Showing papers on "Sperm motility published in 1982"

Normal development following in vitro fertilization in the cow.

Abstract: A repeatable procedure for fertilization of bovine ova in vitro is described. Oocytes were recovered from ovarian follicles or from oviducts near the time of ovulation following treatment of donors with pregnant mare's serum gonadotropin (PMSG) and prostaglandin F2 alpha (PGF2 alpha). For in vitro capacitation semen was incubated, then high ionic strength treated and subsequently incubated in defined medium prior to insemination of oocytes. In one experiment frozen bull semen was successfully used. In experiments with 4 bulls (B, C, D, F), 34 (43.6%) of 78 ova and 13 (19.7%) of 66 follicular oocytes were fertilized in vitro. In the last series (spermatozoa from Bull F) the fertilization of 22 (62.9%) of 35 tubal ova was achieved. In vitro development proceeded to the 8-cell stage. No fertilization in vitro followed use of one male (Bull E), even though his spermatozoa could penetrate zona-free hamster ova in vitro, and higher than usual bacterial contamination of his semen was implicated as the probable cause. Findings suggested vigorous progressive sperm motility and acrosome integrity to be important features of good sperm samples. In one experiment a 4-cell stage embryo was transferred with the result that the recipient gave birth to a normal bull calf on June 9, 1981. The first calf resulting from in vitro fertilization has been found to be completely normal.

Reproductive functions in young fathers and grandfathers.

Abstract: Testicular functions were investigated in 23 grandfathers [60--88 yr old; 67 +/- 7.8 (mean +/- SD)], i.e. men with fertility proven earlier in life. They were recruited from a nonpatient population and led an active life, most of them with a permanent partner. The grandfathers were compared with a group of 20 unrelated healthy fathers, 24--37 years old (29.2 +/- 3.2). Whereas sperm density was higher in the older group, there were no significant differences in ejaculate volume and sperm morphology between the younger and older men. Sperm motility and seminal fructose, however, decreased with age. The fertilizing capacity of sperm as assessed in the heterologous ovum penetration test using zona pellucida-free hamster eggs did not decrease significantly with age. Whereas the basal serum testosterone and estradiol levels were not different between the younger and older men, the response to 2 days of hCG stimulation decreased significantly with age. This decrease was observed in older men whether they had frequent or infrequent sexual activity. Basal serum LH and FSH levels were elevated in the older men. The LH response to GnRH stimulation relative to basal; values was significantly reduce, while FSH responses did not change with age. We conclude that sperm counts and fertilizing capacity of sperm are not negatively influenced by old age, at least not in men with sustained sexual activity. However, the pituitary as well as the testis show signs of decreased endocrine reserve capacity in old age.

Spontaneous lipid peroxidation in rabbit epididymal spermatozoa: its effect on sperm motility.

Abstract: Rabbit spermatozoa released from the cauda epididymidis into Tris phosphate medium contain- ing KCI or NaCl and 0.4 mM EDTA underwent spontaneous lipid peroxidation during aerobic incubation at 37#{176}C. In the medium containing 130 mM Kand 0 mM Na� (KTP), the rate of lipid peroxidation, as measured by malonaldehyde production, proceeded at a linear rate of 0.045 nmol malonaldehyde/h per 108 cells for 22 h. The motility of these spermatozoa declined with time in medium KTP, with 40% initial forward motility decreasing to zero in 4 h and initial 60% flagellar beating ceasing after 12 h. The percent inert spermatozoa showing no flagellar motion in KTP increased linearly with production of malonaldehyde; all flagellar activity stopped at 0.5 nmol malonaldehyde/10 cells. In the Tris phosphate medium containing 120 mM Naand 10 mM K� (NTP), the percentage of sperm showing forward motility was close to 100% and this declined to 60% after 16 h aerobic incubation. Flagellar beating was not observed. In medium NTP, the rate of lipid peroxidation was 0.0056 nmol malonaldehyde/h per 10 cells, eightfold lower than that observed in KTP. The same linear correlation between malonaldehyde production and percent inert sperm was found as for KTP: 0.5 nmol malonaldehyde/108 cells also corresponded to cessation of flagellar motion. The dependence of motility maintenance on K+ concentration in Tris phosphate medium containing (Na+ + K+)=1 30 mM showed maximal maintenance at 10 mM K+, with a decline at 0 mM K+ and steep decline at K+ concentrations greater than 30 mM. This strong dependence of rabbit sperm peroxidation on ionic composition of the medium is suggested to involve perturbation of the equilibrium between O9 and its conjugate acid HO2', the latter species being the agent of peroxidation.

Promotive effect on human sperm progressive motility by prostasomes.

Abstract: Seminal plasma constituents were separated on Sephadex G200 gel columns. The column eluate was analysed with regard to protein content, ATPase activity and promotive activity on sperm progressive motility. Two different chromatographic fractions were also subjected to electron microscopy after sedimentation by preparative ultracentrifugation. A maximum promotive value on sperm progressive motility coincided with a maximum ATPase activity value in a single peak from seminal plasma eluted first on the column and containing less protein than the other peaks appearing later in the chromatogram. This first peak was the only one containing ATPase activity and membrane-surrounded organelles named prostasomes. Other peaks, rich in protein but lacking ATPase and prostasomes, displayed a moderate and rather irregular pattern in reference to promotive activity on sperm progressive motility. Evidence is given that the positive effect by prostasomes is specific on sperm progressive motility. Hence, procedures aiming at a change of membrane integrity of the prostasomes resulted in diminished effects on sperm progressive motility. This could be explained by a probable dissipation of the electrochemical gradient of calcium ions.

Caenorhabditis elegans spermatozoan locomotion: amoeboid movement with almost no actin.

Abstract: The pseudopods of Caenorhabditis elegans spermatozoa move actively causing some cells to translocate when the sperm are dissected into a low osmotic strength buffered salts solution. On time-lapse video tapes, pseudopodial projections can be seen moving at 20-45 micrometers/min from the tip to the base of the pseudopod. This movement occurs whether or not the cell is attached to a substrate. Translocation of the cell is dependent on the substrate. Some spermatozoa translocate on acid-washed glass, but a better substrate is prepared by drying an extract of Ascaris uteri (the normal site of nematode sperm motility) onto glass slides. On this substrate more than half the spermatozoa translocate at a velocity (21 micrometers/min) similar to that observed in vivo. Translocating cells attach to the substrate by their pseudopodial projections. They always move toward the pseudopod; changes in direction are caused by changes in pseudopod shape that determine points of detachment and reattachment of the cell to the substrate. Actin comprises less than 0.02% of the proteins in sperm, and myosin is undetectable. No microfilaments are found in the sperm. Immunohistochemistry shows that some actin is localized in patches in the pseudopod. The movement of spermatozoa is unaffected by cytochalasins, however, so there is no evidence that actin participates in locomotion. Fertilization-defective mutants in genes fer-2, fer-4, and fer-6 produce spermatozoa with defective pseudopodial projections, and these spermatozoa are largely immotile. Mutants in the spermatozoa do not translocate. Thus pseudopod movement is correlated with the presence of normal projections. Twelve mutants with defective muscles have spermatozoa with normal movement, so these genes do not specify products needed for both muscle and nonmuscle cell motility.

Correlation between regional specificity of antisperm antibodies to the spermatozoan surface and complement-mediated sperm immobilization.

Abstract: Sera from men at risk for immunity to spermatozoa were screened for antisperm antibodies by immunobead binding following passive antibody transfer to antibody-free sperm of fertile donors. The percent motile sperm after incubation in diluted antibody positive serum in the presence of complement was compared with the regional distribution of immunoglobulins bound to the sperm surface. The extent of complement-mediated sperm immobilization varied with immunoglobulin class and with the location of antibody bound to the sperm surface. Tests utilizing complement-mediated immobilization of sperm are insensitive to the presence of antibodies of IgG and IgA classes that are directed against the head, the distal one-fifth of the sperm tail principal piece, or the tail end piece. A high degree of immobilization was found only when IgG binding occurred on the distal two-fifths to three-fifths of the principal piece of the tail or when IgM bound to the sperm tail end piece. (Am J Reprod Immunol. 2:222-224.)

Development of the ability to bind to zonae pellucidae during epididymal maturation: reversible immobilization of mouse-spermatozoa by lanthanum.

Abstract: Mouse spermatozoa recovered from the caput, corpus, or cauda epididymidis were examined for their ability to bind the vitro to zonae pellucidae. Since spermatozoa from the caput epididymidis do not display progressive motility as compared with more mature spermatozoa, direct comparison of the different sperm populations may not measure zona binding ability validly. To equalize the motile properties of the spermatozoa, a method was developed to immobilize vigorously motile corpus and cauda spermatozoa. Reversible immobilization was achieved by incubation in 25 microM La3+ which resulted in a twitching, nonprogressive type of motility. La3+ incubation did not appear to affect the spermatozoa adversely, since vigorous motility (equivalent to the controls) of corpus and cauda sperm was displayed upon subsequent incubation in standard La3+-free culture medium. Moreover, cauda spermatozoa preincubated for 90 min in La3+ displayed levels of fertilization in vitro equivalent to their control counterparts. Using this La3+-immobilization technique, the zona binding ability of the different sperm population could be assayed. Gamete collision was insured under these conditions by shaking the gamete-containing dishes at 100 cycles/min. Regardless of the extent of sperm motility, a similar zona-binding pattern emerged: cauda sperm bound in high numbers, corpus sperm bound at some intermediate level (an average of 24% of cauda binding level), and caput sperm bound rarely (2% of cauda binding level). Thus it appears that, for mouse spermatozoa, the onset of fertility during epididymal transit parallels the ability to bind to zonae pellucidae. Unlike the interaction between spermatozoa and zonae, La3+ was unable to support sperm binding in to egg plasma membrane, supporting the view that mouse sperm may have different sites for interaction with the zonae pellucida and the egg plasma membrane.

Bovine serum albumin, sperm motility, and the “dilution effect”

Abstract: Epididymal spermatozoa from rabbit and ram were washed either once or twice using an efficient washing procedure and were then diluted in various media to a final concentration of approximately 1.4 x 10(7) cells/ml and incubated at 30 degrees C for up to 12 hours. Bovine serum albumin (BSA), either untreated or defatted, was found to be better than polyvinylpyrrolidone, ovalbumin, or alpha-lactalbumin, both at stimulating and maintaining motility levels and at reducing the tendency of the washed spermatozoa to stick to glass. BSA was effective in all media tested, being independent of Ca2+, PO4(3-), HCO3-, and ionic strength). BSA has a reversible stimulatory effect on motility. If BSA was added to sperm suspensions 3 1/2 hours after they had been washed and diluted in protein-free medium, motility was stimulated to levels not significantly lower than those observed in samples that had been washed and diluted in the presence of BSA. However, samples washed into BSA and then washed free of it behaved essentially as though they had never been in contact with protein. The motility, survival, and response to BSA of twice-washed spermatozoa were the same as those of once-washed spermatozoa, showing that epididymal plasma factors are not required for survival in vitro. It was concluded that dilution is not essentially detrimental to rabbit and ram spermatozoa. However, severe dilution of semen may result in levels of male reproductive tract fluids insufficient either to stimulate motility or to prevent sticking of motile cells to container surfaces. Few motile spermatozoa are recovered from samples of such diluted semen.

Taxol inhibits the nuclear movements during fertilization and induces asters in unfertilized sea urchin eggs.

Abstract: Taxol blocks the migrations of the sperm and egg nuclei in fertilized eggs and induces asters in unfertilized eggs of the sea urchins Lytechinus variegatus and Arbacia punctulata. Video recordings of eggs inseminated in 10 microM taxol demonstrate that sperm incorporation and sperm tail motility are unaffected, that the sperm aster formed is unusually pronounced, and that the migration of the egg nucleus and pronuclear centration are inhibited. The huge monopolar aster persists for at least 6 h; cleavage attempts and nuclear cycles are observed. Colcemid (10 microM) disassembles both the large taxol-stabilized sperm aster in fertilized eggs and the numerous asters induced in unfertilized eggs. Antitubulin immunofluorescence microscopy demonstrates that in fertilized eggs all microtubules are within the prominent sperm aster. Within 15 min of treatment with 10 microM taxol, unfertilized eggs develop numerous (greater than 25) asters de novo. Transmission electron microscopy of unfertilized eggs reveals the presence of microtubule bundles that do not emanate from centrioles but rather from osmiophilic foci or, at times, the nuclear envelope. Taxol-treated eggs are not activated as judged by the lack of DNA synthesis, nuclear or chromosome cycles, and the cortical reaction. These results indicate that: (a) taxol prevents the normal cycles of microtubule assembly and disassembly observed during development; (b) microtubule disassembly is required for the nuclear movements during fertilization; (c) taxol induces microtubules in unfertilized eggs; and (d) nucleation centers other than centrioles and kinetochores exist within unfertilized eggs; these presumptive microtubule organizing centers appear idle in the presence of the sperm centrioles.

Sperm transport in the human female reproductive tract in relation to semen analysis characteristics and time of ovulation.

Abstract: Laparoscopic sperm recovery from the pouch of Douglas and tubal fimbriae was performed in 64 infertile couples. Spermatozoa were recovered from 16/35 couples investigated after AIH, and from 13/29 couples post coitum. The method of insemination had no effect on the result, which was positive in 45.3% of all couples, although AIH did result in significantly larger numbers of peritoneal spermatozoa. The number of peritoneal spermatozoa did not show any direct correlation with the number inseminated, but there were reductions along the tract of 5.83 (+/- 1.4 s.d.) orders of magnitude for total sperm count, and 5.52 (+/- 1.21 s.d.) for the number of motile spermatozoa. Only sperm motility had a significant influence on the success of sperm transport; spermatozoa were recovered from patients with sperm densities as low as 3.0 and 3.5 x 10(6)/ml, but with 56 and 44% motile spermatozoa. No influence of cycle day within the range +/- 4 days of ovulation on sperm transport was found. In 45 couples, routine semen analyses were apparently completely normal, but the incidence of sperm recovery was still only 49% (22/45), suggesting that a failure of sperm transport may have been a significant causative factor in their infertility.

Effects of gossypol on reproductive and endocrine functions of male cynomolgus monkeys (Macaca fascicularis).

Abstract: Adult male cynomolgus monkeys were treated orally with 5 (n=4) or 10 (n=3) mg per kg per day of gossypol acetic acid (gossypol) for 6 months. A significant decrease in sperm concentration and motility,determined by evaluating semen ejaculates,was observed without any significant decrease in circulatinglevelsof testosterone (T) among 10 mg per kg per day gossypol-treated animals. Similarly, there was no significant difference in plasma T levels after an intravenous bolus injection of luteinizing hormone releasing hormone (LHRI-l) (50 �sg/animal) between control and gossypol-treated animals, further suggesting an adequate release of pituitary luteinizing hormone (LH) and normal Leydig cell function. Transient azoospermia was observed in 1 out of 4 and in 2 out of 3 animals after 4 months of gossypol treatment at 5 and 10 mg per kg per day, respectively. The effects of gossypol (10 mg per kg per day) were more dramatic and consistent on sperm motility. No striking abnormality of spermatozoa was observed by light microscopy, although there was an increase in the number of sperm with coiled or broken tail pieces and an occasional detached head and tail. However, at the ultrastructural level disruption of the axial complex was commonly observed with gossypol treatment. The effect was manifested at 5 mg per kg per day as a disruption of radial arms. At 10 mg per kg per day the entire axial complex was frequently destroyed, suggesting an impairment of sperm motility. No serious clinicopathologic side effects were observed except temporary diarrhea and anorexia among 10 mg per kg per day gossypoltreated animals during the initial stages of treatment. In addition, gossypol had a hypolipidemic effect which is a new significant entity for this compound. In conclusion, it is suggested that gossypol may be interfering with spermatogenesis and/or directly acting on the sperm itself within the testis or during its passage through the male reproductive tract in ways thataffectboth sperm production and sperm motility.

An investigation of sperm migration into the oviducts of the mare

Abstract: A total of 23 mares were inseminated once within 0-6 h after clinical detection of ovulation, 14 with fresh and 9 with deep-frozen semen containing 0.1 x 10(9) to 4.7 x 10(9) motile spermatozoa. Within these two groups, the mares were slaughtered 2, 4 or 6 h after insemination and their genital tracts removed. The utero-tubal junction, isthmus and ampulla ipsilateral to the ovary in which ovulation occurred were flushed separately for sperm recovery. In 1 or 2 mares of each group, the uterine horn and corpus uteri, the cervix and vagina were also flushed. Tissue samples were collected from the contralateral oviduct and the other genital regions and prepared for scanning electron microscopy to show spermatozoa distribution in situ. Flushings were also prepared for scanning electron microscopy. There were no significant differences in the extent of sperm migration and in the number of spermatozoa in the different regions of the oviduct 2, 4 or 6 h after insemination of fresh or frozen semen. There was, however, a striking difference in sperm number within the time intervals examined; the numbers were greatest at 4 h after insemination. SEM of spermatozoa in the various regions of the oviducts failed to indicate any alterations to the sperm-head membranes that could be associated with sperm capacitation. The majority of spermatozoa found in the uterotubal junction, isthmus and ampulla showed morphological integrity.

Method of increasing the incidence of female offspring

Abstract: The likelihood of conceiving a female fetus is substantially enhanced by promoting ovulation in a fertile female mammal with an ovulation-inducing agent and artificially inseminating the female mammal during the period of expected ovulation with a sperm fraction of enhanced sperm motility from which immotile sperm and non-sperm seminal components have been separated and which is suspended in serum albumin or like physiologically acceptable vehicle.

Flow cytometry-fluorescence measurements for characterizing sperm

Abstract: A process for characterizing sperm motility and viability by staining a sperm sample with Rhodamine 123 and ethidium bromide, and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm, the green counts being correlated with sperm motility and the red counts being correlated with putative dying or dead cells. Additionally, a sample of sperm can be characterized as to type and normality by staining a sample of sperm with acridine orange and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm.

Angiotensin I converting enzyme in rat testis, epididymis and vas deferens under different conditions.

Abstract: Angiotensin I converting enzyme (ACE) was found in the testis, epididymis and vas deferens of rats. In tissue specimens without a prior washout of seminal fluid the highest specific ACE activity was measured in the testis. The enzyme activity was significantly lower towards the end of the excurrent ducts, suggesting that the enzyme is synthesized in the testis and secreted into the seminal fluid there. An ACE inhibiting substance may be secreted by the epididymal epithelium. Enzyme synthesis and enzyme inhibition are probably under simultaneous endocrine control. In-vitro inhibition, pH- and temperature-dependence of gonadal ACE correspond with that of lung and blood plasma. However, the physiological function of the enzyme on sperm motility and fertility remains unsolved.

Ultrastructural Studies of Spermatozoa and the Epithelial Lining of the Epididymis and Vas Deferens in Rats Treated with Gossypol

Abstract: This study was undertaken to determine the effects at early time intervals of gossypol on sperm motility and on the ultrastructure of rat epididymal and vasal sperm and epididymal and vasal epithelium. Rats were treated by gavage with 20 or 30 mg/kg/day of gossypol for 7 weeks; control animals were unfed or received the vehicle alone. The results confirm and extend earlier observations demonstrating that epididymal sperm of gossypol-treated rats exhibit distinct ultrastructural changes under the experimental conditions employed. The severity and frequency of the degenerative changes appear to increase with dose and duration of treatment. Striking ultrastructural defects can be seen as early as 3 weeks after 20 mg/kg/day of gossypol. By the fifth week of either 20 or 30 mg/kg/day of gossypol, significant damage to virtually all sperm flagella is observed throughout the epididymal duct of all treated rats. The initial and predominant defect is degeneration of the midpiece mitochondria but additional flagell...

Stimulation of Sperm Progressive Motility by Organelles in Human Seminal Plasma

Abstract: A simple method for objective determination of sperm progressive motility is described. Comparisons were made between results with this method and subjective grading of motility. Although congruence between the two methods was demonstrated, the subjective evaluation had obvious disadvantages. In suspensions of NaCl-washed spermatozoa, seminal plasma was required for progressive motility. Various isotonic salt solutions without seminal plasma were ineffective in this respect. Washed spermatozoa, however, were always motile in a medium consisting of pellet II, obtained from ultracentrifugation of human seminal plasma, thoroughly mixed with isotonic NaCl solution. The progressive sperm motility was two to three times as great in that medium as in seminal plasma mainly devoid of organelles. Magnesium, calcium and zinc ions, in the presence of seminal plasma, could exert stimulatory and inhibitory effects on sperm progressive motility, depending on the concentration of each divalent cation. Magnesium gave the ...

Sperm Motility of Northern Pike and Chain Pickerel at Various pH Values

Abstract: The activity of sperm from northern pike Esox lucius and chain pickerel Esox niger was observed at pH 3.9, 4.5, 5.4, 6.0, 6.4, 6.9, 7.4, and 7.9. The sperm of the northern pike remained motile for shorter periods of time than that of the chain pickerel (P = 0.01). Northern pike sperm was not observed swimming actively at pH values less than pH 5.4 and showed a strong trend toward increased time of activity with increasing pH. For this species, minimum motility occurred at pH 5.4 (28 seconds) and maximum motility was found at pH 7.9 (67 seconds). In contrast, chain pickerel sperm was acid-tolerant, swimming actively for periods of over 90 seconds at pH 3.9 and 4.5. Maximum sperm motility for this species was at pH 6.9 (137 seconds). Motility time for chain pickerel sperm was reduced at the two most basic levels tested: 109 seconds at pH 7.4, and 81 seconds at pH 7.9. Substantial variation in sperm motility time occurred between the two chain pickerel males tested, but not between the two northern ...

Regulation of Sperm Motility in Starfish. I. Initiation of Movement

Abstract: Starfish seminal plasma has such characteristics as higher concentration of potassium ions, higher osmolality, and lower pH compared with those of sea water. These factors independently inhibited the movement of spermatozoa. Sperm movement was recorded by taking photographs of the swimming paths under a dark field microscope. Spermatozoa either did not move or swam with decreased beat frequency in isolated seminal plasma or in the presence of certain fractions of seminal plasma obtained by gel-filtration or by partitions. In normal sea water (containing calcium ions), a low pH of less than 6 resulted in a decrease in the number (%) of swimming spermatozoa, though the speed of propulsion and the beat frequency was not affected. On the other hand, in a calcium-free sea water, a large proportion of spermatozoa moved in both low and high pH conditions, but in low pH the speed of propulsion was reduced and many spermatozoa (31 %) swam in smaller circles.