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PubMed1990

Expression of villin in the mouse oviduct and the seminiferous ducts.

Villin, a 95-kD cytoskeletal protein selectively expressed in the microvilli of some absorptive cells was localized immunohistochemically in the oviduct and the seminiferous excretory ducts of the mouse. Villin was found in the proximal part of the oviduct, comprising the preampulla, ampulla, and part of the isthmus. Distal to the isthmus the oviductal cells lining the junctura and the intrauterine colliculus tubaris were devoid of villin. No villin could be detected in the uterine cells. Ductuli efferentes, connecting the rete testis with the epididymis were the only portion of the male seminiferous ductal system expressing villin. The cells lining the epididymis and the vas deferens were devoid of villin. These data show that villin is selectively expressed in male and female reproductive systems and that it is limited to anatomically defined proximal portions of the reproductive ducts.

Animals↗
PubMed1994

Distribution of endogenous and exogenous 5'-nucleotidase on bovine spermatozoa.

A polyclonal rabbit antibody against 5'-nucleotidase purified from bull seminal plasma was used to localize the antigen on bovine spermatozoa. Spermatozoa taken from the ampulla of the vas deferens showed strong immunofluorescence at the anterior rim of the head portion. Evaluation of spermatozoa prepared from different segments of the seminal pathway indicated the presence of the antigen already in rete testis and epididymal spermatozoa. On cryostat sections of testis tissue a positive immunoreaction was found in the anterior head portion of elongated spermatids, but not in earlier forms of sperm development. This distribution corresponded with the enzyme activity and results of Western blotting in extracts of testicular and epididymal spermatozoa. Immunoelectron microscopy of ampullary spermatozoa using antibody detection with gold-labelled anti-rabbit IgG showed a clear-cut labelling of the plasma membrane in the acrosome region. Treatment of ampullary spermatozoa with 0.1% Triton X-100 did not completely remove the immunoreactive material from the acrosome, showing a very stable linkage of the protein to the plasma membrane. Treatment with phospholipase C from Bacillus thuringiensis, however, removed immunoreactive material from the plasma membrane, indicating its binding by a phosphoinositol anchor. Our findings show that endogenous 5'-nucleotidase is present on the plasma membrane covering the anterior head portion of bovine spermatozoa and indicate specialized functions during the acrosomal reaction. Soluble enzyme derived from seminal vesicle secretion covers the whole sperm surface during emission, but is not covalently bound. It provides generalized enzyme activity to the sperm surface in addition to the specialized area of the sperm head.

5'-Nucleotidase↗
PubMed1995

Immunohistochemical localization of spermadhesin AWN in the porcine male genital tract.

Boar spermadhesin (AWN) is a 14-kDa multifunctional protein, attached to the surface of the spermatozoa and involved in sperm capacitation and zona pellucida binding. The cellular origin of AWN was previously unknown. Moreover, the region of the male genital tract in which AWN becomes attached to the surface of spermatozoa was also uncertain. By using monospecific polyclonal antibodies against AWN, the immunohistochemical distribution pattern of AWN epitopes has been investigated in tissue sections of the porcine male genital tract. Our study has revealed that AWN is synthesized in the rete testis and in the epithelium of the seminal vesicles. The latter are also the major contributors of seminal plasma AWN. In addition, immunoblotting analysis has shown that AWN is present on epididymal spermatozoa. Our results indicate that the cellular origin of spermadhesins is species-specific. The attachment of AWN to epididymal spermatozoa is probably important in developing the capacity for fertilization.

Animals↗
PubMed1978

Histochemical demonstration of disulfide-groups in the lamina propria of human seminiferous tubules.

The distribution of disulfide-groups was investigated in the tunica propria of human seminifersou tubules by means of a thiosulfation/Alcian Blue + 0.8 Mol MgCl2-staining reaction. Controls had shown the absence of significant amounts of sulfhydryl- or sulfate-groups in the lamina propria, which groups would also be demonstrated by the method employed. The lamina propria of human seminiferous tubules is rich in disulfide groups. The staining reaction decreases in the region of the tubulus rectus, is only faint in the connective tissue which underlies the epithelium of the rete testis, and is absent in the lamina propria of efferent ducts. It is suggested that microfibrils and type IV collagen (both rich in cystine) are the materials responsible for the histochemical reaction described. The occurrence of multiple layers of basal lamina material (type IV collagen) and bundles of microfibrils is shown in comparative electron microscopic studies.

Disulfides↗
PubMed1985

Heterotopic adrenal tissue.

In a group of 102 adult patients operated upon for cryptorchism heterotopic adrenal tissue was found in two cases. In one case it was present in the inguinal canal and in another case it was situated at the border between the testicle and rete testis.

Adrenal Cortex↗
PubMed2000

[Microsurgical vasovasostomy in the age of modern reproduction medicine. A cost-benefit analysis].

WS represents the standard procedure of choice for the treatment of obstructive azoospermia following vasectomy. However, recently, ICSI has been suggested by some to represent the solution for all cases of male factor infertility regardless of its etiology based on its success rates. Therefore, we compared VVS to MESA/TESE and ICSI in terms of pregnancy, complications, and costs. Between 1/93 and 6/98 157 VVS was performed microsurgically using the 2-layer technique in 157 patients following prior vasectomy. Between 9/94 and 9/97 69 couples underwent MESA/ICSI for epididymal obstruction not amenable to micro-surgical reconstruction such as post-inflammatory obstruction and congenital absence of the vas deferens; in the same time period 42 couples underwent TESE/ICSI for azoospermia of testicular origin due to cryptorchidism, testicular atrophy, obstruction of the rete testis. In most cases MESA or TESE and ICSI were performed metachronously. Mean intervall of vasal obstruction was 7.6 (0.5-18) years; patency after VVS was 77%, pregnancy rate was 52%. Local complication rate was 4.7%, no major complications were observed. Costs per life birth after VVS were as high as 5,447,-DM or 2,800 Euro. Pregnancy rates after MESA/TESE and ICSI were 22.5% and 19.5%, respectively with 16 singletons, 3 twins and 3 abortions; local complications occurred in 3.9% of the men. Multiple birth were noticed in 15.8% following ICSI, but only in 0.7% following VVS. 5.7% and 1.4% of the female partners experienced serious complications as a mild or severe ovarian hyperstimulation-syndrome, respectively. Costs per life birth after MESA/TESE cycle were as high as 28,804,-DM or 14,100 Euro. Even in the era of ICSI microsurgical vasovasostomy represents the standard approach for obstructive azoospermia following vasectomy. Based on a cost-benefit analysis VVS is more successful in terms of pregnancy rates (52% vs. 22.5%). We conclude that MESA/ICSI should be reserved for patients not amenable for microsurgical reconstruction.

Adult↗
PubMed1987

Adoptive transfer of murine autoimmune orchitis to naive recipients with immune lymphocytes.

A protocol was developed for reproducibly transferring experimental autoimmune orchitis (EAO) to naive recipient mice. Cell donors were (C57BL/6 x A/J)F1 mice immunized about 14 days earlier with mouse testicular homogenate with Freund's adjuvant and an extract of Bordetella pertussis. Lymphocytes from lymph nodes and spleens were equally capable of transferring disease. As few as 5 X 10(6) cells were able to transfer EAO, which began on Day 5-7 after transfer. Infiltrate of lymphocytes and macrophages in the region of the rete testis and straight tubules was the most reproducible early lesion, suggesting that this is the initial site of T cell-antigen interaction. It was not necessary to use both Mycobacteria and B. pertussis adjuvants in donor immunization to achieve transfer of EAO. Disease transfer was antigen specific since only cells from donors immunized with TH could transfer disease. In vitro stimulation of the cells with testicular antigens and/or concanavalin A was a prerequisite to successful transfer of EAO, which was dependent on the presence of L3T4+ T cells since depletion of these cells greatly diminished EAO in recipients and the lymphocyte proliferation response to testicular antigens. Disease did not depend on an antibody response by the recipients. The results imply that effector cells, once generated by immunization and fully activated or selected by in vitro stimulation, can home to specific locations in the testis, locate relevant autoantigens, and cause disease.

Adjuvants, Immunologic↗
PubMed1989

Biochemical and structural evidence for ethanol-induced impairment of testicular development: apparent lack of Leydig cell involvement.

The present study was conducted to provide biochemical and morphological evidence for ethanol-induced impairment of testicular development. The specific activities of testicular postmeiotic enzyme markers--sorbitol dehydrogenase (SDH), lactate dehydrogenase (LDH), and alpha-glycerophosphate dehydrogenase (GDH)--increased with age in CFW mice from ages 23 to 60 days, providing a biochemical measure of testicular development during puberty. Chronic ethanol treatment via liquid diets from ages 20 to 55 days resulted in decreased activities of SDH and LDH at ages 40 and 44 days, and of GDH at ages 34, 40, and 44 days. These decreases were consistent with an arrest in the developmental increase in SDH, LDH, and GDH at ages 31 +/- 0.6, 31 +/- 2.6, and 24 +/- 0.5 days, respectively. After 29 days of ethanol treatment (age 50 days), testicular weights, epididymal sperm content, and sperm motility were reduced, relative to controls, by 37, 83, and 60%, respectively (p less than 0.05). Epididymal weights were unaffected. Light microscopic evaluation of testes revealed disorganization of spermatogenesis, germ cell degeneration, decreased tubular luminal diameter, and vacuolation of Sertoli cells in ethanol-treated mice at age 50 days. Electron microscopic analysis showed that germ cell degeneration was not restricted to a specific cell type. Stage IX-XI tubules were observed in which spermatids had been retained and underwent phagocytosis within the Sertoli cell. Sertoli cells showed evidence of atypical nuclear invaginations. Sertoli cells underwent degenerative changes and were sloughed into the rete testis. However, relative Leydig cell size, as well as fractional volume occupied by the nucleus, mitochondria, and endoplasmic reticulum were unaffected by ethanol. The data (1) confirm previous findings suggesting ethanol-induced delayed testicular development; (2) suggest that certain aspects of testicular development are arrested relatively early in ethanol treatment (4-11 days); and (3) indicate that the Sertoli cell, rather than the Leydig cell, is the primary target with regard to the deleterious effect of chronic ethanol treatment on testicular maturation.

Administration, Oral↗
PubMed1992

Adenomatoid tumor of testicle.

A rare case of intratesticular adenomatoid tumor is described. Pathologically and clinically this tumor is associated with the seminiferous tubules and the rete testis. The patient underwent left inguinal exploration to confirm the testicular tumor. A radical orchiectomy was performed. We believe this is the first case of documented intratesticular adenomatoid tumor that did not arise from either tunica albuginea or epididymis. The cytologic origin of adenomatoid tumors and an unusual finding of decreased spermatogenesis with marked atrophy of the testes are also presented.

Epididymis↗
PubMed1983

The absence of specific interactions of Sertoli-cell-secreted proteins with antibodies directed against H-Y antigen.

Radiolabeled proteins secreted into the medium by rat Sertoli cells in primary culture have been examined for specific interactions with polyclonal and monoclonal antibodies directed against serologically detectable H-Y antigen(s). None of the proteins secreted by Sertoli cells reacted specifically with H-Y antibodies, as determined with immunoprecipitation procedures and immunoabsorbent affinity chromatography, followed by SDS gel electrophoresis. Radioactivity profiles of proteins obtained after reaction with H-Y antibodies were similar to those observed after treatment with nonimmune sera or with irrelevant antibodies. We obtained comparable findings with proteins secreted by the mouse cell line TM4, which is of presumptive Sertoli cell origin, and with proteins present in ram rete testis fluid. These and other findings presented do not support the contention that Sertoli cells secrete a protein having the properties of serologically detectable H-Y antigen as previously described.

Animals↗
PubMed1994

Experimental allergic orchitis in mice. VII. Preliminary characterization of the aspermatogenic autoantigens responsible for eliciting actively and passively induced disease.

Experimental allergic orchitis (EAO) can be induced actively and passively in mice by either immunization with mouse testicular homogenate (MTH) in conjunction with the appropriate adjuvants or by transferring CD4+ T cells isolated from sensitized donors into non-immunized, naive recipients. The distribution of inflammatory lesions seen in active and passive EAO are markedly different. In active EAO maximal disease is observed in the seminiferous tubules, whereas in passive EAO lesions occur primarily in the straight tubules, rete testis, and ductus efferentes. These observations suggest that different immunopathogenic mechanisms and/or aspermatogenic autoantigens may be responsible for the distinct histopathologic profiles. Two murine testis-specific aspermatogenic autoantigens (mAP1 and mAP2) were partially purified from MT acetone powder by extraction in 7-M urea under reducing conditions, gel filtration, ion-exchange chromatography, and preparative isoelectric focusing from pH 3 to 10. In gel filtration on Sephacryl S-400 in 7-M urea, mAP1 is confined to the V0 peak, while mAP2 is in the major included peak. mAP1 has an isoelectric point of 4.4-4.9, is sensitive to both pronase and DNase but not RNase, and is active at a minimal dose of 250-500 micrograms (dry wt). Dose-response bioassays for active and passive EAO revealed that mAP1 preferentially elicits active disease, whereas mAP2 is most effective at eliciting passive disease. These results support the concept that the different histopathologic profiles seen in active and passive EAO are, in part, the result of different immunopathologic responses elicited by separate aspermatogenic autoantigens.

Animals↗
PubMed1991

Testicular toxicity and mutagenicity of steroidal and non-steroidal estrogens in the male mouse.

The mutagenicity and toxicity of diethylstilbestrol (DES), 17 beta-estradiol and zeranol on the male mouse germ cells were investigated with meiotic micronucleus assays in vivo and in vitro, sperm-head abnormality test and morphometry. Further, the developmental effects of DES on testicular morphology were explored. Micronucleus induction was observed at 10(-7) M concentration of DES and 17 beta-estradiol in vitro, but other treatments yielded negative results. The micronucleus assay in vivo revealed a small number of micronuclei in early haploid spermatids 17 days after a single subcutaneous injection of DES 50 mg/kg, whereas estradiol and zeranol gave negative results. The sperm-head abnormality rates were significantly elevated 5 weeks after treatments with high doses of DES, 17 beta-estradiol and zeranol, and testicular morphometry revealed transient changes in the volume densities of testicular tissue components. Prenatal and neonatal estrogen administration resulted in permanent alterations in seminiferous epithelium and dilatation of the rete testis, but did not affect micronucleus or sperm-head abnormality rates. The mutagenicity and toxicity of hormones in the mouse testis paralleled the hormonal activity of these compounds. Early estrogenization was the most sensitive toxicity test, followed by in vitro meiotic micronucleus induction, whereas the sperm-head abnormality assay and morphological analysis did not reveal subtle changes.

Animals↗
PubMed1996

Astrocytes synthesize and secrete prostaglandin D synthetase in vitro.

Prostaglandin D synthetase [PGD-S, prostaglandin-H2 D-isomerase, (5Z, 13E)-(15S)-9alpha, 11 alpha-epidioxy-15-hyrdroxyprosta-5,13-dienoate D-isomerase, EC 5,3,99,2], an enzyme that catalyzes the formation of prostaglandin D2, was originally isolated from homogenates of rat brain and spleen and is known to be a membrane-bound enzyme. Subsequent immunohistochemical studies have shown that PGD-S is associated with neurons in the brain of immature rats, whereas in adult rats it is associated with oligodendrocytes. Several recent studies have shown that the beta-trace protein isolated from human cerebrospinal fluid (CSF), the second most abundant protein in human CSF after albumin, is equivalent to PGD-S. In this paper, we report the preparation of a monospecific polyclonal antibody against purified PGD-S isolated from human CSF and the establishment of a specific radioimmunoassay for this protein. Using this radioimmunoassay in conjunction with immunoblot analysis, PGD-S was detected in various biological fluids including serum, aqueous humor, and rete testis fluid. In addition, an antibody prepared against human PGD-S partially cross-reacted with the PGD-S in the rat and ram. Using a monospecific polyclonal antibody prepared against purified rat PGD-S isolated from rat CSF in conjunction with [35S]methionine incorporation and immunoprecipitation techniques, it was shown for the first time that PGD-S is actively synthesized and secreted by astrocytes cultured in vitro, suggesting the astrocyte is the cellular origin of PGD-S in the CSF. The identification of the astrocyte as the cellular origin of this unique enzyme will allow the use of an in vitro system to study its regulation.

Aging↗
PubMed2005

The management strategies for stage I seminoma.

About 80% of men with seminomatous testicular germ-cell cancer are diagnosed with stage I disease. For many years, the standard treatment for this patient group has been radiation to para-aortic and iliacal lymph nodes at the same side as the orchiectomy. However, iliac radiotherapy is unnecessary in patients without prior inguinal or scrotal surgery. Furthermore, in recent years, other treatment modalities for this patient group have evolved. The use of single-agent carboplatin has shown promising results, similar to the effects obtained by radiotherapy. In addition, surveillance after primary orchiectomy with no additional treatment is found to be a safe follow-up for many of these patients. On the basis of new knowledge about primary tumour risk factors, it is now possible to identify patients at a particular high risk of relapse (rete testis invasion, primary tumour size > 4 cm, or both). This will be a helpful tool to identify patients who can be safely included into a surveillance strategy, and those who could have adjuvant treatment. The final decision about treatment will depend on risk factors, capacity of the healthcare service to carry out frequent follow-up examinations and the patient's own preferences. In this paper, we will discuss advantages and disadvantages of the various treatment options in the management of stage I seminoma.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed2004

Spermatocytic seminoma: a review.

OBJECTIVE: Spermatocytic seminoma is a rare testicular tumour that has an extremely low rate of metastasis. We present a review of the management of this malignancy at our institution. METHOD AND MATERIALS: Between 1981 and 1999, 771 patients were treated at our institution for testicular seminoma. Of these, 13 had spermatocytic seminoma; one was excluded as he had treatment elsewhere. All patients were initially diagnosed at other hospitals and subsequently referred for management and had their pathology reviewed locally prior to any treatment. RESULTS: All patients had stage I disease, 5 patients received radiotherapy to the para-aortic and pelvic nodes, the other 7 were followed on a surveillance program. The median age was 62 years. With a median follow-up of 8.5 years no relapses were observed. Some patients exhibited adverse histological features associated with increased risk of relapse in seminoma including rete testis invasion and large primary tumour size. CONCLUSIONS: Spermatocytic seminoma may occur in younger patients and may not be restricted to the older population as commonly reported. Surveillance following orchidectomy is the preferred management option.

Adult↗
PubMed2004

Stage II testicular seminoma: patterns of recurrence and outcome of treatment.

OBJECTIVES: To review treatment outcome and patterns of failure for patients with stage II testicular seminoma and to identify prognostic factors for relapse. METHODS: From 1981 to 1999, 126 men with stage II seminoma were treated at Princess Margaret Hospital. Of these, 95 were treated with radiotherapy (RT) and 31 with chemotherapy (ChT). Patient and tumour characteristics were analyzed for prognostic significance for subsequent relapse. RESULTS: At median follow-up of 8.5 years, the 5- and 10-year overall survival were both 93%, the 5- and 10-year cause-specific survival were both 94% and the 5- and 10-year relapse-free rates were both 85%. Patients with stage IIA and IIB disease treated with RT and stage IIB treated with chemotherapy had 5-year relapse-free rates of 91.7%, 89.7% and 83.3%, respectively. Seventeen percent of patients treated with radiotherapy and 6% of those treated with chemotherapy have relapsed. Of the RT patients the commonest sites of relapse were left supraclavicular fossa, lung/mediastinum, bone, para-aortics and liver; nine patients had a solitary site of relapse. Two patients treated with chemotherapy had recurrence in the para-aortic and iliac nodes. For RT patients, larger primary tumour size was associated with a reduction in relapse rate. Age, rete testis invasion and lymphovascular invasion were found not to be of prognostic significance. CONCLUSIONS: In stage IIA/B seminoma, radiotherapy continues to provide excellent results, as the majority of patients will be cured with this treatment alone. Chemotherapy is the treatment of choice for stage IIC seminoma.

Adult↗
PubMed2005

Long-term outcome of postorchiectomy surveillance for Stage I testicular seminoma.

PURPOSE: To examine the long-term outcome and patterns of relapse in clinical Stage I testicular seminoma managed with surveillance alone after radical inguinal orchiectomy. METHODS AND MATERIALS: This was a prospective, single-arm study. Patients with Stage I testicular seminoma were treated with surveillance alone in accordance with regular, predefined, schedules and investigations. RESULTS: The study accrued a total of 88 patients between 1985 and 1996. The median age at diagnosis was 34 years. The median tumor size was 3.5 cm. The median follow-up as of June 2003 was 12.1 years. Only 3 patients were lost to follow-up. Of the 88 patients, 71 remained free of relapse and 17 did not. The actuarial relapse-free rate was 83%, 80%, and 80% at 5, 10, and 15 years, respectively. Most relapses (15 of 17) were below the diaphragm. Of the 17 patients with relapse, 14 were treated with radiotherapy and 3 with combination chemotherapy. Only 1 had a second relapse and was further salvaged by chemotherapy. All 17 relapsed patients remained free of recurrence after salvage treatment. None died of seminoma. The statistically significant predictive factor for relapse on the Cox proportional hazards model was the presence of rete testis invasion (hazard ratio 3.5, p = 0.03). CONCLUSION: Surveillance with the reservation of radiotherapy or chemotherapy for salvage of relapse is a safe alternative to upfront postoperative adjuvant therapy for Stage I testicular seminoma.

Adult↗
PubMed2005

Immunisation of the male tammar wallaby (Macropus eugenii) with spermatozoa elicits epididymal antigen-specific antibody secretion and compromised fertilisation rate.

Immunocontraception has been proposed as an effective and humane means of controlling overabundant kangaroo populations in Australia. We have examined the feasibility of using a sperm-based vaccine for this purpose using a model macropod species, the tammar wallaby (Macropus eugenii). This study has demonstrated immunocontraception in a marsupial species following immunisation of males with homologous spermatozoa. Serum anti-sperm IgG titres were associated with a significant reduction in fertilisation rates following mating with superovulated female wallabies. Antigen-specific IgG penetrated the reproductive tract at the rete testis and bound spermatozoa in vivo. IgG was detected bound to the acrosome and midpiece regions of both epididymal and ejaculated spermatozoa. The absence of adverse testicular pathology and sperm movement effects suggests that contraception may have been achieved by antibody-mediated blocking of sperm surface antigens essential for fertilisation. This study demonstrates that a contraceptive vaccine targeting sperm antigens has potential for fertility control in male macropods.

Acrosome↗