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PubMed1995

Expression of mRNA and immunocytochemical localization of inhibin alpha- and inhibin beta A-subunits in the fetal sheep testis.

In order to investigate the ontogeny of gonadal inhibin production in the male fetal sheep, testes were collected from male fetuses at days 70, 100, 130 and 140 of gestation (term = 145 days). The expression and localization of inhibin alpha- and inhibin beta A-subunit mRNA and protein were evaluated using in situ hybridization and immunocytochemistry. The expression of inhibin alpha-subunit mRNA was localized within the seminiferous cords of the developing fetal testis and progressively increased with gestational age. Immunostaining corresponding to immunoreactive inhibin alpha-subunit was detected in Sertoli cells within the seminiferous cords at days 100, 130 and 140 of gestation. In addition, immunostaining was detectable in a small proportion of Leydig cells. No expression of inhibin beta A-subunit mRNA or immunoreactivity was detected in any testicular tissue at any stage of gestation. These data show that the Sertoli cells of the developing fetal sheep testis have the capacity to produce inhibin alpha-subunit by day 100 of gestation and that production increases during late gestation.

Animals↗
PubMed1989

Corticosteroid 11 beta-dehydrogenase in rat testis.

Corticosteroid 11 beta-dehydrogenase, the enzyme that catalyzes the oxidation of the biologically active steroid cortisol to its inactive metabolite cortisone, is present in testis. Since excess cortisol in men and other mammals and excess corticosterone in rodents cause physiological abnormalities including abnormal testicular function, it was pertinent to study the cellular distribution of 11 beta-dehydrogenase in the testis. Purified antiserum directed against homogeneous rat 11 beta-dehydrogenase was used to localize the enzyme in the developing rat testis. With immunofluorescence, the enzyme was not detectable in fetal testis or in the testis of young male rats until the 26th day of development. A few interstitial cells were stained in the testis of 26-day-old animals. In the testis of 31-day-old rats many cells in the interstitium were positive. In adult animals the entire interstitial region displayed bright fluorescence. Depleting animals of germ cells did not abolish the fluorescence. The appearance of this enzyme correlates temporally with the postnatal increase in Leydig cell number and the developmental rise in serum testosterone. We suggest that 11 beta-dehydrogenase of Leydig cells protects the testis from the deleterious effects of cortisol.

11-beta-Hydroxysteroid Dehydrogenases↗
PubMed1999

Developmental expression of steroidogenic factor 1 in a turtle with temperature-dependent sex determination.

A variety of reptiles possess temperature-dependent sex determination (TSD) in which the incubation temperature of a developing egg determines the gonadal sex. Current evidence suggests that temperature signals may be transduced into steroid hormone signals with estrogens directing ovarian differentiation. Steroidogenic factor 1 (SF-1) is one component of interest because it regulates the expression of steroidogenic enzymes in mammals and is differentially expressed during development of testis and ovary. Northern blot analysis of SF-1 in developing tissues of the red-eared slider turtle (Trachemys scripta), a TSD species, detected a single primary SF-1 transcript of approximately 5.8 kb across all stages of development examined. Analysis by in situ hybridization indicated nearly equivalent SF-1 expression in early, bipotential gonads at male (26 degrees C)- and female (31 degrees C)-producing incubation temperatures. In subsequent stages, as gonadal sex first becomes histologically distinguishable during the temperature-sensitive period, SF-1 expression increased in gonads at a male-producing temperature and decreased at a female-producing temperature, suggesting a role for SF-1 in the sex differentiation pathway. SF-1 message was also found in adrenal and in the periventricular region of the preoptic area and diencephalon, but there was no apparent sex bias in these tissues at any stage examined. The overall developmental pattern of SF-1 mRNA expression in T. scripta appears to parallel that found in mammals, indicating possible homologous functions.

Animals↗
PubMed1983

Sexual development, maturation, and behavior.

The Y chromosome directs the primitive gonad to develop into a testis. Without a Y chromosome an ovary will develop, but that ovary will not be normal unless two X chromosomes are present. Active intervention is needed for male differentiation; internal male structures will not be found unless sufficient and effective testosterone is secreted by the immature testis to develop the wolffian duct system into the internal male reproductive tract. The testis must also secrete mullerian regression factor to cause the demise of the internal female duct structures. Finally, enough 5-alpha-reductase activity must be present to convert testosterone into dihydrotestosterone for the normal virilization of the male external genitalia. Without testosterone or its receptor sites, without dihydrotestosterone, and without mullerian regression factor, the reproductive system is female. Early in life, a child assumes both a gender identity (an awareness of what sex he or she belongs to) and a gender role (behavior deemed to be more or less characteristic of one sex or the other). As puberty is passed, sexual orientation becomes more obvious, although the development of that orientation has probably been in the making since early childhood. Early developmental hormone milieu and social environment undoubtedly all play a role in subsequent sexual behavioral patterns, but the extent to which each of these impacts upon that behavior still remains unknown.

Adolescent↗
PubMed1995

Development of the gubernaculum and processus vaginalis in freemartinism: further evidence in support of a specific fetal testis hormone governing male-specific gubernacular development.

BACKGROUND: Freemartinism occurs in some species of ruminants and affects most female bovine fetuses in heterosexual, multiple pregnancies owing to fusion of the chorionic blood circulations soon after implantation. Maldevelopment of the ovaries and Müllerian ducts have been described and recognized as resulting from exposure of their respective primordia to an excess of anti-Müllerian hormone. The present study aimed to analyse the prenatal growth and development of the gubernaculum in freemartins to find out its possible affliction through foetal testis hormones derived from their male co-twin. METHODS: Histological sections of young and drawings and photographs of further developed freemartins and control male and female bovine foetuses were analysed. The specimens had been collected earlier for analysis of the time course of male and female gonadal and genital development and its impairment associated with freemartinism. RESULTS: The gubernaculum of 35-40-day-old male and female fetuses was in the initial stage of development and of similar appearance in all specimens. Gubernacula of 60-70-day-old male fetuses differed from those of females of similar age in various respects: the male gubernaculum size was larger and extension of the processus vaginalis was deeper. Freemartins showed an intermediate development with some individuals resembling male and others resembling female agemates. During further development, gubernacula in males developed into muscular cremaster sacs, whereas those in females generally did not develop beyond the size and structural complexity of 70-day-old foetuses. Beyond day 70 of fetal life, gubernaculum development in freemartins definitely showed male characteristics with respect to size and growth of a processus vaginalis with a cremaster muscular wall. The male-like pattern of the outgrowth of the processus vaginalis changed during the second half of prenatal life. Rather than its further deepening as in males, this structure became inverted to become emerging as a papilla-like structure from the inguinal abdomen bottom. An explanation is proposed for this unprecedented inversion, taking into account: (1) the faster and higher reaching rightsided ascent of the kidneys and gonads, (2) the femalelike outgrowth of the cranial gonadal suspensory ligaments, and (3) the absence of scrotum development. The ovaries and mesonephric remnants in developing freemartins, during their ascent together with the kidneys while remaining attached to the bottom of the developing processus vaginalis sacs via the gubernaculum ligament, are proposed to act together to pull up the bottom of the processus vaginalis sacs. From this action, "inverted hernia sacs" result as the irreversible consequence. CONCLUSION: The data support the concept that foetal testes act, via as an yet unidentified third hormone, to establish malelike development of gubernacula into muscular cremaster sacs. Further work is required to reveal the identity of this hormone. Furthermore, the apparent similarity of the freemartins' inverted processus vaginalis sacs and the fetal rodents' gubernacular cones suggests that the ruminants' and rodents' processus vaginalis are essentially similar structures. Thus there is no longer an urgent need to distinguish between two different types of gubernaculum development and testis descent in rodents and ruminants, respectively, and involving or not fetal gubernacular cones. The present observations may thus contribute to the development of a unified hypothesis for sexually dimorphic development of the gubernaculum throughout the mammalian class.

Androgens↗
PubMed1985

Canine cryptorchism and subsequent testicular neoplasia: case-control study with epidemiologic update.

A retrospective study of 2,912 cryptorchid dogs identified 14 breeds with significantly high risk. Among six distinct closely interrelated breed groups (e.g., toy, miniature, and standard poodles), the risk in the smaller breed was always greater than that in the larger relative, suggesting that genetically influenced maldescent could be, in part, related to physical size or the rate of growth of the involved structures. Testicular tumors were diagnosed in 5.7% of the cryptorchid dogs; half had only Sertoli cell tumors, one-third had only seminomas. The relative risk for Sertoli cell tumor or seminoma was not directly related to a familial risk for cryptorchism. Using the health experience of a control population composed of male dogs with anal sac disease (N = 4,184), there is an estimated relative risk of 9.2 in cryptorchid dogs to develop a testis tumor (95% confidence interval, 5.9-14.3) and 4.2 in dogs with inguinal hernia (95% confidence interval, 1.8-9.5). Considering that the anatomical development of the genital tract, testis descent, and tunic relationships in dog are very similar to that in man, and that the associations of cryptorchism and inguinal hernia with testis neoplasms are also similar, the dog should be an excellent model system to further investigate the causes of human cryptorchism.

Animals↗
PubMed1997

Cytoplasmic localization of cyclin D3 in seminiferous tubules during testicular development.

Using a newly developed polyclonal antibody against murine cyclin D3, we have found that protein levels of cyclin D3 were highly detectable only in thymus and testis in rats. Since testis offer unique opportunities to examine the cell cycle in vivo, we examined the temporal and spatial expression of cyclin D3 and the DNA synthesis indicator, proliferating cell nuclear antigen (PCNA), in the rat testis during development. The protein levels of cyclin D3 protein in testis from 7 days to 3 months old were almost constant and then decreased gradually thereafter. The protein levels of cyclin D1 and PCNA were high in the testis of 7- and 14-day-old rats and decreased during testicular development. In the seminiferous tubules of 7-day-old newborns, cyclin D3 was surprisingly located in cytoplasm of stem cells that had bigger nuclei than the nuclei of surrounding cells. Interestingly, cyclin D3 immunopositive cells did not immunostain with PCNA in nuclei. In the adult testis, anti-cyclin D3 antibody strongly stained the cytoplasm of early stage primary spermatocytes, lightly stained pachytene spermatocytes, but did not stain elongated spermatids. There was no detectable cyclin D3 in Sertoli cells, interstitial cells, or fibroblasts within seminiferous tubules, or in blood vessels within the interstitial matrix. The known cyclin D3 partner, cyclin dependent kinase 4, was located mainly in nuclei of spermatogonia and in early stage primary spermatocytes. Strong PCNA immunopositive staining was located in the nuclei of spermatogonia in adult testis. These results indicate that cyclin D3 is detectable in meiotically active male germ cells (PCNA-negative cells), but is conspicuously absent from mitotically active spermatogonia (PCNA-positive cells). Moreover, in contrast to in vitro reports, cyclin D3 is not located in the nucleus, but rather in the cytoplasm of male germ cells in vivo. Taken together, the presence of cyclin D3 in spermatocytes and its location in the cytoplasm lead us to speculate that cyclin D3 may have functions in male germ cells other than mitosis.

Aging↗
PubMed1994

Mouse A-myb encodes a trans-activator and is expressed in mitotically active cells of the developing central nervous system, adult testis and B lymphocytes.

C-myb encodes a transcriptional activator that is essential for the development of the hematopoietic system but appears to lack major roles in non-hematopoietic cells. The identification of two conserved myb-related genes, designated A-myb and B-myb, has raised the possibility that these genes are functional equivalents of c-myb in non-hematopoietic cells. Here, we report the isolation and preliminary characterization of the mouse A-myb gene. Mouse A-myb maps to the proximal region of chromosome 1 and encodes a transcriptional activator with properties similar to those of the c-myb and v-myb proteins. During embryo-genesis A-myb is predominantly expressed in several regions of the developing central nervous system (CNS) and the urogenital ridge. Expression in the CNS is confined to the neural tube, the hindbrain, the neural retina and the olfactory epithelium, and coincides with the presence of proliferating immature neuronal precursor cells. In the adult mouse, A-myb is expressed during the early stages of sperm cell differentiation and in B lymphocytes located in germinal centers of the spleen. Taken together, these results suggest a role for A-myb in the proliferation and/or differentiation of neurogenic, spermatogenic and B-lymphoid cells.

Amino Acid Sequence↗
PubMed2003

SOX8 is expressed during testis differentiation in mice and synergizes with SF1 to activate the Amh promoter in vitro.

Sox8 is a member of the Sox family of developmental transcription factor genes and is closely related to Sox9, a key gene in the testis determination pathway in mammals. Like Sox9, Sox8 is expressed in the developing mouse testis around the time of sex determination, suggesting that it might play a role in regulating the expression of testis-specific genes. An early step in male sex differentiation is the expression of anti-Müllerian hormone (AMH) in Sertoli cells. Expression of the Amh gene during sex differentiation requires the interaction of several transcription factors, including SF1, SOX9, GATA4, WT1, and DAX1. Here we show that SOX8 may also be involved in regulating the expression of Amh. Expression of Sox8 begins just prior to that of Amh at 12 days post coitum (dpc) in mouse testes and continues beyond 16 dpc in Sertoli cells. In vitro assays showed that SOX8 binds specifically to SOX binding sites within the Amh minimal promoter and, like SOX9, acts synergistically with SF1 through direct protein-protein interaction to enhance Amh expression, albeit at lower levels compared with SOX9. SOX8 and SOX9 appear to have arisen from a common ancestral gene and may have retained some common functions during sexual development. Our data provide the first evidence that SOX8 may partially compensate for the reduced SOX9 activity in campomelic dysplasia and substitute for Sox9 where Sox9 is either not expressed or expressed too late to be involved in sex determination or regulation of Amh expression.

Animals↗
PubMed1987

Hyperplasia of spermatic cord nerves: a sign of testicular absence.

The comparative histologic study of the spermatic cord in the absence of testis, epididymis-testis separation, and normal development of both testis and epididymis, revealed that there is nerve trunk hyperplasia and hypertrophy in absence of the testis. This finding may greatly aid the diagnosis of testicular absence in the management of impalpable testes.

Cryptorchidism↗
PubMed1988

Autosomal genes involved in mammalian primary sex determination.

Beginning with findings made during the late 1950s and early 1960s, evidence continues to accumulate in support of the hypothesis that the mammalian Y chromosome carries a gene that induces the undifferentiated foetal gonad in XY individuals to develop as a testis. Recently a DNA sequence has been isolated from the human Y chromosome that appears to be the hypothesized Y-linked testis-determining gene, and advances have also been made toward identifying genes that interact with the Y-linked testis-determining (Tdy) gene to initiate testis formation. These loci have been identified in specific stocks of mice carrying the mutant Thp or TOrl allele at the T locus located on chromosome 17, and in crosses involving the transfer of a Y chromosome from two populations of Mus domesticus into the genomes of specific inbred strains of mice. The data in both cases support the hypothesis that there are several loci involved in testis determination and that abnormal interaction of these loci disrupts initiation of testis determination, resulting in development of ovarian tissue in XY individuals.

Animals↗
PubMed1995

The effect of ultrasound exposure in utero on the development of the fetal mouse testis: adult consequences.

The effects of exposure in utero to 1 MHz, continuous-wave ultrasound on adult growth and testicular development in the mouse was investigated. The spatial peak temporal average intensity (ISPTA) employed ranged from 1 to 10 W/cm2, with exposure durations (t) of 200 s to 20 s. Exposures were made on days 9, 12 or 15 of gestation. Results showed an increase in postpartum deaths, an increase in the number of stillbirths, and a decrease in litter size when I2 t > or = 1125 W2 s/cm4, such that there was significant loss of pups. Birthweights of pups from nearly all dosage groups was significantly lower than that of the sham or cage control groups. Results also showed that males exposed to ultrasound in utero had decreased testis size and decreased daily sperm production ranging from 9% to 30%. This study showed that ultrasound exposure in utero is capable of disrupting fetal development and having potential subsequent effects on fertility in the adult male.

Animals↗
PubMed2003

Molecular cloning and expression of Sox17 in gonads during sex reversal in the rice field eel, a teleost fish with a characteristic of natural sex transformation.

The Sox is a large family of genes which encode transcription factors with high-mobility-group DNA binding domain related to the SRY, with diverse roles in development, and a few of them are involved in sex determination and differentiation. We report here the identification of Sox17 gene of the rice field eel, a teleost fish with a characteristic of natural sex transformation. This gene, located on chromosome 5, consists of two exons which encode a 399-amino acid protein with a conserved HMG box. Phylogenetic analysis shows that the rice field eel Sox17 fits within the Sox17 clade of vertebrates. The rice field eel Sox17 was dominantly expressed in gonads of male, female, and intersex, besides in brain and spleen. Gene expression analysis by in situ hybridization showed its expression in testis, ovary, and ovotestis, and specifically in the gonadal lamellae of ovary, ovotestis, and testis and developing spermatogenic cells of testis, suggesting that they have potentially important roles in gonadal differentiation during sex reversal in this species.

Amino Acid Sequence↗
PubMed2001

The histopathology of iatrogenic cryptorchid testis: an insight into etiology.

PURPOSE: Iatrogenic undescended testis may develop after inguinal hernia repair, presumably as a result of mechanical tethering of the testis or cord in scar tissue. Because some true cryptorchid testes appear to be completely descended at birth and later ascend during childhood, some iatrogenic undescended testes may be low lying undescended testes. To determine whether iatrogenic undescended testes may be unrecognized cryptorchid testes at herniorrhaphy we examined biopsies of iatrogenic undescended testes and the corresponding contralateral descended testis. MATERIALS AND METHODS: Between 1985 and 1999 bilateral testis biopsies were obtained at orchiopexy in 37 boys 1.5 to 11.8 years old who previously underwent inguinal hernia correction. Histomorphometric analysis of germ cell counts was performed on the undescended and contralateral descended testes, and compared to the count in bilateral biopsies of 37 age and position matched patients with true unilateral cryptorchidism. RESULTS: There were no significant differences in volume or total and differential germ cell counts in the undescended and contralateral descended testes in the study groups and age matched controls with primary unilateral cryptorchidism. The mean number of germ cells per tubule in the undescended testis in patients with a greater than 5-year interval from herniorrhaphy to orchiopexy was significantly decreased compared to those with an operative interval of less than 5 years (0.27 +/- 0.33 versus 0.93 +/- 1.4, p = 0.026). CONCLUSIONS: Some patients with iatrogenic undescended testis may have an unrecognized low cryptorchid testis. Careful physical examination before and after inguinal surgery is recommended. The early repair of iatrogenic undescended testis is warranted to prevent further damage.

Child↗
PubMed1985

[Effect of an analog of proline (L-azetidine-2-carboxylic acid) on in vitro differentiation of the rat fetal testis].

The initial stages of the development of the seminiferous cords involve the differentiation and the aggregation of primordial Sertoli cells opposite to cells which acquire a mesenchymal-like aspect. The hypothesis that the development of the seminiferous cords depends on epithelial-mesenchymal relations between the two cell types was submitted to experimental test. Male gonadal primordia of rat fetuses were cultured in vitro in a synthetic medium containing the proline competitor, L-Azetidine-2-Carboxylic Acid. This drug is known to disturb the synthesis and secretion of collagen and proline-containing proteins. It prevents testicular organogenesis or destroys it if it has begun. It suppresses the expression of laminin and fibronectin in the gonadal primordium. These observations are taken as evidence that cellular correlations of the epithelial-mesenchymal type play a role in the development of the testis as they do in that of other organs.

Animals↗
PubMed2001

MT1-MMP in rat testicular development and the control of Sertoli cell proMMP-2 activation.

Metalloproteases (MMPs) are likely to be involved in the restructuring events occurring in the testis throughout development. We here demonstrate that membrane-type 1 (MT1)-MMP, a physiological activator of proMMP-2 under TIMP-2 control, is present within the testis together with MMP-2 and TIMP-2. In the prepubertal testis MT1-MMP immunoreactivity was uniformly distributed, whereas in the adult it was confined to the apical compartment of the tubules, where meiosis and spermiogenesis occur. We further showed that the two cell lineages (somatic and germinal) expressed MT1-MMP and TIMP-2, whereas MMP-2 was of somatic origin. To get a better picture into proMMP-2 activation, use was made of a model of cultured Sertoli cells treated with FSH or co-cultured with germ cells to mimic an immature or a mature developmental period, respectively. We found that follicle-stimulating hormone enhanced the expression of MMP-2 and TIMP-2 but not of MT1-MMP, and promoted the activation of proMMP-2. In co-cultures, a tremendous elevation and activation of MMP-2 was observed, which might relate to the processed MT1-MMP form solely detected in germ cells. That MMP-2 synthesis and activation are under local (germ cells) and hormonal (follicle-stimulating hormone) regulation emphasizes the importance of MMPs in testicular physiology.

Animals↗
PubMed1986

[Seminoma of testis. Analysis of failures and development of therapeutic strategies. Apropos of a Lyons series of 117 cases].

From Jan.61 to Dec.81, 117 patients with seminoma of testis were treated in the Leon Berard Centre, Lyon. All had undergone lymphography during investigation of possible extension, 19 were treated with 200 KV up 1966, 64 with Cobalt up to 1978 and 29 with photons x of 18 MV since that date. From 1979 adjuvant chemotherapy has always included cisplatinum. The 5 years survival rate was 95% of stage I (51/54 cases), 72% of stage II (26/36 cases) and 1/7 of stage III. Unsuccessful treatment of neoplasm was noted in 23 patients, in 80% of cases during the first two years and involving mainly pulmonary metastases. Three patients had mediastinal metastases while recovery surgery was possible in 4 cases. Three fatal iatrogenic complications were observed. Since the use of high energies, particularLy x beams of 18 MV there has been almost total absence of radic complications. Therapy now proposed is as follows: stage I: surgery plus radiotherapy; stage II A-B: surgery and irradiation avoiding mediastinum; stage II C and III: primary chemotherapy.

Adult↗
PubMed1994

Carcinoma in situ of the testis followed by an overt malignant teratoma of the testis within 12 months.

A case of bilateral metachronous testicular non-seminomatous germ cell tumour (NSGCT) is presented. The second tumour was preceded by carcinoma in situ, diagnosed at the time of the first orchidectomy. The patient was placed under active surveillance and 1 year later the second testis tumour developed. A second orchidectomy was performed and testosterone replacement begun. Carcinoma in situ of the testis is discussed.

Adult↗