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PubMed2004

[Cystic testicular lesions in infancy].

The present article reports a case 11 month-old infant with a right intratesticular cyst. We analyze the etiology, differential diagnosis and management off all cystic lesions of the pediatric testis. Patient age at presentation, examination features, tumor markers and sonographic appearance may assist in making a presumptive and occasionally definitive diagnosis preoperatively. The differential diagnosis include intratesticular simple cyst, epidermoid cyst, tunica albuginea cyst, testicular teratoma, juvenil granulosa cell tumor-gonadal stromal tumor, cystic dysplasia of the rete testis, cystic lymphangioma, and testicular torsion. Usually enucleation is the best treatment. A thorough understanding of potentially cystic testis lesions in children leads to the best management choices and often to preservation of a substantial portion of the affected testis.

Cysts↗
PubMed2003

Autologous and homologous transplantation of bovine spermatogonial stem cells.

The aim of this study was to develop a method for spermatogonial stem cell transplantation into the bovine testis. Five-month-old Holstein-Friesian calves were used and half of the calves were hemicastrated to allow autologous transplantation and the other half were used for homologous transplantation. Approximately 20 g of each testis was used for cell isolation. On average 106 cells per gram of testis containing about 70% type A spermatogonia were isolated. The cells were frozen in liquid nitrogen until transplantation. Testes were irradiated locally with 10-14 Gy of X-rays to deplete endogenous spermatogenesis. At 2 months after irradiation, cells (approximately 10 x 10(6) were injected into the rete testis through a long injection needle (18 gauge), using ultrasonography and an ultrasound contrast solution. At 2.5 months after transplantation, calves were castrated and samples of testes were taken for histological examination. After 2.5 months in the irradiated non-transplanted control testes, only 45% of the tubules contained type A spermatogonia. However, after autologous spermatogonial transplantation, >80% of the tubule cross-sections contained type A spermatogonia. In addition, only 20% of the tubules of the control testes contained spermatocytes and, except for a few tubules (5%) with round spermatids, no more advanced germ cells were found. After autologous spermatogonial transplantation, about 60% of the tubules contained spermatocytes; 30% contained spermatids and in about 15% of tubules spermatozoa were found. No improvement in spermatogonial repopulation was found after homologous transplantation. The results of this study demonstrate, for the first time, successful autologous transplantation of bovine spermatogonial stem cells resulting in a complete regeneration of spermatogenesis.

Animals↗
PubMed1998

NADPH-diaphorase in glandular cells and nerves and its relation to acetylcholinesterase-positive nerves in the male reproductive tract of man and guinea-pig.

The presence of NADPH-diaphorase activity and acetylcholinesterase in the testis, epididymis, vas deferens, seminal vesicle, pelvic plexus, prostate and urethra of man and guinea-pig was investigated with the nitro blue NADPH technique and the thiocholine method, respectively. In human material NADPH-diaphorase activity was found in the Leydig cells, Sertoli cells and the epithelial linings of the rete testis, the excretory ducts, seminal vesicle, prostate and urethra. The guinea-pig material showed staining of the Leydig cells and spermatozoa and similar epithelial staining of the tract as man. Nerves beneath the epithelium and in the muscle layers of cauda epididymis, vas deferens, seminal vesicle, prostate and urethra were also stained. NADPH-diaphorase-positive nerve cells were seen in the pelvic plexus. Some cells also displayed acetylcholinesterase activity but others showed activity for only one of the enzymes or no activity for either enzyme. In the cauda epididymis, vas deferens, seminal vesicle, prostate and urethra acetylcholinesterase-positive nerve fibres formed a plexus beneath the secretory cells. It is concluded that NADPH-diaphorase, generally accepted as a nitric oxide synthase, is present in glandular cells of the male genital tract. The enzyme is also present in nerves, where it is partly co-localized with acetylcholinesterase.

Acetylcholinesterase↗
PubMed1989

Cysts of the testicular parenchyma and tunica albuginea.

Five cases of cysts of the tunica albuginea and two cases of cysts of the testicular parenchyma are described. The cysts of the albuginea were located in the anterior, lateral, and inferior faces of the testis (distant from the epididymis) and covered by a thin albuginea. They extended toward the testicular parenchyma or toward the tunica vaginalis. Their epithelial lining varied from low cuboidal to columnar and consisted of ciliated or nonciliated cells. These cysts were probably of mesothelial origin. The cysts of the testicular parenchyma were also located distant from the mediastinum testis and were not in contact with the albuginea. Their epithelial lining consisted of flattened, cuboidal, nonciliated cells. The origin of the cysts of the testicular parenchyma is not clear. In one case the occurrence of spermatozoa in the cyst lumen and connections with seminiferous tubules suggests an origin in the rete testis.

Aged↗
PubMedDate not supplied

S-100 protein immunoreactivity in bovine testis.

The present study deals with immunohistochemical localization of S-100 protein in the bovine testis. Immunoreactivity for the protein was seen in Sertoli cells of the seminiferous tubules and as a particularly intense staining in the terminal tubular segment (transitional region, middle portion, and terminal plug), which is mainly composed of modified Sertoli cells. Immunoreactivity was also found in epithelial cells of the straight testicular tubules and rete testis, and in the endothelium of capillaries, veins and lymphatic vessels. Although the functional significance of S-100 protein immunoreactivity in the Sertoli cells remains unclear, the present results suggest that it may be involved in the microtubule assembly-disassembly system. The specificity of the immunolabelling observed should enable the antigen and/or antibody to S-100 to be used as an investigative and diagnostic tool in the study of bovine Sertoli cell function.

Animals↗
PubMed1996

In vivo microperfusion of the ductuli efferentes testis of the rat: flow dependence of fluid reabsorption.

Individual ducts from the initial zone of the efferent ducts of the rat were microperfused in vivo using a double cannulation procedure, which allowed the recovery of perfused fluids for analysis and determination of the rate of fluid reabsorption from the perfused duct. The ducts were perfused at rates from 0.025 to 0.4 microliters min-1 with either Krebs-Ringer bicarbonate (KRB) solution or the native rete testis fluid (nRTF) that perfuses the ducts in situ. Reabsorption of KRB solution increased linearly with a perfusion rate of between 0.025 and 0.1 microliter min-1 (from 17.4 +/- 1.5 to 34.3 +/- 3.2 nl (10 mm duct)-1 min-1), then increased no further. Reabsorption of nRTF increased linearly between 0.025 and 0.2 microliters min-1 (from 17.7 +/- 1.5 to 61.4 +/- 13.5 nl (10 mm duct)-1 min-1) and then declined. The reabsorption rate from nRTF perfusates was significantly higher than from KRB perfusates. As a proportion of the luminal perfusate, reabsorption declined from 73.0 +/- 6.0 to 7.4 +/- 3.0% (10 mm duct)-1 for KRB solution and from 73.1 +/- 6.0 to 4.1 +/- 1.3% (10 mm duct)-1 for nRTF. There was no significant change in the concentration of either Na+ or Cl- in KRB solution or nRTF during perfusion through the efferent ducts, indicating that the reabsorption of these ions was isomolar. However, the reabsorption of K+ from nRTF occurred at a greater rate than that of water, and the initial [K+] declined from 17.2 +/- 0.4 mM in nRTF perfusates to 5.7 +/- 0.5 mM in collectates (perfusion rate, 0.1 microliter min-1) to achieve equilibrium with blood plasma (4.7 +/- 0.4 mM). The osmotic pressure of both KRB and nRTF perfusates equilibrated with blood plasma, indicating a high permeability of the epithelium to water. The results of this study provide further evidence that fluid reabsorption in the efferent ducts is isosmotic, or close to isosmotic, and have shown that, as in the homologous proximal kidney tubule, reabsorption is dependent on luminal flow rate. In contrast to the proximal tubule, however, reabsorption in the efferent ducts is not maintained as a constant proportion of the perfusion load. It is concluded that microperfusion in vivo provides a useful technique for studying fluid reabsorption in the efferent ducts of the rat.

Absorption↗
PubMed1975

Maintenance of rate testis fluid testosterone and dihydrotestosterone levels by pregnenolone and other C21 steroids in hypophysectomized rats.

This study was undertaken to determine whether maintenance of spermatogenesis in hypophysectomized rats by pregnenolone and other C21 steroids may be due to in vivo conversion of these compounds to androgens. Hypophysectomized rats were treated sc with 2 mg of pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone or testosterone propionate in 0.2 ml sesame oil daily for 14 days beginning 2 days after hypophysectomy. Rete testis fluid (RTF), peripheral blood, and testicular venous blood were collected on the day of the last injection. Testosterone (T) and dihydrotestosterone (DHT) were measured by radioimmunoassay after chromatographic separation. Results demonstrate that T and DHT could be found in the RTF of C21 steroid-treated hypophysectomized rats at levels similar to those seen in the intact rat. Results imply that the maintenance of spermatogenesis by C21 steroids is probably due to the conversion of these compounds to T in the testis. Relatively little T was released from the testis into the peripheral circulation of these rats since T levels in testicular venous plasma were low and peripheral plasma T levels were not distinguishable from those seen in untreated hypophysectomized rats. Histological examination of the testes of C21 steroid-treated hypophysectomized rats showed nearly quantitative maintence of spermatogenesis and atrophy of Leydig cells. These findings suggest that most of the conversion of C21 steroids to androgens occurred in the seminiferous tubules.

17-alpha-Hydroxypregnenolone↗
PubMed2000

Oestrogen in fluid transport in efferent ducts of the male reproductive tract.

This review focuses on the importance of oestrogen and oestrogen receptors in the male reproductive system, with a special interest in the newly discovered role of oestrogen in the regulation of fluid reabsorption in the efferent ductules of the testis. Early work on oestrogen synthesis indicated that Leydig and Sertoli cells were the only important cells in the production of this steroid in the adult testis. However, more recent work has shown that germ cells and spermatozoa also contain aromatase and produce oestrogen. The observation that germ cells synthesize oestrogen contributed to a new hypothesis that oestrogen in the lumen of the male reproductive tract targets the epithelial lining of efferent ductules and the epididymis. The location of nuclear oestrogen receptors in the male reproductive tract has also been investigated and it has been found that oestrogen receptor alpha is more abundant in the efferent ductules of the testis than in any other tissue of the male or female. In all species examined to date, oestrogen receptor alpha has been found to be abundant in the efferent ductules. The structure and function of the efferent ductules are taken into account as these tubules are responsible for the reabsorption of almost 90% of the luminal rete testis fluid. Thus, it was logical to hypothesize that oestrogen receptors play a role in the regulation of fluid reabsorption in efferent ductules. The oestrogen receptor alpha knockout mouse was used to help define this role of the receptor in males. In this animal model, the efferent ductules are altered markedly from a reabsorptive epithelium to a squamous epithelium devoid of lysosomes and endocytotic organelles. Although the separate roles for oestrogens and androgens in the regulation of fluid reabsorption are controversial and remain to be resolved, it is now established that loss of oestrogen receptor function in males interferes with the resorptive function of efferent ductules, a function that is essential for fertility. Future studies will focus on the biochemical and physiological mechanisms involved in the regulation of water and ion movement by oestrogen in the male reproductive tract.

Animals↗
PubMed1991

[Unusual tumors of the testis. Apropos of 22 cases].

The authors report their experience of rare non-germ-cell tumours of the testis over a period of 10 years. The criteria of benign disease, justifying testicular preservation in 5 out of 7 cases of epidermoid cysts, are defined. Four cases of Leydig cell tumours, including 3 with gynaecomastia, are described. Three cases of adenomatoid tumours with conservative surgery, 3 lymphomas, 2 mature teratomas and 2 cysts of the rete testis are also reported. The place of ultrasonography is defined and the possibility of conservative surgery is discussed in relation to a review of the recent literature.

Adult↗
PubMed1994

Response of alpha 2-macroglobulin messenger ribonucleic acid expression to acute inflammation in the testis is different from the response in the liver and brain.

Recent studies from this laboratory have shown that Sertoli cells derived from 20-day-old rats and cultured in vitro synthesize and secrete a nonspecific protease inhibitor that is structurally and immunologically similar to serum alpha 2-macroglobulin (alpha 2-MG). In contrast to its serum homologue, the testicular alpha 2-MG is not an acute-phase protein in the rat since its protein concentration in the rete-testis fluid does not increase in response to inflammation. In the present study we examined the expression of alpha 2-MG mRNA in the rat testis in comparison to that in the brain and liver following induced inflammation. alpha 2-MG mRNA in the testis did not respond to induced inflammation, whereas its protein concentration in serum and its mRNA level in the brain and liver increased significantly in 20-day-old inflamed rats. In 8-day-old rat testis, where the blood-testis barrier is not yet formed, alpha 2-MG mRNA expression also did not respond to induced inflammation. The mRNA expression of clusterin, another authentic Sertoli cell protein whose secretion appears to be closely related to cell-cell interactions in the seminiferous epithelium, was shown to be unaffected by induced inflammation in the testis, brain, and liver. In view of the unexpected differential expression of alpha 2-MG mRNA to induced inflammation in the testis and liver, we sought to examine whether Sertoli cell alpha 2-MG would respond to FSH and testosterone (T), the major regulators of testicular function. Interestingly, expression of alpha 2-MG and clusterin mRNA in the Sertoli cell was not regulated by FSH, T, or a combination of FSH and T. Since there is an intimate morphological relationship between Sertoli cells and germ cells, we next examined the effect of germ cell-conditioned medium (GCCM) on Sertoli cell alpha 2-MG and clusterin mRNA expression. It was noted that GCCM caused a dose-dependent stimulation of alpha 2-MG and inhibition of clusterin mRNA expression in Sertoli cells, respectively. Therefore, our studies have shown that the regulatory mechanism that modulates the expression of alpha 2-MG mRNA in the rat testis is different from its counterpart in the brain and liver.

Aging↗
PubMed1991

NCAM in developing mouse gonads and ducts.

The expression of the Neural Cell Adhesion Molecule, NCAM, in mouse gonads and ducts was studied from fetal life to maturity. The methods used were immunocytochemical staining and Western blotting. The immunocytochemical studies showed that the only structures that remain NCAM-positive throughout life were the mesonephric-derived rete ovarii and rete testis. Also in the fetal gonads some somatic cell lining the groups of differentiating germ cells were stained. In the immature as well as in the mature ovary the granulosa cells and oocytes of growing and large follicles--but not of small follicles--were stained. A particularly strong staining of the cytoplasm of the oocyte, healthy as well as atretic, was seen. All cells of the testis remained negative except for weakly stained residual bodies and late spermatids. At all ages the male ducts showed only weak staining, whereas in the female Müllerian duct the epithelium became strongly positive at puberty. The stroma of the Müllerian duct was positive during a transitory period around day 16 of fetal life in both sexes. One-dimensional gel immunoblotting of total protein from gonads, rete and ducts from immature and mature mice showed that only the two largest isoforms of NCAM (NCAM-A and NCAM-B) were present. The gonads and the rete of both sexes and the adult uterus expressed only NCAM-B, whereas NCAM-A was also detected in the adult epididymis. The present findings suggest that NCAM may be involved in the normal development and formation of both the gonads and ducts.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMedDate not supplied

Adenovirus-mediated p53 gene transfer to rat testis impairs spermatogenesis.

The tumor suppressor protein p53 participates in normal cell differentiation as well as induction of programmed cell death. The authors investigated the effect of p53 overexpression on spermatogenesis by transferring p53 gene into the rat testes. Replication-deficient recombinant adenovirus vectors were constructed to include cytomegalovirus (CMV) promoter driving wild-type p53 (Ad-CMV-p53) or beta-galactosidase (Ad-CMV-beta-gal). Virus was delivered to cells of the tubules by slow retrograde injection through the rete testis. At 0, 4, 7, and 14 days, testes were removed, weighed, and analyzed histopathologically, including immunohistochemistry for p53, Bcl-2, Bax, and interleukin-1beta converting enzyme (ICE). Testicular weight was decreased in Ad-CMV-p53 group at 14 days after injection, while no change occurred in phosphate-buffered saline-injected controls or Ad-CMV-beta-gal-infected testes. Beyond 4 days, cell degradation in tubules interfered with immunohistochemical observation in the Ad-CMV-p53 group. At 4 days, p53 was expressed mostly in spermatocytes. Bax showed greater expression in the p53 group than in the control or Ad-CMV-beta-gal group. ICE, expressed mostly in spermatids, was more abundant in the p53 group than in controls. Overall, p53 overexpression in the testis impaired spermatogenesis.

Adenoviridae↗
PubMed2004

Clear cell adenocarcinoma of the tunica vaginalis of the testis with an adjacent uterus-like tissue.

Testicular and paratesticular neoplasms that resemble the common epithelial type of ovarian tumor are quite rare. Paratesticular clear cell carcinoma is very uncommon in the testis, with no reported cases of a tumor arising from the tunica vaginalis in the literature to our knowledge. The present case shows that it is highly malignant and metastatic. The differential diagnosis of the tumor was made after thorough clinical, pathological and immunohistochemical investigations, from the mesothelioma of the tunica vaginalis, paratesticular serous papillary carcinoma, carcinoma of the rete testis, epididymal adenocarcioma, yolk sac tumor of the testis and metastatic carcinoma. The tumor showed Bcl-2 and Her-2/neu immunoreactivity, but was non-reactive for p53. This tumor, with a uterus-like structure as a paratesticular tumor-like mass, was composed of endometrial-type glands and stroma surrounded by bundles of smooth muscle, and is the third example of this kind of structure in English written literature. The patient, having normal external genitalia and fertility, represents the first reported case of paratesticular malignant differentiation of müllerian-type epithelium in the normal gonadal state. Müllerian-type epithelium located in the vicinity of the testis and/or endometriotic metaplasia of the mesothelium of the tunica vaginalis might be the possible origins for this uterus-like structure, and as a result, for this tumor.

Adenocarcinoma, Clear Cell↗
PubMed1989

Papillary serous cystadenoma of the testis.

A case is presented of a 50-year old man with a unilocular cystic intratesticular tumour exhibiting the morphological features demanded from WHO for the diagnosis of serous papillary cystadenoma of the ovary. Keratin filaments could be demonstrated in the cyst lining and papillae covering cells by means of PAP-technique; AFP and SP-1 were lacking. The epithelial cells of the tumour showed a lectin binding pattern (WGA, UEA-I, PNA, Con A, PSA, LCA, RCA) different from the epithelium of rete testis and epididymis. We intend to classify our tumour as the male analogue of the respective ovarian growth.

Cystadenoma↗
PubMed1992

Developmental expression of the S35-S45/SGP-2/TRPM-2 gene in rat testis and epididymis.

Testosterone-repressed prostate message-2 (TRPM-2) was originally isolated and cloned from the regressing ventral prostate of the rat. In this tissue, and in other hormone-dependent tissues such as the mammary gland, this gene is induced in the absence of the appropriate trophic hormone. Sequence analysis of the cDNA and genomic clones of TRPM-2 have demonstrated that the coding sequence of this gene is identical to S35-S45 (also known as SGP-2 and clusterin), which is constitutively expressed by the Sertoli cells of the adult testis. Using Northern, slot blot, S1-nuclease analysis, and in situ hybridization, we have investigated the regulation of TRPM-2 expression in the testis and epididymis during development. Slot blot analysis of RNA extracted from the testis and epididymis of 7-, 14-, 28-, 35-, and 91-day-old rats demonstrates that the gene is induced to detectable levels between days 7 and 14 and that the relative level of expression does not change significantly after day 14. In situ hybridization using frozen sections of testis from day 2-, 7-, 14-, 28-, 35-, and 91-day-old rats confirms that there is little expression of TPRM-2 in the seminiferous epithelium of 7-day-old rats, but this increases considerably after 14 days, primarily in Sertoli cells but also in association with meiotic developing spermatogenic cells. However, TRPM-2 mRNA is expressed in the rete testis at 2 days of age, reaches a peak at 35 days of age, and continues to be expressed in the adult. Slot blot analysis demonstrates that TRPM-2 is also induced in the epididymis between 7 and 14 days of age, although, as has been demonstrated by in situ hybridization, TRPM-2 mRNA is detectable in the epithelial cells in the head of the epididymis but is barely detectable in the midportion or tail regions. Northern analysis suggests that the size of the TRPM-2 transcript in the testis also changes during development. In the early stages of testicular development, the TRPM-2 transcript appears to be a broad band of approximately 1.5 kb, while the transcript in the adult appears to be approximately 1.8 kb in length. S1-nuclease protection assays suggest that this increase in size is not due to differential splicing of the first exon of TRPM-2/SGP-2 and most probably reflects a difference in the polyadenylation of the mRNA in the testis at different times during development.

Animals↗
PubMed1990

Sertoli cell synthesizes and secretes a protease inhibitor, alpha 2-macroglobulin.

The mechanism by which the seminiferous epithelium limits the damaging effects of proteases that are released from degenerating late spermatids does not depend upon protease inhibitors in the systemic circulation since these proteins are excluded from the seminiferous tubule by the blood-testis barrier. The purpose of this study was to identify the major protease inhibitor of the testis and determine its cellular origin. Sertoli cells, the major epithelial component of the seminiferous epithelium, release a protease inhibitor, testicular alpha 2-macroglobulin, in vitro. Immunoprecipitation using [35S]methionine and a monospecific polyclonal antibody prepared against purified testicular alpha 2-macroglobulin establishes that this protein is actively synthesized and secreted by Sertoli cells. Measurements of immunoreactive protease inhibitors in tubular and rete testis fluids collected by micropuncture suggest that alpha 2-macroglobulin rather than alpha 1-antitrypsin is the major protease inhibitor in the seminiferous tubules in vivo. The ability of alpha 2-macroglobulin to inactivate proteases and growth factors such as TGF-beta by a common mechanism suggests that this protein may have a dual function in the testis.

Amino Acid Sequence↗
PubMed1999

Sex-specific localization of laminin alpha 5 chain in the differentiating rat testis and ovary.

The localization of laminin (Ln) alpha 5, beta 1 and beta 2 chains in the differentiating rat testis and ovary was studied by immunolabeling light and electron microscopy. The initial formation of the male and female gonadal blastemas included an emergence of Ln alpha 5 and beta 1 chains, but not of Ln beta 2 chain. The sexual differentiation of the embryonic male gonadal cords included rapid sex-specific disappearance of the incipient Ln alpha 5 chain. The rete testis cords, in contrast, remained positive for Ln alpha 5 chain. In the postnatal testis, the Ln alpha 5 chain reappeared in Ln beta 1 chain-positive cord basement membranes, which also became positive for Ln beta 2 chain. The differentiating myoid cells also gradually became positive for both Ln alpha 5 and Ln beta 1 chains. In the ovary Ln alpha 5 chain persisted in BMs of the cords throughout the fetal phase. Small and newly formed follicles in the early postnatal rat ovary were also positive for Ln alpha 5 chain, whereas growing and large follicles were negative. During the early postnatal phase, Ln beta 1-chain positive follicular BMs became also positive for the Ln beta 2 chain. Basement membranes of testicular and ovarian surface epithelia contained the Ln alpha 5 chain throughout the study. The blood vessels of the male and female gonad showed differentiation-dependent variation in their reactivity for the Ln alpha 5 and beta 2 chains. The present results show that the Ln alpha 5 chain is an early molecular marker for sexual differentiation, which therefore may be regulated by the testis-determining factors. The results also show that in the early postnatal rat ovary, the follicular basement membranes are heterogeneous in their Ln content, which may offer a means to distinguish different follicular populations from each other and to identify the different stages of follicular growth.

Animals↗
PubMedDate not supplied

The origin of some acid hydrolases of the fluid of the rat cauda epididymidis.

The activity of N-acetyl-beta-D-glucosaminidase (NAG, 60.1 units/mg protein) and of acid phosphatase (57.7 units/mg protein) in fluid from the cauda epididymidis formed without any contribution from the testis (fluid obtained from a perfused and isolated cauda epididymidis or from an epididymis whose corresponding efferent ducts had been ligated for 40 days) was significantly higher than the activity of these enzymes in normal fluid (39.6 and 41.2 units/mg protein, respectively). Arylsulphatase activity of the locally formed fluid (11.2 units/mg protein) was lower than that of normal fluid (74.1 units/mg protein). The rete testis fluid was relatively rich in arylsulphatase since the ratio of arylsulphatase to acid phosphatase activity was 17 times higher in this fluid than in locally formed fluid. It is concluded that the activities of NAG and acid phosphatase in normal fluid from the epididymis originate in the epididymal tissue, while most of the arylsulphatase activity comes from the testis.

Acetylglucosaminidase↗