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The maintenance of motility and the surface properties of epididymal spermatozoa from bull, rabbit and ram in homologous seminal and epididymal plasma.

Epididymal spermatozoa from bull, rabbit and ram were incubated in homologous epididymal plasma or seminal plasma in a buffered saline-based medium with or without serum albumin. The spermatozoa were either diluted directly into the medium or were washed first. No effect of washing was observed on the subsequent reaction of the cells to the different media. A considerable proportion of the populations of epididymal spermatozoa survived (i.e. continued to exhibit motility) for up to 22 h at 30 degrees C in the simple saline-based medium. Initially epididymal plasma had a slight stimulatory effect on sperm motility in ram and bull but it had no effect on sperm survival in any of the 3 species. Seminal plasma stimulated motility markedly in ram initially, but in all 3 species seminal plasma was detrimental to survival: in ram even a 15-min exposure to the fluid reduced survival. Serum albumin also stimulated motility; it delayed, but did not prevent, the detrimental effect of seminal plasma, although it had no effect itself on survival. The effects of epididymal plasma, seminal plasma and serum albumin on surface properties of epididymal spermatozoa, i.e. agglutination, sticking-to-glass and eosinophilia, were also noted. These varied between species and there was no correlation between these effects and the effects on motility and survival.

Animals

Effect of alpha-chlorohydrin on epididymal sperm and epididymal plasma in swine.

Investigation of the contraceptive mechanism of alpha-chlorohydrin was done by analyses of epididymal plasma and certain epididymal sperm characteristics after oral administration of 0, 5, 10 or 30 mg/kg, day of the drug to boars for 15 days. Water resorption in caput epididymidis was slightly decreased in all treatment groups. Sodium, potassium, chloride, glycerylphosphorylcholine levels, and seminal antigens in epididymal plasma were not altered significantly by 10 or 30 mg/kg of the drug. The boars on 5 mg/kg exhibited significantly elevated sodium, potassium, or chloride values in various segments. Motility was significantly lower on corpus and proximal cauda epididymal spermatozoa from alpha-chlorohydrin treated boars. Only boars receiving 30 mg/kg exhibited impaired sperm motility in the distal cauda. The movement of the cytoplasmic droplets to the distal position was retarded in boars on the two highest dose levels. The results suggest that the contraceptive effects of alpha-chlorohydrin in the boar is probably not mediated via an impaired epididymal function.

Animals

[Epididymitis and fertility. Treatment results in acute unspecific epididymitis].

Spermiograms of 46 patients were examined 8 days, 4 weeks, 3 months, 6 months and 1 year after acute unilateral non-specific epididymitis. The treatment consisted in 24 cases besides antibiotic treatment with doxycycline in local infiltrations of the spermatic cord with 1% Scandicain solution. Primarily there was found an inhibiting effect on spermatogenesis. In more than 2/3 of the cases a varying oligoasthenospermia was shown 8 days after beginning of the infection. In the further course this disturbance of fertility improves; if local infiltration of the spermatic cord with 1% Scandicain solution is performed, the improvement is more evident. Still, there remain about 20% of the cases in which the fertility disturbance is so severe, that sterility results. One year after the beginning of the infection spermagglutinating antibodies could be shown only in 1 case. The seminal fluid fructose was controlled over 1 year following the beginning of infection and found to be normal in all cases.

Acute Disease

Lectin-binding sites on the plasma membranes of rabbit spermatozoa. Changes in surface receptors during epididymal Maturation and after ejaculation.

MODIFICATIONS IN RABBIT SPERM PLASMA MEMBRANES DURING EPIDIDYMAL PASSAGE AND AFTER EJACULATION WERE INVESTIGATED BY USED OF THREE LECTINS: concanavalin A (Con A); Ricinus communis I (RCA(I)); and wheat germ agglutinin (WGA). During sperm passage from caput to cauda epididymis, agglutination by WGA drastically decreased, and agglutination by RCA(I) slightly decreased, although agglutination by Con A remained approximately unchanged. After ejaculation, spermatozoa were agglutinated to a similar degree or slightly less by Con A, WGA, and RCA(I), compared to cauda epididymal spermatozoa. Ultrastructural examination of sperm lectin-binding sites with ferritin- lectin conjugates revealed differences in the densities of lectin receptors in various sperm regions, and changes in the same regions during epididymal passage and after ejaculation. Ferritin-RCA(I) showed abrupt changes in lectin site densities between acrosomal and postacrosomal regions of sperm heads. The relative amounts of ferritin-RCA(I) bound to heads of caput epididymal or ejaculated spermatozoa. Tail regions were labeled by ferritin RCA(I) almost equally on caput and cauda epididymal spermatozoa, but the middle-piece region of ejaculated spermatozoa was slightly more densely labeled than the principal-piece region, and these two regions on ejaculated spermatozoa were labeled less than on caput and cuada epididymal spermatozoa. Ferritin-WGA densely labeled the acrosomal region of caput epididymal spermatozoa, although labeling of cauda epidiymal spermatozoa was relatively sparse except in the apical area of the acrosomal region. Ejaculated spermatozoa bound only a few molecules of ferritin-WGA, even at the highest conjugate concentrations used. Caput epididymal, but not cauda epididymal or ejaculated spermatozoa, bound ferritin-WGA in the tail regions. Dramatic differences in labeling densities during epididymal passage and after ejaculation were not found with ferritin-Con A.

Animals

Epididymal extravasation following vasectomy as a cause for failure of vasectomy reversal.

Twenty-eight men undergoing vasectomy reversal who were found to have no sperm in the proximal vas fluid on one or both sides underwent microscopic epididymal exploration. In 33 of 39 cases so explored, normal sperm were found in the epididymal fluid of the corpus, despite absence of sperm in the vas fluid. Epididymal histology distal to this site revealed extensive interstitial sperm granulomas resulting from rupture of the epididymal duct. Testicular biopsy revealed normal spermatogenesis. Secondary epididymal obstructions were noted when there was copious fluid in the vas deferens proximal to the vasectomy site as well as when there was scanty fluid. It is concluded that persistent azoospermia after an accurate microscopic vasovasostomy results from the secondary epididymal obstruction induced by rupture of the epididymal duct related to the pressure increase after vasectomy.

Adult

Effect of hyperthermia on epididymal cyclic AMP levels in diabetic non-diabetic and hypophysectomized rats.

The effect of elevated body temperatures on the concentrations of epididymal cyclic AMP levels in non-diabetic, diabetic and hypophysectomized rats was studied. Cyclic AMP levels were increased during hyperthermia in all animals examined. This increase in epididymal cyclic AMP concentration was not seen in animals that had been supplemented with exogenous insulin prior to the experiment. The effect of pituitary lipolytic hormones on epididymal cyclic AMP levels was also investigated. Significant elevations of epididymal cyclic AMP levels were observed in hypophysectomized rats during hyperthermia indicating that pituitary hormones are not essential in causing these increases. Extrapituitary hormones, such as glucagon, might be responsible for epididymal cyclic AMP increases. Increases in epididymal cyclic AMP levels may therefore be the result of the reduction of blood insulin and concomitant increases of lipolytic hormones of both pituitary and extrapituitary origins.

Adipose Tissue

In vitro fertilizing ability of testicular, epididymal, and ejaculated rabbit spermatozoa.

The fertilizing ability of testicular, epididymal, and ejaculated rabbit spermatozoa was evaluated in vitro following in vitro capacitation by high ionic strength treatment. Fewer than 11% of inseminated ova were apparently fertilized (i.e., in pronuclear, two-, and four-cell stages as determined by light microscopy) when testicular sperm treated with caffeine, caput epididymal, or corpus epididymal sperm samples were tested. A greater fertilizing ability, reflected by the percentage of ova fertilized and more normal progression of embryonic development, was exhibited by cauda epididymal sperm. Of 93 ova, 68 (73.1%) were fertilized by cauda sperm, whereas ejaculated sperm from the same 10 bucks fertilized 34 (36.6%) of 93 ova (P is less than 0.005). Ultrastructural examination of selected ova apparently fertilized by sperm from levels of the male reproductive tract proximal to the cauda epididymidis revealed abnormal activation. Authentic fertilization occurred when ova were inseminated with cauda epididymal and ejaculated sperm. An unusual and infrequent form of activation involving failure of cortical granule breakdown in ova penetrated by cauda epididymal and ejaculated sperm was seen. A comparison of fertilizing ability of sperm from first, second, and third ejaculates revealed a significant decrease with the third ejaculate (P is less than 0.01).

Animals

Etiology, manifestations and therapy of acute epididymitis: prospective study of 50 cases.

There were 50 patients with acute epididymitis who were evaluated prospectively by history, examination and microbiologic studies, including cultures for aerobes, anaerobes, Neisseria gonorrhoeae, Chlamydia trachomatis and Ureaplasma urealyticum. Escherichia coli was the predominant pathogen isolated from the urine of men more than 35 years old, while Chlamydia trachomatis and Neisseria gonorrhoeae were the predominant pathogens isolated from the urethra of men less than 35 years old. The etiologic role of Escherichia coli and Chlamydia trachomatis was confirmed by isolation from epididymal aspirates from a high proportion of men with positive urine or urethral cultures for these agents. Chlamydia trachomatis epididymitis accounted for two-thirds of idiopathic epididymitis in young men and often was associated with oligospermia. Of 9 female sexual partners of men with Chlamydia trachomatis infection 6 had antibody to Chlamydia trachomatis, of whom 2 had positive cervical cultures for this organism and 2 others had non-gonococcal pelvic inflammatory disease. Antibiotic therapy with tetracycline was effective for the treatment of men with Chlamydia trachomatis epididymitis and should be offered to the female sex partners.

Adolescent

Chlamydia trachomatis as a cause of acute "idiopathic" epididymitis.

To assess the etiologic role of C. trachomatis and other micro-organisms in "idiopathic" epididymitis, 23 men underwent microbiologic studies, including cultures of epididymal aspirates in 16. Eleven of 13 men under the age of 35 years had C. trachomatis infection whereas eight of 10 over 35 had coliform urinary-tract infection. Cultures of epididymal aspirates yielded C. trachomatis alone in five of six men under 35, and coliform bacteria alone in five of 10 over 35. These results suggest that C. trachomatis is the major cause of "idiopathic" epididymitis, and coliform bacteria the major cause of epididymitis in older men. Expressible urethral discharge and inguinal pain were more common in the chlamydial cases, whereas concurrent genitourinary abnormality and scrotal edema and erythema occurred more commonly in the coliform cases. The morbidity attributable to C. trachomatis is as serious as that attributable to Neisseria gonorrhoeae.

Acute Disease

Acute epididymitis: etiology and therapy.

Fifty patients with acute epididymitis were evaluated prospectively by history, examination, and microbiologic studies, including cultures for aerobes, anaerobes, N. gonorrhoeae, Chlamydia trachomatis, and Ureaplasma urealyticum. E. Coli was the predominant pathogen isolated from the urine of men over 35 years old and C. trachomatis and N. gonnorrheae were the predominant pathogens isolated from the urethra of men under 35. The etiologic role of E. Coli and C. trachomatis was confirmed by isolation from epididymal aspirates from a high proportion of men with positive urine of urethral cultures for these agents. C. trachomatis epididymitis accounted for two thirds of "idiopathic epididymitis" in men, and was often associated with oligozoospermia. Of nine female sexual partners of men with C. trachomatis infection, six had antibody to C. trachomatis, of whom two had positive cervical cultures for this organism, and the others had nongonococcal pelvic inflammatory disease. Antibiotic therapy with tetracycline was effective for the treatment of men with C. trachomatis epididymitis, and should be offered to their female sex partner.

Adolescent

The effect of a synthetic progestine on the fine structure of the epididymal head (stereological analysis).

The alterations induced in principal cells of the epididymal head tissue of the rat by administration of a synthetic progestine (R2323, ethinyl-norgestrienone) were studied by electron microscopy. A total of 3 male, sex mature rats was applicated ethinyl-norgestrienone during a 3 months period in low dosage (60 mug) per day and was studied with 5 control animals. Stereological methods devised by Weibel (1969) and Rohr et al. (1976) were applied to light and electron micrographs. The calculated values are related to 1 cm3 of epididymal tissue, 1 cm3 of epididymal cell and 1 cm3 of epididymal cell cytoplasm. The Golgi apparatus was further subdivided; the values for the Golgi compartments are therefore refered to 1 cm3 of Golgi apparatus was further subdivided; the values for the Golgi compartments are therefore refered to 1 cm3 of Golgi apparatus. In the treated animals there was, related to the unit of epididymal tissue, a significant decrease in the volume density of the interductular tissue, whereas that for the lumina was significantly higher. The volume density of the glandular epithelium remains unchanged. Refered to the unit volume of principal cell cytoplasm the rough endoplasmic reticulum shows a signficant decrease by 6.5%, whereas the smooth endoplasmic retiuclum is increased by 6.3%. The volume density of the lysosomes was calculated to be 4.3% (Control 2.5%). The stereological data of the Golgi apparatus indicate a vacuolar transformation: Increased volume of vacuoles, decreased volume fraction of saccules and vesicles. However, the volume density of the whole Golgi apparatus remains unchanged. An impaired secretory activity of the principal cells could be assumed.

Animals

Testicular torsion and epididymitis demonstrated by radionuclide angiograms and static imaging.

Radionuclide testicular angiography and static images were performed using technetium 99m sodium pertechnetate in an effort to differentiate between testicular torsion and acute epididymitis in 12 patients. The diagnosis of testicular torsion was made in four cases that were confirmed at surgery. Acute epididymitis or acute epididymo-orchitis was diagnosed six times. These patients were treated with broad-spectrum antibiotics leading to a subsidence of their clinical symptomatology during therapy with no sequelae. The radionuclide angiogram and static image changes of chronic epididymitis are also discussed.Radionuclide testicular angiograms and static images can be significant benefits in addition to the clinical and physical findings to distinguish between testicular torsion and acute and chronic epididymitis.

Diagnosis, Differential

In vitro fertilization of hamster eggs by ejaculated or epididymal spermatozoa in the presence of male accessory secretions.

In vitro fertilization of hamster eggs by ejaculated or epididymal spermatozoa in the presence of seminal plasma or male accessory gland secretions was examined. There was no difference in the penetration rates, the time of sperm penetration and the optimal sperm concentration between ejaculated and epididymal spermatozoa. The swelling of the zona pellucida, a high incidence of polyspermy, distortion of the vitellus and the degeneration of eggs were observed after incubation of ejaculated or epididymal spermatozoa in the presence of seminal plasma or accessory gland secretions. Using ejaculated spermatozoa, the fertilization rates of eggs with or without follicular cells were similar but fertilization by epididymal spermatozoa was inhibited by secretion of the seminal vesicle or ventral prostrate gland but not by that of the coagulating gland or the dorsal prostrate.

Animals

Effects of castration and androgen-substitution on the morphology of the epididymal epithelium of the Japanese monkey, Macacus fuscatus, as revealed by scanning electron microscopy.

The caput epididymidis from castrated and androgen-supplemented, castrated Japanese monkeys was observed with the scanning electron microscope. The experimental findings were compared with the normal structures in control animals. The epididymal lumen of control animals was lined by a tall, pseudostratified columnar epithelium possessing long, slender stereocilia which were densely arranged in a tuft-like form. After castration, the epididymal epithelium was decreased in height to one-fifth of controls. The stereocilia were also considerably reduced in length and in number, resulting in a flattened epithelial surface with polygonal boundaries. Frequent projection of a long, single cilium from an epithelial cell into the lumen was also a prominent feature in the epididymal ducts of the castrated animals. Administration of testosterone to the castrated animals resulted in almost complete recovery of the epididymal epithelium as well as regeneration of the stereocilia which regained a tuft-like arrangement.

Animals

Changes in 5alpha- dihydrotestosterone binding to epididymal cytosol during sexual maturation in rabbits: correlation with morphological changes in the testis and epididymis.

We previously demonstrated that the caput epididymis of intact sexually mature rabbits contains a specific high-affinity binding protein for 5alpha-dihydrotestosterone (5alphaDHT). The other anatomical segments (corpus and cauda) of the epididymes of these animals had no detectable 5alphaDHT-binding activity. We have further shown that this binding was due to an androgen-binding protein of testicular origin. In the present study we have investigated 5alphaDHT binding to epididymal cytosol from sexually immature rabbits (20-104 days old). Using sucrose gradient ultracentrifugation, we have detected a unique pattern of binding. The pattern correlated well with testicular and epididymal maturation, but there was little correlation with chronological age or body weight. In the most immature animals (Group I) the seminiferous tubules appeared as solid cords and the epithelium of the ductus epididymis detectable 5alphaDHT-binding activity. In the second group (Group II), there was 5alphaDHT-binding to all three segments. The seminiferous tubules of these rabbits exhibited spermatogenic activity and lumen formation. The height of the epididymal epithelium had increased uniformly throughout the duct. The third group (Group III) had 5alphaDHT-binding only in caput cytosol. Spermatogenesis had progressed to the formation of elongated spermatids in the most immature animals of this group to the release of spermatozoa in the most mature ones. The caput epithelium of this last group of rabbits was fully differentiated. Unilateral orchidectomy of Group II rabbits resulted in a decrease in [3H]5alphaDHT-binding activity on the operated side as compared to the contralateral non-operated control side, suggesting the testicular origin of the binding protein. The failure of cyproterone or cyproterone acetate to inhibit [3H]5alphaDHT-binding to the protein, the lack of effect of N-ethylmaleimide on binding, and the rapid dissociation rate of the [3H]5alphaDHT-binding protein complex suggested that the binding moiety was testicular androgen-binding protein (ABP).

Aging

Activity and androgenic control of enzymes associated with the tricarboxylic acid cycle, lipid oxidation and mitochondrial shuttles in the epididymis and epididymal spermatozoa of the rat.

1. Enzyme activities (units/g wet wt.) were determined in the caput and cauda epididymidis and in epididymal spermatozoa of the rat. 2. The activity of most enzymes in the cauda was between 50 and 100% of that in the caput, except that ATP citrate lyase was barely detectable in the cauda. 3. Spermatozoa, unlike epididymal tissue, contained sorbitol dehydrogenase but lacked ATP citrate lyase. NADP+-malate dehydrogenase, mitochondrial glycerol 3-phosphate dehydrogenase, succinate dehydrogenase, carnitine acetyltransferase and citrate synthase were 5 to 400 times as active in spermatozoa as in epididymal tissue. 4. 2-Oxoglutarate dehydrogenase was the least active member of the tricarboxylic acid cycle in all tissues and most closely matched the measured flux through the cycle. 5. The concentrations of hydroxyacyl-CoA dehydrogenase and carnitine palmitoyltransferase were equivalent to the more active enzymes of the tricarboxylic acid cycle, indicating the capacity for extensive lipid oxidation, and the presence of 3-hydroxybutyrate dehydrogenase suggests that these tissues can also oxidize ketone bodies. 6. Transfer of reducing equivalents from cytoplasm to mitochondrion is unlikely to occur by means of the glycerol phosphate cycle because mitochondrial glycerol 3-phosphate dehydrogenase is relatively inactive in epididymal tissue, whereas the cytoplasmic enzyme has little activity in spermatozoa, but transfer may be accomplished by the malate-aspartate shuttle. 7. Transfer of acetyl units from mitochondrion to cytoplasm could be effected by the pyruvate-malate cycle in the caput of androgen-maintained rats, but not in the other tissues because of the low activity of ATP citrate lyase. Acetyl unit transfer could take place via acetylcarnitine, mediated by carnitine acetyltransferase. 8. Castration resulted in a decrease in the concentration of nearly all enzymes, although subsequent administration of testosterone restored concentrations to values similar to those in animals maintained by endogenous androgen. The extent to which enzyme concentration was changed by an alteration in androgen status was highly variable, but was most marked in the case of pyruvate carboxylase.

Androgens