Correlation between testicular biopsies (prepubertal and postpubertal) and spermiogram in cryptorchid men

Twenty-one young men who underwent testicular biopsy and orchidopexy in infancy consulted owing to infertility and had biopsies again. The first and second biopsy specimens from these patients were compared by means of a semiquantitative study of the seminiferous tubules to evaluate the evolution of germ cells and to correlate these data with spermatozoon numbers. The infant testes showing lesions were classified into 3 types according to the mean tubular diameter and tubular fertility index: (1) slight lesions, (2) marked germinal hypoplasia, and (3) severe germinal hypoplasia. In the adult testes, spermatogenesis was evaluated by calculating the average numbers of spermatogonia, primary spermatocytes, young spermatids, and mature spermatids. These testes were classified as (1) normal; (2) having lesions in the adluminal compartment; (3) having lesions in the basal compartment; and (4) mixed atrophy. The number of differentiated spermatids was correlated with the expected number of spermatozoa in the ejaculate by a power regression curve. The observation of certain histologic lesions in the seminiferous tubules was assumed to indicate excretory duct obstruction: ectasia, indented outline of the seminiferous epithelium, intratesticular spermatocele, apical cytoplasmic vacuolation of Sertoli cells, and mosaic distribution of testicular lesions. There was a correlation between the prepubertal lesions and the degree of spermatogenesis in postpubertal biopsy specimens. The evolution of the 40 testes without regard to their location in infancy (cryptorchid or scrotal) was as follows. The 14 infant testes with a normal histologic pattern (5 testes) or minor lesions (9 testes) evolved to testes with lesions of the adluminal compartment (8 testes), mixed atrophy (4 testes), or lesions of the basal and adluminal compartments (2 testes). The 6 testes with marked germinal hypoplasia evolved to testes with mixed atrophy. The 20 testes with severe germinal hypoplasia evolved to testes with mixed atrophy (17 testes), Sertoli-cell-only tubules (2 testes), or lesions in the basal compartment (1 testis). In the 9 patients with a histologic pattern of obstruction bilaterally (6 men) or unilaterally (3 men), the expected number of spermatozoa according to the correlation curve was much higher than the actual number in the spermiogram. This means that the testes of many azoospermic men produce spermatozoa, and this finding corroborates the importance of testicular biopsy in infertility studies.     

Anti-MIC2 as a tool in examination of testicular biopsies.

MIC2 is a pseudoautosomal gene localized on X and Y chromosomes. The MIC2 gene product is a glycoprotein expressed on the cell membranes of a number of somatic cells, including Sertoli cells of the testis, but not on the cell membranes of germ cells. In cases of cryptorchidism, a testicular biopsy is recommended in order to evaluate future fertility potential. The spermatogonia are identified on histological sections and the number per tubular transverse section is compared with normal values for age. The patient is at 33-100% risk of subsequent infertility when the number of spermatogonia per tubular transverse section is lower than 1% of the lowest normal age-matched value. Besides Sertoli cells the seminiferous tubules in undescended testes contain only a few germ cells, and it may be difficult to pinpoint the germ cells in small biopsies. Especially in nonpalpable testes their number may be heavily reduced. A reliable identification of germ cells may also be difficult in cultures of testicular biopsies from undescended testes. Against this background, we tried the use of an immunohistochemical method with DAKO antibody to the MIC2 gene product (MIC2, 12 E7, code no. M3601) in order to obtain a "negative reaction" of germ cells, contrasting with the stained Sertoli cells. The material comprised: 44 specimens of testicular parenchyma taken at time of surgery for cryptorchidism from 24 cryptorchid boys with nonpalpable testes and 14 testicular biopsies from 13 cryptorchid patients with palpable testes which had been cultured in vitro for 7, 14 or 21 days. In all cases the immunohistochemical method with DAKO antibody to the MIC2 gene product was helpful for identification of Sertoli cells and germ cells, and we therefore recommend the use of anti-MIC2 in all testicular biopsies where it is difficult to pinpoint the germ cells.     

Macro-orchidism: light and electron microscopic study of four cases.

A hormonal and quantitative light microscopy study of one man with macro-orchidism associated with mental retardation and fragile X chromosome (case no. 1) and three men with idiopathic macro-orchidism (cases no. 2 to 4) is reported. Hormonal study revealed slightly increased follicle-stimulating hormone serum levels in cases no. 1 to 3. The testes from cases no. 1 (orchidoepididymoectomy specimen) and 2 (testicular biopsy) presented interstitial edema and three different tubular patterns that were arranged in a mosaic-like manner. Type I tubules had an increased diameter (less than 220 microns), dilated lumen, and thin seminiferous epithelium usually consisting of Sertoli cells, spermatogonia, primary spermatocytes, and sometimes a few spermatids. Type II tubules had a normal diameter (180 to 220 microns) and germ cell development varied between complete spermatogenesis and Sertoli-cell-only tubules. Type III tubules had decreased diameter (less than 180 microns), atrophic seminiferous epithelium, and thickened tunica propria. The appearance of the nuclei of the Sertoli cells in the three types of tubules could be either mature or immature. Some of the mature Sertoli cells presented a granular cytoplasm. A few of these granular cells grouped together, forming nests that protruded into the tubular lumen. The testicular biopsies from cases no. 3 and 4 only presented type II tubules that contained both mature and immature Sertoli cells. Quantitative study revealed that the large testicular size was principally due to an increased tubular length in all four cases. Although the seminiferous tubule lesions and interstitial edema suggest an obstructive process, the testicular excretory ducts (studied in case no. 1) appeared normal or only slightly dilated. It is possible that the seminiferous tubule lesions (dilated lumen and germ cell depletion) might be secondary to the Sertoli cell lesions (granular cytoplasm and nuclear immature-like pattern.     

Focal orchitis in undescended testes: discussion of pathogenetic mechanisms of tubular atrophy.

OBJECTIVE: To evaluate seminiferous epithelium lesions in adult cryptorchid testes showing lymphoid infiltrates in seminiferous tubules and interstitium (i.e., focal orchitis). Also, to consider the possible role of this lesion in the etiology of tubular atrophy. METHODS: We performed a histopathologic study of the cryptorchid testes and adjacent epididymides removed from 50 adult men who had not been previously treated for cryptorchidism. The study included morphologic and semiquantitative evaluation of seminiferous tubule pathology (according to germ cell numbers), Sertoli cell morphology, tubular lumen dilation, rete testis pattern (normal, hypoplastic, or cystic), and epididymal pattern (normal or epididymal duct hypoplasia). The study also included immunohistochemical evaluation of immune cell markers. The results were compared with clinical and laboratory findings. RESULTS: Focal lymphoid infiltrates (mainly lymphocytes) in seminiferous tubules and interstitium were found in 22 patients (44%), all of whom had unilateral cryptorchidism. The course of orchitis was asymptomatic, and laboratory data were normal. According to the seminiferous tubule pathology, a variety of histopathologic diagnoses, were made: (1) mixed atrophy consisting of Sertoli cell-only tubules intermingled with tubules showing maturation arrest of spermatogonia (11 testes, 4 of which also showed hyalinized tubules); (2) Sertoli cell-only tubules plus hyalinized tubules (4 testes); (3) Sertoli cell-only tubules (3 testes); (4) intratubular germ cell neoplasia (2 testes, 1 of which also showed hyalinized tubules); (5) complete tubular hyalinization (1 testis); and (6) tubular hyalinization plus some groups of tubules with hypospermatogenesis (all germ cell types were present although in lower numbers, 1 testis). Dysgenetic Sertoli cells, that is, Sertoli cells that had undergone anomalous, incomplete maturation, were observed in all nonhyalinized seminiferous tubules with inflammatory infiltrates. Tubular ectasia was observed in 13 cases. The rete testis was hypoplastic and showed cystic transformation in 18 testes, and the epididymis was hypoplastic in 15 testes. CONCLUSIONS: The causes of these focal inflammatory infiltrates are unknown. It is possible that tubular ectasia and Sertoli cell dysgenesis are involved and that these alterations cause a disruption of the blood-testis barrier and allow antigens to enter the testicular interstitium, giving rise to an autoimmune process.     

The adolescent varicocele. A histopathologic study of 13 testicular biopsies

Testicular varicocele, the most common cause of male infertility, frequently presents in early adolescence. To determine whether testicular damage occurs early in the natural history of varicocele, testicular biopsy specimens from 13 patients, 13 to 18 years of age (mean age, 15.5 years), were studied. The biopsies were compared with testicular tissue from six normal control subjects 15 to 28 years of age (mean age, 23.2 years). Nine of the patients with varicoceles (69.2%) demonstrated some degree of tubular sclerosis. Ultrastructural study demonstrated that the tubular sclerosis was due to collagen deposition by fibromyocytes in the peritubular sheath. Premature germ cell sloughing was present in greater than 50% of tubules examined in all but one biopsy. Six patients (46%) demonstrated small vessel sclerosis. Quantitation of the germinal epithelium revealed that the mean germ cell/Sertoli cell ratio and the percentage of germ cells present as late stage forms (secondary spermatocytes, spermatids and spermatozoa) were significantly reduced in the varicocele group. The testes of two patients exhibited severe hypospermatogenesis approaching germ cell aplasia. None of these changes were seen in the control group. The authors conclude that pathologic changes in the testes of patients with varicoceles are found at or soon after puberty. The histopathologic features include peritubular sclerosis, small vessel sclerosis, premature germ cell sloughing, and variable degrees of hypospermatogenesis.     

Time response of rat testicular alterations induced by cryptorchidism and orchiopexy

Cryptorchidism is one of the main risk factors for infertility and testicular cancer. Orchiopexy surgery corrects cryptorchidism effects. Different models of cryptorchidism developed in the rat include surgery. We assessed testicular alterations in rats submitted to surgical cryptorchidism and examined their potential for reversibility at different time points in order to verify time dependency effect(s) on the recovery of the undescended testes. Cryptorchidism was induced in 3‐week‐old rats. Animals were euthanized 3, 6 or 11 weeks after surgery to evaluate the morphological progression of cryptorchidism‐induced germinative epithelial alterations. Other groups underwent orchiopexy 3, 5 or 9 weeks after surgical cryptorchidism, before or after puberty. Animals were euthanized 3 or 8 weeks after orchiopexy. Controls underwent sham surgery at the same time points as the surgical groups. Cryptorchid testes showed decreased weight, germinative epithelial degeneration, apoptosis and vacuolation, corresponding to impairment of spermatogenesis and of Sertoli cells. Some tubules has a Sertoli cell‐only pattern and atrophy. The intensity of damage was related to the duration of cryptorchidism. After orchiopexy, spermatogenesis completely recovered only when testicular relocation occurred before puberty and the interval for recovery was extended. These results indicate that age, sexual maturity and extension of germ cell damage were relevant for producing germ cell restoration and normal spermatogenesis. We provide original observations on the time dependency of testicular alterations induced by cryptorchidism and their restoration using morphologic, morphometric and immunohistochemical approaches. It may be useful to study germ cell impairment, progression and recovery in different experimental settings, including exposure to exogenous chemicals.     

Sertoli cell nodules in the undescended testis: a histochemical, immunohistochemical, and ultrastructural study of hyaline deposits

AIMS: To document the morphology, immunohistochemical staining properties, and ultrastructural features of hyaline material in Sertoli cell nodules of undescended testis and contrast them with those of sex cord tumour with annular tubules (SCTAT), which is histologically similar. To highlight the need to distinguish these nodules from other Sertoli cell hyperplasias, such as intratubular Sertoli cell proliferations, which occur in specific clinical contexts. MATERIALS/METHODS: A retrospective study of 46 orchidectomy specimens from cryptorchid testes, 27 of which contained Sertoli cell nodules. Special histochemical stains, immunohistochemical stains for type IV collagen and fibronectin, and ultrastructural examination of the hyaline material were performed using tissue from paraffin wax embedded tissue blocks. RESULTS: The hyaline deposits in SCTAT and Sertoli cell nodules had similar staining patterns-periodic acid Schiff (PAS) and PAS-diastase positivity with variable staining of Martius scarlet blue and Masson trichrome. Type IV collagen immunoreactivity was seen in hyaline areas, although fibronectin was negative. Electron microscopy of hyaline areas confirmed a compact matrix identical to components of the basement membrane in the adjacent seminiferous tubules. CONCLUSION: This study describes an unusual form of Sertoli cell proliferation in undescended testes, which must be distinguished from Sertoli cell tumours and other forms of proliferation. In addition, the hyaline material within Sertoli cell nodules in the cryptorchid testis is histochemically, immunohistochemically, and ultrastructurally consistent with both matrix and fibrous components of seminiferous tubule basement membranes. Increased production of basement membrane material, with subsequent invagination into tubules, is the most likely origin of this material.     

The morphogenesis of cyclohexylamine-induced testicular atrophy in the rat: in vivo and in vitro studies

Male Wistar strain rats were fed a diet providing an intake of 0 or 400 mg cyclohexylamine (CHA)/kg body weight/day for 1, 3, 7, 9, or 13 weeks. At the end of the appropriate feeding period the rats were perfused-fixed with Karnovsky's fixative. The weights of the fixed testes were recorded and the testes, epididymides, and spermatic cord were sampled and processed into methacrylate resin. Histopathological examination of the testes showed changes after 3 weeks of CHA administration. The most frequent and consistent lesion consisted of a focal, basal vacuolation of the Sertoli cell cytoplasm associated with the local loss of spermatocytes and spermatogonia. After a 7-week administration, the Sertoli cell vacuolation was extensive, while the germ cell population showed mild to moderate degeneration and depletion. After longer periods of treatment the lesion was more severe and affected a greater number of tubules leading to general disruption of the germinal epithelium. Cocultures of Sertoli and germ cells were prepared from the testes of Wistar strain rats and exposed to (CHA) or its metabolite 4-aminocyclohexanol (4ACH) at concentrations ranging from 0.1 to 10 mM for periods of 24-72 hr. The cultures were fixed, stained, and examined by light microscopy. Cultures exposed to CHA or 4ACH showed morphological changes comparable with those seen in vivo. Sertoli cell vacuolation was the earliest change with progressive germ cell degeneration and exfoliation from the Sertoli cell monolayer. At equimolar concentrations, CHA produced more marked changes than 4ACH. These results suggest that CHA itself acts directly on the testis and that its primary cellular target is the Sertoli cell.     

Atypical development of Sertoli cells and impairment of spermatogenesis in the hypogonadal (hpg) mouse

Testes of hypogonadal (hpg) mice show arrested postnatal development due to congenital deficiencies of gonadotrophin-releasing hormone (GnRH) and gonadotrophin synthesis and secretion. Follicle-stimulating hormone (FSH), androgen or oestrogen treatment restore qualitatively normal spermatogenesis in hpg testes. Understanding the cellular and molecular changes accompanying hormone-induced spermatogenesis in hpg mice requires detailed morphological analyses of the germ cells and Sertoli cells in the untreated hpg testis. We compared seminiferous epithelial cytology in adult hpg, immature and adult wild-type mice using unbiased optical disector-based stereology, immunolocalization of Sertoli cell microtubules (MT), espin (a component of the blood-testis barrier), markers of Sertoli cell maturity (p27(kip1) and WT-1), and electron microscopy. Hpg testes had marked reductions in weight, seminiferous cord volume and length, and severe spermatogenic impairment with germ cells per testis < 1% of adult wild-type testes. Sertoli cell nuclei expressed WT-1 in hpg testes, but often were centrally located, similar to 9-14-day-old wild-type testes, and they expressed p27(kip1), indicating that hpg Sertoli cells were post-mitotic. Hpg testes had significantly (P < 0.05) reduced Sertoli cells per testis (0.56 million) compared with 10-day wild-type (1.15 million) and adult wild-type testes (2.06 million). Immunofluorescence labelling of normal adult Sertoli cells showed supranuclear MT columns and basally located espin, but these features were absent in 10-day-old and hpg Sertoli cells. Hpg Sertoli cells showed pleomorphic nuclear ultrastructure with mature-type nucleoli, similar to normal adult-type Sertoli cells, but hpg Sertoli cells exhibited incomplete tight junctions that lacked ectoplasmic specializations. We conclude that in hpg mice, chronic gonadotrophin insufficiency restrains Sertoli cell proliferation and maturation, forming pseudo-adult-type Sertoli cells that are incapable of supporting germ cell proliferation and maturation.     

Spontaneous testicular lesions in purpose-bred beagle dogs

Spontaneous testicular lesions were assessed in 50 control purpose-bred male beagle dogs. They were selected from 13 toxicology studies conducted over the period 1988-1999. Age of the dogs at study termination varied from 8-20 months with an average age of 13 months. Regardless of age, the most significant finding was bilateral segmental hypospermatogenesis in 15/50 (30%) of the dogs. Cross sections of tubules with hypospermatogenesis were distributed randomly throughout the testes and were characterized by reduced proportions of germ cells, tubular shrinkage, and Sertoli cell prominence. These changes were occasionally associated with giant cells, with cellular debris, and in 6/15 (40%) with atrophic tubules devoid of germ cells, indicating a degenerative process. Focal subcapsular tubular atrophy or hypoplasia (tubules lined by Sertoli cells only) was also found in 9/35 (26%) of dogs without hypospermatogenesis. Inhibited spermiation with retention of mature sperm in tubules was seen in 6/50 dogs, 3 of which also showed hypospermatogenesis. Other findings of high incidence but low prevalence included tubules with multinucleated giant cells, swollen spermatocytes, or apoptotic germ cells. These latter changes are probably a constituent of normal spermatogenesis. In conclusion, about 30% of control beagle dogs show segmental hypospermatogenesis, which may be associated with degenerative changes, and an additional 18% of the dogs exhibit focal tubular atrophy/hypoplasia in otherwise normal testes. These changes have to be distinguished from compound-related toxic effects.     

Small intestine biopsies in children with malabsorption syndrome. Autoradiographic studies with 3H-thymidine

Bioptic material of the small intestine is suitable for short-time in-vitro incubation in 3H-thymidine labelled medium followed by autoradiography. Histological and autoradiographic investigations of the small intestine of 45 children yielded the following characteristic findings: 1. 93 per cent of the healthy controls (n = 14) showed normal mucous membrane (Type I according to Shmerling) and a mean 3H-thymidine labelling index of 13 per cent. 2. Normal histological findings but a mean autoradiographic labelling index of 22 per cent were recorded from 87 per cent of children with cow's milk or fructose incompatibility (n = 12). 3. Mucous membrane of Shmerling Type I and a labelling index of enterocytes of 32 per cent was recorded from 50 per cent of children with coeliac disease (n = 19) after diet periods of seven or three to five months. Other children with coeliac disease but without therapy up to that time showed in 80 per cent the Shmerlings Type III and in 20 per cent Type II accompanied by the highest labelling index of 35 per cent. The increase of proliferative activity of enterocytes in coeliac disease shown by autoradiography and histological observations (villous atrophy) suggested an increased rate of cell death which led to the need for high cell replacement. This autoradiographic method used complementarily to usual histological diagnosis, has proved to help saving an enormous amount of time in the diagnosis of coeliac disease.     

Distinctive enchondromatosis with spine abnormality, regressive lesions, short stature, and coxa vara: importance of long-term follow-up.

We report a girl with a unique type of enchondromatosis observed from birth to puberty. Radiographic abnormalities documented at the age of 14 months included distinctive spondylometaphyseal enchondromatous types of lesions with minimal involvement of the short tubular and flat bones. Follow-up radiographic examinations documented progressive coxa vara and hypoplasia/dysplasia of the left ulna. At puberty, the short tubular bones appeared normal. There was marked regression of the flat bone, rib, and spinal lesions. This case shows the importance of long-term observation of unclassified forms of skeletal dysplasia.     

Distinctive enchondromatosis with spine abnormality,regressive lesions,short stature,and coxa vara: Importance of long‐term follow‐up

We report a girl with a unique type of enchondromatosis observed from birth to puberty. Radiographic abnormalities documented at the age of 14 months included distinctive spondylometaphyseal enchondromatous types of lesions with minimal involvement of the short tubular and flat bones. Follow‐up radiographic examinations documented progressive coxa vara and hypoplasia/dysplasia of the left ulna. At puberty, the short tubular bones appeared normal. There was marked regression of the flat bone, rib, and spinal lesions. This case shows the importance of long‐term observation of unclassified forms of skeletal dysplasia. © 2001 Wiley‐Liss, Inc.     

Histometric investigations of the effect of cyclosporin A on the testicular tissue of rats

The effect of cyclosporin A (CsA) on rat testes was assessed in Lewis rats which were given 10 mg/kg of CsA im per day. The rats were divided into 13 subgroups, and 1 subgroup was sacrificed each month. The rat testes were measured and examined for histometrical and morphological changes in comparison to controls. Histometrical analysis included testicular cross-sectional surface area, tubular density, tubular diameter, and the amounts of testicular germinal epithelium, lumen, and interstitial tissue. In the parameters examined, there were no overall differences between CsA-treated animals and controls. CsA does not affect rat testicular tissue.     

Effects of oral administration of Euphorbia prostrata extract on the reproductive system of male albino rats: a histometric and biochemical study

Methanolic extract of Euphorbia prostrata when administered orally at the dose level of 400?mg/kg body weight per day for 60?days to male rats did not cause any significant loss in their body weights, but the weights of their reproductive organs, i.e., testes, epididymides, seminal vesicle, and ventral prostate, decreased in a significant manner when compared to the controls. The production of round spermatids was reduced by 73.04% in E. prostrata-treated rats. The population of preleptotene spermatocytes and spermatogonia were reduced by 65.07% and 47.48% and secondary spermatocytes by 73.41%, respectively. The cross-sectional surface area of Sertoli cells, as well as the cell counts, was found to have been depleted significantly. Leydig cell nuclear area and the number of mature Leydig cells decreased by 57.47% and 54.42%, respectively. Sperm motility, as well as sperm density, reduced significantly. E. prostrata reduced the fertility of male rats by 100%. The protein, glycogen, and cholesterol contents of the testes, fructose in the seminal vesicle, and protein in the epididymides significantly decreased. Histoarchitecture of the testes showed vacuolization at the primary spermatocyte stage. Highly reduced seminiferous tubular diameter and increased intertubular space were also observed when compared to the controls.     

Partial obstruction of the seminal path, a frequent cause of oligozoospermia in men

The objective of this clinical study was to determine the real frequency and clinical importance of partial obstruction of the seminal path in patients with oligozoospermia. We have designed a prospective clinical study including men with oligozoospermia seen at an andrological consultation in both private and institutional hospitals. A testicular biopsy was done on all patients under local anaesthesia. A complete study for sterility was also done [hormonal determinations: follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, testicular ultrasound, semen analysis, testicular Doppler ultrasound, etc.]. We have made a quantitative and qualitative evaluation of testicular biopsy (percentage of tubules with Sertoli cell only or with hyalinization; mean tubular diameter; number of spermatogonia, primary spermatocytes, young spermatids, mature spermatids and Sertoli cells; and evaluation of testicular interstitium: number of Leydig cell clusters, presence of angiectasis, presence of perivascular inflammation). Sixty one per cent of all oligozoospermia cases were obstructive. The principal cause of obstructive oligozoospermia was the presence of testicular varicocele. In obstructive oligozoospermia, the tubular diameter and number of mature spermatids are statistically significantly higher than in non- obstructive oligozoospermia. Obstructive oligozoospermia is a frequent condition caused by partial obstruction of seminal path. A quantitative analysis of the testicular biopsy is the only method of diagnosis.      

Androgen insensitivity syndrome: an immunohistochemical, ultrastructural, and morphometric study

OBJECTIVE: To evaluate the morphometric, immunohistochemical, and ultrastructural lesions of the testes in prepubertal and adult patients with androgen insensitivity syndrome. METHODS: We examined the testicular biopsy using immunohistochemistry for vimentin, smooth muscle actin, and collagen IV antigens. Quantification of seminiferous tubules and testicular interstitium was performed in prepubertal and adult patients with androgen insensitivity syndrome and results were compared with normal testes from both infants and adults. RESULTS: The adult testes presented nodular and diffuse lesions that consisted of Sertoli-cell-only seminiferous tubules. Two types of Sertoli cells could be distinguished, namely, immature vimentin-positive Sertoli cells and nearly mature Sertoli cells. In the nodules, the lamina propria was thin and contained a scant number of actin-positive peritubular cells. Leydig cells were hyperplastic. The prepubertal patients showed only diffuse lesions characterized by Sertoli cell hyperplasia, decreased germ cell numbers, and a discontinuous immunoreaction to collagen IV. CONCLUSIONS: The testicular lesions in androgen insensitivity syndrome are probably caused by primary alterations that begin during gestation. These lesions become progressively more pronounced at puberty, when the nodular lesion pattern (adenomas) is completely developed.     

Neonatal exposure of male rats to estradiol benzoate causes rete testis dilation and backflow impairment of spermatogenesis

Estrogens administered to perinatal rodents cause spermatogenesis impairment; this study was undertaken to determine the mechanisms by which estrogens exert this effect. Neonatal male Wistar rats received estradiol benzoate (either 0.5 mg/5g BW or 1 mg/5g BW) and were killed at days 10, 22, 33, 45, and 60. Controls received vehicle. In tubule cross-sections of transverse sections of the right testes, 1) tubular diameter (TD) and seminiferous epithelium height (SEH) were measured, 2) normal and impaired spermatogenesis were classified in terms of the most advanced germ cell type present, including tubules lined by Sertoli cells only. A significant dose-dependent rise in the tubule percentage lined by Sertoli cells only at day 60 reflected spermatogenesis impairment. This was evidenced by the presence of multinucleated germ cells in a thin epithelium and sloughed into an enlarged tubular lumen, which was reflected in a significant dose-dependent increase in TD/SEH values from day 22 onward. TD was significantly greater and SEH significantly lower in tubular segments located at the cranial than the caudal halves of rat testes treated with the high (days 22, 33, and 60) and the low dose (day 33). This indicated distension in cranial tubular segments, perhaps due to the fact that these segments were the closest to the dilated rete testis. Consequently, they showed the highest TD/SEH values and the most regressive features of spermatogenesis (tubules lined by Sertoli cells only). In contrast, caudal segments in rat testes treated with the low dose showing TD/SEH values similar to controls displayed a delayed maturation of spermatogenesis coinciding with the late appearance of mature Leydig cells. Anat. Rec. 252:17–33, 1998. © 1998 Wiley-Liss, Inc.     

Infant feeding with soy formula milk: effects on puberty progression, reproductive function and testicular cell numbers in marmoset monkeys in adulthood

BACKGROUND: This marmoset study addresses concerns about feeding human male infants with soy formula milk (SFM). METHODS: From age 4 to 5 days, seven male co-twin sets were fed standard formula milk (SMA) or SFM for 5-6 weeks; blood samples were subsequently collected at 10-week intervals. Testes from co-twins killed at 120-138 weeks were fixed for cell counts. RESULTS: SFM- and SMA-fed twins showed normal weight gain; puberty started and progressed normally, based on blood testosterone measurements. Body weight, organ weights (prostate, seminal vesicles, pituitary, thymus and spleen) and penis length were comparable in co-twins. All SMA- and 6/7 SFM-fed males were fertile. Unexpectedly, testis weight (P = 0.041), Sertoli (P = 0.025) and Leydig cell (P = 0.026) numbers per testis were consistently increased in SFM-fed co-twins; the increase in Leydig cell numbers was most marked in males with consistently low-normal testosterone levels. Seminiferous epithelium volume per tubule showed a less consistent, non-significant increase in SFM-fed males; raised germ cell numbers per testis, probably due to increased Sertoli cells, conceivably resulted in larger testes. Average lumen size, although greater in SFM-fed group, was inconsistent between co-twins and the difference was not significant. CONCLUSIONS: Infant feeding with SFM has no gross adverse reproductive effects in male marmosets, though it alters testis size and cell composition, and there is consistent, if indirect, evidence for possible 'compensated Leydig cell failure'. Similar and perhaps larger changes likely occur in adult men who were fed SFM as infants.     

A morphological and histochemical study of sertoli cells in normal and XX sex-reversed mice

Morphological and histochemical studies on the testes of mice with XX sex-reversal demonstrates several profound differences in the ultracytology and oxidative enzyme activity of Sertoli cells as compared to the normal. Membranous structures, cytoplasmic inclusions, and certain dehydrogenases in Sertoli cells of normal mice are described in relation to a possible role in steroid synthesis. In XX sex-reversed mice, two types of Sertoli cells are observed in the seminiferous tubules which are devoid of germ cells. One type is the randomly dispersed cells, which reveal the following ultrastructural alterations when compared with the normal cells: large spherical nuclei with prominent nucleoli, reduced smooth endoplasmic reticulum, pleomorphic mitochondria with no apparent cristae, and decreased lipid droplets. The other type is clustered cells which appear similar to immature Sertoli cells. These cells have small nuclei with poorly developed nucleoli, small profiles of rough endoplasmic reticulum, pleomorphic mitochondria, decreased lipid droplets, and an absence of specialized inter-Sertoli cell junctions. The results of this study suggest that functional and ultrastructural alterations of Sertoli cells in XX sex-reversed mice may result from degeneration or absence of germinal cells, and that steroidogenesis by Sertoli cells in mice may be associated with the development of germinal cells. It is also suggested that Sertoli cells may secrete a steroid which controls follicle-stimulating hormone secretion.