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Biomedical subjects

R Rikmenspoel

Publications and source records attributed to R Rikmenspoel.

At least 19 recordsLinked to original sources

Algebraic expressions for the waveforms of sea urchin sperm flagella.

The waveforms of live sea urchin sperm flagella were digitized with video-digitizing apparatus. The flagellar waveforms were expressed by the coordinates of 20 points spaced 2 micron apart along the flagella. The waveforms were condensed into simple algebraic expression with four parameters. Each of these parameters showed a systematic variation with the flagellar frequency. These average trends of the parameters made it possible to define an average, idealized, waveshape as a function of the flagella frequency, in the range 8-80 Hz.

Animals↗

Digitized precision measurements of the movements of sea urchin sperm flagella.

High speed cinemicrographs were made of sea urchin sperm at temperatures varying from 22 to 6 degrees C. Apparatus, combining a television camera and a video digitizer, was constructed to scan individual flagellar images and to digitize the flagellar waveforms. With appropriate smoothing and averaging procedures, the rough data were condensed by a microcomputer into the coordinates of 20 points along a flagellum, spaced 2 microns apart. The curvature of the flagellum at these points was also computed. The coordinates of the flagellar positions were obtained to an accuracy of approximately +/- 0.1 micron, flagellar curvature to an accuracy of approximately +/- 50 cm-1. At all temperatures the amplitude of the flagella was found to vary with time in a purely sinusoidal fashion to within +/- 2%. The local curvature of the flagella had basically a purely sinusoidal time course to within +/- 50 cm-1, but a varying amount of asymmetry was present in the distal and the proximal ends of the flagella. This asymmetry in the curvature was related to the radius of the circular path of the sperm. The flagellar waveforms can probably be summarized in simple algebraic functions.

Animals↗

Time course of the motion of bull sperm flagella.

Detailed measurements were made of the time course of the motion of bull spermatozoa. Fourier analysis of the data showed the time course to be basically sinusoidal within 2% to 3%. An asymmetry in the motion was present, resulting in a second harmonic component in the Fourier spectra of normal sperm of approximately 11% of the main component. When the energy metabolism of the sperm was inhibited or when the external viscosity of the medium was raised, the asymmetry was reduced. When the internal Mg2+ content of the sperm was lowered, the asymmetry was increased. The asymmetries and the corresponding second harmonic components in the Fourier spectra were correlated with the overall bend shape of the sperm and with the curvature of the path in which the sperm were swimming. Model calculations showed that the asymmetry could reside in either the internal active moments in the sperms or in the stiffness of the sperm flagella.

Animals↗

Movements and active moments of bull sperm flagella as a function of temperature and viscosity.

Detailed measurements were made of the movements and waveforms of bull sperm flagella over a temperature range of 5 to 37 degrees C and over a range of external viscosities of 1-3600 X 10(-3) Pas-1. The data obtained were used for an evaluation of the internal active moments in the flagella, using exact, large amplitude, algebra. The results of the calculations show that the axoneme in bull sperm flagella can produce the forces necessary to power the flagellar motion over the entire range of conditions. The calculations further suggest that no other structure in the bull sperm flagella has a force-producing function.

Animals↗

Effects of infrared laser damage to the Euglena photoreceptor on the control of flagellar motility.

When the area of the stigma of Euglena was irradiated with an infrared laser beam at a dose too low to cause permanent loss of motility, a reduction in flagellar motility was observed only when the external medium contained less than 1 mM Mg2+. At these low Mg2+ concentrations, the laser caused a decrease in flagellar frequency and a tendency for the flagellar waveform to shift towards that taken during reversed swimming. This suggests that the effect of the laser irradiation was to deplete the cells of Mg2+. After the laser pulse the reversal response remained sensitive to the wavelength of the illuminating light. In white light (420-700 nm) 60% of the Euglena showed a reversed waveform; in orange light (530-700 nm) this increased to 90%. This shows that the photoreceptor was not destroyed by the laser irradiation. These experiments were performed on cells that had been impaled on a microelectrode. If direct electric current was passed into the laser-irradiated cells, the current necessary to cause flagellar arrest was 2 to 4 times less than that for cells not laser irradiated. It is concluded that an internal Mg2+ store is present in the Euglena, localized in the area of the paraflagellar swelling; and that the laser irradiation eliminates this Mg2+ store, but at the power used it does not destroy the ability of the stigma-paraflagella to control the flagellar activity.

Adenosine Triphosphate↗

Effects of vanadate, Mg2+ and electric current injection on the stiffness of impaled bull spermatozoa.

Measurements were made of the rigor stiffness of impaled bull sperm flagella. Vanadate-free ATP was found to have a stronger plasticizing action than vanadate-contaminated ATP. Trace amounts of Mg2+ were found to be necessary to relieve the rigor condition by ATP. During the injection of negative electric current into the sperm head, the flagella stiffened, presumably by depletion of Mg2+ from the flagellar interior.

Adenosine Triphosphate↗

Motion characteristics of flagellar fragments of long insect sperm.

The flagellar length of cricket spermatozoa was reduced in steps from congruent to 1,000 micrometer (intact length) to 50 micrometer. In intact sperm the flagellar wave properties were largely independent of the viscosity of the external medium. When the flagellar length had been reduced to less than 100 micrometer the flagellar frequency was reduced at a raised external viscosity. Independent motion of different sections of a flagellum was not observed when its length is less than 100 micrometer. It is concluded that in long thin flagella, transverse viscous forces cannot exert a moment beyond a lever length of approximately 100 micrometer. It is shown that the existence of a maximum lever length, beyond which no moment can be transmitted, leads to the absence of a standing active contractile moment in the long insect sperm.

Animals↗

Effects of Mg2+ and Ca2+ on photoinduced Euglena flagellar responses.

The flagellar frequency and waveform of Euglena were analyzed under full illumination (420-700 nm) and in a restricted wavelength band (530-700 nm) when the cells were in a medium containing Mg2+ or had been microinjected with Mg2+, Mn2+, or Ca2+ in solution. Magnesium abolished the change in flagellar frequency and the reversal in waveform that cells exhibit when illuminated by a 530-700 nm wavelength band. Under this restricted illumination, Ca2+ caused an increase in flagellar waveform reversal and a decrease in beating frequency. The flagellar motility of cells impaled on a microelectrode was examined in cells illuminated with various wavelengths.

Animals↗

A selective effect of Ni2+ on wave initiation in bull sperm flagella.

Bull sperm that are extracted with 0.1% Triton X-100 and restored to motility with Mg2+-ATP lose coordination and stop swimming in the presence of 0.5 mM NiSO4. Although spontaneous coordination of flagellar waves is lost after exposure to Ni2+, other functions of the flagellum remain intact. The capacity for wave propagation along the flagellum is maintained together with the capacity for microtubular sliding. Wave motility can be restored to Ni2+-inhibited sperm by inducing a permanent bend onto the flagellum by micromanipulation. In the absence of such intervention, the loss of wave coordination is complete and irreversible. Ni2+-inhibited demembranated cells that are kept active by maintaining a bend in the flagellum exhibit a normal beat frequency. Both intact and demembranated sperm can retain spontaneous wave production at considerably slower rates of motion than Ni2+-inhibited cells. Short segments from the distal tip of the flagellum contain only the 9 + 2 microtubular axoneme. These short segments are able to propagate imposed bends even in the presence of Ni2+. In addition to wave propagation Ni2+-treated sperm can be shown to exhibit a normal sliding tubule phenomenon by direct assay. Although Ni2+-treated cells have a functional sliding tubule mechanism, and consequently the axoneme can propagate bends, it appears that these retained functions are not sufficient to cause spontaneous bend initiation. Our findings show that bend initiation is inhibited by Ni2+, and therefore is an independent process separate from the sliding tubule mechanism responsible for wave propagation.

Animals↗

Electrical control of flagellar activity in impaled bull spermatozoa.

The control of bull spermatozoon flagellar activity has been investigated using direct current injection into the cells through an impaling glass microelectrode. Negative current injection results in a decrease in the flagellar frequency. Flagellar frequencies can be decreased to zero with high negative currents. This current injection response is dependent on the magnesium concentration available to the spermatozoon interior. The current injection response is nearly independent of ATP concentrations. Resistance measurements indicate that the current injection pathway has a resistance of about 200 +/- 300 k omega, and that the current flowing through the cell membrane is not exceedingly large. Measurements of the induced potentials indicate transmembrane potentials during current injection of about -35 +/- 30 mV per microA of injected current. The results are compatible with an active transport process in bull spermatozoa that controls the flagellar activity in response to current injection by decreasing the internal Mg2+ concentrations during the injection of current.

Action Potentials↗

The equation of motion for sperm flagella.

The equation of motion for sperm flagella, in which the elastic bending moment and the active contractile moment are balanced by the moment from the viscous resistance of the surrounding fluid, is solved for a wave solution that superimposes partial solutions. Substitution of the expression for the wave solution into the equation leads to an expression for the active contractile moment. This active moment can be decomposed into two parts. The first part describes an active moment that travels over the flagellum with the mechanical flagellar wave, the second part represents a moment in phase over the entire length of the flagellum, which decreases linearly towards the distal tip. The linear synchronous moment, to which an amount of traveling moment has been added as a perturbation, leads to wave solutions that closely resemble flagellar waves. Properties such as wavelength and wave amplitudes and also the shape of the waves in sea urchin sperm flagella at different frequencies are accurately described by the theory. The change in wave shape in sea urchin sperm flagella at raised viscosity is predicted well by the theory. The different wave properties caused in bull sperm flagella by different boundary conditions at the proximal junction are explained. When only a traveling active moment is present in a flagellum, the wave solutions describe waves of a small wave length in a long flagellum. Some properties of the wave motion of sperm flagella are derived from the theory and verified experimentally.

Animals↗

Movement of sea urchin sperm flagella.

The motion of the sea urchin sperm flagellum was analyzed from high-speed cinemicrographs. At all locations on the flagellum the transversal motion and the curvature were found to vary sinusoidally in time. The curvatures of the flagella increase strongly near the proximal junction. Two sperm are described in transient from rest to normal motion. The full wave motion developed in both sperm within 40 ms.

Animals↗

Mg2+-dependent electrical control of flagellar activity in Euglena.

When a Euglena gracilis in a Ca2+-containing medium is impaled with a microelectrode, the flagellum is instantly ejected. In a Ca2+-free medium to which 1 mM EGTA has been added, the flagellum remains attached to the organism, but it loses activity upon impalement. Externally added ATP at a concentration of 10 mM will sustain normal flagellar activity (at approximately 20 Hz) of an impaled Euglena. If negative direct current of several tenths of a microamp is injected through the impaling microelectrode, the flagellar activity is stopped or much reduced. When the current injection is turned off the flagellum returns to its initial activity. This cycle can be repeated many times on the same animal, independent of whether Mg2+ is present in the external medium or not. If 1 micrometer of gramicidin is added to Ca2+-free medium containing 1 mM EGTA and 10 mM ATP, the flagellar activity becomes dependent on external Mg2+. Without external Mg2+ no flagellar activity is present after one or two current injection cycles as described above. With 1 mM Mg2+ present in the external medium many cycles (up to 10) can be produced. This Mg2+-dependent flagellar activity shows a smooth dependence on the amount of current injected. Observations taken by high speed cinemicrography show that in the third injection cycle the average frequency of the flagellar motion is 16-3 Hz at 0 muA, is 8 Hz at 0-2 muA, and is approximately 0 at 0-6 muA of negative current. The injection of positive current results in an increase in flagellar frequency dependent on the amount of current injected. The data indicate that the control of motility of Euglena flagella is dependent on an electrically activated Mg2+ pump.

Adenosine Triphosphate↗