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PubMed2005

TGF-beta3 regulates anchoring junction dynamics in the seminiferous epithelium of the rat testis via the Ras/ERK signaling pathway: An in vivo study.

Recent studies have shown that transforming growth factor (TGF)-beta3 regulates blood-testis barrier (BTB) dynamics in vivo, plausibly by determining the steady-state levels of occludin and zonula occludens-1 (ZO-1) at the BTB site via the p38 MAP kinase signaling pathway. Since BTB is composed of coexisting TJs and basal ectoplasmic specializations [ES, a testis-specific adherens junction (AJ) type] in the seminiferous epithelium of the rat testis, we sought to examine if TGF-beta3 would also regulate anchoring junction dynamics. Using an in vivo model in which rats were treated with AF-2364 [1-(2,4-dichlorobenzyl)-indazole-3-carbohydrazide] to perturb Sertoli-germ cell AJs without affecting the integrity of TJs at the BTB, it was noted that the event of germ cell loss from the epithelium was associated with a transient surge in TGF-beta3. Furthermore, it was also associated with a surge in the protein levels of Ras, p-ERK, and the intrinsic activity of ERK, illustrating TGF-beta3 apparently regulates Sertoli-germ cell ES function via the Ras/MEK/ERK signaling pathway. Indeed, pretreatment of rats with TbetaRII/Fc chimera, a TGF-beta antagonist, or U0126, a specific MEK inhibitor, could significantly delay and partially block the disruptive effects of AF-2364 in depleting germ cells from the epithelium. While the protein levels of the cadherin/catenin complex were significantly induced during AF-2364-mediated germ cell loss, perhaps being used to retain germ cells in the epithelium, this increase failed to reverse the loss of adhesion function between Sertoli and germ cells because of a loss of protein-protein interactions between cadherins and catenins. Collectively, these results illustrate that the testis has a novel mechanism in place in which an agent that primarily disrupts TJs can induce secondary loss of AJ function, leading to germ cell loss from the seminiferous epithelium. Yet an agent that selectively disrupts AJs (e.g., AF-2364) can limit its effects exclusively at the Sertoli-germ cell adhesive site without perturbing the Sertoli-Sertoli TJs.

Animals↗
PubMed2006

Identification of a novel population of adrenal-like cells in the mammalian testis.

Steroidogenic cells of the adrenal and gonad are thought to be derived from a common primordium that divides into separate tissues during embryogenesis. In this paper, we show that cells with mixed adrenal and Leydig cell properties are found dispersed in the insterstitium of the embryonic and adult mouse testis. They express the adrenal markers Cyp11b1 and Cyp21 and respond to ACTH. Consistent with these properties, we show that the embryonic testis produces the adrenal steroid corticosterone. These cells also express Cyp17 and respond to hCG stimulation but do not express the Leydig specific marker Insl3 showing that they are a population of steroidogenic cells distinct from Leydig cells. Based on their properties, we refer to these cells as adrenal-like cells of the testis and propose that they are the mouse equivalent of the precursors of human adrenal rests, tumors found primarily in male patients with congenital adrenal hyperplasia. Organ culture studies show that ACTH-responsive cells are present at the gonad/mesonephros border and seem to migrate into the XY but not the XX gonad during development. Consistent with this, using transgenic Cyp11a1 reporter mice, we definitively show that steroidogenic cells can migrate from the mesonephros into the XY gonad. We also show that the region between the mesonephros and the gonad harbors steroidogenic cell precursors that are repressed by the presence of the mesonephros. We propose that this region is the source of the adrenal-like cells that migrate into the testis as it develops and are activated when Leydig cells differentiate. These studies reveal the complex nature of steroidogenic cell differentiation during urogenital development.

Adrenal Glands↗
PubMed2005

Effects of cadmium exposure on testis apoptosis in the marine teleost Gobius niger.

It is known that heavy metals can accumulate in tissues during aquatic organism growth (bioaccumulation) and often biomagnify up the food chain interfering with the health and reproduction of both wildlife and humans. Recently, cadmium (Cd) was included in the endocrine disruptors list, exerting its effect on gametes quality and reproductive functions; in addition, its role as apoptotic factor was evidenced in different cell types and tissues. In the present study, the effects of two different Cd doses on testis and liver of the black goby Gobius niger were analyzed. Cd concentration in the water and its uptake by the gills were measured by differential pulse anodic stripping voltammetry. Toxic, apoptotic, and stressor Cd effects were analyzed using metallothionein (MTT), caspase 3 and heath shock protein 70 (HSP70), respectively, as bioindicators. The results of the present study suggested that, in the gills, the saturation of all specific metal sites was reached only with the highest Cd dose exposure. Either testis and liver showed an increase of MTT gene expression and protein synthesis in addition to HSP70 gene expression, related with Cd concentration in the water indicating that both tissues were affected by Cd exposure. In conclusion, the present study, not only shows the toxic effect of Cd on hepatic tissue, but also indicates its potency as apoptotic factor in the testis. This is supported by the increase of caspase 3 gene expression and the presence of its active form in testis of exposed fish.

Animals↗
PubMed2002

Distribution of dynamins in testis and their possible relation to spermatogenesis.

Dynamin 2 and dynamin 3 are highly expressed in testis. However, their functions in the tissue remain unclear. Considering that dynamin 1, neuron-specific isoform of dynamin, plays a pivotal role in endocytosis, functions of dynamin 2 and dynamin 3 in testis must be essential. Cellular expression and subcellular localization of dynamin 2 and dynamin 3 in testis were investigated. Dynamin 2 and dynamin 3 were highly expressed in germ cells and Sertoli cells, constituents of seminiferous tubules. By immunofluorescence it was revealed that dynamin 2 colocalizes with clathrin both at the plasmamembrane and at Golgi in a cell line of Sertoli cells. Immunoreactivity for dynamin 3, on the other hand, appeared as finer puncta, which did not colocalize with clathrin, suggesting that these two dynamins have distinct functions in Sertoli cells. In the klotho deficient mouse testis, which demonstrates disorder in spermatogenesis, expression of dynamin 2 and dynamin 3 was drastically reduced indicating possible association of these proteins with spermatogenesis.

Animals↗
PubMed1991

Atrial natriuretic factor in mammalian testis: immunological detection in spermatozoa.

Immunoreactive atrial natriuretic factor (ANF) was localized by immunochemistry and radioimmunoassay in mouse and rat testes. The analyses of acid extracts of testes by gel filtration and reverse phase high pressure liquid chromatography (HPLC) revealed the presence of a processed 31-residues peptide and the precursor form of 126-residues pro-ANF molecule corresponding to a molecular weights (Mr) of 3,300 and 18,000, respectively. The concentration of ANF in mice testis averaged 12 +/- 3 ng and in rat testis 8 +/- 2 ng per g of tissue. Specific immunochemical staining was localized in the spermatids and elongating spermatozoa of mammalian testis. The demonstration of immunoreactive ANF in testis and specific localization in spermatids reveals a new site at which ANF may be actively synthesized and regulate paracrine and/or autocrine function(s) during spermiogenesis, suggesting a broader spectrum of ANF action in addition to its known regulatory role in the control of blood pressure homeostasis.

Animals↗
PubMed1994

Immunocytochemical localization of epidermal growth factor receptors in human testis from infertile subjects.

OBJECTIVE: To investigate the immunolocalization of the epidermal growth factor receptor (EGFR) in normal and pathological human testis by immunocytochemical technique. DESIGN: Cytologic specimens were obtained by bilateral fine needle aspiration (FNA) of the testis and stained in May Grünwald-Giemsa for the cytologic analysis; immunolocalization of EGFR was analyzed on duplicate slides from each testis using two anti-EGFR monoclonal antibodies and peroxidase-antiperoxidase technique. SETTING: Infertility center of an academic unit. PATIENTS: A total of 42 infertile patients, affected by various testicular diseases. The control group was made up of 10 normal sperm patients with autoimmune infertility and cytologic picture of normal spermatogenesis. INTERVENTIONS: Exogenous FSH was administered 75 IU IM on alternate days for 3 months on 16 of the infertile patients who showed oligospermia and normal FSH plasma levels. Semen analysis and testicular FNA (and after cytologic and immunocytochemical studies) were repeated at 3 months of treatment. MAIN OUTCOME MEASURES: Luteinizing hormone and FSH plasma levels were determined by RIA methods; qualitative and quantitative parameters for the cytologic evaluation are reported in our previous works. RESULTS: The cytologic analysis permitted identification of seven classes of infertile subjects, characterized by different cytologic pictures. Epidermal growth factor receptor immunostaining evidenced weak positivity on Sertoli and germ cells (with the exception of spermatozoa) in the presence of normal germ line and normal FSH plasma levels and strongly intense positivity in the presence of serious hypospermatogenesis, spermatogonial or spermatocytic arrest, and Sertoli cell-only syndrome. These conditions were characterized by higher FSH plasma levels than normal controls. All of the subjects who received exogenous FSH, with moderate hypospermatogenesis or spermatidic arrest, showed on Sertoli and germ cells a weak EGFR immunostaining before the treatment and intense immunostaining after the treatment. CONCLUSIONS: These results confirm recent demonstrations of EGFRs in human testis and evidence different EGFR immunostaining in the presence of various degrees of testicular damage, suggesting a role of this growth factor in growth and differentiation of the germ cells throughout spermatogenesis. The observation that intense EGFR immunostaining was found in subjects showing high FSH plasma levels and in all of the patients who received exogenous FSH, supports a possible role of this gonadotropin in the modulation of the EGFR expression.

Adult↗
PubMed1995

The clinical characteristics of 82 patients with Sertoli cell-only testis histology.

OBJECTIVE: To determine the clinical and laboratory features associated with Sertoli cell-only testis histology. DESIGN: Case-controlled retrospective analysis. SETTING: University-based male infertility clinic. PATIENTS: Eighty-two patients with germ cell aplasia histology on testis biopsy from 1984 to 1994. Hormonal findings from a control group of 34 fertile men with normal semen analyses were used for comparison. MAIN OUTCOME MEASURES: History, physical examination, semen analysis, and endocrine findings. RESULTS: Sertoli cell-only histology was observed in 82 of 652 (13%) biopsies performed in infertile men with azoospermia. A major medical illness or genital anomaly (hypospadias, cryptorchidism) was found in 31 men (38%). On physical examination, 75% of patients had small testis (< 18 mL volume) and 45% had abnormally soft testis. On hormone evaluation only serum FSH concentration was elevated significantly (33 mIU/mL [33 IU/L]) compared with fertile controls (7.1 mIU/L [7.1 IU/L]). Patients (6%) with confirmed elevations in serum PRL had lower FSH values than those men with normal PRL levels. Normal ejaculate volumes were observed in all patients. CONCLUSIONS: In approximately 38% of patients, germ cell aplasia is associated with underlying disease states; in the remainder it is largely idiopathic. The physical examination is often characteristic, along with an elevated serum FSH concentration. Normal FSH levels tend to be associated with hyperprolactinemia in these men.

Case-Control Studies↗
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↗
PubMed1982

Protection of the solitary testis.

In 15 negative explorations for unilateral cryptorchidism, inspection of the contralateral scrotum revealed a "bell-clapper" deformity in 13 patients. Since the descent of the testis is influenced by the gubernaculum and gubernacular abnormalities are usually bilateral, the data suggest that an inadequate gubernaculum is associated with most instances of antenatal and postnatal torsion. Unilateral absence of the testis is a form of this syndrome; congenital monorchidism is probably due to antenatal torsion rather than hypoplasia or agenesis. When monorchidism is confirmed at operation, exploration and suture fixation of the contralateral testis at the same procedure is recommended to protect the solitary testis from future torsion.

Adolescent↗
PubMed1989

Characterization and localization of in vivo phospholipid methylation in the hamster testis.

Although previous studies have demonstrated that phospholipid methylation occurs in the testis and may be involved in Leydig cell function, phospholipid methylation in spermatogenic cells has not been characterized. In this study we describe the occurrence, time course, and localization of phospholipid methylation in the hamster testis following intratesticular injection of radioactive methyl precursor. Adult and pubertal (seven day old) hamsters were injected intratesticularly with [3H-methyl]-methionine and sacrificed 10 min. to 31 hours thereafter. The testes were then removed and homogenized or dispersed into cell suspensions. Spermatogenic cell and Leydig cell enriched preparations were isolated from the dispersed cell preparations using elutriation and Percoll gradient centrifugation and assayed for methylated phospholipids and proteins. These experiments demonstrated that 1) phospholipid methylation occurs in the hamster testis at a level seven-fold greater than protein methylation, 2) the incorporation of radioactivity associated with phospholipid methylation is progressive over time, and 3) in vivo, spermatogenic cell preparations enriched with pachytene spermatocytes have an almost four-fold higher level of measurable phospholipid methylation when compared to whole testis preparations. Taken together, these results suggest that phospholipid methylation may play an important stage-specific role in spermatogenesis.

Animals↗
PubMed1987

Testicular cancer risk in boys with maldescended testis: a cohort study.

Testicular maldescent is considered as a predisposing condition for development of testicular malignancy. Male subjects with a history of cryptorchidism have been suggested by some authors to have a 40 to 50 times increased risk of testis cancer. However, the magnitude of this risk is a point of considerable disagreement. Therefore, we studied the records of 506 consecutive patients hospitalized for maldescended testis from January 1949 to December 1960. Testis cancer developed in 6 patients, which when compared to the 1.3 expectant Danish incidence rate, yielded a statistically significant relative risk of 4.7 (95 per cent confidence interval 1.7 to 10.2). Thus, our study confirmed that male subjects with a history of testicular maldescent have an increased risk for testis cancer, although the magnitude of this risk was lower than suggested previously.

Adult↗
PubMed1987

On unilateral testicular and epididymal torsion: no effect on the contralateral testis.

It is often stated that unilateral testicular torsion results in damage to the contralateral testis; however, there are a growing number of experimental and clinical papers which suggest this is not so. Conflicting results from experimental studies confuse the issue and may be due, among other things, to some specifics of the experimental model. In the present paper, we have examined bilateral rat testes 30 and 60 days after 720 degrees torsion to determine 1) the effect of unilateral testicular torsion with and without the inclusion of epididymal torsion, 2) the effect of relatively chronic torsion (24 hr., 10 day) versus relatively acute torsion (two hr., four hr.), and 3) the effect of establishing the model using scrotal surgery versus using an abdominal approach. Bilateral testicular histology, testis wt. (gm.), cauda epididymal sperm concentrations (sp./ml.), and cauda sperm motility scores (0-4) were examined. Ipsilateral testicular torsion or testicular plus epididymal torsion of two hr. or four hr. duration significantly reduced (p less than .05) ipsilateral testis weights, sperm concentrations, and motility scores, and disrupted normal tissue histology. Contralateral testicles were not altered. Epididymal ischemia alone produced no significant ipsilateral or contralateral effects. Chronic torsion (one day, 10 days) also destroyed ipsilateral testis function without altering the contralateral testicles. The occult cryptorchidism associated with the scrotal approach to establishing the torsion model had no effect on contralateral testicles. In no group, using either Lewis rats or Sprague-Dawley rats, were contralateral testicles altered by unilateral testicular torsion. These results plus recent clinical reports indicate that contralateral testicular damage due to ipsilateral torsion is hardly a proven phenomenon, let alone a significant factor contributing to male infertility.

Animals↗
PubMed1985

Crossed ectopic testis with common vas deferens.

Crossed ectopic testis is an uncommon anomaly. A 10-month-old boy with a crossed ectopic testis and a common vas deferens is presented. At operation 2 testes were in the left inguinal canal. The seminal vesiculogram showed that both vasa deferentia fused to form a common lumen, and then formed a common stem on the left side and opened into the posterior urethra. The spermatic arteriogram revealed that each testis was supplied from the right and left sides, respectively. No testis or vas deferens was located in the right retroperitoneal space. These features suggested that the testes developed from both sides and subsequently descended to the left side.

Choristoma↗
PubMed1998

DNA adducts in mouse testis and lung after inhalation exposure to 1,3-butadiene.

1,3-Butadiene is a widely used industrial chemical and also an environmental contaminant. Recent findings have shown that butadiene can also be a male germ cell mutagen. In this study, DNA adduct formation in testis and lung has been explored by using N7-alkylated guanines as a marker of biological effective dose. The adducts measured were the four structurally different guanine N7-adducts alkylated by butadienemonoepoxide, the main metabolite of 1,3-butadiene. This study demonstrates the dose-dependent adduct formation in lung and testis. At lower exposures (50 and 200 ppm) the adduct levels were about the same in the two organs, but at 500 ppm the adduct level was significantly (p < 0.03) higher in testis than in lung. The enantiomeric composition of the adducts detected was also different. In lung, all 4 possible adducts were present (S-C-1" dominating, 49%), but in testis only two out of four adducts were detected (S-C-2" being the most abundant adduct, 71%). These novel observations indicate that the DNA repair is different in these two organs studied and that heritable genetic effects observed may be mediated through the DNA adducts.

Alkylation↗
PubMed1995

Building a testis.

Specific cellular, subcellular and acellular components of the rat testis including the capsule, the peritubular tissue (tunica propria) and the lymphatic endothelium were analyzed using morphometric techniques at cellular and subcellular levels to yield volume and surface area data. These data were integrated with previously published data for other cellular components of the rat testis to provide information about the volumetric composition for virtually every component of this organ. For major cell types (Leydig, Sertoli, myoid cells and germ cells) the data are expressed to the subcellular level in terms of volume and, in some instances, surface area. Graphic portrayals of testis constituents are used for rapid visual understanding of testis structure. The data presented herein are useful in conjunction with biochemical data to describe physiological properties of cells and cell components and also for understanding how structure differs under experimental and in pathological situations.

Animals↗
PubMed1996

Cystic transformation and calcium oxalate deposits in rete testis and efferent ducts in dialysis patients.

The histological study of the testes and epididymides obtained from autopsies of 24 men with chronic renal insufficiency revealed bilateral cystic transformation of the rete testis and efferent ducts in patients who underwent hemodialysis or peritoneal dialysis, but not in patients who did not receive this treatment. The lesion was associated with an accumulation of crystalline calcium oxalate deposits in the lumen of the rete testis and efferent ducts, and in the connective tissue adjacent to these excretory ducts. The rete testis epithelium showed columnar transformation with occasional papillary proliferations. Neither atypias or mitoses were observed. In three specimens, fibrosis and giant cell reactions was also present in the rete testis at the level of crystalline deposits. In three specimens, the caput epididymidis was enlarged, and the efferent ducts showed an increase in both tubular diameter and epithelial height, irregular outline, and development of diverticula. The lesions appeared within 30 months after the onset of dialysis.

Adult↗
PubMed1994

Peptidergic innervation of the human testis and epididymis.

Neuropeptidergic innervation of the human testis and epididymis was investigated by immunohistochemical methods. The innervation of the epididymis was more dense than that of testis. In the testis only tyrosine hydroxylase- and neuropeptide Y-positive nerves could be found between seminiferous tubules and around blood vessels. In the connective tissue capsule of the testis also small calcitonin gene-related peptide- and metenkephalin-containing nerve fibres were seen. The epididymis was densely innervated by nerve fibres immunoreactive to tyrosine hydroxylase, neuropeptide Y, vasoactive intestinal polypeptide, calcitonin gene-related peptide, galanin, peptide histidine isoleusine and substance P.

Aged↗