Sertoli-stromal cell tumor of the ovary: immunohistochemical, ultrastructural, and genetic studies

The Sertoli-stromal cell tumor (SSCT) of the ovary shows a histologic resemblance to developing or adult testes and is often associated with virilization caused by tumor-produced androgenic hormone. In spite of the unique manifestation of SSCT, detailed characteristics of this tumor are still obscure. The mechanism by which SSCT occurs has not yet been determined. Six SSCTs were studied immunohistochemically, ultrastructurally, and by polymerase chain reaction (PCR) for the presence of sex-determining region Y (SRY) gene and the X chromosome activation state. Immunohistochemically, Sertoli-like cells of SSCT were positive not only for alpha-inhibin but also low-molecular-weight cytokeratin. In control testes, the expression of alpha-inhibin and cytokeratin was limited to a Sertoli cell component and rete testis, respectively. Ultrastructurally, tumor cells composing hollow tubules had an elongated nucleus with deep indentation and annulate lamellae, which are characteristic structures of mature Sertoli cells. In addition, they had studded microvilli on the apical surface and frequent desmosomes, which are structures noted in the cells of rete testis. Histologically, tumor cells of hollow tubules sometimes pouted into the lumen, as did the cells of tubulae rete, entrance into rete testis from seminiferous tubules. All of these findings indicate that some tumor cells of a SSCT show simultaneous differentiation into both Sertoli cells and cells of rete testis. SRY gene was not detected in any cases, and the X chromosome activation pattern was the same as that of the female control.     

Annulate lamellae of the jejunal absorptive cells in 21-day-starved rats

The ultrastructure of annulate lamellae of the jejunal absorptive cells in control and 21 d starved rats was investigated. Annulate lamellae were only rarely encountered in the jejunal absorptive cells of control rats, and then frequently in small stacks continuous with the rough-surfaced endoplasmic reticulum. In contrast, there was a relatively frequent incidence of annulate lamellae in the jejunal absorptive cells of 21 d starved rats, and larger stacks of annulate lamellae were also observed in spite of marked ultrastructural changes of these cells. The annulate lamellae were also continuous with the rough-surfaced endoplasmic reticulum, which was degenerating. The degenerative process of the absorptive cells following starvation might be related to the origin and function of the annulate lamellae.     

The influence of vasovasostomy on testicular alterations after vasectomy in Lewis rats

The occurrence of alterations in testicular weight and morphology after vasectomy and vasectomy reversal by vasovasostomy was studied in Lewis rats. Animals were studied 3, 4, and 7 months after bilateral vasectomy or a vasectomy followed 3 months later by vasovasostomy. Other rats served as sham-operated controls. The weights of the testes in vasectomy and vasovasostomy animals fell into two groups-small testes weighing less than 0.88 g and normal-sized testes of 1.2 g or more. When the extent of testicular alterations was estimated in sections for light microscopy by use of a semiquantitative testicular biopsy score count (TBSC), the morphology of the testes corresponded closely to the testis weight (r = .94), small testes having correspondingly low TBSC scores. In severely altered small testes, the seminiferous tubules were narrower than in sham-operated rats, and numbers of germ cells were greatly depleted. Many tubules contained only Sertoli cells and spermatogonia, although spermatocytes were present in a minority of tubules. A few seminiferous tubules contained multinucleate spermatids. Electron microscopy of severely altered tubules revealed closely apposed processes of Sertoli cells, which contained filaments, microtubules, and endoplasmic reticulum. In contrast, testes with normal weight in vasectomy and vasovasostomy groups resembled those of the sham-operated animals. Comparison of distributions of testicular biopsy score counts demonstrated differences between vasectomy and vasovasostomy groups as time after operation increased. At the 3-4-month intervals, approximately one-third of the testes were severely altered in both vasectomy and vasovasostomy groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Fine structure of Sertoli and Leydig cells in azoospermic human testis.

Testicular biopsy specimens taken from seven patients with azoospermia were studied by electron microscopy. We considered the possibility of a relationship between the annulate lamellae in the Sertoli cells and azoospermia. In the Leydig cells, we observed intranuclear and intracytoplasmic paracrystalline inclusions and considered their relationship with the Reinke crystalloid.     

Maxillary Adenoid Cystic Carcinoma with Annulate Lamellae

Of 29 reviewed ultrastructural studies of adenoid cystic carcinoma, only 1 mentions the presence of annulate lamellae. The present report describes a 65-year-old woman who presented with an adenoid cystic carcinoma in her left maxillary sinus. At the light microscopic level, most of the connective tissue was loose and myxomatous and contained fine strands, cords, and islands of epithelium that in places had a cribriform pattern. The ultrastructural appearance was similar to that described by others. Two epithelial cell types predominated. The cytoplasm of one type contained mainly ribosomes, and that of the other type also had peripheral microfilaments. Also present were pseudocysts, lumina with microvilli, and replicated basal lamina. In addition, a number of both types of cells contained parallel arrays of cisternae that had regular small pores. These were annulate lamellae.     

Human 'testicular dysgenesis syndrome': a possible model using in-utero exposure of the rat to dibutyl phthalate

BACKGROUND: The disorders comprising human 'testicular dysgenesis syndrome' (TDS) may be increasing in incidence. TDS originates in fetal life but the mechanisms are not known, and discerning them requires an animal model. METHODS AND RESULTS: The study investigated whether male rats exposed in utero to dibutyl phthalate [DBP; 500 mg/kg on gestational days (GD) 13-21] would provide a suitable model for human TDS. DBP induced a high rate (>60%) of cryptorchidism (mainly unilateral), hypospadias, infertility and testis abnormalities, similar to those in human TDS. Cell-specific immunohistochemistry and confocal microscopy were used to track development of Sertoli [anti-Müllerian hormone (AMH), Wilm's tumour (WT-1) protein, p27(kip)], Leydig [3beta-hydroxysteroid dehydrogenase (3beta-HSD)], germ (DAZL protein) and peritubular myoid (smooth muscle actin) cells from fetal life to adulthood. In scrotal and cryptorchid testes of DBP-exposed males, areas of focal dysgenesis were found that contained Sertoli and Leydig cells, and gonocytes and partially formed testicular cords; these dysgenetic areas were associated with Leydig cell hyperplasia at all ages. Suppression ( approximately 90%) of testicular testosterone levels on GD 19 in DBP-exposed males, coincident with delayed peritubular myoid cell differentiation, may have contributed to the dysgenesis. Double immunohistochemistry using WT-1 (expressed in all Sertoli cells) and p27(kip) (expressed only in mature Sertoli cells) revealed immature Sertoli cells in dysgenetic areas. DBP-exposed animals also exhibited Sertoli cell-only (SCO) tubules, sporadically in scrotal and predominantly in cryptorchid, testes, or foci of SCO within normal tubules in scrotal testes. In all SCO areas the Sertoli cells were immature. Intratubular Leydig cells were evident in DBP-exposed animals and, where these occurred, Sertoli cells were immature and spermatogenesis was absent. Abnormal Sertoli cell-gonocyte interaction was evident at GD 19 in DBP-exposed rats coincident with appearance of multinucleated gonocytes, although these disappeared by postnatal day 10 during widespread loss of germ cells. CONCLUSIONS: Abnormal development of Sertoli cells, leading to abnormalities in other cell types, is our hypothesized explanation for the abnormal changes in DBP-exposed animals. As the testicular and other changes in DBP-exposed rats have all been reported in human TDS, DBP exposure in utero may provide a useful model for defining the cellular pathways in TDS.     

Maxillary Adenoid Cystic Carcinoma with Annulate Lamellae

Of 29 reviewed ultrastructural studies of adenoid cystic carcinoma, only 1 mentions the presence of annulate lamellae. The present report describes a 65-year-old woman who presented with an adenoid cystic carcinoma in her left maxillary sinus. At the light microscopic level, most of the connective tissue was loose and myxomatous and contained fine strands, cords, and islands of epithelium that in places had a cribriform pattern. The ultrastructural appearance was similar to that described by others. Two epithelial cell types predominated. The cytoplasm of one type contained mainly ribosomes, and that of the other type also had peripheral microfilaments. Also present were pseudocysts, lumina with microvilli, and replicated basal lamina. In addition, a number of both types of cells contained parallel arrays of cisternae that had regular small pores. These were annulate lamellae.     

Pure Sertoli cell tumour of the ovary: report of a case with ultrastructural observations

A case of pure Sertoli cell tumour of the ovary is described. Unusual features were its great size, lack of clinical hormonal activity and the presence of granulomas in its interstitium. Mitochondria with tubular christae are described for the first time in such a tumour. These, together with lipid droplets and plentiful smooth endoplasmic reticulum, suggest a potential for steroid hormone synthesis which was not expressed clinically. Dense membrane-bound granules of undetermined type were present in large numbers. The presence of cilia and microvilli, and the absence of annulate lamellae and Charcot-Böttcher crystalloids, suggests that the tumour cells may not be truly of Sertoli type.     

Annulate lamellae in hepatic and pancreatic beta cells of the chick embryo

This study is concerned with the finding of cytoplasmic annulate lamellae in normally developing avian somatic tissues. Annulate lamellae were observed in hepatic parenchymal and pancreatic beta cells of the chick embryo on days five and eleven, respectively, of egg incubation. The presence of this cytoplasmic membrane system in normal embryonic tissues supports the view that annulate lamellae may represent normally occurring, transitory organelles common to virtually all cells during early embryonic differentiation.     

The effect of phorbol myristate acetate on human lymphocytes. An ultrastructural study.

Phorbol myristate acetate (PMA), a tumour promoter, acts as an agglutinin and as a stimulator of human lymphocytes. Agglutination begins within 1 hr and is macroscopically evident within 24 hr. The ultrastructural findings in lymphocytes stimulated with PMA after 48 and 72 hr show blast transformation of the nucleus, increase in cytoplasm and in ribosomes, and occasional “annulate lamellae.” Apart from the finding of “annulate lamellae” the changes are generally similar to those found in PHA stimulated lymphocytes. The function and significance of the “annulate lamellae” are not known but as they have been most often seen in malignant and embryonal cells they may be representative of a more fundamental dedifferentiation of the cells with PMA as opposed to PHA.     

Nuclear and cytoplasmic annulate lamellae in trophoblast giant cells of rat placenta

Single stranded profiles of nuclear annulate lamellae were identified in giant cells of rat trophoblast from the day when the chorioallantoic placenta first becomes vascularized, viz., day 12 post coitum, until the day before term, viz., day 22. Cytoplasmic annulate lamellae were observed only in giant cells from placentas at day 12. Occasionally cytoplasmic annulate lamellae were found in parallel array. Often the lamellar membranes were continuous with both granular and agranular membranes of endoplasmic reticulum; they closely resembled doubled outer nuclear membrane. Nuclear annulate lamellae resembled doubled inner nuclear membrane; and often the two were found in continuity. In addition, at later gestational ages (17 and 22 days), nuclear lamellae often were related anatomically to the variety of nuclear inclusions which characterize giant trophoblast cells during late pregnancy. A possible relationship of annulate lamellae to the synthesis of DNA, RNA and protein is considered.     

Ultrastructure of the seminiferous tubules in human testes before and after varicocelectomy

Ultrastructure of the membrana propria and the seminiferous epithelium was studied in infertile human testis both before and 3–6 months after varicocelectomy. The frequent alterations, observed before and after the operation, were extremely thickened membrana propria, deep invaginations, multilamination and knob-like formation of basal laminae and formation of multinucleated spermatids, which were all considered as the common response of the testis to different noxious agents. Although the cells of the seminiferous epithelium were clearly affected by varicocele before varicocelectomy, many areas exhibited normal features after the operation. Furthermore, multinucleated cells, sharing common features of Sertoli cell and spermatogonium, were observed, as well as presence of well-developed annulate lamellae in the Sertoli cells, exhibiting centrioles in the vicinity of their nuclei after varicocelectomy. These multiple ultrastructural observations indicate that Sertoli cell division takes place. This study suggests that if the observation period of the tissue samples after varicocelectomy is long enough, the reversible changes of the tubular cells would be seen much more frequently.Part of this study was submitted as a PhD thesis by Asst. Prof. Hülya Özgür and presented at the 12th National Congress on Electron Microscopy held in Antalya, Turkey, 1995     

A scanning electron microscopic study of the Sertoli cell and spermiation in the syrian hamster

The scanning electron microscope was used to examine the Sertoli cells in normal and germ-cell-depleted testes. The Sertoli cells appear to attain characteristic configurations in the various stages of the cycle of the seminiferous epithelium. In tubules containing maturation-phase spermatids, Stages I-IV, the Sertoli cells exhibit column-like bases which give rise to lamellae measuring 25–30 μm in width which ensheath the spermatids and residual cytoplasmic bodies. In the Stages VIII-IX long flat sheets of Sertoli cytoplasm rest atop the step-8 and -9 spermatids. These sheets are oriented parallel to the basement membrane of the tubule with their long axis parallel to the long axis of the tubule. In Stages V-VI the head and proximal portion of the tail of the maturation-phase spermatids are ensheathed in sleeve-like Sertoli cell processes. Cuff-like terminations demarcate the terminus of these sleeves that surround maturing spermatids up to spermiation. In tubules undergoing spermiation, the sleeves retract so that only the tip of the spermatid head remains in the sleeve. Appendicular processes extend from the dorsum of the Sertoli cell sleeves in the tubules undergoing spermiation. After spermiation the appendix elongates while the sleeve evaginates until the everted sleeve is a finger-like process that extends into the tubular lumen. In tubules in which the seminiferous epithelium has been depleted with epinephrine injections the Sertoli cells attain two configurations. The first is characterized by having long attenuated lamellar processes that orient perpendicular to the basement membrane, with numerous ramifying processes arising from the lamellae. In the second configuration the Sertoli cells' lamellae orient parallel to the basement membrane of the tubule and lack the elaborate ramifications seen in the first configuration.     

Viruses and annulate lamellae in Friend erythroleukemia cells

Virus formation in a clone of murine undifferentiated Friend erythroleukemia cells was examined by electron microscopy. Budding C-type particles were present at the cell surface. The principal site of intracisternal particle production was in elements of rough-surfaced endoplasmic reticulum disposed about the periphery of stacks of annulate lamellae. Serial sections demonstrated that these virus-laden cisternae were in direct continuity with the annulate lamellae. In addition, intracisternal particles occurred in membranous honeycomb structures present in the cytoplasm of many cells. Viral elaboration also was associated with stacks of cisternae of endoplasmic reticulum that were devoid of ribosomes, but that were coated with an extensive and continuous layer of dense material. In some instances, the outer nuclear membrane was coated with the same dense substance. It appears that in Friend erythroleukemia cells, a very substantial portion of their cytomembranes is devoted to synthesis of intracisternal particles.     

Morphological profiles of cryptorchid XXY mouse testes

Two mice with an XXY karyotype and cryptorchid testes appeared spontaneously in a colony. The animals were H-Y antigen-positive, and had elevated serum levels of follicle-stimulating (FSH) and luteinizing (LH) hormones. Testes of the affected mice were atrophic, containing a few solid seminiferous cords surrounded by vast amounts of compact interstitial material. The cords were delimited by a broad tunica propria in which the basal lamina was irregularly thickened and stratified into a number of alternating dense and less dense layers. Most sex cords were populated by mature Sertoli cells and small pleomorphic elements resembling monocytic-derived macrophages. Within some cords, the macrophages aggregated into a central mass with which identifiable Sertoli cells and (PAS) periodic acid Schiff-positive fragments of basal lamina were associated. In more severely damaged cords, the basal lamina and peripheral carpet of Sertoli cells were totally missing. Such cords were populated only by the central macrophages with fragments of basal lamina and degenerating Sertoli cells. Finally, a few collapsed remnants of cords contained compact nodules of macrophages surrounded by what appeared to be the outer part of the tunica propria. The interstitial area, as well as the outer walls of the seminiferous cords were also heavily infiltrated by macrophages. Overall, the morphological picture was one of severe immunological injury. We do not know what role, if any, the genetic constitution and/or intra-abdominal environment may play in the expression of these bizarre pathologies. However, such severe changes have not been reported in either Klinefelter's syndrome or the undescended testes of any human or subprimate species.     

Vimentin expression during altered spermatogenesis in rats

The collapse of vimentin caused by some xenobiotics correlates with the loss of structural integrity of the seminiferous epithelium. In this study, we investigated the effect of busulphan (an anticancer drug with toxic effects on dividing germ cells) on vimentin filament distribution in rat seminiferous epithelium and compared it with changes found in testes of unilaterally cryptorchid rats. In the seminiferous epithelium, the vimentin labelling was observed only in the Sertoli cells, showing a stage-specific arrangement of the filaments. Both busulphan treatment and cryptorchism caused altered distribution of vimentin filaments in the Sertoli cells. In both models, the apical vimentin filaments collapsed towards the nuclei and were disorganized in the basal region of the Sertoli cells while the germ cells were diminished in the epithelium. After the busulphan effect subsided (4 weeks after administration), spermatogenesis began to restore and vimentin filaments began to organize in basal and perinuclear regions of Sertoli cells among the spermatogonia and spermatocytes. Vimentin labelling of the sloughed material in the lumen of cryptorchid testes (but not in busulphan treated animals) was observed. We conclude that the Sertoli cell vimentin filaments play an important role in the maintenance of spermatogenesis, their damage is associated with the seminiferous epithelium disintegration and their restoration with a recovery of spermatogenesis after the unfavourable conditions subside.     

Meiosis and spermiogenesis in the testis of Salamandra salamandra (L.) (Amphibia, Urodela)

Meiosis and spermiogenesis in the fire salamander, Salamandra salamandra, take place in an intermediate zone of the testis between the cephal immature and the more caudal mature part. Primary spermatocytes in zytogene and pachytene are characterized by synaptonemal complexes, flattened vesicles at the periphery of the cytoplasm and mitochondria with dilated cristae. Mitochondria in primary spermatocytes during meiosis, in secondary spermatocytes and early spermatids are typically arranged beneath the plasmalemma. Secondary spermatocytes are provided with pro-acrosomal granules, nucleolus-like bodies and complexes of annulate lamellae. Cytoplasmic parts with numerous vesicles seem to become extruded from secondary spermatocytes and spermatids. In testicular lobules containing pachytene spermatocytes the normally fibroblast-like follicle cells transform into glandular Sertoli cells. Already after the second meiotic division of germ cells lobule boundary cells show morphological features of steroid hormone secreting cells.     

Carcinoma in situ of the testis in infertile men. A histological, immunocytochemical, and cytophotometric study of DNA content

Of 723 infertile men (128 with a history of cryptorchidism) whose testes were biopsied at the outer lateral face of the testis, five presented carcinoma in situ (CIS) in one testis. These testes were removed, serially sectioned, and examined by light microscopy. In order to evaluate whether only one or two biopsies are sufficient to diagnose CIS, before sectioning the testes four biopsies were taken at the anterior face, posterior face, superior pole, and inferior pole of the testis, respectively. Two of the five men had undergone orchiopexy in infancy and the testis contained tubules with Sertoli cells and isolated spermatogonia. CIS was also present in some tubules that were principally located near the rete testis. Of the four simulated biopsies, only that performed at the posterior face of the testis revealed CIS. The other three infertile men showed tubules with complete, although reduced, spermatogenesis, and tubules lined by Sertoli cells only. CIS was found in both types of tubules. These tubules with CIS formed lobules that extended throughout the testicular parenchyma. Most simulated biopsies performed in these three testes showed CIS. The average nuclear DNA content of CIS cells was about 4c in all testes. This content was similar both in tubules with complete spermatogenesis and in tubules with Sertoli cells only.     

Annulate lamellae in human malignant tumors: report of three cases

During the course of applying electron microscopy to diagnostic surgical pathological specimens, three malignant tumors (malignant melanoma, fibrous mesothelioma, lymphoblastic lymphoma) were found to contain annulate lamellae, distinctive intracytoplasmic organelles composed of membrane stacks interrupted by constrictions or pores. In one case both annuli and lamellae were present, a combination rarely described in human tissue and in animal models. In this material, the annuli of the annulate lamellae were structurally similar to nuclear pores. It is postulated that the abundant fibrils are probably related to the unusual configuration of the annulate lamellae. A morphologic relationship of the annulate lamellae to both the endoplasmic reticulum (cases 1 and 2) and the nuclear membrane (case 3) supports the theory that annulate lamellae may be related to both of these structures.