Testes Of Men Research Articles

Levels of liver X receptors in testicular biopsies of patients with azoospermia

To determine whether the transcription factors liver X receptors (LXRs) and their downstream genes, which are involved in the regulation of several testicular functions in mouse models, are differentially expressed in testes of men with nonobstructive azoospermia (NOA) or obstructive azoospermia (OA). Prospective study. University hospital. Patients with various types of NOA (n=22) and with OA (n=5). Human testicular biopsies. Transcript levels were measured in testicular biopsies with the use of quantitative polymerase chain reaction. Correlations of LXR mRNA levels with the number of germ cells, the expression of proliferation and apoptosis markers, and the amount of intratesticular lipids and testosterone were evaluated. The localization of LXRα was analyzed by immunofluorescence. LXR mRNA levels were decreased by 49%-98% in NOA specimens and positively correlated with germ cell number. Accumulations of IDOL and SREBP1c (LXR targets involved in lipid homeostasis) were 1.8-2.1 times lower in NOA samples and mRNA levels of the SREBP1c target gene ELOVL6 were increased 1.9-2.4-fold. Interestingly, the amount of triglycerides and free fatty acids were higher in NOA testes (3.4-12.2-fold). LXRα was present in Leydig cells. Accumulations of LXR downstream genes encoding the steroidogenic proteins StAR and 3βHSD2 were higher in NOA testes (5.9-12.8-fold). Knowledge of changes in the transcript levels of LXRs and some of their downstream genes during altered spermatogenesis may help us to better understand the physiopathology of testicular failure in azoospermic patients.

Anti-atherogenic Effects of 17β-Estradiol

Estrogens are secreted primarily by the ovaries and placenta, by the testes in men and also produced by peripheral steroidogenic conversion. The 3 major naturally occurring estrogens are: 17β-estradiol (E2), estrone and estriol, of which E2 is the predominant and most active. The actions of E2 are mediated by at least 3 different receptors - the classical ERs (ERα and ERβ) and G-protein coupled receptor 30 (GPR30). E2 signaling in cardiomyocytes involves ERα- and ERβ-independent pathways, and treatment with the E2 receptor antagonists (Selective Estrogen Receptor Modulators- SERMs), which are agonists of GPR30, inhibits cardiac cell growth. Effects of E2 in preventing endothelial dysfunction, a prerequisite of atherosclerosis, are well recognized. Atherosclerosis involves interaction between the cells of the arterial wall endothelial cells (EC) and vascular smooth muscle cell (VSMC), as well as migration of macrophages into wall tunica media. It is predominantly developed at sites with abnormally high shear stress, such as bifurcations or branching of arteries, initiated by an injury to the endothelium and exposure to atherogenic lipids and toxins, such as those contained in tobacco smoke or infectious agents. Animal studies have shown effects of E2 in preventing atherosclerosis, inflammation and endothelial or vascular dysfunction. Gender differences along this pathogenic pathway have been also described. We review the data from the available animal and human studies, which focus on anti-atherogenic effects of E2. These studies represent evidence, albeit indirect, for an inhibitory effect of E2 on the progression of coronary artery atherosclerosis.

English

Teratomas are the germ cell tumors composed of multiple cell types derived from one or more of the three germ layers.Teratomas range from benign well differentiated (mature) cystic lesions to those that are solid and malignant (immature).Additionally, teratomas may be monodermal and highly specialized. Rarely, within some mature teratomas, certain elements like squamous component undergo malignant transformation.Teratomas are known to occur at birth and are derived from pleuripotent germ cells and embryonic cells. Teratomas of embryonic origin are congenital and are seen in the midline of the body - skull, neck, mediastinum, retroperitoneum, and coccyx.Teratomas derived from the germ cells are seen in testis in men and ovaries in women. AIMS: This prospective and retrospective study was conducted to determine the frequency of the teratomas occurring in various sites along with their histomorphological presentations. MATERIALS AND METHODS : The study was conducted at Rajarajeswari Medical College and Hospital between January 2010 to December 2012 in the Department of Pathology. Morphological and histopathological features of teratomas were studied. RESULTS: Teratomas were more common in females between 25- 35yrs.Most of the teratomas were seen in ovaries followed by retroperitoneum and sacrococcygeal region.87% of teratomas diagnosed were of mature cystic teratoma with histologic grading 0 and remaining 13% showed malignant transformation. CONCLUSION: Mature teratomas are typically benign and found more frequently in women and there is risk of malignant transformation.

Estrogen receptor alpha localization in the testes of men with normal spermatogenesis

It is known that estrogens act on the male reproductive tract by binding to estrogen receptors (ER) α and β. However, studies on ER localization in the human testis are discordant. The aim of this study was to investigate the localization of ERα in the testes of adult men with normal spermatogenesis. Semen analysis of ten adult men revealed azoospermia. FSH, LH and testosterone serum concentrations were within normal values, and the volume of the testes was normal, hence obstructive azoospermia was suspected. The tissues from testicular surgical biopsies were fixed in Bouin's fluid and embedded in paraffin. Assessments of the seminiferous epithelium (scoring 10 to -1), the number of Leydig cells (scoring 1 to 5), the areal fraction of intertubular space (IS), measurements of seminiferous tubule diameter, and the thickness of the tubular wall, were performed on microscopic sections. Immunohistochemical staining was applied with monoclonal antibodies against ERα. The mean spermatogenesis score was 10 points; IS - 30.6 ± 8.1%; seminiferous tubule diameter - 193.9 ± 19.4 μm; thickness of tubular wall - 7.44 ± 1.1 μm; number of Leydig cells - 1.6 ± 1.1 points. Immunohistochemical staining showed the localization of ERα to be in the Sertoli and Leydig cell cytoplasm, while ERα was absent in germ cells. The results of testicular tissue analysis confirmed its normal structure and normal, full spermatogenesis. The presence of ERα in Sertoli and Leydig cells in normal human testis demonstrated in this study suggests that estrogens may affect testicular function.

Estrogen receptor alpha localization in the testes of men with normal spermatogenesis

It is known that estrogens act on the male reproductive tract by binding to estrogen receptors (ER) a and b. However, studies on ER localization in the human testis are discordant. The aim of this study was to investigate the localization of ERa in the testes of adult men with normal spermatogenesis. Semen analysis of ten adult men revealed azoospermia. FSH, LH and testosterone serum concentrations were within normal values, and the volume of the testes was normal, hence obstructive azoospermia was suspected. The tissues from testicular surgical biopsies were fixed in Bouin’s fluid and embedded in paraffin. Assessments of the seminiferous epithelium (scoring 10 to –1), the number of Leydig cells (scoring 1 to 5), the areal fraction of intertubular space (IS), measurements of seminiferous tubule diameter, and the thickness of the tubular wall, were performed on microscopic sections. Immunohistochemical staining was applied with monoclonal antibodies against ERa. The mean spermatogenesis score was 10 points; IS — 30.6 ± 8.1%; seminiferous tubule diameter — 193.9 ± 19.4 μm; thickness of tubular wall — 7.44 ± 1.1 μm; number of Leydig cells — 1.6 ± 1.1 points. Immunohistochemical staining showed the localization of ERa to be in the Sertoli and Leydig cell cytoplasm, while ERa was absent in germ cells. The results of testicular tissue analysis confirmed its normal structure and normal, full spermatogenesis. The presence of ERa in Sertoli and Leydig cells in normal human testis demonstrated in this study suggests that estrogens may affect testicular function.

Expression of BET genes in testis of men with different spermatogenic impairments

To characterize the BET gene expression in human testis with spermatogenetic impairments; to examine BRDT protein expression in testis and semen. Prospective study. Fertility clinic. Azoospermic men (n = 120) who underwent testicular sperm extraction and who were classified as either normal spermatogenesis, mixed atrophy, spermatocyte maturation arrest, or Sertoli cells only according to their combined histologic and cytologic testicular findings and three normozoospermic men who donated sperm. Evaluation of testicular biopsies by qualitative and quantitative reverse transcriptase-polymerase chain reaction, immunohistochemical staining, and analysis of spermatozoa by immunofluorescence. Expression of the four BET genes in testis and localization of BRDT protein in testicular tissue and ejaculated spermatozoa. The BRDT gene was not expressed in testicular tissue from patients with Sertoli cells only, whereas the other three genes of the BET family retained expression in all the pathologies. The BRDT protein was localized in the nuclei of spermatocytes, spermatids, and ejaculated spermatozoa. Expression of BRDT protein was almost nil in testicular tissue specimens with spermatocyte maturation arrest despite normal transcript levels. Human BRDT expression pattern differs from mouse BRDT expression. In human, BRDT is the only BET gene expressed exclusively in testicular germ cells. Its expression in elongated spermatids and ejaculated spermatozoa raises the possibility that it is involved in unidentified additional functions.

Histopathologic analysis in testicular azoospermia

Azoospermia,the absence of spermatozoa in semen, currently affects 4–10% of infertile-couples. The objective of this study was to determine spermatogenesis status in azoospermic men and its correlation to demographic factors. Descriptive report of testicular biopsies in 1767 azoospermic men. Azoospermic men, ages 20-55 (mean 28.5) underwent testicular fine needle aspiration. Samples were obtained, 5 from right and 5 from left testis, using 27 gauge needles. Patients were classified according to presence or absence of sperm and quantity into the following stages: I) Sertoli cells only, II) early stage spermatogenesis arrest, III) late stage spermatogenesis arrest, IV) sperm heads only, V) hypospermatogenesis and VI) obstructive azoospermia. The testis was considered heterogeneous when 5 testicular sites or less showed cytomorphological changes of the highest stage of spermatogenesis diagnosed in that particular patient. A significant correlation between degree of heterogeneity and stage was observed (Spearman rho = 0.48, P<0.001) with stages IV and V having greatest heterogeneity (% total patients, % heterogeneity within stage) as follows: Stage I (35%, 0.6%), Stage II (16.1%, 21.4%), Stage III (10.8%, 60.4%), Stage IV (4.0%, 91.4%), Stage V (30.4%, 71.4%) and Stage VI (8.3%, 8.8%) (x2 = 847.4, df = 5, P<0.001). There was no significant correlation between azoospermia stages and STD, smoking, varicoceles or prior surgery. There was a significant decrease in size of the testes in men with Sertoli cells only syndrome (mean 58+3% of normal size). Men with sperm heads only had significantly smaller testes than all stages except for Sertoli cells only (mean 68+3% of normal size). The largest testes were observed in men with obstructive azoospermia (mean 98+2% of normal size). There was a significant correlation between degree of heterogeneity and histopathologic stage as well as testes size and stage, while no correlation was observed with common demographic factors.

New perspectives on the renal slit diaphragm protein podocin

Podocin is a critical component of the glomerular filtration barrier, its mutations causing recessive steroid-resistant nephrotic syndrome. A GenBank analysis of the human podocin (NPHS2) gene resulted in the possible existence of a new splice variant of podocin in the kidney, missing the in-frame of exon 5, encoding the prohibitin homology domain. Using RT–polymerase chain reaction and immunoblotting followed by sequence analysis, we are for the first time able to prove the expression of a novel podocin isoform (isoform 2), exclusively and constitutively expressed in human podocytes. Furthermore, we reveal singular extrarenal podocin expression in human and murine testis. Our data show the Sertoli cells of the seminiferous tubules to be the origin of testicular podocin. Confocal laser microscopy illustrates the co-localization of podocin with filamentous actin within Sertoli cells, suggesting a role of podocin in the blood/testis barrier. These results led to the rationale to examine podocin expression in testes of men with Sertoli cell-only syndrome, a disorder characterized by azoospermia. Interestingly, we observed a complete down-regulation of podocin mRNA in Sertoli cell-only syndrome, indicating a possible role of podocin in the pathogenesis of this germinal aplasia. Men with Sertoli cell-only syndrome show normal renal podocin expression, suggesting an alternate regulation of the testicular promoter. Our findings may change the perception of podocin and give new insights into the ultrastructure of glomerular slit diaphragm and the blood/testis barrier.

Apoptosis-inhibitor Aven is downregulated in defective spermatogenesis and a novel estrogen target gene in mammalian testis

To study the expression and localization of Aven in rat and human testis from azoospermic patients with different etiologies and its regulation by estrogens. Experimental study. University research center and private IVF clinic. Six men with obstructive azoospermia, five with hypospermatogenesis, and six with Sertoli cell-only syndrome; male Wistar rats. Testicular biopsies and rat seminiferous tubules (SeT) cultured in the presence or absence of 17β-estradiol (E(2)). Testicular cell localization of Aven protein was analyzed by immunohistochemistry. Expression levels of Aven in testicular biopsies and cultured SeT, in the presence or absence of 17β-estradiol, were determined by quantitative reverse transcription-polymerase chain reaction and Western blot. Aven is expressed in Sertoli cells, spermatocytes, and spermatogonia of both rat and human testis. Aven is underexpressed in the testis of men with nonobstructive azoospermia, and its expression levels correlate with severity of spermatogenic status. Aven expression is regulated by E(2) in rat SeT cultured exvivo. The results suggest that deregulation of the expression of the apoptosis inhibitor Aven may be related to male factor infertility. Moreover, Aven is an estrogen target gene and may be involved in the mechanism of testicular apoptosis control by estrogens.

Early Versus Late Maturation Arrest: Reproductive Outcomes of Testicular Failure

There is a paucity of data characterizing infertile men with maturation arrest. We hypothesized that men with early stage maturation arrest could be clinically distinguished from men with late maturation arrest and would have worse reproductive outcomes. We retrospectively reviewed the records of all patients with nonobstructive azoospermia and cryptozoospermia who underwent testis mapping and sperm extraction from 2002 to 2009 and for whom histopathological findings were available. Patients had uniform maturation arrest if multiple biopsies revealed maturation arrest at the spermatogonia/spermatocyte (early maturation arrest) or the spermatid (late maturation arrest) stage. Clinical parameters and pregnancy outcomes of in vitro fertilization/intracytoplasmic sperm injection were examined. Statistical analysis consisted of univariate and multivariate analysis. Uniform maturation arrest was identified in 49 of 219 men (22.3%) undergoing testicular sperm extraction. On multivariate analysis men with maturation arrest had significantly larger testes (p=0.01), decreased follicle-stimulating hormone (p=0.05) and more detectable genetic abnormalities (p=0.01) than men with other histopathological conditions. Men with late maturation arrest had decreased follicle-stimulating hormone (p=0.02), increased testosterone (p=0.03) and a higher sperm retrieval rate at testicular sperm extraction (p=0.01) than men with early maturation arrest. Predictors of successful sperm retrieval were larger testes, cryptozoospermia, late maturation arrest and hypospermatogenesis (each p≤0.05). Pregnancy outcomes for men with maturation arrest were not significantly different from those for men with other histopathological conditions. Maturation arrest is a common, diverse histopathological subtype of severe male infertility. Compared to men with late maturation arrest those with early maturation arrest have increased follicle-stimulating hormone, decreased testosterone and a decreased probability of mature spermatozoa. In vitro fertilization/intracytoplasmic sperm injection outcomes were similar when spermatozoa were discovered during testicular sperm extraction.

Mast cells in human testicular biopsies from patients with mixed atrophy: increased numbers, heterogeneity, and expression of cyclooxygenase 2 and prostaglandin D2 synthase

To determine intratesticular abundance and distribution of tryptase-positive mast cells (MCs) and to examine the expression of key enzymes of prostaglandin (PG) synthesis, cyclooxygenase 2 (COX2), and PGD2 synthase in the testes of men with mixed atrophy (MA) syndrome and in normal samples. Retrospective study. Academic research institute and andrology practice. Nineteen men. Testicular biopsies. Immunohistochemistry and evaluation of COX2 and tryptase-positive MCs, laser microdissection of immunoreactive cells followed by reverse transcriptase polymerase chain reaction for COX2 and PGDS-H mRNA, and transmission electron microscopy. In line with previous studies, few tryptase-positive MCs, but no COX2-positive cells, were observed in testes with normal spermatogenesis. In MA samples, the number of tryptase-positive MCs was significantly increased and the cells accumulated in the walls of the seminiferous tubules. In 11 of 13 MA samples, COX2 protein was detected. In 2 cases, Leydig cells were positive; however, in all 11 of 13 cases, COX2 was localized to MCs, coexpressing tryptase. The proportion of MCs coexpressing COX2 varied from 4% to 35%. Laser microdissection of tryptase/COX2-positive MCs followed by reverse transcriptase polymerase chain reaction revealed PGDS-H mRNA. Transmission electron microscopy identified typical MCs with abundant granules and another subtype with only a few granules, implying that MCs may differentiate in the testes. In patients with MA, testicular MC numbers and phenotypes change with respect to the ability to express COX2 and synthesize PGs. MCs and PGs have emerged as players in spermatogenic dysfunction.

CR16 forms a complex with N-WASP in human testes

The expression of corticosteroids and regional expression16 (CR16) and neural Wiscott-Aldrich syndrome protein (N-WASP) was studied in the testes of men with idiopathic azoospermia by (1) immunohistochemistry and Western blot to detect the expression of CR16 and N-WASP protein, (2) reverse transcription with the polymerase chain reaction (RT-PCR) to determine the amounts of CR16 and N-WASP mRNA, and (3) double-staining immunofluorescence and laser scanning confocal microscopy (LSCM) to demonstrate the co-localization of CR16 and N-WASP proteins. Immunohistochemistry revealed CR16 and N-WASP proteins in the Sertoli cells, specifically in the Sertoli cell-spermatogenic cell junctions (SspJs) in the epithelium of the seminiferous tubules. RT-PCR indicated that the relative amounts of CR16 and N-WASP mRNA were significantly decreased in men with idiopathic azoospermia, although the ratio of CR16 and N-WASP mRNA was approximately 1:1 in both groups of men. Western blot showed the presence of the CR16 monomer and N-WASP in all tissues examined, but the relevant bands were paler in men with idiopathic azoospermia than those in healthy men. The distribution of CR16 and N-WASP was similar in the epithelium of seminiferous tubules based on double-staining immunofluorescence and LSCM, thereby suggesting that CR16 and N-WASP were co-localized in the SspJs as a CR16/N-WASP complex. The decreased levels of CR16 and N-WASP in the testes of men with idiopathic azoospermia compared with healthy men and the co-localization at the SspJs as a CR16/N-WASP complex in the epithelium of seminiferous tubules suggest that CR16 and N-WASP play a role in the pathogenesis of azoospermia.

2015 THE DISCOVERY OF TESTICULAR PODOCIN REVEALS A NEW APPROACH FOR THE PATHOPHYSIOLOGY OF SERTOLI CELL ONLY SYNDROME

You have accessJournal of UrologyInfertility: Physiology, Pathophysiology, Basic Research1 Apr 20112015 THE DISCOVERY OF TESTICULAR PODOCIN REVEALS A NEW APPROACH FOR THE PATHOPHYSIOLOGY OF SERTOLI CELL ONLY SYNDROME Hannes Cash, Stefan Hinz, Christoph Brochhausen, Tom Florian Fuller, Kurt Miller, and Manfred Relle Hannes CashHannes Cash Berlin, Germany More articles by this author , Stefan HinzStefan Hinz Berlin, Germany More articles by this author , Christoph BrochhausenChristoph Brochhausen Mainz, Germany More articles by this author , Tom Florian FullerTom Florian Fuller Berlin, Germany More articles by this author , Kurt MillerKurt Miller Berlin, Germany More articles by this author , and Manfred RelleManfred Relle Mainz, Germany More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.2243AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Podocin represents a critical structural protein of the glomerular filtration barrier. Until now the existence of extrarenal podocin was never proven. The objective of this study was to examine possible extrarenal expression of podocin. Since we were able to prove podocin expression in the human testis we further investigated the cellular expression profile of podocin in normal testicular tissue, sertoli cell tumors and sertoli-cell-only syndrome (SCOS). The pathophysiology of SCOS remains fairly unknown. METHODS A display of human tissues was tested for extrarenal podocin expression by RT-PCR and immunoblot. Testicular podocin expression pattern was determined by immunohistochemistry and confocal laser scanning microscopy. Podocin expression in testes of healthy men and of men with SCOS was quantified by quantitative Real-time PCR. RESULTS We reveal human testis to be the only extrarenal tissue to express podocin. Podocin is co-localized with f-actin in the seminiferous tubules. The immunohistochemical staining patterns of healthy testis and sertoli cell tumor show membranous podocin expression between interdigitating Sertoli cells. These results suggest an association of podocin with the cytoskeleton and hence a role within the blood/testis barrier. Real-time-PCR analysis in testis of men with SCOS showed a complete absence of podocin compared to healthy men. CONCLUSIONS For the first time we were able to demonstrate an extrarenal podocin expression. Sertoli cells seem to be the origin of testicular podocin, where it is co-localized with filamentous actin in the intercellular space. We thus hypothesize an association of podocin with the blood/testis barrier, in analogy to other structural proteins (densin, nephrin) both expressed in podocytes and in testis. Interestingly, in testes of men with sertoli-cell-only syndrome (SCOS) podocin was not expressed. Thus our findings reveal a new approach for the pathophysiology of SCOS, indicating a possible dysfunction of the blood/testis barrier. Where Podocin is associated with the blood/testis barrier and further functional data remains of interest. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e806 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hannes Cash Berlin, Germany More articles by this author Stefan Hinz Berlin, Germany More articles by this author Christoph Brochhausen Mainz, Germany More articles by this author Tom Florian Fuller Berlin, Germany More articles by this author Kurt Miller Berlin, Germany More articles by this author Manfred Relle Mainz, Germany More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Screening for tumours in paraneoplastic syndromes: report of an EFNS Task Force

paraneoplastic neurological syndromes (PNS) almost invariably predate detection of the malignancy. Screening for tumours is important in PNS as the tumour directly affects prognosis and treatment and should be performed as soon as possible. an overview of the screening of tumours related to classical PNS is given. Small cell lung cancer, thymoma, breast cancer, ovarian carcinoma and teratoma and testicular tumours are described in relation to paraneoplastic limbic encephalitis, subacute sensory neuronopathy, subacute autonomic neuropathy, paraneoplastic cerebellar degeneration, paraneoplastic opsoclonus-myoclonus, Lambert-Eaton myasthenic syndrome (LEMS), myasthenia gravis and paraneoplastic peripheral nerve hyperexcitability. many studies with class IV evidence were available; one study reached level III evidence. No evidence-based recommendations grade A-C were possible, but good practice points were agreed by consensus. the nature of antibody, and to a lesser extent the clinical syndrome, determines the risk and type of an underlying malignancy. For screening of the thoracic region, a CT-thorax is recommended, which if negative is followed by fluorodeoxyglucose-positron emission tomography (FDG-PET). Breast cancer is screened for by mammography, followed by MRI. For the pelvic region, ultrasound (US) is the investigation of first choice followed by CT. Dermatomyositis patients should have CT-thorax/abdomen, US of the pelvic region and mammography in women, US of testes in men under 50 years and colonoscopy in men and women over 50. If primary screening is negative, repeat screening after 3-6 months and screen every 6 months up till 4 years. In LEMS, screening for 2 years is sufficient. In syndromes where only a subgroup of patients have a malignancy, tumour markers have additional value to predict a probable malignancy.

The predictive value of testicular ultrasound abnormalities for carcinoma in situ of the testis in men at risk for testicular cancer

Testicular microlithiasis (TM) is sometimes observed during scrotal ultrasound examinations in men. It has been suggested that TM is more prevalent in testes of men at risk for testicular carcinoma in situ (CIS), the precursor cells of all testicular germ cell tumours (TGCT). We have performed a retrospective analysis of ultrasound images and additional clinical data of a selected cohort of men and have determined the risk factor of TM and other ultrasound abnormalities for testicular CIS. Between 2002 and 2007, 176 testicular biopsies were performed in men with abnormalities found on the scrotal ultrasound. TM was found in 76/176 men (43.2%) and CIS was diagnosed in 20 of these men (26.3%). Here, we focused on the group of 76 men with TM to determine additional risk factors, besides TM, for CIS. In both groups, those with and without CIS, reproductive hormones, scrotal ultrasound images and patient history were compared. Predictive ultrasound findings for CIS were TM (sensitivity 100%, 95% CI: 0.8-1.0; specificity 64.1%, 95% CI: 0.6-0.7; PPV 26.3%, 95% CI: 0.2-0.4) and within this group an inhomogeneous testicular parenchyma (OR 16.1, 95% CI 2.4-106.8; sensitivity 75.0%, 95% CI: 0.5-0.9; specificity 79.0%, 95% CI: 0.7-0.9; PPV 50.0%, 95% CI: 0.3-0.7). Other significantly ultrasound characteristics for CIS in this population with TM were clusters of TM (p = 0.02) and intra-testicular lesions (p = 0.01). Men with CIS were found to have significantly lower values of inhibin-B (p = 0.02). Clusters of TM, intra-testicular lesions and lower values of inhibin-B were not significantly different in logistic regression analysis. TM on scrotal ultrasound of men with risk factors for TGCT and men with clinical signs of testicular maldevelopment has a high predictive value for CIS. However, the predictive value of an inhomogeneous testicular parenchyma, besides TM, for CIS is much higher.

Expression of the proto-oncogene c-fos and the immunolocalization of c-fos, phosphorylated c-fos and estrogen receptor beta in the human testis.

Spermatogenesis is under the control of a complex endocrine and paracrine system, including estrogen receptor (ER) signaling. In many target cells, ER promotes the transcription of c-fos and other proto-oncogenes to regulate cell growth and differentiation. Thus, in this study we evaluated the expression of the proto-oncogene c-fos and the immunolocalization of c-fos, phosphorylated c-fos and ERbeta proteins in the human testis. Testis tissue samples were obtained from 12 men undergoing orchiectomy as adjuvant treatment for prostate cancer, and were stained by immunohistochemistry for c-fos, phosphorylated c-fos and ERbeta localization. Both forms of c-fos proteins were immunoreactive, mainly in germ cells (spermatogonia, spermatocytes and spermatids) and Sertoli cells, while ERbeta was primarily present in somatic cells (Leydig, Sertoli and myofibrillar cells). In addition, testicular biopsies obtained from infertile men with obstructive azoospermia/normal spermatogenesis (n=8) or non-obstructive azoospermia/severely impaired spermatogenesis (n=12) were evaluated for c-fos and ERbeta mRNA levels using real time polymerase chain reaction. The expression of c-fos mRNA was significantly lower (fold change = 0.08, p<0.05) whereas that of ERbeta mRNA was higher (fold change = 9.43, p<0.05) in the testis of men with non-obstructive azoospermia compared to those with obstructive azoospermia. These findings suggest a complex interrelation between estrogen signaling and c-fos transcriptional activity within the human testis, with the increase of ERbeta mRNA being putatively a compensatory mechanism for lower c-fos expression in infertile men with damaged spermatogenesis.

On the morphology of the transitional zone of the seminiferous tubule and the rete testis in man.

Die Morphologie der Übergangszonen zwischen Samenkanälchen und Rete testis beim Menschen Die Übergangszonen zwischen den Tubuli seminiferi und den Tubuli recti des Rete testis beim Menschen wurden licht-und elektronenmikroskopisch untersucht. Es ließen sich drei Anastomosierungstypen darstellen: End-zu-End-Anastomosen, End-zu-Seit-Anastomosen und Seit-zu-End-Anastomosen. Im Terminalsegment besteht das Keimepithel aus modifizierten Sertolizellen, zwischen denen vereinzelt Spermatogonien liegen. Das Tubuluslumen ist eng, die Lamina propria ist dick. Am Abschluß des Tubulus seminiferus sind die modifizierten Sertolizellen lippenartig in das Lumen des Tubulus rectus vorgewölbt. Die Sertolizellen des Terminalsegments sind gegenüber den normalen Sertolizellen charakteristisch verändert: die Zellkerne sind oval und glatt begrenzt, perinukleäre Einschlüsse fehlen, im apikalen Bereich des Zytoplasmas befinden sich zahlreiche Lipidtropfen und Residualkörper. Im gesamten Zytoplasma sind Mikrotubuli und Mikro-filamente vermehrt. Die Lamina propria besteht aus einer Basalmembran und aus unregelmäßig angeordneten Myofibroblasten, zwischen denen dicke Bündel kollagener Fibrillen das Terminalsegment ringförmig umschließen. Am Anfang der Tubuli recti findet man häufig ektopische Inseln des modifizierten Keimepithels, die um ein bindegewebiges Zentrum angeordnet sind.

Pituitary gonadal function in infertile men with varicocele.

Infertile men with varicocele or idiopathic infertility were compared with a control group. Spermocytograms were taken and the following radioimmunological plasma analyses carried out: testosterone, FSH and LH before and after 50 micrograms LRH, Prolactin (PRL) before and after 200 micrograms TRH; in addition, 8 patients with varicoceles and 3 controls received LRH intravenously (0.4 microgram/min.) for 4 hours. The binding of [125I] human chorionic gonadotrophin (hCG) to testicular tissue obtained by biopsy from 10 infertile men was also investigated. Of the parameters studied, no differences were found between the unilateral or bilateral varicoceles. In the two groups of infertile men, sperm motility and percentage normal forms were similar and significantly lower than in controls. As compared to the controls, in the groups of infertile men, basal LH and testosterone levels were no different but basal FSH levels was increased, basal PRL was higher (p less than 0.05) in the varicocele group. Responses of the LH, FSH and PRL to LRH and TRH stimulations were generally higher in infertile men than in controls. As compared to the idiopathic infertile men, testosterone levels and responses of plasma FSH to LRH injection were lower in varicocele group. Moreover, in infertile men with varicocele, age was correlated negatively with sperm motility and testosterone level and it was correlated positively with LH response to LRH injection. For each patient, testicular tissue was able to specifically bind [125I]hCG, but in some cases of varicoceles, hCG binding capacity was different in the two testes and seemed higher than that observed in men with obstructive azoospermia. These results suggest: 1) dysfunction in both spermatogenesis and Leydig cells with a compensatory hyperfunction of the pituitary gland in infertile men with varicocele; 2) worsening in Leydig cells and tubular lesions with longer duration of varicocele; and 3) absence of any gross abnormality in hCG binding to its specific receptors in the testis of men with varicocele. These data suggest varicoceles may play a causal role in infertility.

On the expression of protamine genes in the testis of man and other mammals

Protamines are low molecular weight, highly basic nuclear proteins involved in the condensation of sperm chromatin. cDNA clones for human protamine 1 and 2 (PRM1 and PRM2) were used for Northern blot experiments with RNA from different human tissues. Protamine transcripts, 0.6 kb and 0.9 kb in lengths for PRM1 and PRM2, respectively, were detected only in testicular RNA. The hybridization signals though did not produce sharp bands but enclosed a minor fraction of significant smaller transcripts. When polysomal RNA fractions were used in the hybridization, these shorter transcripts, 0.45 kb in length for PRM1 and 0.7 kb for PRM2, were specifically enriched. As demonstrated by in situ hybridization on human testis sections, the transcripts of both protamine genes are restricted to the central cell layer of the tubuli seminiferi corresponding to the spatial arrangement of postmeiotic cells. This result indicates that the protamine genes in the human are postmeiotically and haploid expressed. When cDNA clones of both protamines of the boar (BPrm-1 and BPrm-2) were used for hybridization experiments with the testicular RNA of those mammalian species which lack protamine 2 in their spermatozoa, the presence of transcripts for both protamines was detected. It can be assumed that mammals in general are endowed with at least two protamine genes which are both transcribed but are translationally regulated in a species-specific manner.

Comparing Expression of Progesterone and Estrogen Receptors in Testicular Tissue From Men With Obstructive and Nonobstructive Azoospermia

The objective of this study was to identify and compare the expression profiles of progesterone receptor (PR) and estrogen receptor alpha (ERalpha) in the testes of men with obstructive azoospermia (OA), maturation arrest (MA), and Sertoli cell-only (SCO) histology. Testicular biopsies were obtained from 10 patients with OA, 10 patients with MA (either early or late arrest), and 8 patients with SCO who did not have hormonal abnormalities and varicoceles. Expression of PR and ERalpha was detected by immunofluorescence and Western blot. PR was expressed in the spermatogenic, Leydig, and Sertoli cells in the testes of OA patients. In the MA and SCO patients, the expression of PR was reduced in all cell types as compared with that in the OA patients. Western blot demonstrated that both the full-size (120 KDa) and the truncated (52 KDa) isoforms of the PR were expressed in the OA and MA testes. However, in the SCO testes, only the truncated isoform of PR (52 KDa) was expressed. ERalpha (66 KDa) was expressed principally in the spermatogenic and Leydig cells in the OA testes. By immunohistochemistry staining, expression of ERalpha was decreased in the spermatogenic and Leydig cells of the MA testes, whereas its expression was enhanced in the Leydig cells of the SCO testes. However, by Western blot, expression of ERalpha was significantly reduced in the SCO testes as compared with that in the OA and MA testes. We conclude that PR and ERalpha may play a role in the pathogenesis of the MA and SCO phenotype in patients with infertility.