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PubMed2003

Differentiation of adult-type Leydig cells occurs in gonadotrophin-deficient mice.

During mammalian testis development distinct generations of fetal and adult Leydig cells arise. Luteinising hormone (LH) is required for normal adult Leydig cell function and for the establishment of normal adult Leydig cell number but its role in the process of adult Leydig cell differentiation has remained uncertain. In this study we have examined adult Leydig cell differentiation in gonadotrophin-releasing hormone (GnRH)-null mice which are deficient in circulating gonadotrophins. Adult Leydig cell differentiation was assessed by measuring expression of mRNA species encoding four specific markers of adult Leydig cell differentiation in the mouse. Each of these markers (3beta-hydroxysteroid dehydrogenase type VI (3betaHSD VI), 17beta-hydroxysteroid dehydrogenase type III (17betaHSD III), prostaglandin D (PGD)-synthetase and oestrogen sulphotransferase (EST)) is expressed only in the adult Leydig cell lineage in the normal adult animal. Real-time PCR studies showed that all four markers are expressed in adult GnRH-null mice. Localisation of 3betaHSD VI and PGD-synthetase expression by in situ hybridisation confirmed that these genes are expressed in the interstitial tissue of the GnRH-null mouse. Treatment of animals with human chorionic gonadotrophin increased expression of 3betaHSD VI and 17betaHSD III within 12 hours further indicating that differentiated, but unstimulated cells already exist in the GnRH-null mouse. Thus, while previous studies have shown that LH is required for adult Leydig cell proliferation and activity, results from the present study show that adult Leydig cell differentiation will take place in animals deficient in LH.

17-Hydroxysteroid Dehydrogenases↗

PubMed1991

Hypoxanthine phosphoribosyl transferase deficiency, haematopoiesis and fertility in the mouse.

We have looked for effects of deficiency in hypoxanthine phosphoribosyl transferase (HPRT) in the mouse comparable to non-behavioural consequences of HPRT-deficiency in humans. HPRT-deficient humans show abnormalities in haematopoiesis and, in heterozygotes, there is strong selection in haematopoietic tissues against HPRT-deficient cells arising as a result of X-chromosome inactivation. We have examined two situations in mice in which HPRT- and HPRT+ cells occur in the same individual. First, in chimaeras resulting from the injection of HPRT- embryonal stem cells into HPRT+ blastocysts the fate of HPRT- and HPRT+ cell populations was monitored by their expression of different isozymes of glucose phosphate isomerase and also, in those chimaeras that resulted from injecting the male ES cells into female blastocysts, by in situ hybridisation using a Y-chromosome-specific repetitive DNA probe. There was a small statistically significant selection against the HPRT- population in haematopoietic tissues in both XX in equilibrium with XY and XY in equilibrium with XY chimaeras. Second, in female mice doubly heterozygous for HPRT-deficiency and for an electrophoretic variant of the X-linked enzyme phosphoglycerate kinase, there was a similar small statistically significant selection against the HPRT- population in haematopoietic tissues. While further work is required to establish whether this selection is a consequence of the HPRT mutation, it is clear that any selection against cells in the haematopoietic system as a consequence of HPRT-deficiency is at most small compared with the effect seen in humans. In HPRT-deficient human males surviving beyond the normal age of puberty, there is testicular atrophy. However, we find no effect of HPRT-deficiency on the fertility of either male or female mice. Thus, as with effects on behaviour, the consequences of HPRT-deficiency for haematopoiesis and testis development in the mouse are at most small compared with those in the human. We conclude that the reason for the difference in effects between the two species lies in a difference in purine-related intermediary metabolism per se, rather than in its interaction with brain amine biochemistry.

Animals↗

PubMed1990

[A case of mixed gonadal dysgenesis: examination of chromosomes in various parts of the body].

We report a 17 year old patient with mixed gonadal dysgenesis (MGD) raised as a female. The patient had undergone amputation of the phallus at 10 months of age and showed short and masculinized characteristics with ambiguous external genitalia. Abdominal exploration revealed a hypoplastic uterus with a fallopian tube and streak gonad on the right side and a poorly developed testis and epididymis with vas deferens on the left side. Following chromosomal analysis, the cultured peripheral lymphocytes and bone marrow cells showed 45,X in karyotype. Although the mosaicism of 45,X/46,X+mar was revealed by both G- and Q-banding methods in the cultured skin fibroblasts and the cells of various gonadal organs examined, the rate of 45,X karyotype was high (77-97%) in the cells of each organ tissue. Endocrine examinations showed plasma testosterone levels to be slightly higher than those of a normal female. The response of plasma testosterone and estradiol to HCG was low. Hypergonadotropic hypogonadism was revealed.

Adolescent↗

PubMed2006

Duplicated insertion mutation in the microtubule-associated protein Spag5 (astrin/MAP126) and defective proliferation of immature Sertoli cells in rat hypogonadic (hgn/hgn) testes.

Male rats with hypogonadism (hgn/hgn) experience sterility from testicular dysplasia, which is controlled by a single recessive gene, hgn. The postnatal growth of the seminiferous tubules was severely affected. In this study, we localized the hgn locus to a 320 kb region on rat chromosome 10 and detected the insertion of a 25 bp duplication into the sixth exon of the sperm-associated antigen 5 (Spag5/astrin/MAP126) gene, which codes for a microtubule-associated protein. This mutation results in a truncated Spag5 protein lacking the primary spindle-targeting domain at the C terminus. Immunological staining with antibodies to markers for Sertoli and germ cells during the early postnatal period indicated that the abnormal mitosis with dispersed chromosomes in hgn/hgn testes occurs in proliferating Sertoli cells. Therefore, apoptotic Sertoli cell death would result from the disorganization of the spindle apparatus caused by defective Spag5. These findings suggested that the Spag5 is essential for testis development in rats and that the hgn/hgn rat is a unique animal model for studying the function of Spag5.

Amino Acid Sequence↗

PubMedDate not supplied

A case of mixed gonadal dysgenesis (MGD)--with a review of MGD patients reported in Japan.

A 17-year-old patient with mixed gonadal dysgenesis (MGD) showing ambiguous genitalia and hypergonadotropic hypogonadism was described. By intraabdominal exploration, a poorly developed uterus with a fallopian tube and a streak gonad was found on the right side and a poorly developed testis with epidydimis and vas deferens on the left. Chromosomal analysis on cultured peripheral lymphocytes and bone marrow cells showed 45,X karyotype, while among the majority of 45,X cells small numbers of 46,X+ mar cells (3-23%) were found in cultured fibroblasts from the abdominal skin and various organ tissues. We compared our patient with the Japanese patients with MGD reported in the literatures.

Adolescent↗

PubMed1999

Campomelic syndrome and deletion of SOX9.

The human SOX9 gene, located in chromosome region 17q24.1-25.1, encodes a transcription factor involved in chondrogenesis and testis development. Mutations in this gene cause campomelic syndrome (CMPS) with autosomal sex reversal. Here we describe an infant girl with CMPS and an interstitial deletion on the long arm of chromosome 17 (46,X,del(17)(q23.3q24.3). The extent of SOX9 deletion on one chromosome 17 was defined using unique sequence fluorescent in situ hybridization probes. This is the first report of a patient with CMPS bearing a complete deletion of one SOX9 gene, and as such is the strongest evidence to date for dose-dependent action of the SOX9 protein in normal chondrogenesis.

Bone Diseases, Developmental↗

PubMedDate not supplied

Testicular changes associated with the initiation of spermatogenesis.

Testicular changes preceding and accompanying the initiation of spermatogenesis are reviewed. The complex cellular changes occurring in the developing testis are found to be particularly well defined with the use of electron microscopy. The findings suggest that electron microscopic examination may be useful in the evaluation of male infertility, especially in those cases in which clinical and laboratory findings are negative or inconclusive. The amount of tissue required is minimal and can be taken as part of the regular testicular biopsy. The additional information provided by electron microscopy can be used to clarify the observations made on routine light microscopic study and may also lead to better understanding of the pathologic changes occurring in some of the less well understood forms of male infertility.

Animals↗

PubMed2000

[Laparoscopy in the diagnosis, classification and therapy of nonpalpable undescended testes].

OBJECTIVE: The use of laparoscopy in children for the diagnosis, classification and treatment of non-palpable testes. High forms of undescended testes in an abdominal or high inguinal position are a diagnostic and therapeutic problem in paediatric surgery. METHOD: The authors evaluate retrospectively the laparoscopic approach used in 68 boys for the diagnosis, classification and treatment of 79 undescended testes. The mean age of the operated boys was 4.58 years. In 22 (27.9%) agenesis or atrophy of the testis was found (Type I). Thirty-two (40.5%) testes in an abdominal position (Type II) were mobilized laparoscopically, moved to the groin and the operation was completed by open orchidopexy. In 25 cases (31.6%) the laparoscopic method described by Fowler-Stephens was used treatment was completed by orchidopexy in the second stage (Type III and IV). RESULTS: No serious complications were recorded during or after operation. In two instances (2.5%) after the two-stage Fowler-Stephens operation atrophy of the testis developed. CONCLUSION: Miniinvasive laparoscopy is considered at present the standard method for the localization and treatment of non-palpable testes.

Child↗

PubMed2001

[Genotypic sex and phenotypic sex: clinical, biochemical and molecular aspects in a patient with male hypogonadism and 46XX-45XO karyotype].

We report here the case of a patient with primary male ipogonadism, with small testes and deficient virilization of the external genitalia, but with 46XX, 45X0 karyotype. Hormonal determinations showed high LH and FSH and low testosterone levels. Ultrasonography confirmed the presence of small testes within the scrotum. Cytogenetic analysis demonstrated a female karyotype, with 90% 46XX, 10% 45X0 mosaicism. Using DNA probes for genes located on the Y chromosome, the presence of the "Sex-Determining Region" of the Y chromosome (SRY) was evidenced in the genomic DNA of this patient. By Fluorescence in Situ Hybridization (FISH), SRY locus was localized in the p terminal region of an X chromosome. SRY is the primary inducer of testis development; it acts as a transcription factor leading to a sequence of gene activations critical in the process of testicular differentiation and morphogenesis. A condition characterized by testicular development in subjects who lack a normal Y chromosome has been described; most of these patients are carriers of the short arm of the Y chromosome transferred to one of the two X chromosome, suggesting a form of X-Y paternal interchange. In our patient, the development of male gonade in the absence of an Y chromosome was explained by the demonstration of the SRY gene in an X chromosome.

Aged↗

PubMed2002

[P.H.S. (Prolene Hernia System) mesh--initial experience].

In 197 among plastic operations of the groin without tension, using a mesh and new method was added: P.H.S.--Prolene Hernia System--mesh. At the Surgical Clinic of the General Faculty Hospital and First Medical Faculty Charles University in Prague this method is used for plastic operations of the inguinal canal since October 1999. Every year some 300 patients with inguinal hernia are operated, incl. 18.7% operations or multiple relapses. 91% of the group are men and the mean period of hospitalization is 4.9 +/- 3 days. The method "under tension" (McVay, Lotheissen) is used on average in 56.2% patients, in the remainder we use methods "without tension": laparoscopic hernioplasty (TAPP) in 26.7%, Lichtenstein's method in 2.4% and hernioplasty by means of PHS in 14.7% of the patients. Since October 1999 we used it so far in 88 male patients, mean age 55.3 years (2-90 years) with 97 primary or recurrent inguinal hernias. In these patients in 36 instances an indirect hernia was involved (37.1%), a direct hernia was present in 59 patients (60.8%). In two patients we found a saddle hernia (2.1%) and 11 patients (11.4%) had a bilateral hernia. In 12 (12.4%) patients we observed haematoma in the subcutaneous layer in the area of the wound and on the scrotum. In 4 (4.1%) patients there was a seroma in the wound, and subfebrile temperatures up to 38 degrees C were recorded in 9 (9.3%), in 4 patients (4.1%) pain or oedema of the testis developed but receded very rapidly. Postoperative hospitalization lasted on average 2.4 days (1-6 days) and the period before return to normal physical activity was 9.9 days (7-16 days). The patients are checked after regular intervals: 2 weeks, 6 weeks, 6 months, 1 and 2 years. Between October 1999 and February 2002 in the group only one relapse occurred after 6 months.

Adolescent↗

PubMed1985

The differentiation of germ cells and gonads during development of carp (Cyprinus carpio L.). A study with anti-carp sperm monoclonal antibodies.

Gonadal development, germ cell differentiation and the appearance of membrane antigenic determinants, specific for male and female germ cells during gonadogenesis, was studied in larval and juvenile carp (Cyprinus carpio L.) until 25 weeks after fertilization. Indirect immunofluorescence studies with four monoclonal antibodies raised against carp spermatozoa revealed that monoclonal antibody WCS 29 stained the outer membranes of primordial germ cells in larvae from 3 days after fertilization. The monoclonal antibodies WCS 3 and 17 reacted with the outer membranes of germ cells from 7 weeks after fertilization onwards, simultaneously with the onset of germ cell proliferation. With monoclonal antibody WCS 28 germ cell membranes were clearly stained from 18 weeks after fertilization. Similar reactions were observed in both sexes, however, female germ cells reacted at an earlier developmental stage with the monoclonal antibody WCS 28 than male germ cells. In the developing testis the monoclonal antibodies stained all types of spermatogenic cells. In the ovary, however, only oogonia and early prophase oocytes showed a positive reaction with the four monoclonal antibodies. The results indicate that germline-specific antigens are present on the outer membranes of primordial germ cells and their male and female descendants, with the exception of elderly oögenic stages. It is assumed that the appearance and disappearance of these membrane antigens reflect differentiation steps of germ cells during gonadogenesis.

Animals↗

PubMedDate not supplied

True cryptorchidism and retractile testes in infertile men.

The present study entails clinical evaluation of 77 infertile men with testicular descent disturbances. The results summarize as follows: The incidence of disturbance in testicular descent among infertile men was 15.5%, 15.5% and 4.9% in azoo-, oligozoo- (less than or equal to 10 mill/ml) and normozoo- (greater than 10 mill/ml) spermia, respectively. Out of the 77 men investigated, 50 had a history of true cryptorchidism and 27 retractile testes. 31 of the patients with cryptorchidism had previously been subjected to orchidopexy. In the group of infertile men with a history of retractile testes, 20 subjects still retained palpable pathological changes of the testes. Testicular biopsies (from men with retractile testes and sperm count below 10 mill/ml) demonstrated hypospermatogenesis (germ cell arrest to aplasia). 6) 53% of the subjects exhibited elevated serum FSH. Two men had developed testis tumours (seminoma/teratoma). It is advocated that early hormonal therapy and/or surgical procedures should be initiated in an attempt to improve the chances of future fertility both in cryptorchidism and retractile testes.

Adult↗

PubMed1979

[Malignant tumor originating from the interstitial cells of the testis (leydigoma)].

The authors describe a very rare and very interesting case of malignant tumour, originating from the interstitial cells of the testis, developed in a male of 53, with multiple metastases. The patient developed chronic hypocorticism, owing to bilateral metastases in adrenal glands and finally followed by an Addison coma. The case is of interest because no data are available in literature about metastases in adrenals with and adrenal insufficiency advancement.

Acute Kidney Injury↗

PubMed1993

[Molecular genetic analysis on six cases of sex reversal syndrome].

Southern blot hybridization using Y-specific DNA probes and electrophoretic analysis of the SRY gene fragment amplified through polymerase chain reaction (PCR) have been performed in six cases of sex reversal syndrome (three 46, XX males and three 46, XY females). The result showed that the Y-specific DNA hybridization signal and SRY sequence were absent in one XX male and present in the remaining cases. This finding indicates that testis development of this XX male was initiated in the absence of the SRY gene.

Adult↗

PubMed1995

Steady state steroid 5 alpha-reductase messenger ribonucleic acid levels and immunocytochemical localization of the type 1 protein in the rat testis during postnatal development.

Steroid 5 alpha-reductase is the rate-limiting enzyme in the production of 5 alpha-reduced steroids in many tissues. Developmental changes in 5 alpha-reductase activity play an important role in regulating the amount of testosterone that is secreted by the testis. To date, the regulation of testicular 5 alpha-reductase has been studied extensively at the level of enzyme activity. Regulation at the messenger RNA (mRNA) and protein levels, however, has not been investigated. The objectives of the present study were to determine the steady state mRNA levels for the 5 alpha-reductase isozymes, types 1 and 2, and to immunolocalize the 5 alpha-reductase type 1 protein in the developing rat testis (7-91 days postpartum). Consistent with previously reported enzyme activity studies, type 1 5 alpha-reductase mRNA levels were most abundant in the immature animal (days 21-28). Unlike 5 alpha-reductase activity, however, type 1 mRNA levels did not decline thereafter to reach nearly undetectable levels in the adult (day 91). In contrast, 5 alpha-reductase type 1 mRNA levels remained relatively constant between days 42-91. The 5 alpha-reductase type 1 transcript size did not remain constant during postnatal testicular development. The characteristic 2.5-kilobase type 1 transcript size was detected in immature rats (days 21-28), whereas in the adult (day 91), a slower migrating 2.7-kilobase type 1 mRNA species was observed. An antipeptide antiserum specific to rat 5 alpha-reductase type 1 was used to immunolocalize the 5 alpha-reductase type 1 protein. At all ages examined, the immunoperoxidase reaction was localized predominantly to the interstitial tissue of the testis. On postnatal day 7, clusters of interstitial cells resembling fetal Leydig cells were clearly immunoreactive. The staining intensity increased steadily from day 7 onwards, so that by days 21 and 28, interstitial cells with the appearance of immature Leydig cells were intensely immunoreactive (peak expression). This was followed by a progressive decrease in staining intensity between days 28-91, so that by day 91 (adult) Leydig cell immunoreactivity was barely detectable. Immunocytochemical staining revealed a predominantly cytoplasmic localization; significant nuclear staining was not evident. We conclude that the expression of the 5 alpha-reductase type 1 protein is primarily found in the cytoplasm of Leydig cells, is dependent on age, and that this expression closely parallels 5 alpha-reductase enzyme activity.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

PubMed1987

Postnatal development of the testis in the Mongolian gerbil, Meriones unguiculatus.

The testicular development in gerbils was studied over 16-week periods starting from birth. Testicular weight and seminiferous tubule diameter increased considerably between 1 and 11 weeks of age. At 16 weeks the testicular weight was about 540 mg. Spermatogenesis commenced at about 2 weeks when mitoses first appeared in spermatogonia. Spermatozoa appeared in a few of the seminiferous tubules by 7 weeks and consistently so in all of the tubules at 10 weeks. Epididymal spermatozoa appeared first in the cauda epididymis at 10 weeks and were consistently present at 12 weeks. Formation of mature Leydig cells with a grouped perivascular arrangement appeared by 3 weeks and continuously so thereafter. From these results, it is evident that the male Mongolian gerbil is almost sexually matured by 10 to 12 weeks of age.

Animals↗

PubMed1989

Immunolocalization of type IV collagen and laminin during rat gonadal morphogenesis and postnatal development of the testis and epididymis.

The distribution of type IV collagen and laminin was studied by immunocytochemistry during rat gonadal morphogenesis and postnatal development of the testis and epididymis. Immunostaining appeared as early as the 12th day of gestation along the basement membranes of the mesonephric-gonadal complex. The connection between some mesonephric tubules and coelomic epithelium was seen between the 12th and 13th day of gestation. Discontinuous immunostained basement membranes delineated the differentiating sexual cords in 13-day-old fetuses; this process probably began in the inner part of the gonadal ridge. The seminiferous cords surrounded by a continuous immunoreactive basement membrane are separated from the coelomic epithelium by the differentiating tunica albuginea in 14-day-old fetuses. During the postnatal maturation of epididymis and testis, the differentiation of peritubular cells is accompanied by a progressive organisation of the extracellular matrix into a continuous basement membrane. This change is associated with a gradual condensation of peritubular cells inducing an increase of immunostaining. In adult animals, the tubular wall of epididymis is thicker than the lamina propria of seminiferous tubules. Both type IV collagen and laminin immunostaining paralleled during ontogenesis at the light-microscope level.

Aging↗

PubMed2003

Expression and localization of Smad1, Smad2 and Smad4 proteins in rat testis during postnatal development.

AIM: To study the expression and regulation of Smad1, Smad2 and Smad4 proteins (intracellular signaling molecules of transforming growth factor-b family) in rat testis during postnatal development. METHODS: The whole testes were collected from SD rats aged 3, 7, 14, 28 and 90 (adult) days. The cellular localization and developmental changes were examined by immunohistochemistry ABC method with the glucose oxidase-DAB-nickel enhancement technique. Quantitative analysis of the immunostaining was made by the image analysis system. The Smads proteins coexistence in the adult rat testis was tested by the double immune staining for CD14-Smad4 and Smad2-Smad4. The protein expression of Smad during rat testicular development was examined by means of Western blots. RESULTS: Smad1, Smad2 and Smad4 were present throughout testicular development. The immunostaining of Smad1 and Smad2 were present in spermatogenic cells. A positive immunoreactivity was located at the cytoplasm, but the nucleus was negative. Smad1 was immunolocalized at the d14, d28 and adult testes, while Smad2, at the d7, d14, d28 and adult testis. There was positive immunoreaction in the Sertoli cells and Leydig cells as well. The immunolocalization of Smad4 was exclusively at the cytoplasm of Leydig cells and the nuclei were negative throughout the testicular development. No expression was detected in the germ cells. The results of image and statistical analysis showed that generally the expression of Smad1, Smad2 and Smad4 in the testis tended to increase gradually with the growth of the rat. CONCLUSION: The present data provide direct evidences for the molecular mechanism of TGF-bgr action in rat testes during postnatal development and spermatogenesis.

Animals↗