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PubMed2001

Transient disruption of spermatogenesis by deregulated expression of neurturin in testis.

Two related ligands, glial cell line-derived neurotrophic factor (GDNF) and neurturin (NRTN), are expressed by Sertoli cells, but their cognate ligand-binding co-receptors, GDNF family receptor alpha1 and alpha2, are displayed by different germ cells suggesting different targets for the ligands. GDNF regulates cell fate decision of undifferentiated spermatogonia 'Science 287 (2000) 1489'. The role of NRTN was now approached by targeted overexpression in mouse testis. Between 3 and 5 weeks of age, transient degeneration of spermatogenic cells was observed in approximately 20% of all five transgenic lines generated. Spermatids and pachytene spermatocytes underwent segmental degeneration, if the rete testis was undilated. When it was dilated, the spermatids and spermatocytes were more generally depleted. After 5 weeks of age, spermatogenic defects were no more observed and the NRTN overexpressing mice were fertile. The data suggest that NRTN might regulate survival and differentiation of spermatocytes and spermatids, but the low penetrance indicates that either the transgene expression has not been high enough or NRTN is not as essential as GDNF for spermatogenesis.

Animals↗
PubMed2000

Immunohistochemical localization of taurine in the male reproductive organs of the rat.

The amino acid taurine has been implicated in several aspects of reproductive system physiology. However, its localization in these organs has not been previously analyzed. The aim of this study was to characterize its distribution in male rat reproductive organs by immunohistochemical methods. Taurine was localized in the smooth muscle cells of the tissues studied and in the skeletal fibers of the cremaster muscle. In the testis, taurine was found in Leydig cells, vascular endothelial cells, and other interstitial cells. No immunoreactivity was observed in the cells of the seminiferous tubules, either in germ cells at all spermatogenic stages or in Sertoli cells. However, peritubular myoid cells were immunostained. Most epithelial cells of the efferent ducts were immunolabeled, whereas the epithelial cells of the rete testis (extratesticular segments), epididymis (caput, corpus, and cauda regions), and ductus deferens were unstained. However, most epithelial cells from the intratesticular segments of the rete were immunopositive. Some cells identified as intraepithelial macrophages and lymphocytes, apical cells, and narrow cells were intensely immunolabeled. Regional differences in the distribution of these cell types along the ducts studied were also noted. The possible functional roles for taurine in these cells are discussed.

Animals↗
PubMed2002

Aspermia and chronic testicular pain after imperforate anus correction. Cryopreservation of sperm cells extracted from whole orchiectomized testis: case report.

This paper describes an unusual association of aspermia and untreatable, chronic testicular pain in a young man who underwent 14 surgical interventions for an imperforate anus. Physical examination and ultrasonography revealed left epididymal and vas enlargement, normal-sized testes, tubular ectasia of the left rete testis and a small intraprostatic paramedian left cyst. Retrograde ejaculation and urogenital infections were excluded, and the FSH and karyotype results were normal. The patient gave his consent to an exploratory intervention with possible radical left orchiectomy. The patency of the left distal seminal duct was unexpectedly normal, and no sperm were found in the epididymis or vas deferens despite their obstructive appearance. Sperm were only found in a 'testicular touch' preparation. The removed testis was immediately opened and most of the testicular lobules were removed, thus allowing the extraction of 25 x 10(6) sperm, which were cryopreserved in 35 straws. An 8-month follow-up examination documented the complete absence of pain and, during the next few months, it is planned to use the thawed sperm for ICSI. Radical orchiectomy plus the cryopreservation of sperm extracted from the whole testis must be considered in the case of the co-existence of chronic unilateral testicular pain and aspermia.

Adult↗
PubMed1997

Immunohistochemistry of unclassified sex cord-stromal tumors of the testis with a predominance of spindle cells.

Unclassified sex cord-stromal tumors (SCSTs) of the testis comprised predominantly of spindle cells can be difficult to classify. To achieve better definition of these tumors, we examined the histologic, histochemical, and ultrastructural features of four unclassified SCSTs with spindle-cell features, and compared their immunohistochemical features with those of 24 other SCSTs of the testis and ovary. Three of the spindle-cell tumors were composed of relatively short spindled cells with prominent nuclear grooves and intermixed epithelioid cells. All of the three were located adjacent to the rete testis. The fourth case consisted of elongate spindle cells that were reminiscent of smooth muscle. In all of the four cases, reticulin enveloped aggregates of cells of various sizes but not individual cells. Ultrastructural analysis of two of the spindle-cell tumors revealed desmosomes, numerous thin filaments, and focal dense-bodies. Immunohistochemically, all of the four tumors were reactive for S-100 protein and smooth muscle actin. Staining for S-100 protein and smooth muscle actin was also observed in three of six granulosa cell tumors and both juvenile granulosa cell tumors. Although variable staining for S-100 protein was found in 5 of the 12 other SCSTs (4 Leydig cell, 6 Sertoli-Leydig cell, and 2 unclassifiable ovarian SCSTs), reactivity for smooth muscle actin was present in only 1 Sertoli-Leydig cell tumor. In contrast, all of the four ovarian fibromas/thecomas were reactive for smooth muscle actin but failed to stain for S-100 protein. Taken together, the histologic, histochemical, immunohistochemical, and ultrastructural features of the spindle-cell tumors are similar to those of granulosa cell tumors. Reactivity for S-100 protein and smooth muscle actin is characteristic of these tumors. These tumors should be distinguished from other unclassified SCSTs.

Actins↗
PubMed1982

[Correlation between serum FSH and histological findings on testicular biopsy in men with azoospermia and severe oligozoospermia].

In 51 infertile men with azoospermia or severe oligozoospermia the histological findings on testicular biopsy were compared with the preoperative serum FSH level. In cases of absence of spermatogenesis on testicular biopsy the FSH showed the expected highly significant "feed back" increase, whereas in cases of functioning, but highly pathological spermatogenesis the serum FSH level gave no indication of the type of dysspermatogenesis. Normal FSH levels were found in over 90% of men showing normal spermatogenesis with obstructive azoospermia. Hence, complete inspection of the testis, with attempted demonstration of spermatozoa in the epidydimis, is essential in these cases during testicular biopsy in order to avoid an unnecessary secondary operative refertilizing procedure (epidydimovasostomy) when the site of obstruction is the rete testis.

Adult↗
PubMed1987

Immunocompetent cells in human testis in health and disease.

The authors have investigated lymphocyte subpopulations and macrophages in normal human testes and the testes of patients under investigation and treatment for subfertility. Specific monoclonal antibodies were used in an indirect immunoperoxidase technique. In normal tissues, T lymphocytes (Leu 4-positive cells) were present in the rete testis with a preponderance of cells of the suppressor/cytotoxic phenotype. In contrast, no lymphocytes were detected within the peripheral portions of the testis. Cells reacting with the anti-Leu M3 monoclonal antibody, which defines monocytes/macrophages, were detected in appreciable numbers in peripheral testis with a specific location around the seminiferous tubules. HLA-DR-positive cells (human leukocyte antigens--class II [DR] determinants of the major histocompatibility complex) also were identified and showed a similar pattern of distribution to that of the Leu-M3 positive cells. While no lymphocytes were seen in the normal peripheral testis, T lymphocytes were detected in testicular biopsies from subfertile patients. Suppressor/cytotoxic T cells (Leu 2a-positive) predominated in patients with oligozoospermia and obstructive azoospermia while T cells of the helper/inducer phenotype predominated in patients with unilateral testicular obstruction and in postvasectomy patients. Sperm antibody measurements correlated with these findings.

Adult↗
PubMed1985

Lymphocyte sub-populations in the male genital tract.

A series of monoclonal antibodies that react with human lymphocyte subsets was used in an indirect immunoperoxidase technique to study representative blocks from normal human testis, epididymis, vas deferens, prostate and seminal vesicles. Biopsies of testis, epididymis and vas obtained during surgical procedures directed at the investigation and treatment of infertile males were also studied. In all normal tissues, apart from the peripheral testis where no lymphocytes were identified, T lymphocytes were the predominant cell type (Leu 4+). These lymphocytes were largely of the suppressor/cytotoxic phenotype (Leu 2a+) and were more abundant in between the epithelial cells in the rete testis, epididymis, vas deferens, seminal vesicles and prostatic acini. Cells of the helper/inducer phenotype (Leu 3a+) were identified mainly within the interstitium of the epididymis and the prostate. B-lymphocytes (Leu 12+) were few in number and were mainly in the stroma of the prostate. In each organ the ratio of the T-cell subsets was determined and changes in this ratio were observed in epididymal and vasal biopsies from some infertile males. Finally, in testis biopsies from infertile men, suppressor/cytotoxic T-cells were demonstrated between the germinal epithelium and the fibrous tunica of the seminiferous tubules and as focal aggregates in the interstitium.

Antibodies, Monoclonal↗
PubMed1999

Testicular biopsy in patients with obstructive azoospermia.

The present report studies the testicular biopsy lesions (histologic and semiquantitative) in a series of 48 patients with obstructive azoospermia of known etiology (vasectomy, congenital absence of vas deferens, herniorrhaphy, hydrocelectomy, Young's syndrome, and ejaculatory duct obstruction) in order to establish objective testicular data that permit the pathologist to diagnose an obstructive process, which should not be mistaken with a primary testicular lesion. The semiquantitative study included determinations of the average numbers of spermatogonia, primary spermatocytes, young spermatids (Sa + Sb), and differentiated spermatids (Sc + Sd). According to this study, the testes were classified into the following groups: (1) normal testes whose germ cell numbers were within normal limits (27 testes); (2) testes with lesions in the adluminal compartment; these lesions comprise two subgroups: (2a) late sloughing of primary spermatocytes (both spermatid types were greatly reduced in number while the other germ cell types were in normal numbers) (45 testes); and (2b) early sloughing of primary spermatocytes (normal spermatogonial number, reduced number of spermatocytes, and scanty spermatids) (9 testes); and (3) lesions in the basal compartment; these lesions comprise two subgroups: (3a) pure hypospermatogenesis (a proportionate decrease in the numbers of all germ cell types) (8 testes); and (3b) hypospermatogenesis associated with sloughing of primary spermatocytes (decreased numbers of all germ cell types with a very scanty number spermatids) (4 testes). Two testes appeared hyalinized and one testis was removed owing to cryptorchidism. The most frequent testicular lesion observed (alteration in the adluminal compartment of seminiferous tubules) seems to be related to the increase in hydrostatic pressure in the tight compartment formed by seminiferous tubules, rete testis, efferent ducts, the epididymal duct, and the initial portion of the vas deferens. The severity of the lesions is probably related to the cause and span of the obstruction. In addition, two azoospermic men without obstructive azoospermia and whose testicular biopsy study revealed meiotic anomalies (with the subsequent bad prognosis) were also studied for comparison. The semiquantitative study of these patients permitted the differential diagnosis between two lesion types. Testes with meiotic anomalies had a disproportionately elevated number of primary spermatocytes, and an extremely low number of young spermatids.

Adult↗
PubMed1979

Measurement of the motility of rat spermatozoa collected by micropuncture from the testis and from different regions along the epididymis.

Spermatozoa from the testis and various regions along the epididymis of the rat were collected by micropuncture and their motility after dilution was estimated over a 15-min period by using a Quantimet image analyser. The motility of sermatozoa from the rete testis and seminiferous tubules was too low to be measured. The estimate of motility of spermatozoa from the proximal caput epididymidis was much lower than that of spermatozoa from the other regions. Spermatozoa from the distal part of the caput showed sustained motility for 15 min, whereas those from the caudal region and ductus deferens, although active initially, became less active during this period.

Animals↗
PubMed1991

Risk-related adjuvant chemotherapy for stage I non-seminoma of the testis.

Seventy-Two patients with stage I testicular non-seminoma presenting between January 1984 and December 1988 were managed either by adjuvant chemotherapy in the presence of histological adverse prognostic factors or by surveillance if none of these factors were present. The determining factors were vascular invasion, lymphatic invasion, involvement of the epididymis, involvement of the rete testis. Thirty patients were treated with three courses of platinum, vinblastine and bleomycin (PVB) and 42 were managed by surveillance. All 72 patients are alive and have been free of disease from 12-60+ months. One of 42 patients managed by surveillance relapsed 16 months after orchidectomy and he has been disease-free for 32+ months after chemotherapy. No relapses occurred in patients treated with adjuvant chemotherapy. The results confirm the suggestion by Peckham (Hoskin et al., 1986) that histopathological analysis of the primary tumour can provide the basis for subsequent management either by surveillance or chemotherapy.

Adolescent↗
PubMed2000

Immunocytochemical localization of lipocalin-type prostaglandin D synthase in the bull testis and epididymis and on ejaculated sperm.

Previously, we identified a 26-kDa fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase. The objective of the present study was to immunohistochemically localize this enzyme to the various cell types within the bull testis and seven subsegments of the epididymis, and on ejaculated sperm in order to gain further insight into its potential function in male reproduction. In the testis, immunoperoxidase staining was localized within the elongating spermatids and Sertoli cells of the seminiferous tubules, varying with the stage of the spermatogenic cycle. The highest level of staining occurred during stages III-VII. The cuboidal epithelial cells of the rete testis and efferent ducts were also immunoreactive. Expression of lipocalin-type prostaglandin D synthase was not uniform in the seven epididymal subsegments, suggesting a possible role in sperm maturation. In all epididymal regions, expression was limited to the epithelial principal cells; no immunoreactivity was apparent in other cell types. Lipocalin-type prostaglandin D synthase was strikingly localized in the caput epididymidis, while moderate to weak staining was observed in the remainder of the epididymis. Droplets of reaction product observed within the lumen increased progressively from the caput to cauda. Using fluorescence microscopy, we also localized lipocalin-type prostaglandin D synthase to the apical ridge of the acrosome on ejaculated sperm.

Animals↗
PubMed1988

Immunocytochemical localization of actin and tubulin in rat testis and spermatozoa.

Using commercial monoclonal antibodies against actin and tubulin (alpha and beta), the respective antigens were localized on semithin and ultrathin sections of the rat testis. Tubulin immunofluorescence was found in the socalled manchette surrounding the heads of the maturating spermatids as well as the sperm tail. The distribution pattern varied with sperm development. Modified Sertoli cells found at the transition between the seminiferous tubules and the rete testis displayed much filamentous tubulin-reactive material. The immunofluorescence findings could be confirmed at the ultrastructural level using the indirect immunogold method. Actin immunofluorescence was demonstrated in vascular smooth muscle cells, interstitial macrophages and - most intensely - in peritubular cells. Inside the seminiferous tubules the Sertoli cell junctions and the ectoplasmic specializations of the Sertoli cells that follow the outer contour of spermatid heads displayed distinct actin immunofluorescence. In addition to the locations mentioned, actin-like immunoreactivity was visualized at the ultrastructural level in the chromatoid body and the subacrosomal space of spermatids as well as on the outer dense fibers of the sperm tail. Immunoblotting experiments with actin antibodies showed that in extracts from testicular spermatozoa, intact or fragmented into heads and tails, from isolated Sertoli cells grown in vitro, and from testis tissue in addition to authentic actin a protein was present in sperm tail extracts that strongly bound the actin antibody. This protein may be an actin-related protein and may be responsible for the actin-like immunoreactivity of the outer dense fibers of the sperm tail.

Actins↗
PubMed1987

Structural differentiation and fluid reabsorption in the ductuli efferentes testis of the rat.

The ductuli efferentes testis of the rat form a cord which is embedded in adipose tissue. The cord is anatomically differentiated into a proximal cylindrical region, the initial zone, and an ampulla, the coni vasculosi. The initial zone contains six or seven ductuli which leave the rete testis and run in a sinuous path, roughly parallel with one another. However, the ductuli in the coni vasculosi are more sinuous than in the initial zone and they anastomose; pairs join together to form ultimately a single, common ductulus efferens. Stereological studies of paraffin sections and electron micrographs showed that the differentiation of the ductuli into two parts can be recognized at tissue and cellular levels of organization. Stereological and micropuncture studies showed that the ductuli efferentes reabsorb most of the fluid leaving the testis and it was concluded that most reabsorption occurred in the initial zone. It was estimated that the rate of fluid absorption is greater in the ductuli efferentes than in the proximal convoluted tubules of the kidney. The mechanism of fluid transport across the mucosa of the ductuli is considered in the Discussion. It is concluded that transport in vesicles and vacuoles could not account for the rate of fluid reabsorption and that the main mechanism of transport probably involves the coupling of water and active salt transport.

Absorption↗
PubMed2003

Immunohistochemical distribution of S-100 protein and subunits (S100-alpha and S100-beta) in the swamp-type water buffalo (Bubalus bubalis) testis.

The distribution and localization of S-100 protein (S-100) and its subunits (S100-alpha and S100-beta) in the testis of swamp-type water buffalo were investigated using immunohistochemistry. S-100 was detected in the Sertoli cells in the convoluted seminiferous tubules, modified Sertoli cells lining the terminal segment of the seminiferous tubules and in the intratesticular excurrent ducts (straight tubules and rete testis). S100-beta showed the same distribution and localization with that of S-100. However, the cytoplasmic extension of the Sertoli cells in S100-beta staining showed less staining intensity compared with that of S-100. S100-alpha showed a positive staining only in the modified Sertoli cells of the terminal segment of the seminiferous tubule. Endothelial cells of blood vessels were also positive with the proteins while the Leydig and spermatogenic cells showed a negative reaction. The localization of S-100 in the testis of the water buffalo was in parallel with that of other artiodactyls which supports the hypothesis that this protein is a multifunctional protein. S100-beta in the Sertoli cells suggests that this protein is involved in establishing blood-testis barrier. Its presence in the modified Sertoli cells and in the epithelium of the excurrent ducts suggest secretory and absorptive function, respectively. Meanwhile, S100-alpha, which was detected only in the modified Sertoli cells, is involved in the secretory activity of these cells that are related to exocrine function.

Animals↗
PubMed1989

Evaluation of the human testis and its age-related dysfunction.

The human testis has been evaluated by its endocrine function, daily sperm output in ejaculates, general appearance of seminiferous tubules, differential cell counts in the testis, and daily sperm production. Within-subject variation for total sperm count in ejaculates is extremely high at 42% to 75% coefficient variation. This variation can be reduced to 12% by averaging the counts obtained for the last three of five daily ejaculates. Plasma FSH concentrations are particularly useful in assessing the status of seminiferous epithelium and/or its Sertoli cell function in infertile men. In aged men, plasma LH, FSH, and estradiol concentrations are higher while plasma testosterone, free testosterone, and the ability of the testis to secrete testosterone following stimulation are reduced. Other age-related changes in human testes include a high incidence of azoospermia, reduced sexual activity, reduced testicular size, impaired spermatogenesis, reduced tubular length, increased thickness of tubular boundary tissue, sclerosis, focal mononuclear orchitis, and dilation of the rete testis. Due to the long duration of the spermatogenic cycle and low numbers of germ cells in human testis, daily sperm production per g parenchyma (efficiency of spermatogenesis) is much lower in humans than in other species. Testicular parenchymal weight, proportion of testis occupied by seminiferous epithelium, volume of seminiferous epithelium, and daily sperm production are significantly reduced in aged men. In various species, including man, germ cell degeneration occurs during spermatocytogenesis, meiosis, and/or spermiogenesis. Germ cell degeneration plays a pivotal role in spermatogenesis, but the mechanisms of degeneration, its etiology, and approaches for its prevention remain unclear.

Adult↗
PubMed1976

Studies on the testis of the camel (Camelus dromedarius). III. Histochemical observations.

The histochemical localization of carbohydrates and lipids and some oxidative, hydrolytic and steroid-linked enzymes has been studied in the testis of the camel with particular reference to the effect of the season on the distribution of these substances. PAS-positive, but diastase-resistant, material was seen mainly in the wall of blood vessels and in the boundary tissues of the seminiferous tubules, tubuli recti and rete testis. Clear cyclical changes were seen for glycogen in the lining epithelium of the seminiferous tubules. Glycogen was most abundant in early stages and very scanty or absent in the late stages of the cycle of the seminiferous epithelium. Numerous small lipid droplets were seen in the interstitial cells and towards the lumen of the seminiferous tubules that contain elongate spermatids or spermatozoa. Large lipid droplets were also demonstrable in the basal layer of the seminiferous epithelium and in the cytoplasmic debri. Alkaline phosphatase was demonstrated in the boundary tissues of the seminiferous tubules, tubuli recti and reti testis and in the cells bordering the lumen of the seminiferous tubules. Succinate and lactic dehydrogenases showed similar patterns of distribution in the interstitial elements and intratubularly. delta5-3beta hydroxysteroid dehydrogenase was exclusively demonstrated in the interstitial cells. 17beta-hydroxysteroid dehydrogenase could not be demonstrated. The season seems to have no effect on the distribution of all these substances. The possible significance of all these findings is discussed.

Alkaline Phosphatase↗
PubMed2002

Mucinous cystadenoma of the testis.

A 35-year-old man complained of a painless enlargement of the right testis. Imaging diagnostic procedures demonstrated a multiloculated cystic tumor, 9 cm in maximal diameter, in the right testis with hydrocele. Orchidectomy specimen showed that the tumor was confined within the testis and separated from the epididymis. The locules of the tumor were lined by single-layered columnar epithelium, intermingled with MUC2 immunopositive goblet and chromogranin-A immunopositive neuroendocrine cells, exhibiting intestinal differentiation. No ciliated cell, teratomatous element or intratubular germ cell neoplasia were seen. Channels of rete testis were compressed peripherally by the tumor but there was no connection with the tumor locules. The tumor was diagnosed as mucinous cystadenoma of the testis. This seems to be the first published case of benign mucinous cystadenoma occurring within the testis. This intratesticular tumor with intestinal differentiation may represent a benign monodermal teratoma.

Adult↗
PubMed1997

Immunolocalization of androgen receptor and estrogen receptor in the developing testis and excurrent ducts of goats.

BACKGROUND: Because of the significance of androgens and estrogens in prenatal and postanatal differentiation of the testis and excurrent ducts, it is important to understand the developmental pattern of androgen receptor (AR) and estrogen receptor (ER) in these organs. METHODS: Tissues from 1-23-week-old goats were fixed in 4% paraformaldehyde and embedded in Paraplast-plus. Antigenic sites for AR and ER were immunolocalized using the PG-21 rabbit anti-rat/human antibody and the H-222 rat anti-human monoclonal antibody, respectively. The avidin-biotin horseradish peroxidase procedure was used to identify positive immunoreactivity. Controls included incubation of sections with irrelevant IgG in place of primary antibody. RESULTS: Within the testis, immunostaining for AR in the nuclei of Sertoli cells increased gradually from mild at week 1 to strong at week > or = 19. In contrast, nuclei of peritubular myoid cells and Leydig cells exhibited moderate to strong reaction for AR in all animals. Germ cells were negative. Within the rete testis, efferent ductules, regions I-V of the epididymis, and ductus deferens, nuclei of all epithelial cells, peritubular myoid cells, and intertubular connective tissue cells expressed moderate to strong staining for AR at all ages. ER were confined to nonciliated cells of the efferent ductules, which displayed moderate staining in all animals, beginning from week 1. CONCLUSIONS: Nuclear AR staining, found in all testicular cells (except germ cells) and excurrent duct cells examined, was observed to change in an age-related manner only in Sertoli cells, where staining intensity increased between week 1 and week 19. Staining for ER, confined to nonciliated epithelial cells of the efferent ductules, was not affected by postnatal age.

Animals↗