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PubMed1976

Effects of alpha-chlorohydrin on the metabolism of testicular and epididymal spermatozoa of rams.

Spermatozoa were collected from the rete testis and vas deferens of conscious rams. The endogenous oxygen uptake of the spermatozoa was unaffected by alpha-chlorohydrin added in vitro, although this compound abolished the stimulation of oxygen uptake caused by the addition of glycerol. The metabolism of [14C]glycerol by testicular and epididymal spermatozoa was markedly reduced by alpha-chlorohydrin, CO2 production and lactate accumulation being almost totally inhibited. These effects were dependent upon a period of preincubation of the spermatozoa with alpha-chlorohydrin alone, since the presence of glycerol protected the spermatozoa from its action. Longer exposure and a higher concentration of alpha-chlorohydrin were needed with testicular than with epididymal spermatozoa to achieve a maximal effect. The metabolism of [14C]glucose by both sperm types was also inhibited by alpha-chlorohyrin. Spermatozoa of the ram are therefore susceptible to the action of alpha-chlorohydrin throughout the epididymis, although more mature spermatozoa are more affected. It is suggested that alpha-chlorohydrin is converted to an intermediate which is the agent responsible for the inhibition of glycolysis in spermatozoa.

Animals↗
PubMed1992

Immunocytochemical localization of sulfated glycoprotein-1 (SGP-1) and identification of its transcripts in epithelial cells of the extratesticular duct system of the rat.

The localization of sulfated glycoprotein-1 (SGP-1) in the extratesticular duct system was analyzed using an affinity purified antibody raised against the protein in conjunction with light (LM) and electron (EM) microscope immunocytochemistry. In the LM an intense immunoperoxidase reaction product was observed over the cytoplasm of Sertoli cells as well as over the tails of late spermatids. The rete epithelial cells and nonciliated cells of the efferent ducts also showed an intense uniform reaction over their entire cytoplasm. In the EM, immunogold labeling was noted over the entire endocytic apparatus of these cells including coated pits, endosomes, multivesicular bodies, and secondary lysosomes. Since there was no labeling of the luminal contents including sperm along the epididymis, it was concluded that the Sertoli-derived SGP-1 must dissociate from the sperm and be taken up by epithelial cells at the level of the rete testis and efferent ducts. In all regions of the epididymis, except the cauda, the principal cells showed, in the LM, an intense reaction over bodies of various shapes and sizes in their supranuclear region; this corresponded in the EM to a strong immunogold labeling of secondary lysosomes. No labeling was noted, however, over coated pits, endosomes, or pale multivesicular bodies, suggesting that SGP-1 was not being endocytosed from the lumen. Similar observations were noted for the epithelial clear cells along the entire epididymis. In the cauda epididymidis, principal cells presented a weak immunolabeling of their secondary lysosomes. Northern blot analysis revealed a strong 2.6 Kb band corresponding to the mRNA of SGP-1 in the efferent ducts and all regions of the epididymis with the exception of the cauda. Coincident with the mRNA expression of SGP-1 it was found that small clusters of gold particles representing anti SGP-1, presumably membrane bound, were associated with the Golgi apparatus as well as in close proximity to secondary lysosomes. There was, however, no evidence for the secretion of SGP-1 into the lumen. These results suggest that SGP-1 is synthesized by the epithelial cells of the male duct system and ferried by small vesicles derived from the Golgi apparatus to secondary lysosomes. Because SGP-1 has recently been shown to have substantial sequence similarity to prosaposin, it may be speculated that SGP-1 is instrumental in the degradation of membrane glycolipids present within secondary lysosomes of epithelial cells of the extratesticular duct system.

Animals↗
PubMed1976

Steroids in fluids and sperm entering and leaving the bovine epididymis, epididymal tissue, and accessory sex gland secretions.

Ten steroids which may have a role in the process of sperm maturation within the epididymis were quantified by competitive protein binding or radioimmunoassay. Rete testis fluid (RTF) carrying testicular sperm into the epididymis was rich in dehydroepiandrosterone and testosterone (21 +/- 2 and 33 +/- 3 ng/ml) while cauda eipididymal plasma (CEP) around sperm which have completed maturation had high levels of progesterone, dihydrotestosterone, 3beta-androstanediol, dehydroepiandrosterone and testosterone (7.4 +/- 0.8, 20.3 +/- 1.1, 6.5 +/- 0.4, 8.0 +/- 0.7 and 11.5 +/- 0.7 ng/ml). About 4 mug of steroids enter the epidymis daily in RTF, but less than 1% was found in CEP; the balance presumably was absorbed by the epithelium in the proximal caput epididymidis. Nevertheless, tissue levels of total 17beta-OH androgens were lower in the proximal caput than in the distal caput or corpus epididymidis. In all zones of the epididymis, dihydrotestosterone accounted fro about 70% of the total 17beta-OH androgens found in the nuclear fraction. In the cytoplasmic fraction, however, dihydrotestosterone predominated only in the distal caput and corpus epididymidis. In the cauda epididymidis, CEP and sperm probably accounted for less than 35% of the total 17beta-OH androgens and less than 25% of the dihydrotesterone. The progesterone concentration of the cauda than in the caput epidymidis. Twice washed testicular sperm contained more testosterone than cauda epididymal or ejaculated sperm (16.6 +/- 1.9, 1.6 +/- 0.2 and 1.5 +/- 0.3 ng/10(9) sperm, respectively), but less progesterone (0.5 +/- 0.1, 1.3 +/- 0.2 and 1.0 +/- 0.4 ng/10(9) sperm, respectively). As a consequence of mixture with estrogen-rich prostatic fluid (150 +/- 9 pg/ml), ejaculated sperm contained a relatively high amount of estrogens (112 +/- 15 pg/10(9) sperm). These studies revealed marked differences in steroid profiles of fluids entering and leaving the epididymis and of infertile testicular and fertile cauda epididymal sperm.

Androstane-3,17-diol↗
PubMed2005

Germ cell transplantation and testis tissue xenografting in domestic animals.

Transplantation of germ cells from fertile donor mice to the testes of infertile recipient mice results in donor-derived spermatogenesis and transmission of the donor's genetic material to the offspring of recipient animals. Germ cell transplantation provides a bioassay to study the biology of male germ line stem cells, develop systems to isolate and culture spermatogonial stem cells, examine defects in spermatogenesis and treat male infertility. Although most widely studied in rodents, germ cell transplantation has been applied to larger mammals. In domestic animals including pigs, goats and cattle, as well as in primates, germ cells can be transplanted to a recipient testis by ultrasonographic-guided cannulation of the rete testis. Germ cell transplantation was successful between unrelated, immuno-competent pigs and goats, whereas transplantation in rodents requires syngeneic or immuno-compromised recipients. Genetic manipulation of isolated germ line stem cells and subsequent transplantation will result in the production of transgenic sperm. Transgenesis through the male germ line has tremendous potential in domestic animal species where embryonic stem cell technology is not available and current options to generate transgenic animals are inefficient. As an alternative to transplantation of isolated germ cells to a recipient testis, ectopic grafting of testis tissue from diverse mammalian donor species, including horses and primates, into a mouse host represents a novel possibility to study spermatogenesis, to investigate the effects of drugs with the potential to enhance or suppress male fertility, and to produce fertile sperm from immature donors. Therefore, transplantation of germ cells or xenografting of testis tissue are uniquely valuable approaches for the study, preservation and manipulation of male fertility in domestic animals.

Animals↗
PubMed1976

[Spermatogenesis after testosterone supplementation in the hypophysectomized ram].

Six adult rams were hypophysectomized and immediately injected with 0,5 g (group A) or 2 g (group B) of testosterone per day for 2 weeks. Three normal rams (group C) received the solvent only. The testosterone concentrations in the testis (ng/g) were the same in groups A (23 +/- 7) and C (22 +/- 4) and 3 times higher in group B (72 +/- 18). There was an uptake of testosterone by the testis of the two supplemented groups. In the Rete Testis Fluid, the testosterone concentration was normal with 0,5 g/day (29 +/- 0.6 ng/ml). The testicular weight was maintained by the two treatments. However, spermatogenesis was abnormal in both supplemented groups; meiosis and spermiogenesis occurred normally, but the efficiency of spermatogonial divisions, as shown by the number of leptotene and zygotene primary spermatocytes, was greatly reduced (85%). The results indicate that testosterone alone in the Ram is unable to support complete spermatogenesis, which is contrary to that found in the Rat.

Animals↗
PubMed1999

Fas and Fas-ligand expression in seminomatous testes.

Sixteen seminomas with surrounding tissue containing normal and precancerous (cis) seminiferous tubules were examined for the expression of Fas (CD95, APO-1) and Fas ligand (FasL) (CD95L). This was done by analyzing frozen specimens using immunohistochemistry with antibodies directed against Fas and FasL. The study showed that varying numbers (mean approx. 20%) of Fas-positive lymphocytes were present among tumor-infiltrating lymphocytes, but very few FasL-positive lymphocytes. Fas was not expressed by normal seminiferous tubules and only occasional Fas-positive epithelial cells were seen in cis tubules. FasL was expressed in 9 out of 10 cases in virtually all normal seminiferous tubules, mainly as a thin layer at the base of the seminiferous epithelium. In precancerous tubules, this layer was discontinuous and less pronounced. Rete testis expressed FasL in 2 out of 2 cases with rete present and Fas in 1 out of 1 case. Invasive tumor cells did not express Fas or FasL. The data are discussed in relation to immune reactions to seminomas and to the concept of the testis being an immunologically privileged area.

Antigens, Neoplasm↗
PubMed2002

Germ cell transplantation in pigs.

Spermatogonial stem cells form the foundation of spermatogenesis, and their transplantation provides a unique opportunity to study spermatogenesis and may offer an alternative approach for animal transgenesis. This study was designed to extend the technique of spermatogonial transplantation to an economically important, large-animal model. Isolated immature pig testes were used to develop the intratesticular injection technique. Best results of intratubular germ cell transfer were obtained when a catheter was inserted into the rete testis under ultrasound guidance. The presence of infused dye or labeled cells was confirmed in the seminiferous tubules from 70 of 89 injected isolated testes. Infusion of 3-6 ml of dye solution or cell suspension could fill the rete and up to 50% of seminiferous tubules. The technique was subsequently applied in vivo. Donor cells included testis cells from 1- or 10-wk-old boars (from the recipients' contralateral testis or unrelated donors) and those from mice carrying a marker gene. Porcine testis cells were labeled with a fluorescent marker before transplantation. Testes were examined for the presence and localization of labeled donor cells immediately after transplantation or every week for 4 wk. Labeled porcine donor cells were found in numerous seminiferous tubules from 10 of 11 testes receiving pig cells. These results indicate that germ cell transplantation is feasible in immature pigs, and that porcine transplanted cells are retained in the recipient testis for at least 1 mo. This study represents a first step toward successful spermatogonial transplantation in a farm animal species.

Alkylating Agents↗
PubMed1999

T-lymphocyte subsets in the contralateral testis after unilateral blunt testicular trauma in pre-pubertal mice.

Although spermatozoa express antigens, they normally do not produce an immunological response because of the blood-testis barrier and the predominance of CD8(+) T-lymphocytes in the rete testis. Unilateral blunt testicular trauma (UBTT) has been reported to decrease fertility. The present study was designed to evaluate the sub-populations of T-lymphocytes in mice with testicular trauma. Twenty male mice aged 20 days were randomized into control and test groups. At about 70 days of age the contralateral testis was harvested, cell suspensions were prepared, and immunofluorescence staining was performed for detection of CD4(+ )and CD8(+) T-lymphocytes by flow-cytometry. The ratio of CD8(+) and CD4(+) lymphocytes was significantly higher (P < 0.001) in the control mice compared to the UBTT group (1.3 +/- 0.3 vs 0.5 +/- 0.01). The results suggest that UBTT alters the CD8(+)/CD4(+) ratio in the contralateral testis, which may have an important bearing on the pathogenesis of infertility in cases of testicular injury.

Animals↗
PubMed1989

Testicular carcinoma in situ associated with rhabdomyosarcoma of the spermatic cord.

A 12-year-old boy had an embryonal rhabdomyosarcoma in the distal portion of the spermatic cord. The tumor partially surrounded the testis, infiltrated the testicular tunics and formed an intratesticular nodule near the rete testis. The unaffected testicular parenchyma exhibited the characteristic germ cells of carcinoma in situ. We describe an association between these 2 types of tumors.

Carcinoma in Situ↗
PubMed1982

The contribution of the mesonephros to the development of the sheep fetal testis.

In this study, which was performed on 52 sheep fetuses aged 24 to 85 days, we examined the relationship between testicular development and the "giant" mesonephric nephron, a peculiar structure consisting of a large glomerulus and multiple tubules. The development of the testis occurred in two phases. The preparatory phase began at day 24 of gestation, evolved simultaneously with the involution of the glomerulus of the giant nephron, and was characterized by mobilization of the glomerular cells and by their colonization of the genital ridge. By day 31, a prominent mass of migrating mesonephric cells had developed; it extended uninterrupted from the giant glomerulus into the gonad where the mesonephric cells associated with the germinal cells forming the cellular template from which the testicular cords later became assembled. At complete involution of the glomerulus (day 52), the migratory mass implanted on the tubules of the giant nephron, which thus became continuous with the gonad. During the organizational phase, which began at day 29 with the formation of the tunica albuginea, the various components of this continuum became progressively organized along a testis-to-mesonephros direction into seminiferous cords, tubuli recti, the cords of the rete testis, and the ductules efferentes. These observations show that, in the sheep, the precursors of the Sertoli cells are mesonephric in origin, and that the genital tract proximal to the epididymis differentiates from a single mesonephric nephron.

Animals↗
PubMed2001

A 22-amino acid synthetic peptide corresponding to the second extracellular loop of rat occludin perturbs the blood-testis barrier and disrupts spermatogenesis reversibly in vivo.

When Sertoli cells were cultured in vitro on Matrigel-coated bicameral units, the assembly of the inter-Sertoli tight junction (TJ) permeability barrier correlated with an induction of occludin expression. Inclusion of a 22-amino acid peptide, NH(2)-GSQIYTICSQFYTPGGTGLYVD-COOH, corresponding to residues 209-230 in the second extracellular loop of rat occludin, at 0.2-4 microM into Sertoli cell cultures could perturb the assembly of Sertoli TJs dose-dependently and reversibly. This peptide apparently exerts its effects by interfering with the homotypic interactions of two occludin molecules between adjacent Sertoli cells at the sites of TJs, thereby disrupting TJs, which, in turn, causes a decline in transepithelial electrical resistance across the Sertoli cell epithelium. When similar experiments were performed using a 22-amino acid myotubularin peptide, NH(2)-TKVNERYELCDTYPALLAVPAN-COOH (residues 156-177), no effects on the assembly of inter-Sertoli TJs in vitro were noted. When a single dose of this synthetic occludin peptide was administered to adult rats intratesticularly at 1.5-10 mg/testis, germ cells began to deplete from the seminiferous epithelium within 8-16 days. By 27 days, virtually all tubules were devoid of germ cells. This antispermatogenic effect was reversible, because germ cells progressively repopulated the epithelium thereafter. Treated testes were indistinguishable from normal or control testes by 68 days post-occludin peptide treatment when assessed using histological analysis. In contrast, control rats receiving either no treatment, vehicle alone, or a 22-amino acid synthetic peptide of myotubularin displayed no changes in the testicular morphology at all time points. The occludin peptide-induced germ cell depletion was also accompanied by a disruption of the blood-testis barrier (BTB) when assessed by micropuncture techniques quantifying [(125)I]-BSA in rete testis fluid and seminiferous tubular fluid following i.v. administration of [(125)I]-BSA through the jugular vein. These results illustrate that the occludin peptide-induced disruption of the BTB may possibly affect the underlying adherens junctions, which causes premature release of germ cells from the epithelium and reversible infertility.

Amino Acid Sequence↗
PubMed2002

Cystic dysplasia of the testis associated with ipsilateral renal agenesis and contralateral crossed ectopia.

Cystic dysplasia is a benign congenital lesion of the rete testis often associated with ipsilateral wolffian duct and ureteral abnormalities. We present a case of cystic dysplasia and associated anomalies in a 12-year-old boy. The workup revealed right testicular cystic dysplasia, ipsilateral renal agenesis, contralateral crossed renal ectopia, and vesicoureteral reflux. We performed right testicular-preserving cyst enucleation and ureteroneocystostomy. No testicular atrophy or new cyst formation was evident at 1 year of follow-up. This is the first reported case of cystic dysplasia associated with ipsilateral renal agenesis and contralateral crossed renal ectopia.

Child↗
PubMed1987

Distribution of histopathology and Ia positive cells in actively induced and passively transferred experimental autoimmune orchitis.

Histopathology in testes from mice with actively induced experimental orchitis (EAO) (active EAO) and those from recipients of testis-sensitized lymphocytes (passive EAO) had different distributions. In passive EAO, maximum orchitis existed in the straight tubules, rete testis, and ductus efferentes, obstruction of which led to extreme dilatation of seminiferous tubules. Unusual intralymphatic granulomata also resulted in dilated testicular lymphatics. In active EAO, maximum orchitis affected seminiferous tubules under the testicular capsule, away from the rete testes. Vasitis was common and occurred in both active and passive EAO. In normal testes, IA+ F4/80+ cells were sparse but formed a cuff around the straight tubules. After immunization with testis in adjuvant or with adjuvant alone, the number, size, and staining intensity of IA+ cells increased dramatically beginning on day 5, 7 days before disease onset. Simultaneously, epithelial cells confined to the ductus efferentes became Ia+. Although recipients of sensitized lymphocytes also developed epithelial Ia in the ductus efferentes, they did not show changes in testicular interstitial Ia+ cells. Our findings indicate that testicular autoantigens are not completely sequestered, but are accessible to and can react with passively transferred immune lymphocytes in well-defined regions of the germ cell compartment. These regions coincided to a large extent with maximum expression of periductal or epithelial Ia. Changes in Ia+ cells in the testis, which are inducible by adjuvants and precede orchitis, may account in part for the different distribution of histopathology of active EAO.

Animals↗
PubMed2005

Primary signet-ring stromal tumor of the testis.

We present, in a 47-year-old man, the first case of the signet-ring stromal tumor of the testis. The tumor was located beneath the tunica albuginea surrounded by the testicular tubules and rete testis. It was sharply circumscribed by a thin and irregular fibrous capsule. Histologically, it was composed of cells with a widespread signet-ring cell change separated by fibrous stroma. In some places, the signet-ring cells formed vague Indian files, thus resembling metastatic carcinomas with signet-ring cell morphology. Under high magnification, most of the cytoplasm of the tumor cells was seen to be replaced by an empty clear vacuole which pushed the nuclei to the periphery of the cells. Some of the nuclei were indented by the cytoplasmic vacuoles, others were without indentation. Only in a small area did the tumor show cells without a signet-ring cell change. They looked like epithelioid fibroblasts forming abortive and vaguely tubular structures. Mitoses and necroses were absent. Mucicarmine and PAS stains were negative. Immunohistochemically, the tumor was vimentin positive and it was negative with antibodies to cytokeratins, inhibin, prostatic acid phosphatase, prostate-specific antigen, smooth muscle actin, S-100 protein, EMA and calretinin. The signet-ring stromal tumor of the testis is thus similar morphologically and immunohistochemically to the signet-ring stromal tumor of the ovary. The patient was free of recurrence and metastasis 3 years after the excision.

Biomarkers, Tumor↗
PubMed1998

Androgen receptor expression in the testes and epididymides of prenatal and postnatal Sprague-Dawley rats.

The androgen receptor (AR) plays a critical role in sexual differentiation and in the virilization of the male reproductive system. A clear understanding of AR expression at the early stages of sexual development will help elucidate the sensitivity of perinatal animals to endocrine modulation by external agents, such as some environmental chemicals. Immunohistochemistry was used in this study to localize the AR in the differentiating testis and epididymis of Sprague-Dawley rats starting from gestation day 15 until postnatal day 21. Positive AR staining was found on gestation day 15 in the mesenchymal as well as in the epithelial cells in the mesonephros. Weak staining was also observed in a small number of interstitial cells in the primordial testis at this age. The fetal interstitial and peritubular myoid cells showed positive AR immunoreactivity early in development, but the Sertoli cells did not overtly express the receptors until postnatal day 5. The intensity of staining and number of AR-positive cells in the testis and epididymis increased over time. The epithelium in the mesonephros-derived tissues, including rete testis and epididymis, appeared to exhibit a higher capacity to express AR than the rest of the testicular tissue. The results demonstrate that AR expression in the primordial male reproductive system is highly specific to time and cell type and modify previous understanding on the timing of AR expression in the testicular tissue. Since AR-positive cells at various developmental stages may be potential sites of interaction with chemicals that adversely affect sexual differentiation, improved understanding of AR ontogeny will help in investigating the effects of AR-reactive agents, such as environmental antiandrogens, with respect to specific windows of sensitivity.

Animals↗
PubMed1998

The Parkes Lecture. Heat and the testis.

The evidence for the lower temperature of the testes of many mammals is summarized, and the reasons suggested for the descent of the testes into a scrotum are discussed. Descriptions are given of the various techniques used for studying the effects of heat on the testis, whole body heating, local heating of the testes (by inducing cryptorchidism, scrotal insulation or immersion of the scrotum in a water bath), and heating of tissue or cell preparations in vitro. The effects of heat are discussed, effects on the testis (weight, histology, physiology, biochemistry and endocrinology), on the numbers and motility of spermatozoa in rete testis fluid and semen, on fertilizing ability of spermatozoa and on the subsequent development of the embryos produced when spermatozoa from heated testes are used to fertilize normal ova. The possible mechanisms for the damaging effects of heat are discussed, as well as the importance of heat-induced abnormalities in male reproduction in domestic animals and humans.

Animals↗
PubMed2005

Effect of epidermal growth factor on spermatogenesis in the cryptorchid rat.

PURPOSE: Epidermal growth factor (EGF) is secreted mainly from the submandibular glands. Submandibular gland ablation causes a marked decrease in male fertility, which suggests that EGF influences spermatogenesis. We investigated the effect of EGF in combination with orchiopexy on cryptorchid rat testes in which tubular deterioration had become partially irreversible. MATERIALS AND METHODS: Unilaterally cryptorchid rats were obtained by daily administration of 7.5 mg flutamide (Nihonkayaku, Tokyo, Japan), an androgen receptor antagonist, to pregnant rats. At age 10 weeks the unilaterally cryptorchid rats underwent orchiopexy with or without EGF administered into the cryptorchid testis. EGF solution (10 microg/ml) was delivered into the seminiferous tubules by retrograde perfusion through the rete testis. At 14 days testicular recovery was assessed based on the maturity of spermatogenesis using a modified Johnsen score and from the number of apoptotic germ cells per seminiferous tubule. RESULTS: Mean Johnsen score +/- SEM was significantly higher in the orchiopexy with EGF than in the orchiopexy without EGF group (7.85 +/- 0.12 vs 7.12 +/- 0.13, p <0.001). The number of apoptotic germ cells tended to be smaller in the orchiopexy with EGF group than in the orchiopexy without EGF group (0.16 +/- 0.05 vs 0.28 +/- 0.08). CONCLUSIONS: Although orchiopexy for cryptorchidism partly improved spermatogenesis, recovery was limited. EGF administered in combination with orchiopexy was more effective for spermatogenesis than orchiopexy alone. This may be applicable in patients with cryptorchidism.

Analysis of Variance↗
PubMed2003

[Cystic dysplasia of the testis].

Cystic dysplasia of the testis is an exceptional benign tumour resulting from a functional abnormality of the connection between the rete testis and the efferent ductules of the epididymis. The authors report a new case observed in a child. The frequent association with a renal abnormality is highly suggestive. Treatment usually consists of orchidectomy, but can be conservative with long-term surveillance. The prognosis is usually excellent.

Cysts↗