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PubMed1999

Exposure with the environmental estrogen bisphenol A disrupts the male reproductive tract in young mice.

Environmental estrogens (endocrine disruptive chemicals) have been shown to affect reproduction in wild life and it has been reported that maternal exposure with those chemicals have adverse effects on the male reproductive tract. However, little is known about the potential effects of prepubertal or pubertal exposure with environmental estrogens on the male reproductive tract. Here we examine plasma hormone levels and histology in the testis of mice following either 4- or 8-week oral administration of bisphenol A. Plasma free testosterone levels were dramatically decreased following 8 weeks of bisphenol A treatment compared with control group and morphologically multinucleated giant cells having greater than three nuclei were found in seminiferous tubules in the testis following the 8-week bisphenol A treatment. No differences in plasma corticosterone and luteinizing hormone levels were seen between bisphenol A and control groups. Thus, exposure with bisphenol A around pubertal period may directly disrupt the male reproductive tract. These facts suggest that more detailed studies will warrant the assessment of the risk to the developing human testis from exposure to bisphenol A and other environmental estrogens in prepubertal and pubertal period.

Administration, Oral↗
PubMed1990

Subunit structure of casein kinase II from bovine testis. Demonstration that the alpha and alpha' subunits are distinct polypeptides.

The relationship between the alpha and alpha' subunits of casein kinase II was studied. For this study, a rapid scheme for the purification of the enzyme from bovine testis was developed. Using a combination of chromatography on DEAE-cellulose, phosphocellulose, hydroxylapatite, gel filtration on Sephacryl S-300 and heparin-agarose, the enzyme was purified approximately 7,000-fold. The purification scheme was completed within 48 h and resulted in the purification of milligram quantities of casein kinase II from 1 kg of fresh bovine testis. The purified enzyme had high specific activity (3,000-5,000 nmol of phosphate transferred per min/mg protein) when assayed at 30 degrees C with ATP and the synthetic peptide RRRDDDSDDD as substrates. The isolated enzyme was a phosphoprotein with an alkali-labile phosphate content exceeding 2 mol/mol protein. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis three polypeptides were apparent: alpha (Mr 45,000), alpha' (Mr 40,000), and beta (Mr 26,000). Several lines of evidence conclusively demonstrated that the alpha and alpha' subunits are distinct polypeptides. Two-dimensional maps of 125I-tryptic peptides derived from the two proteins were related, but distinct. An antipeptide antibody was raised in rabbits which reacted only with the alpha subunit on immunoblots and failed to react with either the alpha' or beta subunits. Direct comparison of peptide sequences obtained from the alpha and alpha' subunits revealed differences between the two polypeptides. The results of this study clearly demonstrate that the alpha and alpha' subunits of casein kinase II are not related by post-translational modification and are probably encoded by different genes.

Amino Acid Sequence↗
PubMedDate not supplied

Cytologic diagnosis of rhabdomyosarcoma in a patient with germ cell tumor. A case report.

The development of sarcomas in patients with germ cell tumors is rare but has been reported previously. Theories about its pathogenesis include derivation of the tumor cells from pluripotential germ cells and malignant transformation from teratomatous elements. This report describes the occurrence of rhabdomyosarcoma in a patient with a history of mixed germ cell tumor of the testis who developed a malignant pleural effusion. Cytologic evaluation of the fluid revealed single malignant cells with high nuclear/cytoplasmic ratios and prominent nucleoli. While these features initially suggested a diagnosis of metastatic seminoma, careful cytologic and immunohistochemical examination revealed features consistent with metastatic rhabdomyosarcoma. The exfoliative cytologic findings of other germ cell tumors are also discussed.

Adult↗
PubMed1997

The immunohistochemical localization of transforming growth factor-beta 2 in the fetal and neonatal rat testis.

The localization of transforming growth factor beta-2 (TGF beta 2) in the fetal and neonatal testis (from day 13.5 of fetal life to postnatal day 9) was investigated by an immunohistochemical staining method employing a specific polyclonal antibody. Immunostaining appeared on fetal day 13.5 in primitive Sertoli cells as they begin to come in contact with each other and surround the germ cells to form the seminiferous cords. Staining in Sertoli cells was still clearly observed until fetal day 16.5 and became faint or undetectable from fetal day 18.5 onwards. In fetal-type Leydig cells, a positive reaction for TGF beta 2 appeared on day 16.5 and became very intense from day 18.5 onwards. In the germ cells, immunoreactivity for TGF beta 2 appeared on fetal day 20.5, rose to a maximum on postnatal day 4 and decreased thereafter. On postnatal day 9, staining was still present in type A spermatogonia and absent in type B spermatogonia. No immunoreactivity was detected in peritubular cells on any day studied. In conclusion, our results are in favour of an autocrine/paracrine role of TGF beta 2 in the differentiation of the testis during the perinatal period. It may be involved in the organization of the seminiferous cords, the regulation of testosterone production and the regulation of the number of germ cells. When compared with the immunolocalization of TGF beta 1 that we have previously reported [1], the present study suggests that the roles of TGF beta 2 in the developing rat testis can be specific but also overlap from those of TGF beta 1.

Animals↗
PubMed2000

Pattern of gonadotropin secretion and ultrasonographic evaluation of developmental changes in the testis of early and late maturing bull calves.

This was a study that retrospectively analyzed serum gonadotropin secretion and the ultrasonographic appearance of the testis during development in prepubertal bull calves to determine whether there were differences between early and late maturing bulls. Blood samples were taken every other week from 2 wk of age until puberty. Samples were also taken at 12 minute intervals for 12 hours at 4, 10, 20, 25, 30, 35, 40 and 45 wk of age. The GnRH treatment was administered 10 hours after the start of each period of frequent blood sampling. Bull calves fell into two distinctive groups, with one group maturing between 36.6 and 44.2 wk (n = 12) and the other between 46.4 and 48.9 wk of age (n = 8). In samples taken every other week mean serum LH concentrations were greater in early maturing bulls than in late maturing bulls at 12, 14 and 16 wk of age (P<0.05). In blood samples taken every 12 minutes for 10 hours early maturing bull calves had higher mean serum LH concentrations at 4 and 10 wk of age (P<0.05) and higher LH pulse frequency at 10 and 20 wk of age (P<0.05). Mean serum LH concentrations at 4, 10 and 40 wk of age and LH pulse frequency at 10 and 20 wk of age were negatively correlated with age at puberty in bull calves. Mean pixel units of the right and left testis were higher from 34 to 40 wk of age in early maturing than in late maturing animals (P<0.05). It seems possible that hormone measurements and ultrasonographic characteristics of the testes could be developed into powerful tools for studies on the regulation of reproductive development and may aid in the prediction of reproductive potential.

Animals↗
PubMed1984

[Histological characteristics of the testis and epididymis of calves during postnatal development].

A histologic investigation was carried out of the testis and epididymis of a total of 42 calves of the Black-and-white grouped in 14 test groups. The animals were currently castrated after the age of 2 months and were raised in the open. Material for investigation was taken on the 1st and 15th day, and once in every following month up to the completion of 12 months of age. It was found that up to the 3rd month there was no lumen in the testicular tubules, but only differentiating Sertoli cells and gonocytes were seen. Spermatogonia and spermatocytes were first observed at the age of 4 months, and prespermatids and sporadic spermatids--in the 8th month. Single spermatozoa in the phase of final shaping appeared in the 10th month, and completed spermatogenesis--in the 11th month. The histogenesis of the wall of the tubules of the epididymis completed earlier as compared to the histogenesis process in the testis--by the age of 8 months. Later on there were morphologically in the epididymis some proliferation and folding of the tubules with widening of the lumen, whereupon the amount of the surrounding connective tissue relatively decreased.

Animals↗
PubMed1976

Cancer of the undescended or maldescended testis.

An analysis of 45 cryptorchids (by history or examination) with a testicular cancer treated at Memorial Hospital, between 1934 and 1973, is presented. Twenty-five patients had the cryptorchid state repaired at ages four to 27 years, either spontaneously or by orchiopexy or hormonal therapy. Ipsilateral (24) or contralateral (one) intrascrotal testis tumors developed four to 47 years later. Twenty cryptorchid patients presented with ipsilateral inguinal (eleven), abdominal (seven), or contralateral intrascrotal (two) tumors. There were 18 pure seminomas, 17 embryonal carcinomas, nine teratocarcinomas, and one reticulum cell sarcoma. Five year survival rates as estimated by the product-limit method were 60% for the unrepaired cases and 41% for the repaired cases. The survival seems to follow histologic type and anatomical stage, whether the testis is within the scrotum or not. Five year survival similarly estimated was 78% in the seminomas and 29% in the other tumors. Twelve of thirteen survivors (including nine with seminoma) received postoperative irradiation to the regional lymphatics and eleven were without recurrent tumor for periods ranging from six to 28 years.

Adolescent↗
PubMed1985

Newborn and immature rat testes contain gonadotropin-releasing hormone (GnRH) receptors, and their testosterone production is stimulated by a GnRH agonist in vitro.

The possibility of direct gonadal effects of GnRH in the newborn and immature rat testis was studied by two approaches: the presence of specific high-affinity receptors for a GnRH agonist analogue (Buserelin) was studied in animals between the ages of 1 and 60 days; and the effect of Buserelin on testicular testosterone production was studied in vitro. GnRH receptors were found in the testis tissue at all ages studied, and there was a gradual increase in the number of binding sites per testis with advancing age. The highest concentration of GnRH receptors was found in the rat testis tissue on day 1 of life, 3.2 +/- 0.4 fmoles/g tissue (mean +/- SE, n = 5). The binding declined to a nadir, 1.5 +/- 0.5 fmoles/g tissue (in = 10), on day 15 post-partum (p less than 0.05), and thereafter increased again gradually to a level of 2.7 +/- 0.3 fmoles/g (n = 5) on day 60 of life (p less than 0.05). The biphasic concentration curve of GnRH receptors followed closely the volume density of Leydig cells in the developing rat testis. Buserelin at a concentration of 10(-7) M was able to stimulate significantly (p less than 0.01), by 98-146%, the testosterone production of decapsulated testes at all ages studied between 1 and 60 days of life. These results indicate the presence of functional GnRH receptors in the testis tissue of newborn and immature rats.

Animals↗
PubMed1997

Suppressive effect of perinatal testes on the differentiation of fetal ovaries transplanted into adult males in the rat.

A 14 d ovarian primordium was transplanted with a fetal testis (13-18 d and 21 d of gestation) or a neonatal testis (15, 20, 30 and 45 d after birth) into the renal subcapsular position of an adult male rat. Two weeks after transplantation, transplants were examined as to the degree of ovarian and testicular differentiation. In the combination of a 14 d ovary and a 13 d testis, there were 3 types of result: either the ovary or the testis alone developed or both gonads developed well. Ovaries transplanted in union with 15-18 d testes did not develop, although the testes developed normally. Some ovaries in union with 21 d testes developed normally. In combination with infantile testes, the incidence of developed ovaries increased as the age of testes advanced. These results suggest that the 13 d fetal testes begin to suppress the development of cotransplanted 14 d ovaries, that 14-18 d fetal testes maintain such suppressive effects and that this effect gradually diminishes in infantile testes as they progress toward 45 d after birth.

Animals↗
PubMed1999

The prepubertal testis: a quiescent or a silently active organ?

The development of the testis is characterised by dramatic changes between birth and adulthood. The most conspicuous changes take place during puberty, when the seminiferous tubule diameter increases significantly owing to an important proliferation of germ cells, giving rise to spermatozoa, and to the development of a tubular lumen; in the interstitial tissue characteristic Leydig cells appear and secrete high levels of testosterone. These pubertal changes of the testis can be detected clinically since they result in testicular volume and serum androgen level increments. The prepubertal testis has classically been defined as a quiescent organ. However, since adequate stereological methods for microscopic analysis are available, it has been shown that the male gonad triplicates its volume between birth and the onset of puberty. Sertoli cells and spermatogonia proliferate intensely; this is critical for the development of quantitatively normal adult spermatogenesis. Seminiferous tubule volume increases owing to an increment in tubular length, not diameter. Sertoli cells are also functionally active during childhood: they produce high amounts of anti-Müllerian hormone during the whole prepubertal period, and inhibin B until the age of 2-4 years. Anti-Müllerian hormone and inhibin seem to play a role as modulators of the proliferation and differentiation of Leydig cell precursors.

Adult↗
PubMed2005

A reference map and identification of porcine testis proteins using 2-DE and MS.

The development of the testis is essential for maturation of male mammals. A complete understanding of proteins expressed in the testis will provide biological information on many reproductive dysfunctions in males. The purposes of this study were to apply a proteomic approach to investigating protein composition and to establish a 2-D PAGE reference map for porcine testis proteins. MALDI-TOF MS was performed for protein identification. When 1 mg of total proteins was assayed by 2-D PAGE and stained with colloidal CBB, more than 400 proteins with a pI of pH 3-10 and M(r) of 10-200 kDa could be detected. Protein expression varied among individuals, with CV between 4.7 and 131.5%. A total of 447 protein spots were excised for identification, among which 337 spots were identified by searching the mass spectra against the NCBInr database. Identification of the remaining 110 spots was unsuccessful. A 2-D PAGE-based porcine testis protein database has been constructed on the basis of the results and will be published on the WWW. This database should be valuable for investigating the developmental biology and pathology of porcine testis.

Animals↗
PubMedDate not supplied

Immunocytochemical localization of bioregulatory peptides in marmoset testes.

Immunocytochemical localization of neuropeptides (beta-endorphin, substance P, arginine vasopressin, oxytocin), pituitary hormones (adrenocorticotropin, prolactin, growth hormone, follicle stimulating hormone (FSH), gonadal inhibin, gastrin, and human chorionic gonadotrophin (hCG)) was carried out in marmoset testis during development. Both intensity of immunostaining and distribution of these peptides in testicular compartments viz. seminiferous tubules and Leydig cells changed dramatically during development. In vitro biosynthesis of inhibin and FSH was increased by hCG, whereas prolactin (5 micrograms) and prostatic inhibin peptide suppressed the synthesis of these hormones.

Aging↗
PubMedDate not supplied

Related function of mouse SOX3, SOX9, and SRY HMG domains assayed by male sex determination.

Sox genes encode proteins related to each other, and to the sex determining gene Sry, by the presence of a DNA binding motif known as the HMG domain. Although HMG domains can bind to related DNA sequences, Sox gene products may achieve target gene specificity by binding to preferred target sequences or by interacting with specific partner proteins. To assess their functional similarities, we replaced the HMG box of Sry with the HMG box of Sox3 or Sox9 and tested whether these constructs caused sex reversal in XX mice. Our results indicate that such chimeric transgenes can functionally replace Sry and elicit development of testis cords, male patterns of gene expression, and elaboration of male secondary sexual characteristics. This implies that chimeric SRY proteins with SOX HMG domains can bind to and regulate SRY target genes and that potential SRY partner factor interactions are not disrupted by HMG domain substitutions. genesis 28:111-124, 2000.

Amino Acid Sequence↗
PubMed1987

Structure of the right testis of sexually mature genetically female fowl experimentally masculinized during embryonic life and submitted to a posthatching left castration.

Posthatching left castration of genetically female fowl, Gallus domesticus, preceded, during embryonic life, by a masculinizing treatment associating a testis graft and an antiestrogen resulted in the development of the right rudimentary gonad into a testis. Examined after the sexual maturity, the right testis of most treated animals was entirely composed of seminiferous tubules possessing a spermatogenic cell complement. Spermiogenesis proceeded to the stage of spermatozoon in 4 out of 17 treated animals and was almost as well organized as in a normal cock testis in 3 of them. Testis development appeared then as clearly improved, compared to that described previously in only left-castrated, with or without treatment with an antiestrogen, or only sex-reversed female fowl. The possible mechanism of this improvement is discussed.

Animals↗
PubMed2006

Gonadal changes and blood sex steroids levels during natural sex inversion in the protogynous Mediterranean red porgy, Pagrus pagrus (Teleostei: Sparidae).

Changes in gonadal structure and serum levels of sex steroids were investigated during natural sex inversion from female to male in reared populations of the protogynous Mediterranean red porgy, Pagrus pagrus. Four developmental phases were identified by histological observation: female, early transitional (ETr), late transitional (LTr), and male phases. At female phase, a few nests of spermatogonia were observed at the posterior-ventral part of the gonad mainly in females out of the breeding season. At ETr phase, spermatogonial proliferation occurred while perinucleolar oocytes showed signs of degeneration. At LTr phase, seminiferous lobules were formed and spermatogonial proliferation expanded along the ovary which degenerated. All types of male germ cells could be found. At male phase, functional testis underwent active spermatogenesis while small ovarian remnants associated to fat tissue could be detected. Both 17beta-estradiol (E2) and estrone (E1) blood levels were significantly lower in fish at transitional and male phases in comparison to breeding females, while levels of 11-ketotestosterone (11-KT) and testosterone (T) gradually increased in the transitional and male phases. In conclusion, the protogynous P. pagrus possess a delimited type bisexual gonad with a medio-dorsal ovarian area and a latero-ventral testicular zone. Sex inversion starts mainly after the female breeding season with an active spermatogonial proliferation. The testis tissues develop while ovarian tissues regress to disappear completely in the functional male. This process is accompanied by a sharp decrease of estrogens levels and a progressive increase of androgens levels. The physiological significance of such endocrine changes is discussed.

Animals↗
PubMed1986

A vector-primer-cloner-sequencer plasmid for the construction of cDNA libraries: evidence for a rat glyceraldehyde-3-phosphate dehydrogenase-like mRNA and a ferritin mRNA within testis.

We have developed a vector-primer-cloner-sequencer (VPCS), based on the pUC plasmids, which is easy to prepare. Stable cDNA libraries have been generated from human, rat, and bull testis. The inserts are of various size classes, including high-molecular-weight reverse transcripts which can be easily sequenced. The utility of this VPCS has been demonstrated by isolating a rat ferritin and glyceraldehyde-3-phosphate dehydrogenase-like clone.

Animals↗
PubMed2000

Effects of thyroid and luteinizing hormones on the onset of precursor cell differentiation into leydig progenitor cells in the prepubertal rat testis.

Leydig cells in the adult rat testis differentiate during the neonatal-prepubertal period. However, the stimulus for the initiation of their differentiation is still not clear. In the present study our objectives were to test the effects of thyroid hormone and LH on the initiation of precursor cell differentiation into Leydig cells in the prepubertal rat testis. Four groups of Sprague-Dawley rats were used. All treatments began at postnatal Day 1. Rats in groups I, II, and III received daily s.c. injections of saline (200 microl, controls), triiodothyronine (T(3), 50 microg/kg body weight, hyperthyroid), and LH (ovine LH 10 microg/rat/day), respectively. Rats in group IV were made hypothyroid from postnatal Day 1 by adding 0.1% propylthiouracil (PTU) to their mother's drinking water. Testes of rats were collected at 7, 8, 9, 10, 11, 12, 16, and 21 days of age, fixed in Bouin's solution, and embedded in paraffin for immunocytochemical studies. Immunoexpression of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and LH receptors (LHR) in testicular interstitial cells (other than the fetal Leydig cells) was observed using the avidin-biotin method. In control rats, out of all spindle-shaped cell types in the testis interstitium, only the peritubular mesenchymal cells showed positive immunolabeling for 3beta-HSD, beginning from the postnatal Day 11. However, positive immunolabeling for LHR was first detected in these cells at Day 12, i.e., after acquiring the steroidogenic enzyme activity. In T(3)-treated rats 3beta-HSD positive spindle-shaped cells were first observed at Day 9 (i.e., 2 days earlier than controls), and LHR-positive cells were first observed on Day 11 (2 days later than obtaining 3beta-HSD immunoactivity); they were exclusively the peritubular mesenchymal cells. The 3beta-HSD- and LHR-positive spindle-shaped cells were absent in the testis interstitium of LH-injected rats from Days 7 through 12 but were present at postnatal Day 16. In addition, more fetal Leydig cell clusters and fetal Leydig cells in mitosis were present in LH-treated rats compared to rats in all other treatment groups. Following their first detection, the number of positive cells for each protein continued to increase at each subsequent age in controls, T(3)-, and LH-injected groups. In PTU rats, 3beta-HSD and LHR-positive spindle-shaped cells were absent throughout the experimental period. From these observations, it is possible to suggest the following regarding the developing rat testis interstitium. 1) The precursor cells for the adult generation of Leydig cells in the postnatal rat testis are the peritubular mesenchymal cells. 2) Luteinizing hormone does not initiate the onset of mesenchymal cell differentiation into Leydig cells, instead it delays this process. However, daily LH treatment causes mitosis in fetal Leydig cells and increase in fetal Leydig cell clusters. 3) Thyroid hormone is critical to initiate the onset of mesenchymal cell differentiation into adult Leydig cells.

3-Hydroxysteroid Dehydrogenases↗
PubMed1993

Cellular location and age-dependent changes of the regulatory subunits of cAMP-dependent protein kinase in rat testis.

This study was undertaken to examine the expression and cellular location of the various cAMP-dependent protein kinase (PKA) subunits in different testicular cell types, using cDNA probes, isoenzyme-specific antibodies and activity measurements. Amounts of mRNA and protein were examined in cultured Sertoli cells, cultured peritubular cells, germ cells (pachytene spermatocytes, round spermatids), Leydig cell tumours as well as whole testes from rats of various ages. In Sertoli cells, there was a good correlation between the amount of mRNA and the respective immunoreactive proteins. In other types of cell, such as germ cells and Leydig tumour cells, this was not always the case. Large amounts of RII beta mRNA were found in Leydig tumour cells, whereas the amount of immunoreactive protein was low. Furthermore, large amounts of small-sized, germ cell-specific mRNAs for RI alpha (1.7 kb) and RII alpha (2.2 kb) were also found in the developing rat testis after 30 to 40 days of age, but the large amounts of mRNA were only partially reflected at the protein level. Pachytene spermatocytes and round spermatids were practically devoid of both RII alpha and RII beta protein. During spermatid differentiation, there was a decrease in RI alpha and an increase in RII alpha protein. Cell specific distribution of the various PKA subunits in testicular cell types is described. In some types of cell, discrepancies between mRNA and protein were demonstrated, which clearly suggest cell specific differences in translational efficiencies for some of these mRNAs, particularly the small-sized mRNAs for RI alpha and RII alpha in meiotic and post-meiotic germ cells.

Aging↗