Expression of cyclin-dependent kinase 5 and associated cyclins in Leydig and Sertoli cells of the testis

Sixty patients with a sudden onset of motor disability were assessed for illness behavior and depression. In 30 of the patients, etiology was attributed to a definite structural lesion. The remaining 30 patients were diagnosed as having conversion disorder. The Illness Behaviour Questionnaire (IBQ) and the Hamilton Rating Depression Scale (HRDS) were used as instruments for assessment. The mean HRDS score was significantly higher in the conversion group, indicating a higher degree of affective disease in these patients. According to the results of the IBQ, the patients with conversion disorder showed a higher degree of irritability, disease conviction, and phobic preoccupation, and also, to a greater extent, rejected psychological explanations for their symptoms. Denial was high in both patient groups, coexisting with affective symptoms in the conversion patients but not in the neurological patients. Although valuable information could be extracted from the IBQ, it was not found to be a reliable instrument for distinguishing between psychogenic and organic causes of motor disability.     

Low density lipoprotein receptor-related protein-1 expression in the testis: regulated expression in Sertoli cells

The low density lipoprotein receptor-related protein (LRP-1) is a multiligand receptor capable of mediating endocytosis of a wide array of ligands that relate to both lipoprotein metabolism and proteinase regulation. Many of its ligands are proteinases, proteinase-inhibitor complexes, and lipoproteins known to be contained within the luminal fluid of the seminiferous tubules or in the interstitial spaces of the testis. Immunocytochemical analysis was performed to characterize the pattern of expression of LRP-1 in cells of the rat testis. Immunoperoxidase staining localized LRP-1 to the cytoplasm of Sertoli cells. The distribution and intensity of the Sertoli cell staining was found to vary according to the stages of the cycle of the seminiferous epithelium. Staining was weak in the basal cytoplasm of Sertoli cells during stages II-VIII and strong and granular in the supranuclear cytoplasm during stages XII-XIV and stage I of the cycle. Immunogold labeling showed gold particles associated with the basal and adluminal plasma membranes, with endocytic vesicles, and with endosome membranes. Labeling was also observed on the plasma membrane and membranes of the endocytic apparatus in macrophages and Leydig cells in the interstitial space. Infusion of 125I-Labeled LRP-1 antibody into seminiferous tubules followed by radioautography showed silver grains overlaying the ad-luminal plasma membrane of Sertoli cells at time 0 and in endocytic vesicles and endosomes in the supranuclear region of Sertoli cells 10-minutes postinjection. When the 125I-Labeled LRP-1 antibody was injected into the interstitial space, silver grains overlayed the basal plasma membrane and coated endocytic pits of Sertoli cells at time 0 and, at 10 minutes, the grains labeled endosomes located in the basal pole of Sertoli cells. 125I-Labeled LRP-1 antibody also labeled the plasma membrane and the endocytic apparatus of macrophages and Leydig cells. The absence of immunogold labeling and radioautographic silver grains within lysosomes of Sertoli cells, Leydig cells, and macrophages suggests that internalized LRP-1 is recycled back to the cell surface. The presence of LRP-1 in the endocytic compartment of these cells is consistent with its functioning in the clearance of proteases involved in seminiferous tubule remodeling and/ or the uptake of cholesterol-bound lipoproteins necessary for the biosynthesis of testosterone. In conclusion, the results of these studies demonstrated for the first time the presence of LRP-1 receptor in the endocytic compartments of Sertoli cells and interstitial cells of the rat testis.     

DNA synthesis in cells of mouse corneal epithelium]

Method of autoradiography was used in order to study the kinetics of population of the cornea epithelium cells of mice. Intervals of different duration were found to exist in the DNA synthesis within the limits of S-period of one mitotic cycle. On the basis of personal and literature data a hypothesis has been put forward of a successive pattern of replication in the cells of eukaryots according to which synthesis of a fragment of the DNA daughter thread (or a chromosome subunit) occurs at each moment in a restricted site of a single matrix thread of DNA (matrix chromosome subunit). No DNA synthesis takes place at this moment in the complementary site of the second matrix thread (matrix chromosome subunit), the fragment (chromosome subunit) of one matrix thread being somewhat larger than the complementary fragment (chromosome subunit of the other matrix thread.     

The postnatal development of the junctional complexes of the mouse Sertoli cells as revealed by freeze-fracture

Mouse testes of newborn to adult were examined by freeze-fracture. Between the newborn Sertoli cells, gap junctions consisting of aggregations of the intramembranous particles (about 8 nm in diameter) are frequently found. Some of the junctions are about 1 mum in diameter and show particle-free regions in the aggregation. Linear arrangements of a few particles, which appear to be the initial formation of the occluding junctions, are seen in the newborn sertoli cells. The occluding junctions are arranged in a meshwork, in which the gap junctions are situated between the stages of newborn to six days of age. The particles of the occluding junctions are predominantly located on the B face in the center of the groove instead of the A face of the ridge. The occluding junctions do not appear to surround the entire circumference of the Sertoli cell of the 6-day-old mouse. The gap junctions decrease in size. In later stages, many parallel occluding junctions (up to forty in number) are found over one Sertoli cell surface and are distributed circumferentially around the entire cell surface, indicating establishment of the blood-testis barrier. The occluding junctions dominate and the gap junctions diminish in number as development proceeds.     

Changes in the distribution of intermediate filaments in rat Sertoli cells during the seminiferous epithelium cycle and postnatal development

BACKGROUND: Intermediate filaments (IFs) are components of the cytoskeleton. In mammalian Sertoli cell, IFs are formed by vimentin. Previous studies have shown some characteristics of its distribution in Sertoli cells, however, very little is known of its distributional changes during the seminiferous epithelium cycle and during postnatal development. METHODS: Immunohistochemical and electron microscopic methods were used to determine the distribution of vimentin-type IFs in rat Sertoli cells during the seminiferous epithelium cycle and postnatal development. RESULTS: The distribution of IFs in adult rat Sertoli cell showed distinct cyclic changes during the seminiferous epithelium cycle. At stages I-VI, bundles of IFs extend from the perinuclear region to the supranuclear and apical regions of the Sertoli cell. These apical extensions became shorter at stage VII, and at stages VIII-X IFs were observed only in the perinuclear region. Short apical extensions reappeared at stages XI-XII; and at stages XIII-XIV, they extended again into the apical region. During this cycle, IFs were always closely associated with the heads of elongate spermatids. IFs were also shown to be in close apposition to some specialized structures on the cell membrane, such as the ectoplasmic specialization between adjacent Sertoli cells. During postnatal (p.n.) development, IFs were mainly observed at the basal nuclear region on p.n. day 7. The IFs in the supranuclear or apical regions first appeared at p.n. day 14 and gradually increased during the development. The perinuclear IFs network was fully established by p.n. day 28 and the adult distribution pattern of the IFs was established by p.n. day 42. CONCLUSIONS: Vimentin-type IFs in rat Sertoli cells are a delicate endocellular network, which is centered in the perinuclear region and extends to the apical region of the cell. During the seminiferous epithelium cycle, the distribution of IFs changes in a stage-dependent manner and is closely related to the location of the heads of elongate spermatids. During postnatal development, IFs gradually increase in numbers and the main distribution area is transferred from the basal nuclear to the perinuclear and supranuclear regions.     

Observations on rat Sertoli ectoplasmic ('junctional') specializations in their association with germ cells of the rat testis

The ectoplasmic ('junctional') specialization, a subsurface modification of the Sertoli cell that is often seen facing germ cells, was studied in relation to the development and maturation of these germ cells. This structure is composed of subsurface bundles of filaments and more deeply placed endoplasmic reticulum. The data indicate that these subsurface modifications of Sertoli cells are reutilized in a cyclic fashion, being transferred from their position facing late spermatids to one opposing less mature germ cells. Ectoplasmic specializations appeared to function mechanically in grasping the heads of the spermatids which are undergoing the elongation and maturation phases of spermiogenesis rather than in actually attaching Sertoli cells to these germ cells. It is postulated that the ectoplasmic specialization imparts rigidity to that area of the Sertoli cell that surrounds the head region of the germ cell, forming a recess and a mantle by which the germ cell may be moved toward the base or toward the surface of the seminiferous epithelium. The observed linkage of microtubules to the cisternae of the complex provided a morphological basis for the changes in the cytoarchietecture of the Sertoli cell, which must accompany these movements.     

Endocytosis during postnatal differentiation in superficial cells of the mouse urinary bladder epithelium

This electron microscopical study was performed in order to follow the endocytic pathway of horseradish peroxidase and colloidal gold tracers and to determine the involvement of endocytosis in postnatal differentiation in superficial cells of the mouse urinary bladder epithelium. Morphometric analyses of late endosomes/multivesicular bodies from day of birth to day 25 were performed. The internalisation and intracellular transport of luminal plasmalemma to multivesicular bodies via endocytic vesicles, early endosomes and pleomorphic compartments was established. Dynamic changes in endocytic activity took place within the first few days of postnatal differentiation. During this period the number of multivesicular bodies changed in an inverse ratio to their size. After the third day endocytic activity gradually approached the low rate of adult urothelium.     

Expression of clusterin/sulfated glycoprotein-2 under conditions of heat stress in rat Sertoli cells and a mouse Sertoli cell line

In the present study we investigated the response of monocytes from AIDS patients, susceptible to cryptococcosis (<200 CD4 cells/microl), against Cryptococcus neoformans. Different patterns of response were observed in these cells compared to cells from healthy donors. In particular, fungicidal activity versus this fungus was impaired; this phenomenon could be due to the difficulty of monocytes to internalize C. neoformans in the presence of an intact complement system. Impairment of complement receptor type 3 and direct involvement of this receptor in phagocytosis of C. neoformans were found in monocytes from AIDS patients, which may account for the difficulty in phagocytosis of the fungus. Also, superoxide anion production was dramatically reduced in monocytes from AIDS patients. An increase of spontaneous tumor necrosis factor (TNF) production was evidenced after in vitro addition of C. neoformans. However, this did not activate the antifungal capacity of monocytes from AIDS patients. Moreover, cryptococcus-laden monocytes from AIDS patients were able to induce only a weak response of autologous T-lymphocytes. Hence, monocyte dysfunction could play a part in the progression of cryptococcosis in AIDS.     

Sertoli cell tumor of the testis. Report of a case with imprint cytology findings

BACKGROUND: Sertoli cell tumor of the testis is a relatively rare neoplasm, and few reports concerning its cytologic findings have been previously documented. CASE: A 41-year-old male had Sertoli cell tumor in the left testis. An imprint smear revealed tumor cells showing a less cohesive or singly dispersed cell pattern. The cytoplasm was faintly stained, and most tumor cells were naked. The nuclei were oval or short and spindle in shape, and some of them showed coffee bean nuclei or nuclear indentation. CONCLUSION: Coffee bean nuclei can be observed in Sertoli cell tumor.     

Sertoli cell tumor of the testis: clinical pathologic varieties. Report of a case

Tumours derived from sex cords and primitive gonadal stroma account for 4% of total testicular tumours. The low frequency of Sertoli's cells tumour (SCT) and the uneven study and follow-up of patients makes analysis of this tumoral entity difficult. This paper contributes one case report of a Sertoli's giant cell tumour calcified in a 13-year old patient, and reviews the clinical aspects, clinico-pathological varieties believed to require assessment in patients with this type of disease. This type of tumour is considered benign in its biological behaviour, although some malignant forms have also been described. SCT is actually an heterogeneous tumoral pathogenic entity with regard to pathogenic and prognostic aspects. Our final conclusions show that the clinico-pathological variety, age, size and associated clinical manifestations appear to be related to the prognosis.     

Large-cell calcifying Sertoli cell tumor of the testis: case report and review of the literature

Large-cell calcifying Sertoli cell tumor (LCCSCT) is a rare sex cord-stromal tumor found predominantly in the pediatric population. This tumor has distinctive histopathologic features and clinical associations. LCCSCT has also been noted in association with the Carney complex, and in patients with Peutz-Jeghers syndrome. The propensity to metastasize is low, and radical orchiectomy has traditionally been the treatment of choice.     

Follicle-stimulating hormone (FSH) regulates the expression of FSH receptor messenger ribonucleic acid in cultured Sertoli cells and in hypophysectomized rat testis

FSH acts on Sertoli cells via interaction with a transmembrane receptor (FSHr). Control of expression of the receptor is surely a factor in the regulation of the action of FSH. The regulation of FSHr by FSH and testosterone was studied both in culture and in vivo. Sertoli cells from 18- to 20-day-old male rats were cultured in the presence of 25 ng/ml ovine (o) FSH. At 8 h after addition of FSH, expression of FSHr mRNA decreased significantly. Addition of FSH and actinomycin D to cells did not result in a further decrease in FSHr mRNA levels, suggesting that FSH does not alter turnover of FSHr mRNA. Treatment of cells with 40 ng/ml testosterone did not have any significant effect on the expression of FSHr mRNA. Hypophysectomy of 20-day-old male rats resulted in an increase in expression of FSHr mRNA as compared to that in sham-hypophysectomized animals. This increase was measured at 24 h posthypophysectomy and was maintained at 72 h after surgery. Injection of rats with 0.2 U oFSH at 48 h posthypophysectomy resulted in a reduction in FSHr mRNA when compared to the levels in hypophysectomized rats. Treatment with 2 mg testosterone propionate had no effect on FSHr mRNA levels. The findings confirm that FSH plays an important role in regulating mRNA expression of the FSHr in Sertoli cells in culture and show for the first time that FSHr mRNA is regulated in vivo by FSH in the immature rat testis.     

Sertoli cell tumors of the testis, not otherwise specified: a clinicopathologic analysis of 60 cases

Sixty Sertoli cell tumors of the testis, excluding large cell calcifying and sclerosing subtypes, are described. Patient age ranged from 15 to 80 years (mean, 45 years). The initial manifestation was usually a testicular mass; in 14 cases it had been enlarging slowly for a period of up to 14 years (mean 3.7 years). Only five patients had testicular pain. Four patients had metastatic disease at the time of presentation. All the tumors were unilateral and ranged from 0.3 cm to 15 cm (mean 3.6 cm). They were typically well circumscribed. Sectioning usually disclosed firm, tan-gray, white, or yellow tissue with areas of hemorrhage and a minor cystic component in approximately one third. Microscopic evaluation usually revealed diffuse sheets or large, nodular aggregates of tumor cells, within which solid or hollow, sometimes dilated, tubules and, less often, cords were usually at least focally identifiable. A relatively acellular, often vascular, fibrous to hyalinized stroma was frequently conspicuous. The tumor cells typically had moderate amounts of pale to lightly eosinophilic cytoplasm, but 10 tumors had cells with abundant eosinophilic cytoplasm. Large cytoplasmic vacuoles were prominent in 26 tumors. Nuclear atypicality was absent or mild in 54 cases, moderate in 4 cases, and marked in 2 cases. Mitotic rate ranged from less than 1 to 21 per 10 high power fields, with 50 tumors having no or only rare mitoses. Vascular space invasion was present in 11 cases and was prominent in 8. Follow-up of more than five years (average 8.4 years), or until evidence of metastasis was seen, was available for 16 patients. Nine were alive and well with no evidence of disease. Four were alive with disease and three died of disease. The pathologic features that best correlated with a clinically malignant course were as follows: a tumor diameter of 5.0 cm or greater, necrosis, moderate to severe nuclear atypia, vascular invasion and a mitotic rate of more than 5 mitoses per 10 high power fields. Only one of nine benign tumors for which follow-up data of 5 years or more were available had more than one of these features, whereas five of seven malignant tumors had at least three.     

Haptoglobin is a Sertoli cell product in the rat seminiferous epithelium: its purification and regulation

Using multiple HPLC steps, a protein of 67 kDa (estimated by gel permeation HPLC) was purified from Sertoli cell-enriched culture medium that consisted of two dissimilar subunits of 9 (alpha chain) and 24 (beta chain) kDa on SDS-polyacrylamide under reducing conditions. Direct protein sequence analysis of the 9-kDa subunit revealed a sequence of NH2-VELGNDATDIEXD, which is identical to the alpha subunit of the rat haptoglobin (Hp). Hp is a 67-kDa tetrameric serum acute-phase protein consisting of two alpha and two beta subunits (alpha2beta2) of 8.5 kDa and 24.5 kDa, respectively. Using a 351-bp cDNA coding for Hp for northerns and two Hp primers for RT-PCR, we have demonstrated the expression of Hp in Sertoli and Leydig cells, germ cells, and the testis, but not in the epididymis. In contrast to the hepatic haptoglobin, an acute-phase protein whose steady-state mRNA level increased by as much as fivefold during induced inflammation, the testicular homolog reduced by fourfold within 24 hours following induced inflammation, suggesting that this gene is regulated differently in the testis and in the liver. Moreover, the testicular steady-state Hp mRNA level increased steadily after birth during maturation, suggesting its involvement in spermatogenesis. Using primary Sertoli cell cultures in vitro, it was found that the Sertoli cell Hp expression was not regulated by either FSH, testosterone, estradiol, dexamethasone, interleukin-1beta (IL-1beta), IL-6, interferon-gamma (INF-gamma), transforming growth factor-beta (TGF-beta), lymphocyte inhibitory factor (LIF), or germ-cell-conditioned medium (GCCM). Since transferrin secreted by Sertoli cells is an important molecule in maintaining the crucial iron level necessary for spermatogenesis, the identification of haptoglobin as a Sertoli and germ cell product adds a new member to the growing family of metal transporters in the testis that are likely to play an important role in iron metabolism in the testis.     

Ribonuclease inhibitors from mouse ascites cells

Ribonuclease inhibitors were found to be present attached to mouse ascites ribosomes and in the post-ribosomal supernatant. Both inhibitors inhibited pancreatic RNAses A and B and two endonucleases prepared from ascites cells but did not inhibit RNAses T1 or N1. Both inhibitors had the same sedimentation coefficient and this taken with the results above suggest that they are identical. The inhibitor was shown to interact directly with the RNAse itself.