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At least 631 records · Page 35Linked to original sources
PubMed1996

Efficacy of microsurgical epididymal sperm aspiration (MESA) and intracytoplasmic sperm injection (ICSI) in obstructive azoospermia.

PURPOSE: Our purpose was to test whether micromanipulation using subzonal insemination and intracytoplasmic sperm injection could improve the poor fertilization and pregnancy rates obtained when attempting in vitro fertilization in patients with congenital absence of the vas deferens and unreconstructable obstructive azoospermia with microsurgically retrieved epididymal spermatozoa. RESULTS: Conventional in vitro fertilization (group A; 14 cycles), subzonal insemination (group B; 13 cycles), and intracytoplasmic sperm injection (group C; 28 cycles) were carried out in 55 treatment cycles. Fertilization rates for groups A, B, and C were 16.1, 31.4, and 48.6%, respectively (P < 0.05). Clinical pregnancy rates for groups A, B, and C were 7.1, 7.7, and 32.1% (P < 0.05), respectively. In five cycles, intracytoplasmic sperm injection using epididymal sperm from alloplastic spermatoceles was performed and two clinical pregnancies (40%) were obtained. CONCLUSIONS: The combined microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection procedure is highly effective in improving the fertilization and pregnancy rate in congenital absence of the vas deferens and unreconstructable obstructive azoospermia. Furthermore, alloplastic spermatoceles may be useful for repeat sperm aspirations.

Epididymis↗
PubMed1996

Morphometric epididymal epithelium analysis in obstructive azoospermic men: predictive values in fertilization rates.

PURPOSE: Our purpose was to determine the obstructed epididymal epithelium morphometric state and to elucidate the influence of morphometric parameters on the fertilizing ability of aspirated spermatozoa. METHODS: During MESA with IVF, epididymal tissue samples were carefully dissected, fixed, and prepared for morphometric analysis. The epithelium of the three regions was studied: head, body, and tail (when present). The total area/luminal area ratio, epithelium thickness, and stereocilium length were measured. RESULTS: We have found significant differences in the total area/luminal area ratio, epithelium thickness, and stereocilium length in obstructed epithelium length > 4 versus 0.5-2 cm (P < or = 0.01). Epithelium thickness and stereocilium length were significantly lower when no fertilization was observed in IVF. CONCLUSIONS: These results suggest that morphometric data of obstructed epididymis epithelium can be used as a means of explaining the failure of IVF with epididymal spermatozoa.

Cell Separation↗
PubMed1994

The effect of partial zona dissection on the fertilizing capacity of chronically obstructed mouse caput epididymal sperm.

PURPOSE: Having observed that sperm from the chronically obstructed caput epididymis fertilize poorly in vitro, we investigated the effect of partial zona dissection (PZD) on fertilization in a murine model of unilateral proximal epididymal obstruction. Cleavage rates were compared for zona-intact oocytes and PZD oocytes incubated with sperm from the following epididymal segments: the obstructed caput, the contralateral nonobstructed caput, the contralateral cauda, and a sperm-free preparation to control for parthenogenesis. RESULTS: Unilateral epididymal obstruction resulted in significantly higher sperm counts in the obstructed caput compared to the nonobstructed caput, although there was no difference in motility. Cleavage rates for ova incubated with sperm from the obstructed caput and the nonobstructed caput were uniformly poor and did not differ significantly from those for the sperm-free controls. Cauda sperm fertilized significantly better than all other sperm groups (P < 0.05). Partial zona dissection did not improve cleavage rates in any group. CONCLUSION: We conclude that sperm from the chronically obstructed caput epididymis, like sperm from the normal caput, are unable to fertilize ova, and PZD does not enhance this poor fertilizing capacity. Furthermore, the finding that PZD does not improve the fertilizing capacity of the presumably mature cauda sperm in our mouse model suggests that any beneficial effect of PZD may be strain-specific.

Animals↗
PubMed1994

Successful in vitro fertilization and pregnancy by micromanipulation with epididymal sperm.

PURPOSE: To examine the effectiveness of micromanipulation on the treatment of the patients with obstructive azoospermia, subzonal insemination or partial zona dissection was performed using epididymal sperm of the 23 patients, and the in vitro fertilization and embryo transfer (IVF-ET) outcomes were analyzed. RESULTS: Fertilized oocytes were obtained in 11 cycles of the 25 treated cycles, and six pregnancies were established by the subsequent ET. Through the analysis of 13 cycles in which both the micromanipulation and usual IVF insemination were performed, the fertilization rate per oocyte with the micromanipulation [25% (28/113)] was significantly higher than that with the usual insemination method [4% (4/102)]. The micromanipulation was most useful for the treatment of the patients with congenital absence of the vas deferens. Moreover, there was no need to aspirate a large volume of the epididymal fluid, but it was essential to prepare more than 10 mature oocytes retrieved. CONCLUSIONS: These results indicate that micromanipulation with epididymal sperm is an effective treatment for obstructive azoospermia, and careful preparations of sperm and well-controlled ovarian hyperstimulation are necessary for successful IVF-ET.

Adult↗
PubMed2001

An apical membrane Na+/H+ exchanger isoform, NHE-3, is present in the rat epididymal epithelium.

An acidic milieu is required for sperm maturation and for keeping sperm quiescent during storage in the cauda epididymidis. Previous studies have implicated a Na+/H+ exchanger (NHE) in epididymal acidification together with carbonic anhydrase (CA) and vacuolar proton adenosine triphosphatase (H(+)-ATPase). The present studies were undertaken to discover whether the NHE isoform involved is NHE-3, which is known to mediate Na+ and HCO3- absorption in renal tubules. Using the reverse transcription polymerase chain reaction technique (RT-PCR), Northern blot analysis and in situ hybridization, NHE-3 mRNA was detected mainly in the cauda epididymis and to a lesser extent in other regions of the epididymis. Immunohistochemical studies showed that NHE-3 was present in the apical membranes of the epithelial principal cells and confirmed that its expression is strongest in the cauda region, decreasing towards the more proximal regions. Immunoblotting showed a similar expression pattern. These results demonstrate that NHE-3 is expressed in the rat epididymal duct with strongest expression in its cauda region. These findings are thus consistent with the possibility that NHE-3 in the epididymal duct is involved in luminal Na+ and/or HCO3- absorption, as in the renal proximal tubule, and thereby in the regulation of sperm motility and maturation.

Animals↗
PubMed2006

Ectopic epididymal tissue in appendix testis.

Aberrant epididymal tissue is uncommon and may be associated with an undescended testis. To the best of our knowledge, aberrant epididymal tissue has not been described in a testicular appendix. We report the case of a 7-year-old boy with left cryptorchidism who underwent a homolateral orchidopexy. At operation, the surgeon removed a pedunculated appendix testis containing an epididymal heterotopia. This Wolffian-derivative tissue was confirmed by the diffuse strong CD10 reactivity of the luminal border of the epithelial cells. Awareness of this aberrant tissue avoids misinterpretation as a transected functional reproductive structure.

Biomarkers↗
PubMed1991

Lipid changes of goat sperm plasma membrane during epididymal maturation.

Highly purified plasma membranes of maturing goat caput-, corpus- and cauda-epididymal spermatozoa were isolated by aqueous two-phase polymer methods and their lipid constituents were analysed. Phospholipid (approx. 75% w/w), neutral lipid (approx. 15% w/w) and glycolipid (approx. 10% w/w) were the major sperm membrane lipids. There was a significant decrease in the total lipids (approx. 25% w/w), phospholipid (approx. 30% w/w) and glycolipid (approx. 80% w/w) contents of sperm membrane during epididymal maturation. On the contrary, the mature cauda-sperm membrane showed greater (approx. 50% w/w) neutral lipid content than that of the immature caput sperm. Phosphatidylcholine (PC), phosphatidylethanolamine (PE) and sphingomyelin were the phospholipids of the sperm membrane, the former two being the major lipids. Both PC and PE fractions consisted of three species--diacyl, alkylacyl and alkenylacyl forms, the last one being the dominant species in both PC and PE. Of all the phospholipids, diacyl PE decreased most strikingly (approx. 65% w/w) during sperm maturation. The neutral lipid fraction contained sterols, wax esters, 1-O-alkyl-2,3-diacylglycerol, triacylglycerol and fatty acids. Sterols represented nearly 75% w/w of the neutral lipids and cholesterol was the major component (approx. 95% w/w) of the sterol fraction. The sperm maturity was associated with marked increase of sterol (approx. 60% w/w) and steryl ester (approx. 200% w/w) and decrease (approx. 50-65% w/w) of the other membrane-bound neutral lipids. The glycolipid was identified as monogalactosyldiacylglycerol. The fatty acid profile of the various membrane lipids underwent marked alteration during the epididymal transit of the male gametes. Cholesterol/phospholipid and saturated/unsaturated fatty acid ratios increased greatly in the maturing sperm membrane. The altered lipid profile of the mature sperm membrane leads to changes in its fluidity that play an important role in determining the structure and functions of the biomembrane.

Animals↗
PubMed1992

Changes in the nature of calcium transport systems on the porcine sperm plasma membrane during epididymal maturation.

Comparative studies of 45Ca(2+)-transport across the plasma membrane were performed using porcine caput, corpus and cauda epididymal sperm. The Ca(2+)-uptake is dependent on the presence of the substrates for respiration and is sensitive to verapamil. The Ca(2+)-efflux is mediated by both Na(+)-dependent and -independent systems. In the immature sperm in caput epididymis, Na(+)-independent efflux is predominant, but it is gradually replaced by Na(+)-dependent efflux during the epididymal transit. The net activity of Ca2+ accumulation into sperm increases with the epididymal maturation.

Animals↗
PubMed1983

Radioimmunoreactivity and receptor binding activity of specific epididymal proteins of the rat and other species.

Radioimmunoassay (RIA) and radioreceptor assay (RRA) for the 32K Rat Epididymal Protein (REP) have been developed. Washed intact rat epididymal spermatozoa were used as receptors in the RRA. The effects of epididymal and seminal fluids of other species in these assays were studied. The results showed that there is species specificity in the immunoreactivity and receptor binding activity of sperm coating proteins.

Animals↗
PubMed1991

In vitro organ culture of rhesus monkey epididymal tubules.

The caput and cauda epididymal tubules of rhesus monkey were cultured for 5 days using a simple organ culture system. The viability of the tubules in culture was established by assessing: (a) the histology of the epididymis; (b) motility and viability of spermatozoa; and (c) scanning electron microscopic morphology of spermatozoa before and at the end of culture. The efficacy of the culture was evaluated by introducing the antiandrogen, cyproterone acetate, into the culture medium. Cyproterone acetate caused degenerative changes in the histology of the epididymis and coiling of the epididymal spermatozoa which may be due to alterations in epididymal milieu.

Androgen Antagonists↗
PubMed1984

Lactosaminoglycans synthesized by mouse male germ cells are fucosylated by an epididymal fucosyltransferase.

We have studied the synthesis of protein-bound carbohydrates in differentiating male germ cells in the mouse. Spermatocytes and spermatids synthesize asparagine-linked and high-molecular-weight glycopeptides as the major classes of protein bound carbohydrates. Asparagine-linked glycopeptides were found to be mainly composed of the complex bi-antennary type as shown by affinity chromatography on concanavalin-A Sepharose; high-molecular-weight glycopeptides were represented by nonfucosylated lactosaminoglycans since they were metabolically labeled with [14C]glucosamine but not with [3H]fucose, did not bind to DEAE-cellulose, and were susceptible to endo-beta-galactosidase. Labeling with galactose oxidase/Na B3H4 technique demonstrated that lactosaminoglycans were present on the surface of differentiating germ cells and of testicular and epididymal spermatozoa. Since lactosaminoglycans from germ cells and testicular spermatozoa were not retained on a column of fucose-binding lectin, it was concluded that these molecules do not contain fucose. On the other hand, epididymal spermatozoa lactosaminoglycans bound to the lectin and therefore contained fucose. A soluble fucosyltransferase, capable of transferring fucose to germ cell lactosaminoglycans, was found to be present in the epididymis but not in the testis. These data show that developing germ cells synthesize nonfucosylated lactosaminoglycans which are probably preserved throughout spermiogenesis. We suggest that these molecules are fucosylated in vivo by a fucosyltransferase secreted by the epididymal epithelium.

Amino Sugars↗
PubMed1986

A secretory phospholipase C-like enzyme in the bovine epididymal caput.

An enzyme hydrolysing the synthetic substrate p-nitrophenylphosphorylcholine was highly active in the epididymal caput and very low or absent in other parts of the bovine reproductive organs. This enzyme was also found in the secretory fluid obtained from the epididymal caput but much lower activities were encountered in cauda epididymis and seminal plasma. The enzyme displayed an optimum at pH 6.5, an Mr of about 66000, a pI value of 5.0 and was suppressed by chelating agents and reactivated by Co2+ and Zn2+. After partial purification the enzyme also showed hydrolysis of p-nitrophenyl phenylphosphonate, bis-p-nitrophenyl phosphate and hexadecanoyl-p-nitrophenylphosphorylcholine with slightly different pH optima and modifier characteristics. It is concluded that this secretory enzyme is distinct from the membrane-bound phospholipase C and may play a role in processing of the spermatozoan phospholipids during the epididymal maturation.

Animals↗
PubMed1984

Nuclear androgen binding sites in the male rat. III. Late spermatids and spermatozoa in the testis, with an introduction to epididymal spermatozoa.

Nuclear androgen binding sites were examined in late spermatids (stages 12-19) which resisted sonication of homogenized testes of mature male rats. The measurement of unoccupied binding sites in salt extract of purified spermatid heads by nuclear exchange at -10 degrees C was developed and validated. As in the prostate, unoccupied nuclear androgen binding sites in sonicated testes were in low concentration, were not artefactual, and could be occupied both in vivo and in vitro by exogenous androgens, and uniquely in hemicastrated rats by endogenously compensated androgens in the remaining testis. The properties of occupied binding sites in salt extract of purified spermatid heads (measured by nuclear exchange at 4 degrees C for 48 or more hours with 5 nM [3H]dihydrotestosterone) were almost identical to those of occupied binding sites in nuclei of the ventral prostate, except for their concentration. However, levels of specific binding activity approaching 50 fmol/mg DNA could be expected in salt extract of spermatid pellets, by use of a sulfhydryl reducing agent (dithiothreitol) prior to salt extraction, a protease inhibitor (phenylmethylsulfonyl fluoride) in all buffers, and optimization of the sonication protocol. Nuclear androgen binding sites of sonicated epididymal spermatozoa, collected by retrograde perfusion of the cauda epididymidis, were found to be completely salt-resistant. These binding proteins could be extracted by 0.4 M KCl if dithiothreitol and dihydrotestosterone were incorporated into the sonication buffer, if phenylmethylsulfonyl fluoride was added to all buffers, and if the purified epididymal sperm pellet was treated with sarkosyl, a non-ionic detergent, just before salt extraction. The salt extract of epididymal spermatozoa which were treated as described above contained two binding components: a soluble form which was eluted from hydroxylapatite by increasing concentrations of phosphate buffers, and a non-soluble form, free of DNA, which remained in the hydroxylapatite column, and which contained most of the androgen binding sites. Affinity (Kd) of dihydrotestosterone to the soluble and insoluble fractions of the steroid-binding protein complex was determined to be 0.7 and 0.1 nM, respectively. Salt-resistance of binding proteins in germ cells was shown to develop significantly in the last stages of spermiogenesis.

Animals↗
PubMed1995

Epididymal fat depot lipoprotein lipase activity is lower in animals with high endogenous fat preferences.

Adipose tissue lipoprotein lipase (LPL) is a key enzyme responsible for the clearance of circulating triglycerides and has been linked to certain pathologic states such as obesity. In order to investigate whether an animal's endogenous fat-preference is associated with differences in adipose tissue LPL, we measured enzyme activity in epididymal fat from high- and low-fat preferring rats. Utilizing a 24h ad libitum feeding paradigm, four groups of outbred adult male Sprague-Dawley rats were screened separately for their macronutrient preferences. Animals exhibiting high- or low-fat preferences were identified and placed back on standard chow. LPL activity was measured in epididymal fat under chow fed or fasted conditions. Epididymal fat LPL activity was significantly less in the high-fat-preferring animals relative to the low-fat-preferring, in both the standard chow-fed state (p = 0.014) and fasted (p = 0.0007) state. LPL activity in heart ventricle and brown adipose tissue was also measured from the same animals. Activity in heart ventricle and brown adipose tissue was significantly lower in the high-fat-preferring group as compared with the low-fat-preferring only following a 24h fast (p = 0.0012 for heart and p = 0.0085 for brown adipose, high- versus low-fat preferring). The data indicate that differences in tissue LPL activity exist between animals with inherent differences in fat preference. Future comparative studies between the two groups of fat-preferring animals could lead to important clues to the regulation of the LPL.

Animals↗
PubMed1994

Alterations in maternal epidermal growth factor (EGF) effect testicular descent and epididymal development.

OBJECTIVE: (1) To determine the ability of epidermal growth factor (EGF) to reverse antiandrogen-induced cryptorchidism and epididymal abnormalities; (2) to evaluate whether alterations in maternal EGF would result in abnormal testicular descent or mal development of the epididymis. METHODS: Experiment 1: Timed pregnant ICR mice were treated with either flutamide, flutamide plus EGF, or vehicle alone on gestational days 11 through birth. Experiment 2: Maternal EGF was abolished by removing the submandibular glands. Following timed mating, dams were treated with either flutamide, anti-EGF, DHT, or vehicle alone on gestational days 11 through birth. RESULTS: Experiment 1: Treatment with flutamide resulted in a 36 percent (26/72) incidence of undescended testes (UDT), and a 43 percent (31/72) incidence of abnormal epididymides. Rats treated simultaneously with flutamide plus EGF had a reduced incidence of UDT (14%, 6/42) and epididymal anomalies (19%, 8/42); p < 0.01. Experiment 2: The absence of maternal EGF resulted in a significant incidence of cryptorchidism in 11/50 (22%) testes, and epididymal anomalies in 19/50 (38%); p < 0.01. CONCLUSIONS: Our findings suggest that EGF stabilizes the wolffian duct system and partially mediates testicular descent.

Abnormalities, Drug-Induced↗
PubMed1991

A quantitative histological study of testicular and epididymal development in Boran and Boran x Friesian bulls in Ethiopia.

Quantitative histological techniques were used to characterize the pattern of testicular and epididymal development in 42 Boran and 49 Boran x Friesian bulls aged between 6 and 23 mo. Bulls were divided into five age groups having four-month intervals. Between 6 and 23 mo of age, body weight increased from 124 to 293 kg and 179 to 267 kg (P<0.01) in Boran and Boran x Friesian bulls, respectively. Scrotal circumference measurements increased from 6 to 23 mo by 10.2 and 8.9 cm in Boran and Boran x Friesian bulls. Paired testes weight increased significantly (P<0.001) over the same period from 29 to 304 g and 59 to 291 g in Boran and Boran x Friesian bulls, respectively. Both caput and cauda epididymal segment weights also increased significantly (P<0.001) with age. There were significant (P<0.05) genotype differences in seminiferous tubule diameter, which increased by 137 and 90 microm with increasing age in Boran and Boran x Friesian bulls, respectively. Patent seminiferous tubules were first observed at 206 and 188 d of age in Boran and Boran x Friesian bulls, respectively. The appearance of elongated spermatids and spermatozoa in the seminiferous tubules and epididymal tubules occurred at later ages in Boran than in Boran x Friesian bulls.

Journal Article↗
PubMed1994

In vitro fertilization and development of llama (Lama glama ) oocytes using epididymal spermatozoa and oviductal cell co-culture.

A study was designed to determine the feasibility of developing in vitro maturation, fertilization and culture systems utilizing follicular oocytes and epididymal spermatozoa collected from llamas at slaughter. From a total of 1324 cumulus oocyte complexes (COCs) recovered, 972 were cultured in 50-ul drops of TCM-199 medium with 10% heat inactivated steer serum (DBS) and hormones for 30 h. After maturation, the oocytes were randomly allocated into 4 groups in a 2x2 factorial design: cumulus-enclosed oocytes, 2 ug/ml heparin (Group 1); cumulus-enclosed oocytes, 5 ug/ml heparin (Group 2); denuded oocytes, 2 ug/ml heparin (Group 3); and denuded oocytes, 5 ug/ml heparin (Group 4). Denuded oocytes were obtained for groups 3 and 4 by vortexing. Epididymides were also collected at slaugther and fresh spermatozoa (for each replicate) were obtained by mincing the cauda epididymis with a scalpel blade. A total of 721 oocytes were inseminated with 2-3 x 10(6) epididymal spermatozoa/ml in a 50-ul drop of FERT-TALP medium. After 18 h of in vitro insemination, 234 oocytes were placed in a llama oviductal epithelial cell (LLOEC) co-culture in TCM-199 for 9 d. All cultures were done at 38.5 degrees C under 5% CO(2) in air with high humidity. The rate of fertilization, initial cleavage and development in co-culture were evaluated and compared. Of 192 oocytes examined for signs of fertilization, 56 (29.2%) were penetrated by spermatozoa with 57.1% (32 56 ) of the penetrated oocytes having a male and female pronucleus. There were no differences among treatment groups in total fertilization. However, the frequency of oocytes fertilized normally tended to be higher in the denuded oocytes 67.7% (21 31 ) than the oocytes inseminated with cumulus cells 44.0% (11 25 ) independent of heparin concentration (P<0.06). The total embryo development rate to the 2 cells to blastocyst stage was 32.1% (75 234 ). There was no difference in development rate between groups. From the 234 oocytes co-cultured in LLOEC for 9 d, 15.8% developed into 2 to 16 cells, 5.6% into morulae, 6.0% into early/expanded blastocysts and 4.7% into hatching/hatched blastocysts. The results indicate that an in vitro fertilization system is possible in the llama utilizing slaughterhouse material and that llama oocytes can be fertilized in the presence of heparin and epididymal spermatozoa.

Journal Article↗
PubMed1981

A specific cytolytic T cell response induced by subcutaneous challenge with allogeneic rat epididymal spermatozoa.

Groups of virgin female rats were killed on days 2-5 after footpad injection with either thoracic duct lymphocytes or epididymal spermatozoa from allogeneic male rats and the cellular response in the regional lymph nodes determined by the specific cytoadhesion technique. After challenge with both thoracic duct lymphocytes and epididymal spermatozoa a specific cytolytic T cell response to the male strain histocompatibility antigens was evident on the second day and increased until the fifth day. The weight and total lymphocyte content of the nodes were significantly greater than the unimmunized control values throughout the observation period. These results provide further evidence for the expression of histocompatibility antigens by epididymal spermatozoa.

Animals↗