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We have analyzed the mitochondrial DNA of 34 oligoasthenozoospermic men along with 150 normozoospermic fertile men from the Indian subcontinent. A novel missense mutation (C11994T) in the ND4 gene, which replaces threonine with isoleucine, was observed in all of the oligoasthenozoospermic men but not in any of the normozoospermic fertile men.
The existence of a motility-inhibiting substance in human peritoneal fluid (hPF) has been demonstrated. The results of this study indicate that this factor from hPF is a heat-resistant, volatile molecule with a very small molecular mass (e.g., halothane, which is used for general anesthesia in laparoscopic GIFT). This may be the reason for the conflicting results concerning pregnancy rates in gamete intrafallopian transfer.
The angiotensin converting enzyme inhibitor, enalapril, reduced the motility of washed human spermatozoa. The concentrations producing 50% inhibition were: (percentage motility) 11.6 +/- 2.6 mM, (average forward velocity) 8.0 +/- 1.5 mM and (motility index) 8.5 +/- 2.1 mM (mean + s.e.m.). Enalapril 20 mM prevented all forward movement and reduced percentage motility to an extremely low level. Motility was reduced within 20 sec of addition and little further change occurred during a 60-min incubation period. Inhibition of percentage motility by enalapril was reversible with a 5-min washing procedure following a 5-min incubation period, but not following a 60-min incubation period. Enalapril 20 mM did not impair plasma membrane integrity (hypoosmotic swelling test) or viability (nigrosin-eosin stain). It is concluded that the antimotility effects of enalapril are not caused by inhibition of angiotensin converting enzyme, but may be of interest in the search for new spermicidal agents.
Several beta-adrenoceptor blocking agents have been evaluated for spermicidal activity using a transmembrane migration method. The rank order of potency of the active compounds was: penbutolol greater than (+)-propranolol greater than bufuralol greater than (-)-alprenolol greater than oxprenolol greater than metoprolol. Atenolol, pindolol, practolol, tolamolol were without activity. The observed potencies of spermicidal activity are believed to be unrelated to beta-blocking activities, and we have shown that whilst they are not predictable from lipid solubility or nonspecific membrane properties of the compound alone, both these aspects appear to play a role in this pharmacological activity.
The first complete mitochondrial DNA (mtDNA) sequences (approximately 16,569 bp) in 20 patients with asthenozoospermia and a comparison with 23 new complete mtDNA sequences in teratoasthenozoospermic individuals, confirmed no sharing of specific polymorphisms or specific mitochondrial lineages between these individuals. This is strong evidence against the accepted claim of a major role played by mtDNA in male fertility, once supported by haplogroup association studies based on the screening of hypervariable region I. The hypothesis of maternally driven selection acting in male reproductive success must thus be treated with caution.
We examined motility, plasma membrane integrity, and binding capacity to homologous zona pellucidae (ZP) of frozen/thawed epididymal cat sperm as a model species for endangered felines. Epididymal spermatozoa from 20 domestic cats were frozen with freezing egg-yolk extender containing 3.0% glycerol in 0.25-ml straws. Post-thaw motility and plasma membrane integrity of the frozen/thawed spermatozoa were 31.8 +/- 2.4% and 32.2 +/- 4.2%, respectively. The frozen/thawed spermatozoa were co-cultured with frozen/thawed immature homologous oocytes with intact ZP for 3 h to examine their ability to bind to the ZP. Sixteen of the 20 frozen/thawed sperm samples demonstrated the ability to bind to ZP. These results indicated that the freezing system for epididymal sperm used in the present study gives appropriate information for banking the genetic resources of wild felid species.
Hamster spermatozoa were isolated from the caput, corpus and cauda epididymidis. They were observed in culture medium at 37 degrees C with a phase-contrast microscope and their motility recorded cinematographically. About 20 p. 100 of the caput epididymidis spermatozoa were motile and moved in a confined space with no forward progression. 30 p. 100 of the corpus epididymidis spermatozoa were motile, showing increased flagellar activity and moving in wide circles. 90 p. 100 of the cauda epididymidis spermatozoa were motile and moved forward. Forward motility was induced in immotile spermatozoa from the caput epididymidis by adding cyclic 3'-5' adenosine monophosphate (cAMP) phosphodiesterase inhibitors (caffeine, theophylline, IMX) and epididymal plasma. The best stimulation was initiated by 15 mM caffeine with 10 p. 100 of cauda epididymal plasma; a mean of 60 p, 100 of forward motility was obtained which lasted for one hour and then ceased. Cinematographic studies revealed that some induced sperm movements differed from the equivalent natural ones by the amplitude of the head movements. It is shown that during epididymal transit of hamster spermatozoa, the induction of forward motility requires not only an increased cAMP level but also factors from the cauda epididymal plasma. The idea that glycoprotein of epididymal origin initiates forward motility is discussed.
OBJECTIVES: To investigate the influence of cAMP/PKA signal transduction on human sperm motility, and to study the effect of dibutyryl cyclic adenosine monophosphate dibutyryl cyclic adenosine monophosphate (dbcAMP) on human sperm motility in vitro. METHODS: Sperm aseptically obtained by masturbation and prepared by swim-up technique from 10 healthy fertile men were incubated with different concentrations of dbcAMP. Measurement of mobility were carried out at 20, 30, and 60 min in all specimens. RESULTS: The sperm treated with dbcAMP showed a significant increase in sperm progressive motility and the percentage of motile cells. The effect seemed enhanced with the increasing of dbcAMP concentration. VSL and VCL were not affected by dbcAMP. CONCLUSIONS: dbcAMP can activate the mobility of human sperm in vitro.
BACKGROUND: The detection of significant levels of tryptase in human seminal plasma and follicular fluid and of tryptase-positive mast cells (MCs) in the wall of human Fallopian tubes lead us to hypothesize that tryptase may exert regulatory actions on human spermatozoa. METHODS AND RESULTS: Immunoelectronmicroscopy revealed proteinase-activated receptor 2 (PAR-2) in the membranes of the acrosomal region and midpiece of human spermatozoa. These PAR-2 were functional, as exposure of spermatozoa from healthy men (n = 12) with regular standard semen parameters to human recombinant tryptase significantly decreased motility in a dose- and time-dependent fashion. Motile spermatozoa (WHO a + b) were significantly decreased within 10 min of incubation with 1.000 ng/ml tryptase (P = 0.045). After 30 and 60 min, significant reduction of motility was also observed in the presence of lower tryptase concentrations (100 ng/ml, P = 0.037; 10 ng/ml, P = 0.046). The inhibitory effects of tryptase progressed throughout an observation period of 180 min. Furthermore, tryptase effects were reversible after washing procedures and could be inhibited by pretreatment with anti-tryptase antibody or anti-PAR-2 antiserum. CONCLUSIONS: The observations presented raise the possibility that tryptase directly interacts with human spermatozoa during their migration through the female genital tract. Genital tract MCs and their products may be as yet unrecognized factors involved in human fertility/sterility.
The absorption of L-carnitine from the duct of the proximal regions of the rat epididymis was investigated using a stopped-flow, split-droplet microperfusion technique. L-carnitine was absorbed from the duct of the proximal caput epididymidis by a time-dependent and saturable transport system (Km = 25 micron; Vmax = 0.65 pmoles absorbed/min/mm3 tubular volume). Furthermore, absorption appeared to be primarily sodium-independent, although the existence of a minor sodium-dependent pathway cannot be ruled out. A similar transport system was not evident along the distal caput epididymidis, where absorption of L-carnitine was attributable to passive diffusion only. The inward and outward movement of L-carnitine across the epithelium of the proximal and distal caput epididymidis appears to be regulated so that the spermatozoa come into contact with high levels of L-carnitine in the distal caput region.
A 34-year-old male laboratory worker suffered from asthenospermia and fertility problems. He was suspected of having been exposed to solvents used at work due to a malfunction of the ventilation system in his laboratory from August 1996 to April 1997. A laboratory walk-through and air and bulk sample collection were performed to determine the possible exposure levels of chemical hazards in his job. The scenario was reconstructed to simulate the worker's previous exposure during the ventilation shutdown period. It was found that the worker was possibly exposed to chloroform at levels of 10 or 50 times higher than the permissible exposure limit or the threshold limit value of 2 hr/day, 5.5 days/week, and 4.25 weeks/month for 8 months. Because chloroform is known to be spermatotoxic, the possibility of chloroform causing the worker's asthenospermia cannot be ruled out. Further study on spermatotoxicity of chloroform is warranted.
The present investigations evaluate the reversibility of the effect of gossypol on the histoarchitecture of the hamster testis and the motility of the vas deferens spermatozoa. Adult male hamsters of proven fertility were treated orally with gossypol (10 mg/kg/day) for 13 weeks (group III). Another group of animals in the Group IV were given gossypol (10 mg/kg/day) for 13 weeks and were sacrificed 13 weeks after termination of the drug treatment. Animals in groups I and II were given vehicle and served as controls for groups III and IV respectively. To all the animals, gossypol or vehicle was administered by oral intubation. A marked reduction in the spermatozoa motility was observed in the animals of group III. In contrast, the motility of the spermatozoa in the animals of group IV (recovery phase) was in the normal range. A large number of seminiferous tubules were affected by gossypol treatment (group III); these tubules showed degeneration of germ cells, vacuolization and loosening of the germinal epithelium. In the quantitative analysis of the spermatogenic events, the ratio of resting spermatocyte/'A' type spermatogonia was not affected but the ratios of pachytene spermatocytes/resting spermatocytes and stage 19 spermatids/stage 7 spermatids decreased by 21%, 47% and 52% respectively. The tubular diameter and germinal height were decreased by 14% and 33% respectively. In group IV, hamsters were allowed to recover from the gossypol treatment. The seminiferous tubules exhibited normal histoarchitecture and the tubular diameter and germinal height recovered markedly. Recovery in different stages of spermatogenic cycle was evidently clear in this group.(ABSTRACT TRUNCATED AT 250 WORDS)
Boar semen contains a seminal plasma motility inhibitor (SPMI) that blocks the motility of demembranated-reactivated spermatozoa as well as of intact spermatozoa. In this paper, we describe the primary structure of SPMI, the coding of boar SPMI cDNA gene and its expression in various porcine tissues. Nucleotide sequence analysis of the 645-bp SPMI cDNA predicts a coded polypeptide of 137 amino acid residues which includes a 21-residue signal peptide and a 116-residue secreted protein. The amino acid sequence of SPMI was found to be highly homologous to AQN-3, a member of spermadhesin family proteins of boar that bind to spermatozoa. Expression of the boar SPMI gene detected by Northern blot analysis revealed that its expression is very abundant in seminal vesicles and specific to this tissue.
Objective spermatic motility (Hamilton Thorne Research), the rapid progressive spermatozoa (grade A) recovery after swim-up, and the spermatozoa ATP content (bioluminescence) were studied in normoviscous and hyperviscous asthenospermic samples. The amplitude of lateral head displacement (ALH) was significantly lower in hyperviscous semen (normal: 4.6 +/- 0.7 microns [n = 20], high: 3.5 +/- 1.2 microns [n = 16]; p < .05). The grade A recovery percentage after swim-up was significantly higher in semens with high consistency (normal: 71.0 +/- 38.0 [n = 14], high: 181.3 +/- 108.9 [n = 6]; p < .05). The ATP content per living spermatozoa was in the normal consistency group 449.4 +/- 65.1 pmol per million living spermatozoa (n = 29) and in the high consistency batch 605.1 +/- 242.8 (n = 9), p < .05. In asthenospermia, the spermatozoa from hyperviscous samples have minor ALH values, better response to swim-up, and high ATP content than those from normoviscous ejaculates.
The present study proved that desmopressin (DDAVP) (1 microgram DDAVP/12 h/5 días) does not affect ovary, testis and adrenal development in immature Wistar rats (17 days old), because the DDAVP does not modify the weight of the aforementioned organs as compared with the control group. Nevertheless, the male adults Wistar rats (80 days old) showed lower serum testosterone concentrations than the control group, after injection of 4 micrograms/day (2 micrograms/12 h) or 8 micrograms/day (4 micrograms/12 h) of DDAVP during a 5 days period time. Moreover, paradoxical significant lower concentrations of serum testosterone were found in 4 micrograms DDAVP/day-treated rats than in 8 micrograms DDAVP/day-treated ones. The former also showed a decreased number of spermatozoa as compared with the latter and with the control group. The percentage of mobile spermatozoa was lower in rats treated with both concentrations of DDAVP as compared with the control group. Therefore, desmopressin does not delay gonadal and adrenal growth in immature rats, but, at low doses, it affects the testicular function and the mobility of the spermatozoa in male adult rats.
Human seminal plasma contains a protein factor that has the capacity to inhibit the movement of demembranated and intact spermatozoa. This factor 'seminal plasma motility inhibitor' (SPMI) has been shown to originate exclusively from the seminal vesicles. The present results demonstrate that the biological activity of SPMI in semen decreases rapidly from 1000 U/ml, immediately after ejaculation, to 220 U/ml 2 h later. Immunoblots of seminal plasma proteins probed with an antibody against human SPMI, revealed the rapid processing of a predominant 52 kDa SPMI antigen, present in the seminal vesicle secretions. This precursor was degraded initially into intermediate molecular mass fragments of 25-40 kDa, and subsequently into smaller fragments of 17-21 kDa. When seminal vesicle fluid was mixed with prostatic secretions (3:1 v/v), proteases present in prostatic secretions were shown to be responsible for processing of the SPMI precursor. Addition of protease inhibitors such as phenylmethylsulphonyl fluoride (PMSF, 5 mM), benzamidine (100 mM) or ethylenediaminetetraacetic acid (EDTA, 5 mM) to the mixture of seminal vesicle and prostate secretions partially prevented the loss of activity of SPMI by 54%, 27% and 9%, respectively. However, the simultaneous addition of PMSF and benzamidine conferred almost total stability to the SPMI precursor activity. These results demonstrate that SPMI exists as a predominant 52 kDa precursor form in the seminal vesicles and is processed rapidly after ejaculation into less active, lower molecular mass forms by one or more serine proteases and/or metalloproteases of prostatic origin which are present in liquefied semen.
A randomized group of 65 men with oligoasthenozoospermia and left side varicocele was subjected to a controlled study with kallikrein. Kallikrein (600 units orally per day) was given to 38 men for a period of 3 months. The other group of 27 men remained untreated but were followed up during the same period of time. After kallikrein treatment significant improvements in sperm quality could be observed: percentage of motile spermatozoa from 24.47 +/- 9.01 to 35.26 +/- 11.80, percentage of morphologically normal spermatozoa from 58.42 +/- 5.86 to 71.05 +/- 8.12. There was no improvement in sperm parameters in the untreated group of men. The results suggest that kallikrein therapy is useful for those men who refuse an operative or radiologic treatment of their varicocele and also as a primary or associated therapy of patients who underwent ligature or occlusion of spermatic veins.
The study was conducted to evaluate the results of IUI treatment in a homogenous group with male factor infertility, and to assess the correlation of sperm variables, including sperm morphology by strict criteria, with pregnancy achievement after IUI. A total of 108 couples with no apparent female aetiology for infertility underwent 264 intrauterine insemination treatment cycles. A comparison was made between the sperm variables in two groups in which the achievement of pregnancy differed. The percentage of motile spermatozoa, degree of motility and normal morphology (by strict criteria) were significantly higher in the pregnant group compared with that of the nonpregnant group. A significant difference in pregnancy rates per couple after intrauterine insemination was demonstrated among three groups according to the percentage of sperm morphology, i.e. poor (< 4%), fair (4-14%) or good (> 14%) (11.1%; 36.1% and 50.0%, respectively). Intrauterine insemination is a valid mode of treatment in cases with male infertility, provided that normal morphology by strict criteria is higher than 4%.