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Application of the hypo-osmotic swelling test to spermatozoa prepared by swim-up and discontinuous Percoll separation.

The quality of spermatozoa prepared by washing and swim-up or by discontinuous Percoll centrifugation, was compared by applying the hypo-osmotic swelling (HOS) test to semen samples from 116 men of infertile couples. The HOS test performed on 95 normal semen samples showed that the percentage of swollen spermatozoa separated by both techniques was significantly higher than in the initial ejaculate (p < 0.001). The percentage of HOS-positive spermatozoa separated by the Percoll gradient technique was significantly higher than that separated by the swim-up technique (p < 0.001). On the contrary, in 21 abnormal semen samples, there was no significant difference in the percentage of spermatozoa which were positive in the HOS test between the Percoll gradient and the swim-up technique (p = 0.44). It is suggested that the Percoll gradient technique appears to be preferable to the swim-up technique when semen parameters are normal, but there is no significant difference between these two technique in abnormal semen.

Adult↗

The zinc-binding site of a class I aminoacyl-tRNA synthetase is a SWIM domain that modulates amino acid binding via the tRNA acceptor arm.

In its tRNA acceptor end binding domain, the glutamyl-tRNA synthetase (GluRS) of Escherichia coli contains one atom of zinc that holds the extremities of a segment (Cys98-x-Cys100-x24-Cys125-x-His127) homologous to the Escherichia coli glutaminyl-tRNA synthetase (GlnRS) loop where a leucine residue stabilizes the peeled-back conformation of tRNAGln acceptor end. We report here that the GluRS zinc-binding region belongs to the novel SWIM domain family characterized by the signature C-x-C-xn-C-x-H (n = 6-25), and predicted to interact with DNA or proteins. In the presence of tRNAGlu, the GluRS C100Y variant has a lower affinity for l-glutamate than the wild-type enzyme, with Km and Kd values increased 12- and 20-fold, respectively. On the other hand, in the absence of tRNAGlu, glutamate binds with the same affinity to the C100Y variant and to wild-type GluRS. In the context of the close structural and mechanistic similarities between GluRS and GlnRS, these results indicate that the GluRS SWIM domain modulates glutamate binding to the active site via its interaction with the tRNAGlu acceptor arm. Phylogenetic analyses indicate that ancestral GluRSs had a strong zinc-binding site in their SWIM domain. Considering that all GluRSs require a cognate tRNA to activate glutamate, and that some of them have different or no putative zinc-binding residues in the corresponding positions, the properties of the C100Y variant suggest that the GluRS SWIM domains evolved to position correctly the tRNA acceptor end in the active site, thereby contributing to the formation of the glutamate binding site.

Amino Acid Sequence↗

Comparison of a discontinuous Percoll gradient method versus a swim-up method: effects on sperm morphology and other semen parameters.

Increasing the proportion of motile and normal spermatozoa is an important objective for assisted reproductive procedures. The aim of this study was to compare the results of a Percoll and swim-up-method, especially on the sperm morphology. The Percoll technique gave a recovery rate of 81.3% of motile spermatozoa against a recovery rate of 28.98% for the swim-up technique. When the percentage of normal spermatozoa was also brought into calculation the recovery rate with the Percoll technique decreased to 62.03% and increased to 31.81% for the swim-up technique. This was due to a decrease in the resultant morphology of the Percoll technique to a median of 8.0% and an increase to 11.5% for the swim-up technique from a median of 10.5% of the original samples.

Centrifugation, Zonal↗

The activity of cerebellar neurones of the decerebrate dogfish Scyliorhinus during spontaneous swimming movements.

Patterns of activity of cerebellar neurones in response to cutaneous stimulation and during spontaneous, swimming-like movements were examined, using microelectrodes, in decerebrate dogfish (Scyliorhinus canicula). Continuous swimming movements, lasting for several hours, were obtained in fish in which the medial longitudinal fasciculus (m.l.f.) was lesioned in the rhombencephalon. Purkinje cells (P cells) and some stellate cells (S cells) were observed to discharge rhythmically, in phase with swimming movements. These units were distributed throughout the cerebellum, but with no apparent somatotopic distribution. After curarization, rhythmic motor discharges could still be recorded from ventral roots and phase locked P cell discharges were recorded from the cerebellum. P cells that discharged rhythmically during active swimming movements, did not do so when the body was oscillated passively during quiescent periods. Cutaneous stimulation evoked burst discharges in many P cells at long latency (ca. 100 ms) both before and after curarization and whether or not a rhythmic motor output was being generated. In rhythmically discharging units, a similar response was obtained when cutaneous stimulation was applied during that part of a cycle when the unit was most or least active. It was concluded that cerebellar neurones discharged in phase with the output of the spinal locomotory rhythm generators and independently of peripheral sensory feed-back.

Action Potentials↗

Rhizobium meliloti swims by unidirectional, intermittent rotation of right-handed flagellar helices.

The 5 to 10 peritrichously inserted complex flagella of Rhizobium meliloti MVII-1 were found to form right-handed flagellar bundles. Bacteria swam at speeds up to 60 microns/s, their random three-dimensional walk consisting of straight runs and quick directional changes (turns) without the vigorous angular motion (tumbling) seen in swimming Escherichia coli cells. Observations of R. meliloti cells tethered by a single flagellar filament revealed that flagellar rotation was exclusively clockwise, interrupted by very brief stops (shorter than 0.1 s), typically every 1 to 2 s. Swimming bacteria responded to chemotactic stimuli by extending their runs, and tethered bacteria responded by prolonged intervals of clockwise rotation. Moreover, the motility tracks of a generally nonchemotactic ("smooth") mutant consisted of long runs without sharp turns, and tethered mutant cells showed continuous clockwise rotation without detectable stops. These observations suggested that the runs of swimming cells correspond to clockwise flagellar rotation, and the turns correspond to the brief rotation stops. We propose that single rotating flagella (depending on their insertion point on the rod-shaped bacterial surface) can reorient a swimming cell whenever the majority of flagellar motors stop.

Chemotaxis↗

Quorum sensing in Yersinia enterocolitica controls swimming and swarming motility.

The Yersinia enterocolitica LuxI homologue YenI directs the synthesis of N-3-(oxohexanoyl)homoserine lactone (3-oxo-C6-HSL) and N-hexanoylhomoserine lactone (C6-HSL). In a Y. enterocolitica yenI mutant, swimming motility is temporally delayed while swarming motility is abolished. Since both swimming and swarming are flagellum dependent, we purified the flagellin protein from the parent and yenI mutant. Electrophoresis revealed that in contrast to the parent strain, the yenI mutant grown for 17 h at 26 degrees C lacked the 45-kDa flagellin protein FleB. Reverse transcription-PCR indicated that while mutation of yenI had no effect on yenR, flhDC (the motility master regulator) or fliA (the flagellar sigma factor) expression, fleB (the flagellin structural gene) was down-regulated. Since 3-oxo-C6-HSL and C6-HSL did not restore swimming or swarming in the yenI mutant, we reexamined the N-acylhomoserine lactone (AHL) profile of Y. enterocolitica. Using AHL biosensors and mass spectrometry, we identified three additional AHLs synthesized via YenI: N-(3-oxodecanoyl)homoserine lactone, N-(3-oxododecanoyl)homoserine lactone (3-oxo-C12-HSL), and N-(3-oxotetradecanoyl)homoserine lactone. However, none of the long-chain AHLs either alone or in combination with the short-chain AHLs restored swarming or swimming in the yenI mutant. By investigating the transport of radiolabeled 3-oxo-C12-HSL and by introducing an AHL biosensor into the yenI mutant we demonstrate that the inability of exogenous AHLs to restore motility to the yenI mutant is not related to a lack of AHL uptake. However, both AHL synthesis and motility were restored by complementation of the yenI mutant with a plasmid-borne copy of yenI.

Bacterial Proteins↗

Effects of chronic swimming training on cardiac sarcolemmal function and composition.

Cardiac contractile function is dependent on the integrity and function of the sarcolemmal membrane. Swimming exercise training is known to increase cardiac contractile performance. The purpose of the present study was to examine whether a swimming exercise program would alter sarcolemmal enzyme activity, ion flux, and composition in rat hearts. After approximately 11 wk of exercise training, cardiac myosin and actomyosin Ca2+-adenosinetriphosphatase (ATPase) activity was significantly higher in exercised rat hearts than in sedentary control rat hearts. Glycogen content was increased in plantaris and gastrocnemius muscles from exercised animals as was succinic dehydrogenase activity in gastrocnemius muscle of exercised rats in comparison to sedentary rat preparations. Sarcolemmal vesicles were isolated from hearts of exercise-trained and control rats. Sarcolemmal Na+-K+-ATPase and K+-p-nitrophenylphosphatase activities, Na+-Ca2+ exchange, and passive Ca2+ binding did not differ between the two groups. ATP-dependent Ca2+ uptake and 5'-nucleotidase activity were elevated in the cardiac sarcolemmal vesicles isolated from exercised animals compared with sedentary control rats. Sarcolemmal phospholipid composition was not altered by the exercise training. Our results demonstrate that swimming training in rats does not affect most parameters of cardiac sarcolemmal function or composition. However, the elevated sarcolemmal Ca2+ pump activity in exercised rats may help to reduce intracellular Ca2+ and augment cardiac relaxation rates. The enhanced 5'-nucleotidase activity may stimulate adenosine production, which could affect myocardial blood flow. The present results further our knowledge on the subcellular response of the heart to swimming training in the rat.

Animals↗

Activity of myotomal motoneurons during fictive swimming in frog embryos.

1. Dye-filled microelectrodes have been used to identify and to examine the electrical activity of spinal cord motoneurons during fictive swimming in amphibian embryos. 2. Impaled neurons all had ventral cell bodies, dorsal, lateral, or ventral dendrites, and most showed either damaged or complete peripheral axons projecting out to the myotomes. It was rarely possible to identify cells by recording 1:1 motor root spikes evoked by intracellular current pulses. 3. During fictive swimming, motoneurons are tonically depolarized, fire 1 spike per swimming cycle, and are inhibited in phase with motor root activity on the opposite side. Motoneurons can also fire synchronously on the two sides, at double the normal swimming frequency. They occasionally also show a pattern of lower frequency alternating activity in which there is a prolonged burst of discharge on each cycle.

Animals↗

Propulsive impulse as a covarying performance measure in the comparison of the kinematics of swimming and jumping in frogs.

Animals have to modulate their locomotor behavior according to changes in external circumstances. The locomotor requirements are expected to be most extreme for species that move through different physical environments, such as water versus land. In this study, we examine the use of the propulsive impulse as a covariate in the comparison of the kinematics of locomotion of a semi-aquatic frog Rana esculenta, across land and through water. We focused on the propulsive phase because it is functionally the most significant phase of the locomotor cycle in both jumping and swimming, and it is also the most comparable. The frog alters the joint angles of its legs in order to adjust its performance (i.e. impulse) within both locomotor modes. The kinematics and this modulation of the propulsive phase differ between the two modes; however, we found that the impulse ranges of swimming and jumping do not fully overlap. Possible explanations for this include larger lateral forces during swimming, a reduced force transmission due to a lower external load during swimming and reduced muscle recruitment due to differences in coordination patterns.

Animals↗

ASSESSMENT OF MAXIMUM SUSTAINABLE SWIMMING PERFORMANCE IN RAINBOW TROUT (ONCORHYNCHUS MYKISS)

Levels of swimming activity in fishes have been divided into three categories on the basis of the time a given speed can be maintained before the onset of fatigue (Beamish, 1978): sustained (more than 200 min), prolonged (20 s to 200 min) and burst swimming (less than 20 s). The locomotory capacity of a given species reflects both its lifestyle and its body form, although definitions of performance may vary. It is generally accepted that only the aerobic ('red') muscle fibres should be active at truly sustainable swimming speeds, i.e. at speeds that can be maintained indefinitely without fatigue. However, the standard laboratory method of evaluating the maximum sustainable swimming speed (Ucrit; Brett, 1964) almost certainly entails the recruitment of at least some of the rapidly fatigable fast glycolytic ('white') fibres at sub-critical speeds and undoubtedly complicates the evaluation of maximal cardiovascular performance. It would therefore be useful to have an objective and reproducible measure of truly sustainable performance that, by definition, relies solely on aerobic muscle activity. Electromyography (EMG) has been used to examine the pattern of white muscle recruitment following thermal acclimation in striped bass, Morine saxatilis (Sisson and Sidell, 1987). We wished to incorporate this method into a study of the acclimatory responses to chronic changes in environmental temperature of the cardiovascular and locomotory systems in rainbow trout (Wilson and Egginton, 1992). The present communication presents results on the cardiovascular performance and blood chemistry, at rest and during maximal aerobic exercise, of rainbow trout acclimated to 11 &deg;C, as a validation of the methodology currently in use with fish acclimated to seasonal temperature extremes (Taylor et al. 1992). Different acclimation temperatures are known to produce compensatory changes in the relative proportions of red and white muscle mass (Sidell and Moerland, 1989). The aim of these continuing investigations is to compare the anatomical, cardiovascular and locomotory limitations to aerobic exercise over the full temperature range of a eurythermal fish species.

Journal Article↗

STUDIES OF TROPICAL TUNA SWIMMING PERFORMANCE IN A LARGE WATER TUNNEL - KINEMATICS

Yellowfin tuna (Thunnus albacares) swimming kinematics was studied in a large water tunnel at controlled swimming velocities (U). Quantified kinematic variables included the tail-beat frequency, stride length (l), caudal amplitude, yaw, the propulsive wavelength, the speed of the propulsive wave (C) and the sweepback angle of the pectoral fins. In general, all variables, except the propulsive wavelength and consequently C, are comparable to values determined for other teleosts. The propulsive wavelength for the tunas (1.23&shy;1.29 L, where L is fork length) is 30&shy;60 % longer than in other cruise-adapted teleosts such as salmonids. The resulting thunniform swimming mode and the morphological and anatomical adaptations associated with the long propulsive wavelength (e.g. fusiform body shape, rigid vertebral column) act to minimize anterior resistance and maximize caudal thrust. The long propulsive wavelength also increases the maximum l which, in concert with the elevated muscle temperatures of tunas, increases their maximum swimming velocity.

Journal Article↗

Physiology and behaviour of free-swimming Atlantic cod (Gadus morhua) facing fluctuating temperature conditions

Atlantic cod (Gadus morhua L.), acclimated to 5 &deg;C, were equipped with ultrasonic transmitters which allowed the continuous monitoring of their vertical movements and heart rate. Fish were then placed in a 125 m3 tower tank in which the various thermal conditions they encounter in their natural environment were reproduced. Physiological and behavioural responses of cod were followed in parallel to the induced environmental changes. The experimental conditions studied in the tower tank were also reproduced in a swimming respirometer, where oxygen consumption and heart rate could be monitored within the activity range of a free-swimming animal. In a homogeneous water column, a rise in temperature induced marked increases in fish swimming activity, heart rate and heart beat-to-beat variability. In a thermally stratified environment, voluntary activity also increased when the thermal structure of the water column was altered, though no temperature-dependent changes in heart rate were observed. In this case, fish avoided the new temperature conditions, exhibiting distinct thermoregulatory behaviour. Stratification of the water column also prompted daily cyclic changes in fish distribution, animals tending to be in deeper and colder water layers during the day and in shallower and warmer layers at night. Respirometry experiments revealed that the thermoregulatory behaviour observed in free-ranging fish was probably driven by the energetic expedient of maintaining the physiological status quo &shy; i.e. avoiding bioenergically costly reacclimation processes. Indeed, acute temperature increases or decreases of 2.5 &deg;C led to marked differences in oxygen consumption, with metabolic rate changes of 15 and 30 %, respectively. The persistent linear relationship between heart rate and oxygen consumption allowed us to estimate, from the heart rate recorded in free-swimming fish, the entire range of metabolic responses that cod underwent voluntarily while experiencing a thermally stratified water column. The most profound metabolic effect, however, was observed with feeding, when oxygen consumption increased by as much as 80 %, resulting in an estimated 90 % reduction in their subsequent scope for activity.

Journal Article↗

Pectoral fin locomotion in the striped surfperch. I. Kinematic effects of swimming speed and body size

Swimming trials at increasing velocity were used to determine the effects of steady swimming speed on pectoral fin kinematics for an ontogenetic series of striped surfperch Embiotoca lateralis, ranging from 6 to 23 cm in standard length (SL). The fin stroke cycle consisted of a propulsive period, the duration of fin abduction and adduction, and a 'refractory' period, during which the fin remained adducted against the body. Pectoral fin-beat frequency (fp) measured as the inverse of the entire stride period, as in past studies, increased curvilinearly with speed. Frequency, calculated as the reciprocal of the propulsive period alone, increased linearly with speed, as shown previously for tail-beat frequency of fishes employing axial undulation. Fin-beat amplitude, measured as the vertical excursion of the pectoral fin tip during abduction, increased over a limited range of low speeds before reaching a plateau at 0.35&shy;0.40 SL. Pectoral fin locomotion was supplemented by intermittent caudal fin undulation as swimming speed increased. At the pectoral&shy;caudal gait transition speed (Up-c), frequency and amplitude attained maxima, suggesting that the fin musculature reached a physiological limit. The effects of body size on swimming kinematics differed according to the method used for expressing speed. At a given absolute speed, small fish used higher stride frequencies and increased frequency at a faster rate than large fish. In contrast, the relationship between fp and length-specific speed (SL s-1) had a greater slope for large fish and crossed that for small fish at high speeds. We recommend that comparisons across size be made using speeds expressed as a percentage of Up-c, at which kinematic variables influencing thrust are size-independent.

Journal Article↗

Environmental acidity and white muscle recruitment during swimming in the brown trout (Salmo trutta)

Electromyographic recordings show that, for adult brown trout swum up to their critical swimming speed (Ucrit) in a flume at neutral pH, white muscle recruitment occurred when speeds approached 1 body length s-1 (BL s-1) and continued to Ucrit (approximately 2 BL s-1) at both winter (5 &deg;C) and summer (15 &deg;C) acclimation temperatures. However, in the majority of fish swum up to Ucrit at sublethal acidic pH, continuous white muscle recruitment did not occur, although all swam above 1 BL s-1. Any observed electrical activity of the white muscle in these individuals was, at best, intermittent. Consequently, the mean Ucrit of these fish was approximately half that of fish swum at neutral pH. In all fish at sublethal pH, red muscle activity was observed for the whole duration of the exercise period, showing that swimming speeds greater than 1 BL s-1 were achieved largely aerobically. Fish that were chased around a tank at sublethal pH appeared lethargic in their escape response, exhibiting little or no burst swimming. Other observed effects of exposure to sublethal pH, which may have affected swimming capacity, included increases in the resting levels of blood and muscle ammonia, reduced muscle glycogen stores and reduced muscle ion concentrations.

Journal Article↗

What a drag it is getting cold: partitioning the physical and physiological effects of temperature on fish swimming

The influence of temperature-induced changes in water viscosity on the swimming performance and kinematics of larval Atlantic herring (Clupea harengus) was examined using high-speed video recording. The physical effects of viscosity were measured separately from the physiological (Q10) effects of temperature by increasing the viscosity using methyl cellulose. Voluntary swimming speeds of large larvae (18.2 mm total length) were characterized by Reynolds numbers based on length (ReL) between 100 and 500 and varied with temperature and viscosity. Speeds of small larvae (9.6 mm) at ReL between 25 and 125 were strongly affected by viscosity, but virtually unaffected by temperature at equal viscosities. Speeds of large larvae were modulated by transverse tail speed. Small (viscosity-dominated) larvae altered both transverse tail speed and tail amplitude to vary their swimming speed. Stride lengths for both sizes of larvae followed predictions for viscous-regime swimming until ReL>450. The combined data suggest that the viscous hydrodynamic regime for larval herring extends to at least ReL=300 and that viscosity could be important up to ReL of approximately 450. Because the physical effects of viscosity supplement the physiological effects of temperature on locomotor performance (when ReL is below approximately 300), indices such as Q10 can greatly overestimate the dependence of physiological processes on temperature, as demonstrated by an example.

Journal Article↗

Effects of sustained swimming on hepatic glucose production of rainbow trout.

The rate of hepatic glucose production (R(a)glucose) was measured by continuous infusions of 6-[(3)H]glucose in live rainbow trout (Oncorhynchus mykiss) before, during and after swimming for 3 h at 1.5 body lengths s(-)(1) in a swim tunnel. Contrary to expectation, we found that sustained swimming causes a 33 % decline in the R(a),(glucose) of trout (from 7.6+/-2.1 to 5.1+/-1.3 (&mgr;)mol kg(-)(1 )min(-)(1), means +/- s.e.m., N=7), even though exercise of the same intensity elicits a two- to fourfold increase in all the mammalian species investigated to date. Measurements of catecholamine levels show that circulating [epinephrine] decreases by 30 % during exercise (from 4.7+/-0.3 to 3.3+/-0.4 nmol l(-)(1), N=8), suggesting that this hormone is partly responsible for controlling the decline in R(a)glucose. The inhibiting effect of swimming on hepatic glucose production persists for at least 1 h after the cessation of exercise. In addition, rainbow trout can maintain a steady blood glucose concentration throughout sustained exercise by closely matching hepatic glucose production with peripheral glucose utilization, even though this species is generally considered to be a poor glucoregulator. This study provides the first continuous measurements of glucose kinetics during the transition from rest to work in an ectotherm and it suggests that circulating glucose is not an important fuel for aerobic locomotion in trout.

Journal Article↗

A fluid-dynamic interpretation of the asymmetric motion of singly flagellated bacteria swimming close to a boundary.

The singly flagellated bacterium, Vibrio alginolyticus, moves forward and backward by alternating the rotational direction of its flagellum. The bacterium has been observed retracing a previous path almost exactly and swimming in a zigzag pattern. In the presence of a boundary, however, the motion changes significantly, to something closer to a circular trajectory. Additionally, when the cell swims close to a wall, the forward and backward speeds differ noticeably. This study details a boundary element model for the motion of a bacterium swimming near a rigid boundary and the results of numerical analyses conducted using this model. The results reveal that bacterium motion is apparently influenced by pitch angle, i.e., the angle between the boundary and the swimming direction, and that forward motion is more stable than backward motion with respect to pitching of the bacterium. From these results, a set of diagrammatic representations have been created that explain the observed asymmetry in trajectory and speed between the forward and backward motions. For forward motion, a cell moving parallel to the boundary will maintain this trajectory. However, for backward motion, the resulting trajectory depends upon whether the bacterium is approaching or departing the boundary. Fluid-dynamic interactions between the flagellum and the boundary vary with cell orientation and cause peculiarities in the resulting trajectories.

Biological Clocks↗

Low flagellar motor torque and high swimming efficiency of Caulobacter crescentus swarmer cells.

We determined the torque of the flagellar motor of Caulobacter crescentus for different motor rotation rates by measuring the rotation rate and swimming speed of the cell body and found it to be remarkably different from that of other bacteria, such as Escherichia coli and Vibrio alginolyticus. The average stall torque of the Caulobacter flagellar motor was approximately 350 pN nm, much smaller than the values of the other bacteria measured. Furthermore, the torque of the motor remained constant in the range of rotation rates up to those of freely swimming cells. In contrast, the torque of a freely swimming cell for V. alginolyticus is typically approximately 20% of the stall torque. We derive from these results that the C. crescentus swarmer cells swim more efficiently than both E. coli and V. alginolyticus. Our findings suggest that C. crescentus is optimally adapted to low nutrient aquatic environments.

Biomechanical Phenomena↗