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PubMed1979

Micro-stereological study of the avian epididymal region.

Morphometric studies of the epididymal region of three avian species (Gallus domesticus, Coturnix coturnix japonica, Japanese quail, and Numida meleagris, guinea-fowl) were performed using a stereological compensating eyepiece graticule. The proportions of the volume of the region made up of rete testis lacunae, proximal and distal efferent ductules, connecting ducts, ductus epididymidis, blood vessels and connective tissue were determined. The efferent ductules constituted the largest proportion of the tubular components of the organs in the three birds (35.3% for the chicken, 56% for the quail and 61.9% for the guinea-fowl), while the ductus epididymidis formed only 7.6%, 2.4% and 1.8% of the total epididymal volume in the domestic fowl, quail and guinea-fowl respectively. The connective tissue component made up 38.7%, 27.3% and 22.6% respectively. The significance of these results is discussed in relation to the likely functions of the various components of the epididymal region.

Animals↗
PubMed1984

GPLA antigens are not expressed on guinea-pig epididymal spermatozoa.

The expression of guinea-pig major histocompatibility antigens (class I and II) has been investigated on guinea-pig epididymal spermatozoa. Specific alloantisera (anti-B1, anti-B3, anti-Ia2,4 and anti-Ia1,3,7) were obtained by cross-immunization of strains 2, 13 and BIO-AD animals with relevant spleen cell membranes. These sera were tested on spermatozoa from the same three strains by a protein A rosetting assay and by an indirect immunofluorescence test. The results obtained showed non-strain specific reactions of all the alloantisera tested on epididymal spermatozoa of the three strains. These non-strain specific reactions were absorbed by spermatozoa of any strain but they were not absorbed by the splenic cells of the same animals. On the other hand, the alloantisera specific reactivity on peripheral blood cells was not diminished after incubation of the sera with spermatozoa. Furthermore anti-B1 and anti-B3 antibodies eluted from guinea-pig platelets did not react with any spermatozoa but reacted with the relevant peripheral blood cells. These results indicate that the studied guinea-pig sera contained two types of antibodies: anti-MHC antibodies able to react with relevant blood cells but not with spermatozoa and sperm specific antibodies (also observed in untreated and DNP-BGG immunized guinea-pig sera) reacting with all spermatozoa but not with blood cells. They are not compatible with the expression of MHC antigens at the surface of guinea-pig epididymal spermatozoa.

Animals↗
PubMed1984

Characterization of a maturation-associated glycoprotein on the plasma membrane of rat caudal epididymal sperm.

A Mr = 32,000 membrane glycoprotein can be uniquely labeled by galactose oxidase/[3H]sodium borohydride on rat caudal, but not caput, epididymal sperm. It has been suggested that this protein is related to a Mr = 32,000 galactose oxidase-sensitive glycoprotein present in rat caudal epididymal fluid. The tritiated membrane glycoprotein was solubilized and its hydrodynamic properties were determined by conventional gel filtration, high performance gel filtration, sedimentation rate determination in linear sucrose gradients prepared in H2O and D2O, and equilibrium isopycnic centrifugations in CsCl. The Stokes radius and sedimentation coefficient were 4.87 +/- 0.07 nm and 1.73 +/- 0.08 S, respectively. The sedimentation profile in CsCl gradients was asymmetric with a major peak occurring at a density of 1.081 g/cm3 (v = 0.92 cm3/g) and a shoulder at 1.108 g/cm3 (v = 0.90 cm3/g). The glycoprotein did not enter a 5 to 20% linear sucrose gradient prepared in D2O and could be extracted from the intact sperm into acidic chloroform:methanol solutions. These data are consistent with a protein which binds substantial amounts of detergent and/or lipid and has exposed hydrophobic regions. Two-dimensional gel electrophoresis indicated that the membrane protein exhibits charge heterogeneity, with the major components having pI values of 5.4 and 4.9. The fluid glycoprotein was monodisperse on two-dimensional gel electrophoresis having a pI of 3.8. Binding studies failed to demonstrate specific binding of the Mr = 32,000 caudal fluid glycoprotein to caput cells. Moreover, "Western blots" of electrophoretically resolved caput and caudal fluid proteins, followed by immunolabeling with antibodies raised against unfractionated caudal fluid, demonstrated the presence of a Mr = 32,000 protein in caudal fluid which was absent from caput epididymal fluid. Using the same technique, it was shown that antibodies raised against caudal fluid proteins did not cross-react with a Mr = 32,000 caudal membrane glycoprotein. Our data do not support the view that the Mr = 32,000 fluid and membrane proteins are identical.

Animals↗
PubMed1981

[The first cleavage of tubal sheep eggs after fertilization with epididymal or ejaculated spermatozoa].

Prealpes ewes (N = 40) were inseminated with mature (ejaculated or epididymal cauda) or immature (epididymal corpus) spermatozoa deposited in the uterine cavity at the time of an induced ovulation (61 hrs. after PMSG injection). Intratubal eggs, recovered and examined either 38 or 48 hrs. later, showed significant differences in segmentation depending on the degree of epididymal maturation of the spermatozoa. Proportion cleaved: 85% of eggs were cleaved after insemination with mature spermatozoa and only 30% after immature spermatozoa were used. Stage of cleavage: 48 hrs. after insemination with mature spermatozoa, 87% eggs were beyond the 4-cell stage, versus 0% after immature spermatozoa.

Animals↗
PubMed1982

Epididymal maturation of the surface protein structure of mammalian spermatozoa.

The surface proteins of intact rat, ram, boar and bull spermatozoa from caput and cauda epididymidis were radioiodinated and analysed by SDS-polyacrylamide gel electrophoresis. The maturational changes in sperm surface protein patterns of the different species were compared to elucidate any possible common features or differences in the surface protein reorganization mechanism of mammalian spermatozoa during epididymal passage. New labelled proteins appeared on the surfaces of the rat and ram spermatozoa, whereas bull spermatozoa lost some surface protein components during maturation. The surface proteins of boar spermatozoa could be only weakly labelled by radioiodination and no distinct maturational change in the radioactivity patterns of labelled boar spermatozoa could be shown. The analysis of radioiodinated ram spermatozoa also revealed an obvious transformation of the labelled membrane lipids during epididymal transit. The conclusion is that the surface proteins of mammalian spermatozoa undergo some changes during epididymal maturation but there is no uniform mechanism in these changes common to all mammalian species.

Animals↗
PubMed1981

The blood epididymal barrier to [3H]-inulin in intact and vasectomized hamsters.

The net transport of [3H]-inulin into the fluids of the hamster seminiferous and caput, corpus, and cauda epididymal tubules was examined in both intact animals and those vasectomized 10 months previously. Mean isotope concentrations in reproductive tract tubule fluids did not exceeded 10 per cent of blood plasma isotope concentrations during the experiment. There were no significant differences in net transport of [3H]-inulin into any of the tubule fluids sampled. Ten months after vasectomy, the seminiferous tubule, and all regions of the epididymal tubule retain the capacity to exclude [3H]-insulin. Thus in the hamster 10 months after vasectomy, the blood testis and blood epididymal barriers to inulin are intact.

Animals↗
PubMed1981

[Influence of prolonged physical training on the composition of fatty acids of epididymal adipose tissue and of the carcass in the young rat on a dietary regime of sunflower, rapeseed or primor oil].

Weanling SPF male rats were fed for 12 weeks purified diets containing by weight 15 p. 100 sunflower oil, high erucic acid rapeseed oil or low erucic acid rapeseed oil. During the same period of time they were subjected to a moderate treadmill-running program and compared with sedentary animals fed the same diets. At the end of the experiment, epididymal and perirenal adipose tissues were removed and weighed, carcass lipids were extracted and weighed and their fatty acid compositions were determined by GLC on glass capillary columns. Training decreased body weight, fat pad weight and lipid content of the carcass. However the decrease of fat pad weight in response to exercise was more important in the case of perirenal adipose tissue than in that of epididymal adipose tissue. Fatty acid composition was greatly influenced by the nature of the dietary lipids but little affected by training. Palmitic, palmitoleic, oleic, linoleic and eicosenoic acid stores in the epididymal fat of trained rats decreased but not those of stearic, linolenic and erucic acids.

Adipose Tissue↗
PubMed1980

Maturation of equine epididymal spermatozoa.

Spermatozoa from four regions of the epididymis and from ejaculated semen were evaluated for their resistance to cold shock, progressive motility, and structural stability. Spermatozoa were incubated at 38 C and their percentage of eosinophilia was compared with that of spermatozoa cooled to 0 C in 2 minutes, 10 C in 12 minutes, or 4 C in 22 minutes. Spermatozoa motility was estimated visually under phase-contrast microscopy and was recorded by cinematography. Structural stability of spermatozoa incubated in 0.05 M sodium borate buffer, 0.035 M sodium dodecyl sulfate (SDS), 0.002 M dithiothreitol (DTT), or SDS+DDT was evaluated by phase-contrast and electron microscopy. The percentage of eosinophilic spermatozoa did not differ among regions of the epididymis and was not increased by cold shock. Cooling ejaculated spermatozoa to 0 C in 2 minutes increased (P < 0.01) the occurrence of eosinophilia (32% vs 73%). Spermatozoa released from the caput or proximal corpus epididymidis into 0.154 M NaCl were not progressively motile; only 4% of the spermatozoa from the distal corpus were motile Cauda epididymal and ejaculated spermatozoa exhibited similar motility (41% vs 47%). Stability of chromatin was greater in spermatozoa from the distal corpus than those from the caput epididymis. Chromatin of spermatozoa from the distal corpus was resistant to 7.5 minutes of SDS+DTT treatment, whereas virtually all spermatozoal nuclei from the caput were decondensed by SDS alone. Tail organelles of spermatozoa acquired stability between the proximal corpus and the cauda epididymidis. All tail organelles of spermatozoa from the proximal corpus were dissolved by 7.5 minutes of SDS treatment, whereas tail organelles of distal corpus epididymal spermatozoa had dissolved after 7.5 minutes of SDS+DTT treatment. Stability of tail organelles in cauda epididymal and ejaculated spermatozoa was similar. Seemingly, equine spermatozoa are infertile until they enter the cauda epididymidis.

Animals↗
PubMedDate not supplied

Function of human epididymal spermatozoa.

Human epididymal spermatozoa taken from caput, corpus, and cauda were investigated to determine their fertilizing capacity (22 epididymides from 11 patients who had undergone orchidectomy because of prostatic cancer). The following functions, which have been reported to correlate positively with the fertilization rate, were determined: motility and progressive motility, chromatin condensation (assessed by aniline blue staining), acrosin activity, and induction of acrosome reaction by low temperature. In addition, stimulation of motility by pentoxifylline and phosphatidylcholine was examined. The results showed that motility, progressive motility, normal chromatin condensation, and inducible acrosome reaction increased from the caput to the cauda epididymidis, whereas acrosin activity was normal in all sections. Stimulation of progressive motility, especially that of caput spermatozoa, could be achieved by both pentoxifylline and phosphatidylcholine, the latter being definitely superior. In conclusion, our study confirmed that human spermatozoa in physiological status undergo several steps of maturation during the epididymal transit. Stimulation of sperm motility by phosphatidylcholine may be helpful for patients in whom epididymal spermatozoa are used for assisted reproduction.

Acrosin↗
PubMed1995

Management of the epididymal tubule during an end-to-side vasoepididymostomy.

Microsurgical repairs of epididymal obstruction have improved because of specific tubule anastomoses. However, these techniques require considerable microsurgical skills and any modifications to organize and simplify these procedures seem desirable. Four modifications are described for the management of the epididymal tubule during end-to-side vasoepididymostomy, including closed tubule fixation, micro-tubulotomy, micro-suction and placement of double-armed sutures through the epididymal tubule before attachment of the vas. These procedures have been performed on 51 patients. "Mixed procedures," or a modified vasoepididymostomy on 1 side and a vasovasostomy on the other, were done in 32 cases and "pure procedures," which included bilateral modified end-to-side vasoepididymostomy (16) and unilateral procedures (3), were done in 19. The semen and pregnancy data for the 19 cases of pure procedures were patency rate 58% in 11 and pregnancy rate 42% in 8. These modifications appeared to be helpful to organize and simplify several aspects of the procedure, and the results were improved compared to other techniques.

Anastomosis, Surgical↗
PubMed1993

Transepithelial movement of 3H-androgen in rat seminiferous and caput epididymal tubules: saturability and effect of competition with estradiol.

Proluminal movement of 3H-testosterone from peritubular space to intratubular fluids of the adult rat testis and epididymis was studied by using in vivo microperfusion and subsequent micropuncture of seminiferous tubules and caput epididymal tubules. Tubules were perifused with Minimum Essential Medium containing 3H-testosterone. To determine if androgen transport is saturable, 40, 80, 160, and 320 microCi 3H-testosterone were included in the perfusion fluid. Radioactivity of 3H-androgens in the intraluminal fluids was determined at 1 hour after perfusion. Transepithelial 3H-androgen movement in the testis was linear (r = 0.735, p < 0.001, y = 0.17 x +5.6). The movement of 3H-androgen across the epididymal epithelium was saturable (Vmax of 326.5 nM/hr and Km of 77 nM). To determine the effect of estradiol on proluminal androgen movement, estradiol at 10X the cocentration of 3H-testosterone was incorporated in the perifusion fluid. Proluminal 3H-androgen movement into the seminiferous and epididymal tubules was not affected by addition of estradiol to the perifusion fluid. These findings support our previous observations that proluminal transepithelial movement of 3H-androgens could be mediated by its binding to a specific intraluminal androgen binding protein.

Animals↗
PubMed1994

Intraluminal content is required for the maintenance of antigrade proluminal movement of 3H-androgens into rat caput epididymal tubules.

The present study was undertaken to determine whether or not antigrade, proluminal 3H-androgen movement in the caput epididymis occurs in the absence of native lumen content. A single tubule was perfused with artificial caput fluid containing no androgen-binding protein for 30 min and subsequently tubules were perifused with Minimum Essential Medium perifusion fluid containing 26.7 microCi/ml 3H-testosterone and 1.3 microCi/ml 14C-polyethyleneglycol for 1 h. Radioactivity of isotopes in perifusion and intraluminal fluids was determined at 1 h after sustaining perifusion, and the percentage of peritubular isotopes appearing in the intraluminal fluid was determined. Net entry of 3H-androgen into the epididymal tubules in the presence of native intraluminal content was approximately 323%. In contrast, intraluminal 3H-androgen concentrations in the epididymal fluid in the absence of native lumen content were significantly reduced, to 100% of those in the peritubular fluid. Antigrade, proluminal movement of 3H-androgen in the caput epididymis does not occur in the absence of native lumen content. Androgen-binding protein in the epididymal lumen may be required to maintain uphill proluminal movement of 3H-androgen into the tubules.

Androgens↗
PubMed1994

[Epididymal fine needle aspiration replacing vasography].

Epididymal fine needle aspirations (EFNA) were performed on 29 epididymides of 17 infertile, azoospermic men in whom testicular cytology revealed adequate spermatogenesis. Sonograms confirmed the presence of all the excretory male organs. EFNA was performed to diagnose and locate the site of genital tract occlusion. The presence of cuboidal epithelial cells in the aspirate was the sole indication that epididymal content was aspirated. In 10 (34%) of the 29 aspirates there were only cuboidal epithelial cells and no spermatozoa, indicating proximal occlusion in the rete testis, in the canaliculi efferents or in the most proximal segment of the epididymis itself. Occlusion in the distal segments of the epididymis or in the vas deferens was diagnosed in 13 epididymal aspirates (45%) which contained both cuboidal cells and spermatozoa. In 6 epididymides (21%) no conclusion could be reached because of failed aspirations. There are a variety of treatment modalities available for male genital tract obstruction, such as reconstructive surgery or IVF. The success rate of treatment is mainly dependent on the site of obstruction. Since vasography has its own limitations and hazards, diagnostic EFNA should be performed before treatment in men in whom genital obstruction is suspected.

Biopsy, Needle↗
PubMedDate not supplied

The ethane dimethanesulfonate-induced decrease in the fertilizing ability of cauda epididymal sperm is independent of the testis.

Several decades ago it was reported that when adult male rats were exposed to a single injection of 50 mg/kg body weight ethane dimethanesulfonate (EDS) and mated with untreated females, average litter size was significantly reduced as early as 2 weeks later. Recently, we demonstrated that EDS exerts multiple effects in the epididymis of adult rats. Some of these effects were independent of reduced serum testosterone (T) levels. Later we found that EDS has direct effects on epididymal epithelial cells in vitro. Herein, we sought to determine whether EDS perturbs the fertilizing ability of cauda epididymal sperm. Four days after exposure to 50 mg/kg EDS, sperm from the proximal cauda epididymidis were inseminated into adult receptive females in utero; on the next day the percentage of fertilized eggs was determined. Exogenous T administration and castration were used to determine what role, if any, androgen deprivation and the testis had on the fertilizing ability of proximal cauda epididymal sperm. Sperm motion parameters, serum T, T in the caput/corpus epididymidis, and detergent-extracted sperm protein were evaluated and correlated with fertilizing ability. We found that both castration and EDS exposure significantly compromised the fertilizing ability of sperm in proximal cauda epididymidis 4 days after exposure. Exogenous T, sufficient to maintain serum T, completely restored the fertilizing ability of sperm following castration, but not after EDS exposure. Moreover, exogenous T failed to restore fertilizing ability when castrated animals were exposed to EDS. Thus, the effects that EDS exerts on sperm maturation in vivo are independent of the testis. Finally, the only endpoint that was well correlated with fertilizing ability was the relative amount of an acidic 18-kDa sperm protein.

Animals↗
PubMed1993

Modulation in activity of some epididymal glycosidases by prolactin.

Activity of glycosidases in the epididymis was influenced by several factors originating in the testis. Activities of all the three glycosidases studied viz., beta-glucosidase, beta-galactosidase and alpha-mannosidase were found to be significantly lower in the epididymis of orchidectomized animals than in sham operated rats. However, an enhanced activity of epididymal beta-galactosidase and alpha-mannosidase was noticed in prolactin treated orchidectomized rats compared to orchidectomized rats given vehicle alone. On the other hand, activity of these two enzymes in bromocriptine treated orchidectomized rats was even lower than that found in orchidectomized rats given vehicle. Neither prolactin nor bromocriptine treatment had any significant influence on the epididymal beta-glucosidase. The results suggest a selective but definite action of prolactin on epididymal glycosidases.

Animals↗
PubMed1977

[Epididymal tumors (author's transl)].

Six cases of benign primary epididymal tumor have been investigated in our patients. The tumor size fluctuated between 3 and 35 mm in diameter. Frequency, histology and medical development are described. The majority of epididymal tumors (53%) is comprised of so-called adenomatoid tumors of benign nature and their histogenesis is being discussed. The difficulties of differential diagnosis are pointed out. Since malignity exists in 26% of the cases, the surgical exposure and diagnosis by frozen section is being postulated, when suspicion of epididymal tumor becomes evident.

Adenoma↗
PubMedDate not supplied

The use of epididymal fine-needle aspiration for the diagnosis of male genital tract obstruction.

Epididymal fine-needle aspirations (EFNA) were performed on 29 epididymides of 17 infertile, azoospermic men in whom testicular cytology revealed adequate spermatogenesis. Sonograms confirmed the presence of all the excretory male organs. EFNA was performed to diagnose and locate the site of genital tract occlusion. The presence of cuboidal epithelial cells in the aspirate was the sole indication that epididymal content was aspirated. In 10 (34%) of the 29 aspirates there were only cuboidal epithelial cells and no spermatozoa, indicating proximal occlusion in the rete testis, in the canaliculi efferentes or in the most proximal segment of the epididymis itself. Occlusion in the distal segment of the epididymis or in the vas deferens was diagnosed in 13 epididymal aspirates (45%) which contained both cuboidal cells and spermatozoa. From 6 epididymides (21%) no conclusion could be reached because of failed aspirations. There are a variety of treatment modalities available for male genital tract obstruction, such as reconstructive surgery or assisted reproductive technology using sperm aspiration technique. The success rate of treatment is mainly dependent on the site of obstruction. Since vasography has its own limitations and hazards, diagnostic EFNA should be performed before treatment in men in whom genital obstruction is suspected.

Biopsy, Needle↗
PubMed1995

Rat epididymal sperm exhibit on dithiothreitol treatment in vitro quantifiable differences in patterns of light scatter, uptake of 14C-iodoacetamide and binding of ethidium bromide to DNA.

The extent to which chromatin of rat caput (CAP), corpus (COR), cauda (CAU) spermatozoa undergo condensation and compaction is known to be a function of progressive increase in the formation of inter- as well as intra-protamine disulphide bridges during their transit through the epididymis. Relative compaction undergone by the nuclear chromatin of these sperm populations was studied by monitoring their susceptibility to in vitro decondensation induced by varying concentrations (0, 0.01, 1, 5, 10, 50 mM) of disulphide reducing agent, dithiothreitol (DTT) after an initial exposure to 0.01% papain. Following this treatment and staining with the nucleic acid specific fluorochrome, ethidium bromide (EB), it was observed that irrespective of the epididymal region from which they were collected, spermatozoa exhibited DTT dose-dependent (a) increase in nuclear size as seen under fluorescence microscopic examination, (b) decrease in flow cytometrically quantifiable light scatter parameters--forward scatter (FSc, 'nuclear size') and side scatter (SSc, nuclear 'granularity'), (c) increase in individual cell EB binding when analyzed by DNA flow cytometry, and (d) increase in thiol specific 14C-iodoacetamide (14C-IA) uptake. The decrease in both FSc and SSc occurring in spite of actual increase in nuclear size has been attributed to increase in translucency of spermatozoan nuclei consequent to decondensation. The FSc, SSc and EB bindability were studied by monitoring both the channels of maximal cell concentration detected in the flow cytograms as well as by digitally quantitating the numbers of cells within specific channels (1-64, 65-128, 129-192 and 193-256) of the flow cytogram. The latter indicated a measure of the variability in the response of populations of sperm within each sample to DTT induced decondensation. At any given concentration of DTT, especially between 5-10 mM, the differences observed between sperms of different regions were consistent and significant (P < 0.01-P < 0.001), maximal changes being shown by CAP and minimal by CAU sperm, COR sperm appearing in between. The effective concentration of DTT required to elicit 50% of maximal (i.e. that exhibited by CAP sperm when taken as 100%) effect (ED50) varied significantly among CAP, COR and CAU sperms for each of the parameters studied (P < 0.01-P < 0.001). It is concluded that the differences observed among the three epididymal sperm populations are due to differences in the extent of susceptibility to decondensation in vitro and that this is dependent upon the variation in the -S-S-content of their chromatin during different stages of epididymal transit. All the parameters used (with the exception of fluorescence microscopy) can be quantified and as all of them show a similar dose dependency to DTT treatment, any one of these parameters can be conveniently used to determine the mature/immature status of the sperms voided. Application of such a method to determine the quality of sperms voided by man appears feasible.

Animals↗