunction in sperm metabolism, as a result of either impaired enzymes of lack of metabolisable substrate, could be detected in sperm by a non-destructive assay, potentially offering new treatment options to improve overall sperm quality and outcomes for reproduction. STUDY FUNDING AND COMPETING INTEREST(S): This work was supported by the Medical Research Council Grant MR/M010473/1. The authors declare no conflicts of interest
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