Human testicular mast cells contain tryptase: increased mast cell number and altered distribution in the testes of infertile men

OBJECTIVE: To determine whether human testicular mast cells contain the potent fibroblast growth factor tryptase and to examine changes in mast cell morphology and intratesticular distribution in testes with normal spermatogenesis versus abnormal spermatogenesis. DESIGN: Retrospective evaluation of testicular biopsies with the use of immunohistochemistry, morphometry, and electron microscopy. SETTING: University research and clinical institutes. PATIENT(S): Infertile men (total of 24) with severe hypospermatogenesis, germ cell arrest syndrome, or Sertoli cell only syndrome, and men without pathologies. INTERVENTION(S): Diagnostic testicular biopsy. MAIN OUTCOME MEASURE(S): Location, number, and distribution of testicular mast cells. RESULT(S): All groups showed tryptase-positive mast cells. In specimens with normal spermatogenesis, mast cells were round and located mainly in the interstitial spaces close to Leydig cells. In germ cell arrest syndrome, a 2-fold increase was evident, and in Sertoli cell only syndrome, a >3-fold increase of tryptase-immunoreactive mast cells became evident. Moreover, there was a statistically significant shift of the cells from the interstitium to the tubular walls in Sertoli cell only syndrome and germ cell arrest syndrome. Mast cells in specimens of Sertoli cell only syndrome and germ cell arrest syndrome were heterogeneous, with rounded or elongated shapes and signs of degranulation. The thickness of the tubular walls was doubled in specimens of germ cell arrest syndrome and Sertoli cell only syndrome in comparison with normal specimens, and this increase was positively correlated with the number of mast cells in these patients. CONCLUSION(S): Our results suggest that mast cell products, including the potent fibroblast growth factor tryptase, are involved in the thickening of the tubular wall and other changes in infertile testes.     

Results of testicular biopsy in azoospermia]

The causes of male infertility generally fall into three categories: pretesticular, testicular and posttesticular causes. The pretesticular causes are extragonadal endocrine disorders, such as those originating in the pituitary or the adrenals, which have an adverse effect on spermatogenesis. The testicular causes of infertility are conditions in which the primary defects reside in the testes. The posttesticular causes of infertility consist mainly of obstructions of the ducts leading away from the testes. Testicular biopsy is an important method in the diagnosis and management of male infertility.     

Aspiration flow cytometry of the testes in the evaluation of spermatogenesis in the infertile male

Deoxyribonucleic acid (DNA) flow cytometry of testicular tissue has been demonstrated to be a quantitative means of assessing spermatogenesis. This study evaluates testes aspirates and DNA flow cytometry in the evaluation of the infertile male. Testicular tissue obtained from 12 men who underwent bilateral orchiectomy for prostate cancer (group 1) were examined by both flow cytometry and standard histologic technique to assess the correlation between these two modalities. Thirteen men evaluated for infertility (group 2) and requiring histologic evaluation of spermatogenesis underwent both open biopsy and fine needle aspiration of their testes. Histology was independently examined and grouped according to standard nomenclature. Flow cytometric analysis revealed characteristic patterns in the relative numbers of haploid (1C), diploid (2C), and tetraploid (4C) cells. These patterns correlated reproducibly with the histologic diagnoses. DNA flow cytometry of testicular aspirates provides a rapid and reliable quantitative means of assessing spermatogenesis.     

Testicular involution in elderly men: comparison of histologic quantitative studies with hormone patterns

An endocrinologic and quantitative histologic study was carried out in 64 elderly men who underwent orchidectomy owing to prostatic carcinoma. The men were classified into age groups (decade of life), and each group was subdivided into group A (testes with complete spermatogenesis in most tubules) and group B (testes showing maturation arrest of spermatogenesis in most tubules). Up to 80 years of age, men of group A showed hormone levels and testicular parameters similar to those of young control men. From 50 to 60 years of age, men of group B showed a significant decrease in testicular volume, tubular volume, tubular length, number of germ cells, Sertoli cells and Leydig cells per testis, and plasma testosterone levels, whereas the tunica propria thickness and plasma levels of both follicle-stimulating hormone and luteinizing hormone were increased.     

Correlation between spermatozoon numbers in spermiogram and seminiferous epithelium histology in testicular biopsies from subfertile men

A new method for correlating testicular biopsies and spermiograms is proposed. The number of spermatogonia, round spermatids, and elongated spermatids per cross-sectioned tubule were calculated in the testes from 33 subfertile men and in 10 control normal testes. According to these quantitations, the testes in subfertile men were classified as testes with maturation arrest of spermatogenesis, testes with hypospermatogenesis, and testes with associated maturation arrest and hypospermatogenesis. A power regression curve correlating the number of elongated spermatids and sperm numbers in the spermiogram was performed.     

Molecular mechanisms involved in varicocele-associated infertility

Varicocele is a pathologic enlargement of the pampiniform venous plexus within the spermatic cord, a condition that is a common cause of impaired sperm production and decreased quality of sperm. While varicocele is the most common surgically correctable risk factor for male infertility, not all males with varicocele experience infertility. In fact, most men with varicocele have normal spermatogenesis. Despite its prevalence, the molecular mechanisms of varicocele and its effect on testicular function are yet to be completely understood. We postulate that men with varicocele-associated infertility could have preexisting genetic lesions or defects in molecular mechanisms that make them more susceptible to varicocele-mediated testicular injury affecting spermatogenesis.     

Decreased expression of MRE11 and RAD50 in testes from humans with spermatogenic failure

To assess testicular mRNA and protein expression levels of MRE11 and RAD50 in human azoospermia patients. Patients diagnosed with maturation arrest at the spermatocyte stage (MA) and Sertoli cell-only syndrome (SCOS) were recruited through diagnostic testicular biopsy. Patients with normal spermatogenesis were studied as controls. In addition, knockdown of MRE11 and RAD50 was performed in GC-2spd(ts) cells to investigate their roles in cellular proliferation and apoptosis. mRNA and protein expression levels of MRE11 and RAD50 were measured using quantitative polymerase chain reaction, western blotting, and immunohistochemistry, respectively. Knockdown of both MRE11 and RAD50 utilized transfection with small interfering RNAs. Our findings demonstrated altered expression levels of MRE11 and RAD50 in human testes with MA and SCOS, and showed that these alterations might be associated with impaired spermatogenesis. These results offer valuable new perspectives into the molecular mechanisms of male infertility.     

Management of non-obstructive azoospermia

Non-obstructive azoospermia (NOA) is defined as no sperm in the ejaculate due to failure of spermatogenesis and is the most severe form of male infertility. The etiology of NOA is either intrinsic testicular impairment or inadequate gonadotropin production. Chromosomal or genetic abnormalities should be evaluated because there is a relatively high incidence compared with the normal population. Although rare, NOA due to inadequate gonadotropin production is a condition in which fertility can be improved by medical treatment. In contrast, there is no treatment that can restore spermatogenesis in the majority of NOA patients. Consequently, testicular extraction of sperm under an operating microscope (micro-TESE) has been the first-line treatment for these patients. Other treatment options include varicocelectomy for NOA patients with a palpable varicocele and orchidopexy if undescended testes are diagnosed after adulthood, although management of these patients remains controversial. Advances in retrieving spermatozoa more efficiently by micro-TESE have been made during the past decade. In addition, recent advances in biotechnology have raised the possibility of using germ cells produced from stem cells in the future. This review presents current knowledge about the etiology, diagnosis, and treatment of NOA.     

Deoxyribonucleic acid polymerase activity in the testes of infertile men with varicocele

Activities of deoxyribonucleic acid (DNA) polymerase alpha, beta, and gamma were measured in extracts of testicular biopsy specimen obtained from 37 cases of male infertility with left varicocele and compared with those of 6 normal controls. It was observed that levels of DNA polymerase alpha, beta, and gamma were significantly lower in infertile men than normal controls on both sides of testes. Among three DNA polymerases, the level of DNA polymerase beta activity well correlated with the histological findings (Johnsen's score), i.e., the extent of differentiation of germinal cells. DNA polymerase beta activity appeared to be the lowest in the patients whose sperm density was less than 5 X 10(6)/ml. On the other hand, no correlation was apparent between levels of DNA polymerases and other clinical parameters, e.g., testicular volume, sperm motility, grade of varicocele, and serum hormone levels. These results suggest that the combined decrease in the DNA polymerase activities may be one of the factors that have deleterious effects on spermatogenesis in varicocele patients.     

大鼠溶脲脲原体感染与曲细精管微结石形成

４０例ＳＤ雄性大鼠膀胱内接种溶脲脲原体（Ｕｒｅａｐｌａｓｍａｕｒｅａｌｙｔｉｃｕｍ，Ｕ．Ｕ，１０５ＣＣＵ／ｍｌ）。从２５例接种动物的睾丸组织中培养出Ｕ．Ｕ。聚合酶链式反应（ＰＣＲ）扩增得到Ｕ．Ｕ．特异性ＤＮＡ片段（４６０ｂＰ）。４０例接种动物中，有１６例检出膀胱结石，化学分析表明，结石成份为磷酸镁铵（ＭｇＮＨ４ＰＯ４·６Ｈ２Ｏ），矿物学名称为鸟粪石（Ｓｔｒｕｖｉｔｅ）。光镜与电镜观察发现，其中有４例动物的睾丸多数曲细精管管腔内有微结石存在。有微结石处呈现生精细胞与支持细胞脱落或消失，有的曲细精管仅剩下界膜。睾丸间质中未发现结石形成。３０例对照组大鼠睾丸内未检出Ｕ．Ｕ，也未发现膀胱、睾丸内结石形成。提示Ｕ．Ｕ．可导致曲细精管微结石形成并干扰精子发生，因而是造成男性不育的因素之一。     

HIV in testis: quantitative histology and HIV localization in germ cells

The testes of AIDS patients invariably show decreased spermatogenesis and are atrophic. These testicular changes can be grouped into three categories: (1) spermatogenesis present, but decreased; (2) spermatogenic arrest at primary spermatocyte stage; and (3) Sertoli only (or almost Sertoli only). The purpose of this study is 2-fold: firstly, to quantitate the numbers and types of germ cells in these three groups as compared with normals. In addition the presence of HIV-1 DNA positive germ cells was quantitated by PCR in situ hybridization. HIV-1 was identified in 14 of 15 testes from HIV infected adults, and was present in 25-33% of residual germ cells. There was an average of 18.9 HIV infected germs cells/tubule in the spermatogenesis group, 6.3 HIV infected germs cells in the spermatogenic arrest group, and 0.25 HIV infected germ cells in the almost Sertoli only group. HIV-1 DNA was absent in three of the three preadolescent boys testes (HIV acquired in utero). This study quantitates the degree of germ cell loss in AIDS patients and quantitates the degree of HIV positivity of the residual germ cells, thus shedding more light on the testicular HIV burden, with its possible repercussions for sexual transmission of HIV.     

Sertoli cell maturation in men with azoospermia of different etiologies

OBJECTIVE: To evaluate the involvement of Sertoli cell in different spermatogenic disorders. DESIGN: Retrospective case-control study. SETTING: Teaching hospital. PATIENT(S): Azoospermic men who underwent testicular biopsy for sperm recovery in preparation for intracytoplasmic sperm injection. INTERVENTION(S): Testicular biopsy evaluation by quantitative immunohistochemistry for the immature Sertoli cell markers anti-Müllerian hormone and cytokeratin 18 (CK-18). MAIN OUTCOME MEASURE(S): Relative area of immature Sertoli cells in testes with focal spermatogenesis, spermatocyte maturation arrest, or normal spermatogenesis. RESULT(S): The relative area occupied by immature Sertoli cells, as revealed by anti-Müllerian hormone and CK-18 expression, was highest in the 11 men with focal spermatogenesis. In the group representing normal spermatogenesis (obstructive azoospermia, 6 men) and in the group characterized by spermatocyte maturation arrest (6 men), the areas occupied by anti-Müllerian hormone- and CK-18-positive cells were minimal. CONCLUSION(S): Different etiologies underlie the spermatogenic disorders reported in this study. In focal spermatogenesis with high anti-Müllerian hormone and CK-18 expression, the spermatogenic impairment is associated with the presence of immature Sertoli cells. The detection of normal mature Sertoli cells in the spermatocyte maturation arrest group indicates that the spermatogenic defect that is accompanied by an impairment of meiosis is intrinsic to the germ line without affecting Sertoli cell differentiation.     

Fertility preservation for boys with cancer

Childhood cancer is a curable disease due to the development of chemo- and radiation therapies, but long-term survivors suffer late side-effects including infertility. Cytotoxic agents and radiation impair spermatogenesis and cause oligospermia or azoospermia as well as genetic damage in sperm. To date, the only established option to preserve fertility is cryopreservation of sperm before treatment and artificial reproduction techniques, if men with cancer can ejaculate, but only a quarter of men have banked sperm. Lack of information is the most common reason for failing to bank sperm. However, prepubertal patients who have only spermatogonia and spermatocytes in their testes do not benefit from cryopreservation of their sperm and assisted reproductive techniques. Thus, the only available option is to harvest testicular tissues before treatment for cryopreservation, from which immature germ cells can somehow be maturated. Autotransplantation of germ cells into the testis holds promise for fertility restoration, but contamination by malignant cells may induce relapse. Fluorescence-activated cell sorting (FACS) with two surface markers could exclude contaminated leukemic cells from murine germ cells, and transplantation of sorted germ cells successfully restored fertility without transmission of leukemia. Human germ cells could be also isolated from human leukemia and lymphoma cell lines by FACS using surface markers. Before autotransplantation can be applied clinically, some issues, including the risk of contamination by malignant cells and in vitro propagation of spermatogonial stem cells, should be resolved.     

Effect of protopanaxatriol saponin on spermatogenic stem cell survival in busulfan-treated male mice

Background and aims:  Panax ginseng C.A. Meyer is a medicinal herb widely used in Asian countries. Many of its pharmacological actions are attributed to ginsenosides (saponin). However, the pharmacological effects or functions of ginsenosides on mammalian spermatogenesis are unclear.
Methods:  In the present study study, we investigated the therapeutic and prophylactic effects of protopanaxatriol saponin (PT) on testicular organ weight and morphology, testicular germ cells, proliferation, differentiation and spermatogenesis after induction of toxicity by a chemotherapeutic agent, busulfan, in male mice.
Results:  Intraperitoneally (IP) busulfan treatment markedly decreased the organ weight of testis, caput and cauda epididymis. After the treatment, the testes had collapsed seminiferous tubules with incomplete spermatogenesis. However, a single dose of busulfan treatment followed by PT injection showed milder damage on seminiferous tubules than busulfan alone.
Conclusion:  These results suggest that PT is effective in recovery of the male reproductive organ, and induced an increase in the number and viability of germ cells overcoming busulfan toxicity. PT might have applications in the recovery of male infertility arising from azoospermia and oligospermia.     

Testicular mast cell heterogeneity in idiopathic male infertility.

OBJECTIVE: To determine if the mast cell subclass changes in diseased testes. DESIGN: Retrospective. SETTING: University hospital urology clinic and anatomy laboratory. PATIENTS, PARTICIPANTS: Fourteen normal men and 50 patients with idiopathic male infertility. INTERVENTIONS: Histochemical techniques to identify proteoglycans of mast cells were applied to the obtained testicular biopsy specimens. MAIN OUTCOME MEASURES: The numbers of the heparin containing mast cell and the chondroitin sulfate containing mast cell were measured with light microscopy. RESULTS: The total number of mast cells and the ratio of chondoroitin sulfate containing mast cells were significantly increased in the testes from patients with idiopathic male infertility. CONCLUSION: These results suggest that mast cells may play a certain role in the etiology of idiopathic male infertility.     

睾丸的生殖病理学研究(Ⅰ)——40例无精症与少精症患者睾丸的生殖病理观察

本文提出了睾丸生殖病理双重诊断法,即同时作出睾丸病理诊断及睾丸生精障碍分级诊断,对各种病理类型的主要特征作了介绍。另又分析了睾丸各组织成分的病理变化发生率及病理生理意义。