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PubMed2003

Acquisition of arylsulfatase A onto the mouse sperm surface during epididymal transit.

Arylsulfatase A (AS-A) is localized to the sperm surface and participates in sperm-zona pellucida binding. We investigated how AS-A, usually known as an acrosomal enzyme, trafficked to the sperm surface. Immunocytochemistry of the mouse testis confirmed the existence of AS-A in the acrosomal region of round and elongating spermatids. However, immunofluorescence and flow cytometry indicated the absence of AS-A on the surface of live testicular sperm. In contrast, positive AS-A staining was observed in the heads of live caudal epididymal and vas deferens sperm. The results suggested that acquisition of AS-A on the sperm surface occurred during epididymal transit. Immunocytochemistry of the epididymis revealed AS-A in narrow and apical cells in the initial segment and in clear cells in all epididymal regions. However, these epithelial cells are in the minority and are not involved in secretory activity. In the caudal epididymis and vas deferens, AS-A was also localized to principal cells, the major epithelial cells. Because principal cells have secretory activity, they may secrete AS-A into the epididymal fluid. This hypothesis was supported by our results revealing the presence of AS-A in the epididymal and vas deferens fluid (determined by immunoblotting and ELISA) and an AS-A transcript in the epididymis (by reverse transcription polymerase chain reaction). Alexa-430 AS-A bound to epididymal sperm with high affinity (Kd = 46 nM). This binding was inhibited by treatment of sperm with an antibody against sperm surface sulfogalactosylglycerolipid. This finding suggests that AS-A in the epididymal fluid may deposit onto sperm via its affinity to sulfogalactosylglycerolipid.

Animals↗
PubMed1988

Epididymal maturation and the acrosome reaction in mouse sperm: response to zona pellucida develops coincident with modification of M42 antigen.

Development of the sperm's capacity to interact with the zona pellucida was investigated at the stage when the acrosome reaction (AR) is induced. The response of epididymal sperm to agents that affect the occurrence of the AR was used to monitor maturational changes. Despite the finding that sperm from the three main epididymal regions were competent to undergo ARs induced by the divalent cation ionophore A23187 (56% AR, 74% AR, and 83% AR in caput, corpus, and cauda, respectively), the cells' responses to solubilized zonae pellucidae were different. When challenged with 5 zonae equivalents/microliter, both corpus and cauda sperm shed their acrosomes in high numbers (75% AR and 86% AR, respectively), whereas caput sperm did not (23% AR). Previous work has shown that the presence of M42 monoclonal antibody (mAb) during in vitro and in vivo fertilization inhibits sperm penetration through the zona pellucida by specific interference with zonae pellucidae-induced ARs. In this study, presence of the M42 mAb did not affect the incidence of A23187-induced ARs, whereas the zona-induced ARs that occurred in both corpus and cauda sperm were inhibited fully with M42 immunoglobulin (Ig) G. In addition, the antigen recognized by M42 mAb on sperm, termed M42 Ag, was examined during epididymal maturation. Although antigen localization appeared indistinguishable by immunofluorescence on sperm taken from the caput, corpus, and cauda regions of the epididymis, modification of this antigen during epididymal transit was detected. Equilibrium-binding studies using 125I-M42 IgG demonstrated a progressive increase during epididymal transit in the amount of M42 mAb that bound to fixed cells. Corpus and cauda sperm bound 185% and 240%, respectively, of the 125I-M42 IgG detected on caput sperm. These changes in expression of M42 Ag paralleled a structural change: the Mr of the antigen decreased from a 195,000/210,000 doublet in caput sperm to a 185,000/200,000 doublet in corpus and cauda sperm, as determined by immunoblot analysis of sodium dodecyl sulfate (SDS)-extracted sperm. Results presented here demonstrate that mouse sperm develop the capacity to undergo a zona-induced AR during epididymal maturation. The M42 antigen, which is involved in the zona-induced AR, is modified during epididymal transit coincident with development of the sperm's responsiveness to zonae. Our working hypothesis, based on these results, is that development of the sperm's capacity to undergo a physiological AR is related to modification of M42 Ag.

Acrosome↗
PubMed1990

Interactions of labeled epididymal secretory proteins with spermatozoa after injection of 35S-methionine in the mouse.

The sequential interactions of epididymal secretory proteins with spermatozoa during epididymal transit were examined. Mice received injections of 35S-methionine, and the radiolabeled luminal fluid and sperm-associated proteins were analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis at various times after injection. The majority of the luminal fluid and sperm-associated proteins were found in the caput epididymidis at 8 h; by 7 days, many of these proteins had been transported to the cauda epididymidis. Two classes of epididymal protein-sperm interactions were distinguished on the basis of regional synthesis and secretion. The major class consisted of proteins that were synthesized, secreted, and bound to spermatozoa in the caput epididymidis. In this class, however, the binding of proteins to the spermatozoa was variable. For example, a protein of 25 kDa remained associated with spermatozoa in substantial amounts during epididymal transit, while proteins of 40 and 35 kDa decreased in amount. Other proteins such as a protein of 18 kDa did not remain associated with spermatozoa. Another class of proteins (54, 44, 29 kDa) were synthesized and secreted from all epididymal regions but bound only to caput spermatozoa. Most of the epididymal proteins appeared to be tightly bound to the spermatozoa since spermatozoa already saturated with the unlabeled protein in the distal epididymis remained so even though the spermatozoa were surrounded by labeled proteins in the luminal fluid. These studies demonstrate that a variety of specific interactions occur between epididymal secretory proteins and spermatozoa as they migrate and mature in the epididymis.

Animals↗
PubMed1991

Determination of the capacity of ram epididymal and ejaculated sperm to undergo the acrosome reaction and penetrate ova.

An understanding of epididymal maturation of sperm requires descriptions of changes in membrane properties and their relation to changes in cell function. While sperm membranes have been studied in some detail in rams, few reports address associated functional changes. This report provides such data by evaluating (a) the time course of sperm acrosome reaction (AR) induction for cells from each epididymal region; (b) the capacity of epididymal sperm to penetrate ova; (c) differences in physiological AR and general sperm degeneration; and (d) acrosin release of epididymal sperm. Ram epididymal (caput, corpus, and proximal and distal cauda) and ejaculated (EJ) sperm were incubated in vitro to assess their capacity to undergo an AR. Light microscopy revealed that in sperm populations which had traversed the proximal cauda epididymidis, greater than or equal to 50% exhibited an endogenous AR in less time (less than 17 h) than did sperm isolated from more proximal regions of the epididymis (22-greater than 50 h). Heparin added to sperm did not stimulate the AR in epididymal or EJ sperm, whereas addition of a calcium ionophore (A23187) increased AR rates for cauda and EJ sperm, but not caput or corpus sperm. A second experiment evaluating percent AR, percent motile cells, and percent hamster ova penetrated revealed that sperm isolated from regions proximal to the cauda epididymidis failed to penetrate ova. When cauda or EJ sperm exhibited motility greater than 5% and AR greater than 24%, penetration of hamster eggs occurred. Comparisons of acrosomal integrity by electron or light microscopy were not different for sperm at any stage of epididymal maturation, suggesting that minimal nonspecific membrane changes occur and that light microscopy is valid for evaluating the acrosomal status of ram spermatozoa. Acrosin activity (sperm bound and dissociated) also was measured. Both total activity and release of acrosin from sperm to the medium during an 8-h incubation was greater for mature than for immature sperm. Results from these experiments are discussed in relation to the changes that must occur in sperm as they acquire the capacity to undergo an AR and penetrate hamster ova.

Acrosin↗
PubMed1995

Effect of accessory sex gland fluid from bulls of differing fertilities on the ability of cauda epididymal sperm to penetrate zona-free bovine oocytes.

The ability of accessory sex gland fluid to affect the fertility of cauda epididymal sperm was evaluated for 10 bulls that ranged in fertility from 6.2% below to 6.0% above the average fertility of bulls at artificial breeding cooperatives. Cauda epididymal sperm collected from indwelling vasa deferentia catheters and cauda epididymal sperm exposed to accessory sex gland fluid from the same bull were compared on the basis of their rates of in vitro penetration of zona-free oocytes after heterospermic insemination. Incubation of cauda epididymal sperm with accessory sex gland fluid significantly enhanced the ability to penetrate oocytes, and bull fertility affected the magnitude of this improvement. For bulls of average and higher fertility, the positive influence of accessory sex gland fluid on penetrating ability of sperm was highly significant (p < 0.0001). Accessory sex gland fluid from bulls of below-average fertility also improved the penetrating ability of cauda epididymal sperm, although not significantly (p = 0.07). Heterospermic competitions compared the penetrating ability of cauda epididymal sperm exposed to homologous accessory sex gland fluid with a portion of the same sperm population incubated in heterologous accessory sex gland fluid from a bull of contrasting fertility. In experiments involving sperm from 12 different bulls, paired in 42 fertile/subfertile combinations, samples of cauda epididymal sperm mixed with accessory sex gland fluid from the higher-fertility bulls had greater oocyte-penetrating ability than when aliquots of that sample were mixed with accessory gland fluid from lower-fertility bulls (p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1981

Changes in surface protein structure of bull spermatozoa during epididymal maturation.

The surface proteins of bull spermatozoa from caput and cauda epididymis were labelled by lactoperoxidase-catalyzed radioiodination and solubilized and analyzed by SDS-PAG-electrophoresis. The surface protein patterns of caput and cauda epididymal spermatozoa resembled each other but some distinct differences could be found. Caput epididymal spermatozoa revealed a protein peak with molecular weight of 15 000 - 18 000 daltons but this peak was not found on cauda epididymal spermatozoa. On caput epididymal spermatozoa the most intensely labelled protein peak was located between 90 000 and 100 000 daltons but on cauda epididymal spermatozoa the corresponding peak was only weakly labelled and had a molecular weight of 80 000 - 90 000 daltons. Surface protein with molecular weight of 42 000 - 47 000 daltons was dominating on cauda epididymal spermatozoa. The surface protein structure of cytoplasmic droplets did not drastically differ from that of epididymal spermatozoa.

Animals↗
PubMed1993

The use of ELISA to evaluate human antibody binding to epididymal sperm from different species.

PROBLEM: The impact of antibodies to epididymal sperm antigens in human infertility has been poorly understood. Cross-reactivity of human antibodies with animal epididymal sperm has been previously observed, however, only by means of qualitative methods. Moreover, it has been always compared to reactivity against human ejaculated rather than human epididymal sperm. METHOD: Following a screening study of 940 infertility patients, sperm agglutinating and immobilizing sera as well as sperm antibody negative controls were used to standardize an ELISA employing human ejaculated sperm. Nine sera positive in ELISA were further tested against epididymal human, guinea pig, rat, and hamster sperm. Differences among groups were evaluated by factorial analysis of variance (ANOVA). RESULTS: The specificity and sensitivity of ELISA were shown to be 85.1% and 61.18%, respectively. Eight out of nine antisperm antibody-positive sera from infertile subjects reacted relatively stronger with epididymal than with ejaculated human sperm. All tested infertility sera showed strong although variable cross-reactivity with sperm from guinea pig, hamster, and rat. CONCLUSION: ELISA has definite potential in sperm antibody research, allowing quantitative assessment of the results and immotile sperm employment. The suggested predominant role of epididymal sperm antigens in immune responses related to fertility needs further investigation. Some of these antigens are obviously phylogenetically preserved, and possibly in a quantitative aspect present differently on epididymal spermatozoa from various mammalian species.

Agglutination Tests↗
PubMed1987

Further characterization of a model system for the study of human epididymal physiology and its relation to sperm maturation.

Some preliminary speculations about the possible participation of epididymal antigens in sperm function may be supported by the above data. On the one hand, the reduction in the amount of antigens and their abnormal localization on spermatozoa from infertile patients may be coincident with our view about participation of epididymal antigens in the development of zona pellucida binding ability and fertilizing capacity by spermatozoa during maturation. This hypothesis is derived from experiments showing that immature hamster spermatozoa gain their ability to recognize and bind to zona pellucida and to penetrate homologous oocytes when exposed to preparations enriched in androgen-dependent epididymal secretory proteins or preincubated in conditions that favor their interaction with these proteins. Supporting our viewpoint for such a role in humans is evidence showing the progressive development of the ability to interact with hamster denuded oocytes as human spermatozoa pass along the epididymis. On the other hand, the apparent correlation between the loss of epididymal antigens during capacitation and the increased fertilization of human oocytes in vitro may be reminiscent of the removal of a decapacitation or acrosome stabilizing factor known to occur in many species and that must be removed prior to fertilization. Pending further understanding of their physiological role, the androgen-dependent epididymal proteins may become a useful marker of epididymal function and/or of sperm capacitation in humans. Within this context, we wish to stress the potential value of the model system that we have developed for the study of human epididymal physiology.

Epididymis↗
PubMed1982

Structural and functional aspects of cultured epididymal epithelial cells isolated from pubertal rats.

Acidic epididymal glycoprotein (AEG) and androgen-binding protein (ABP) antisera were used to study functional activities of primary cell cultures of the epididymal epithelium of 20--23-day-old rats. Extensive AEG immunoreactivity was associated with almost all epithelial cells of the distal caput, corpus and cauda epididymidis. ABP immunoreactivity was solely confined to some epithelial cells of the caput epididymidis. AEG and ABP immunoreactive cells were identified as principal cells. Morphological studies of enzymically dispersed aggregates of the epididymal epithelial cells showed that stromal cells were satisfactorily removed and that cell aggregates consisted of a predominant population for cells displaying the morphological characteristics of principal cells. Scanning and transmission electron microscopic studies of cultured epididymal epithelial cells in monolayers demonstrated that microvilli and pit-like invaginations of the cell surface were preserved during the first 7--10 days of culture and then gradually disappeared. Other characteristic subcellular structures such as Golgi apparatus and rough endoplasmic reticulum cisterna were preserved. Electrophoretic analysis of [35S]methionine-labelled secretory polypeptides released by epididymal epithelial cells into the culture medium demonstrated a distinct protein band pattern which differed from that observed in the medium of cultured rat Sertoli cells. These results demonstrate that primary cultures of epididymal epithelial cells isolated from sexually immature rats maintain several differentiated characteristics of the intact organ and therefore provide a valuable system for the study of epididymal epithelial cell function.

Androgen-Binding Protein↗
PubMed1997

Regulation of mouse epididymal epithelium in vitro by androgens, temperature and fibroblasts.

The epididymal epithelium provides the microenvironment for sperm maturation. However, the molecular basis of epididymal function is still poorly understood because of the limitations of in vivo systems. For this reason, we have developed an in vitro culture system for mouse epididymal epithelial cells. Cells were purified by enzymatic digestion and centrifugation through a Percoll gradient, and plated on inserts coated with a replacement basement membrane. Cultured cells maintained ultrastructural and immunocytochemical features of epithelia, but did not retain the androgen responsiveness of epididymal cells (as judged by androgen receptor detection and secretion of specific markers) unless cocultured with fibroblasts. The androgen receptor was detected in the nuclei of epididymal epithelial cells only when grown with epididymal fibroblasts in the subjacent chamber. Moreover, specific epididymal secretory proteins were secreted only when epithelial cells were cultured in the presence of both androgens and fibroblasts at 32 degrees C. These results highlight the importance of cell-cell interaction, as well as temperature regulation in the physiology of the epididymis. They also establish the existence of two independent pathways in the differentiation of these cells. The first, leading to the expression of epithelial characteristics, is fibroblast-independent, whereas the second, conferring tissue-specific features, depends upon coculture with fibroblasts.

Androgens↗
PubMed2002

Epididymal anomalies associated with patent processus vaginalis in hydrocele and cryptorchidism.

The epididymal anomalies and patent processus vaginalis are frequently seen in patients with cryptorchidism or hydrocele. We performed a prospective study on the relationship between the epididymal anomalies and the patency of the processus vaginalis in boys with hydrocele (190 cases) or cryptorchidism (89 cases) who were treated from August 1997 to February 2000 (mean age, 51 months; range, 12 to 152 months). The epididymal anomalies were observed with an overall frequency of 48%. Closed, partially closed, and open processus vaginalis were associated with an epididymal anomaly in 14, 38, and 65% of cases, respectively. The epididymal anomalies were more common in association with undescended (61%) than with descended (43%) testes without statistical significance (p=0.415). Incomplete attachment of the caput epididymis was the most common anomaly (35%), followed by detachment of caput and cauda epididymis (31%), cauda epididymis (24%), and long looping epididymis (10%). These data showed that the epididymal anomalies were strongly associated with the patency of the processus vaginalis irrespective of testicular descent (p<0.001), and they provide further evidence for the hypothesis that a common stimulus, possibly androgens, may be required for the epididymal development and obliteration of the processus vaginalis.

Adolescent↗
PubMed1999

[Epididymal cysts in adolescents].

Epididymal tumours are uncommon in children and adolescents and are usually benign. Epididymal cyst is exceptionally reported in the literature, although it is certainly underdiagnosed. The authors report 3 cases of epididymal cyst in 3 children, 12, 14 and 16 years of age. These children presented with an uncomfortable scrotal mass and were treated by excision of the cyst in every case. The aetiology of epididymal cysts is unclear. It is probably a congenital abnormality related to hormonal disorders during embryonic life. Physical examination is very important, but not sufficient for the diagnosis and must be completed by scrotal ultrasonography, which shows an echo-free cystic epididymal structure. Despite ultrasonography, the differential diagnosis of other scrotal cystic masses and even some solid epididymal tumours, which may present all of the sonographic characteristics of a cyst, must be considered. The treatment of symptomatic epididymal cyst in children must be surgical. For asymptomatic cysts diagnosed by sonography, clinical follow-up to document stability of the mass is justified.

Adolescent↗
PubMed1999

Fertilization capability of frozen epididymal sperm for intracytoplasmic sperm injection.

Both microsurgical epididymal sperm aspiration (MESA) and intracytoplasmic sperm injection (ICSI) are great advances in assisted reproductive techniques. By using the ICSI technique, frozen sperm from the epididymis can result in successful fertilization. The epididymal sperm retrieved via MESA can be cryopreserved for an in-vitro fertilization (IVF) procedure, thus, making repeat surgical retrieval of sperm unnecessary. We report a retrospective analysis of 24 ICSI cycles in 16 patients with obstructive or nonreconstructable azoospermia. Fresh epididymal sperm was used in 13 ICSI cycles and frozen-thawed epididymal sperm was used in the other 11. We compared the fertilization capability of ICSI using frozen-thawed epididymal sperm with fresh epididymal sperm. Eleven patients became pregnant and five of these pregnancies resulted from frozen epididymal sperm. The fertilization rate per oocyte was 58% with fresh sperm, and 66% with frozen-thawed sperm. The rate of clinical pregnancy for one embryo transfer was 46% with fresh sperm, and 45% with frozen-thawed sperm. There were no significant differences between fresh and frozen-thawed spermatozoa in the fertilization rate of oocytes or the clinical pregnancy rate. Our results suggest that we should cryopreserve supernumerary spermatozoa during a MESA/ICSI procedure in order to avoid repeated scrotal surgery.

Adult↗
PubMed1998

[Primary epididymal tumors].

OBJECTIVE: To improve the diagnosis of primary epididymal tumors. METHOD: In 23 primary epididymal tumors (22 benign and 1 malignant), 14 were adenomatoid tumors, 6 were leiomyomas, 1 was multiple fibrous pseudomonas and 1 was sclerosing hemangioma. Epididymal adenocarcinoma was seen in a 57-year-old man. RESULT: Epididymal tumors might be easily misdiagnosed as epididymal tuberculosis, chronic epididymis and spermatocele. Apart from certain benign clinical characteristics, benign epididymal tumors could be diagnosed by B-ultrasonography and aspiration biopsy. Most malignant tumors of epididymis presented as rapidly growing scrotal masses and cured by the removal of tumors or the whole epididymis of the same side. 19 of 22 cases were free from recurrence. CONCLUSION: Malignant epididymal tumors should be treated as malignant tumors of testis and their prognosis is extremely poor.

Adenocarcinoma↗
PubMedDate not supplied

A therapeutic alternative in the treatment of epididymal cysts: percutaneous sclerotherapy.

PURPOSE: Epididymal cysts are benign structures commonly seen during urological or ultrasound testicular examinations. They are treated only if symptomatic. Surgery is the standard treatment, although it carries a high risk of complications. We report our experience with sclerotherapy of epididymal cysts with sonographic assistance, and present an analysis of costs. MATERIALS AND METHODS: Between January 1999 and December 2000 we examined 48 epididymal cysts in 45 patients during ultrasound examinations. Except one, all of the patients were symptomatic. We decided to treat 25 symptomatic cysts that were more than 5 cm. in diameter. All of the patients refused surgery as they knew of the existence of the percutaneous method. The procedures were performed on a out-patient basis with ultrasound assistance and using 3% Polidocanol for sclerosis. Follow-up was at 3/6 and 12 months after treatment. If we found persistence of symptoms and/or a cyst was more than 5 cm of diameter, a second session was proposed. RESULTS: Twenty-five epididymal cysts were treated, with a technical success of 100%. Mean fluid evacuation was 36 ml; mean sclerosing agent injected was 4.5 ml. There were no complications. After 3/6 months 17/25 patients were free of symptoms (68%) and the cysts had disappeared in 15 of them (60%). After the repeat procedure, performed on only 4 patients, the number of symptom-free patients was 21/25 (84%). DISCUSSION: Epididymal cysts are often solitary cystic fluid collections that are occasionally reported during physical or ultrasound urological clinical examinations. They present as painless testicular enlargements on palpation and are echo-free at ultrasound examination. Percutaneous sclerotherapy has gained wide acceptance in the therapeutic handling of other pathologies and it appears as an ideal solution for this benign clinical condition of the superficial structures. Polidocanol, widely used on varicose vein therapies for its local anaesthetic properties, is the ideal sclerosing agent for superficial and delicate structures, such as epididymal cysts. The results of the present study have shown that symptomatic cysts may be cured in 84% of cases without complications and with low costs. CONCLUSIONS: Percutaneous sclerotherapy is a valid therapeutic alternative to surgery in the management of epididymal cysts: it is safe, effective, free of complications, less costly and shows good results in the follow-up.

Adolescent↗
PubMedDate not supplied

Direct effects of ethane dimethanesulphonate on epididymal function in adult rats. An in vitro demonstration.

It was recently demonstrated that the Leydig cell toxicant ethane dimethanesulphonate (EDS) produces multiple effects on the epididymis after a single in vivo exposure. To determine whether any of the perturbations were mediated by a direct action of the compound, we used a novel system for the coculture of epididymal epithelial cells and sperm from the caput epididymidis. This system maintains the morphologic integrity and cell polarity of the epididymal epithelial cells before and during coculture, and the sperm recovered after coculture have intact plasma and acrosomal membranes. In addition, several functions required for epididymal sperm maturation are expressed, including the secretion of protein by the epididymal epithelium, the association of secreted protein with the plasma membrane of cocultured sperm, and the acquisition of progressive motility by cocultured sperm. In vitro exposure of epididymal epithelial cells and sperm to EDS results in a significant decline in protein secretion by the epithelial cells during coculture, and in particular, a dose-dependent decline in a 36- to 38-kd protein (PI 4.0 to 4.5) and a 34- to 36-kd protein (PI 4.5 to 5.0). Moreover, these and other proteins are not recovered from the sperm membrane of cocultured sperm after EDS treatment. Finally, EDS results in a dose-dependent decline in the percentage of both motile and progressively motile sperm recovered after coculture compared with that of sperm from untreated cocultures. These effects on sperm motility were not observed when sperm were pretreated with EDS and subsequently cocultured with untreated epithelial cells. We conclude that EDS alters epididymal sperm maturation by acting directly on the epididymal epithelium to mediate changes in sperm membrane protein, and that this may subsequently alter the development of the progressive motility of sperm.

Animals↗
PubMed1989

[A case of primary bilateral epididymal leiomyoma].

Primary epididymal tumors are relatively rare entity. Leiomyoma is the second most common benign epididymal tumor. Recently, we experienced a case of primary bilateral epididymal leiomyoma. A 50-year-old man presented with painless masses in both epididymal tails several years in duration. Solid tumors of both epididymal tails with undetermined characteristics were diagnosed. At surgery both masses were solid and smooth and were found to be well-demarcated. They were removed without difficulty. Pathology was bilateral epididymal leiomyoma. We also reviewed 55 cases of epididymal leiomyoma in the Japanese literature.

Epididymis↗
PubMed1994

Relative force of human epididymal sperm.

OBJECTIVE: To assess the relative escape force of human epididymal sperm using a laser generated optical trap and compare it with that of human ejaculated sperm. DESIGN: Evaluation of the relative force generated by epididymal and ejaculated sperm using an 800-nm laser-generated optical trap system (titanium-sapphire, model 899-01; Coherent Innova, Palo Alto, CA). SETTING: University-based facility at the Beckman Laser Institute and Medical Clinic and Center for Reproductive Health, University of California, Irvine. INTERVENTIONS: A total of 2,720 sperm from 28 samples were randomly analyzed. Fifteen were ejaculated samples (1,650 sperm) obtained from men with proven fertilization, and 13 were epididymal samples (1,070 sperm) aspirated microsurgically from patients with obstructive azoospermia. An optical trap equipped with the 100x Neofluar objective was used to analyze an average of 100 sperm per patient. MAIN OUTCOME MEASURES: Determination of mean relative escape force values in milliwatts for epididymal and ejaculated sperm samples. RESULTS: The mean relative escape force for epididymal sperm was 32.4 mW, significantly lower than ejaculated sperm, which was 85.1 mW. By correlating epididymal sperm relative force with fertilization in vitro at an arbitrary cutoff value of 30 mW, it was found that no fertilization occurred if a sample had < 13% of sperm at that value. CONCLUSIONS: [1] The average relative escape force of the epididymal sperm was found to be 60% weaker than that of ejaculated sperm. [2] It is demonstrated that the noncontact laser optical trap is a sensitive tool that can evaluate single sperm force as a new physiological parameter.

Ejaculation↗