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PubMed1995

Androstenedione metabolism in the indifferent stage of bovine gonad development.

Suspensions of cells obtained from genital ridges and gonads of bovine embryos of 1.0-3.4 cm in crown-rump (C-R) length were used for studies of the metabolism of radiolabeled exogenous steroid precursor. Tritiated androstenedione was employed as precursor and the final products of their metabolism were evaluated after separation by thin-layer chromatography. Genetic sex was determined by the karyotype of hepatocytes from the same embryos. The extent of conversion of tritiated androstenedione was higher in cells from male embryos than cells from female embryos. Furthermore, androstenedione was metabolized mainly to testosterone in male embryos. By contrast, cells obtained from female embryos transformed androstenedione to estrone and 17 beta-estradiol. The onset of this activity was observed at 1.8 cm in C-R length in males and at 2.2 cm in C-R length in females. In all cases, after the onset, the metabolic activity increased in relation to the age of the embryos. These data show that in the bovine embryo gonad, the activity of aromatase and 17 beta-hydroxysteroid dehydrogenase are present when the morphological differentiation of the gonadal sex has not yet been established.

Androstenedione↗
PubMed1994

Effects of pituitary-gonadal suppression with a gonadotrophin-releasing hormone agonist on fetal gonadotrophin secretion, fetal gonadal development and maternal steroid secretion in the sheep.

In order to investigate the regulation of the hypothalamo-pituitary-gonadal axis during fetal development, sheep fetuses at day 70 of gestation were implanted subcutaneously with a biodegradable implant containing the long-acting gonadotrophin-releasing hormone (GnRH) agonist, buserelin. The treatment of fetuses with a GnRH agonist throughout the last half of gestation (term = 145 days) abolished the increase in plasma LH concentrations that was seen in 2-day-old control lambs in response to an injection of GnRH. This attenuated response was associated with corresponding reductions in the pituitary content of LH and FSH. Immunolocalization studies revealed that pituitary glands from newborn lambs implanted with a GnRH agonist during fetal development were devoid of immunopositive LH- and FSH-containing cells. At birth the testicular weights of GnRH agonist-treated ram lambs were significantly decreased by 40% when compared with controls. This was associated with a 45% reduction in the total number of Sertoli cells per testis. In newborn ewe lambs GnRH agonist treatment had no effect on ovarian weight or on the morphological appearance of the ovaries. GnRH agonist treatment had no effect on the plasma concentrations of progesterone and oestrone in the maternal circulation or on the length of gestation. These results show (1) that GnRH positively regulates the synthesis and secretion of gonadotrophins in the fetus, (2) that reduced fetal gonadotrophic support during the last half of gestation results in a reduction in testicular growth, and (3) that fetal gonadotrophins do not affect maternal steroid secretion.

Animals↗
PubMed2002

Early gonadal development: exploring Wt1 and Sox9 function.

Prior to sex determination the gonadal anlage is formed as a bipotential primordium with the capacity to differentiate into either testes or ovaries depending on the presence or absence of the Sry gene. Knockout experiments have implicated five genes in the formation or survival of the gonadal primordium: Wt1, Sf1, Lim1, Lhx9 and Emx2. We are particularly interested in the Wilms tumour suppressor, WT1, which is characterized by complex posttranscriptional modifications. Here we will focus on published in vitro evidence suggesting distinct functions for the various isoforms and present our own results from in vivo experiments. Our data suggest that WT1 is an important regulator of the transcription or stability of the sex-determining gene Sry. One of the first genes expressed after the initial male sex-determining signal is the Sox9 gene. Human SOX9 has been implicated in male-to-female sex reversal. To analyse Sox9 function in mouse development we have performed transgenic experiments and ectopically expressed this gene in XX gonads. Our data indicate that Sox9 is sufficient to induce testis formation in mice. Here we will discuss our new data and present an updated model for Wt1 and Sox9 function in gonad formation and sex determination.

Animals↗
PubMed2001

The possible contribution of pituitary hormones to the heterochronic development of gonads and external morphology in overwintered larvae of Hynobius retardatus.

In Hynobius retardatus, most larvae in regions of low elevation metamorphose by autumn of the same year. However, larvae of some populations found in cold, mountainous ponds cannot metamorphose within the year and become aged, overwintered larvae. Gonadal development in larvae under the age of 1 year (larvae developed from eggs spawned in the same year) and in aged, overwintered larvae (spawned and hatched in previous years) was examined at the same developmental stage (stage 63, full-grown larval stage). The number of germ cells and the cross-sectional areas of the gonads were much larger in 2-season-overwintered (third year) larvae than in larvae under the age of 1 year. To obtain reliable probes for investigating the possible contribution of TSH, FSH and LH to metamorphosis and gonadal development, cDNAs for Hynobius TSHbeta, FSHbeta and LHbeta genes were cloned. Their expressions were analyzed by means of semi-quantitative RT-PCR in larvae under the age of 1 year and in 2-season-overwintered larvae. No differences were observed in expression levels of either TSHbeta or LHbeta between larvae under the age of 1 year and the overwintered larvae. In contrast, expression of FSHbeta was much higher in the overwintered larvae than in larvae under the age of 1 year. These results suggest that gonadal development proceeds gradually with age even in the overwintered larvae, but that metamorphosis is retarded, probably due to the larvae's cold habitat. Heterochronic development of gonads and external morphology has been demonstrated in H. retardatus, suggesting a potency for neotenic reproduction in this species.

Amino Acid Sequence↗
PubMed2005

Male-specific cell migration into the developing gonad is a conserved process involving PDGF signalling.

Male-specific migration of cells from the mesonephric kidney into the embryonic gonad is required for testis formation in the mouse. It is unknown, however, whether this process is specific to the mouse embryo or whether it is a fundamental characteristic of testis formation in other vertebrates. The signalling molecule/s underlying the process are also unclear. It has previously been speculated that male-specific cell migration might be limited to mammals. Here, we report that male-specific cell migration is conserved between mammals (mouse) and birds (quail-chicken) and that it involves proper PDGF signalling in both groups. Interspecific co-cultures of embryonic quail mesonephric kidneys together with embryonic chicken gonads showed that quail cells migrated specifically into male chicken gonads at the time of sexual differentiation. The migration process is therefore conserved in birds. Furthermore, this migration involves a conserved signalling pathway/s. When GFP-labelled embryonic mouse mesonephric kidneys were cultured together with embryonic chicken gonads, GFP+ mouse cells migrated specifically into male chicken gonads and not female gonads. The immigrating mouse cells contributed to the interstitial cell population of the developing chicken testis, with most cells expressing the endothelial cell marker, PECAM. The signalling molecule/s released from the embryonic male chicken gonad is therefore recognised by both embryonic quail and mouse mesonephric cells. A candidate signalling molecule mediating the male-specific cell migration is PDGF. We found that PDGF-A and PDGF receptor-alpha are both up-regulated male-specifically in embryonic chicken and mouse gonads. PDGF signalling involves the phosphotidylinositol 3-kinase (PIK3) pathway, an intracellular pathway proposed to be important for mesonephric cell migration in the mammalian gonad. We found that a component of this pathway, PI3KC2alpha, is expressed male-specifically in developing embryonic chicken gonads at the time of sexual differentiation. Treatment of organ cultures with the selective PDGF receptor signalling inhibitor, AG1296 (tyrphostin), blocked or impaired mesonephric cell migration in both the mammalian and avian systems. Taken together, these studies indicate that a key cellular event in gonadal sex differentiation is conserved among higher vertebrates, that it involves PDGF signalling, and that in mammals is an indirect effect of Sry expression.

Animals↗
PubMed1998

Gonad development: assembling the mammalian testis.

Mammalian primordial germ cells migrate into gonads of either sex indiscriminately and may be functional even across a species barrier; but certain somatic cell lineages are attracted specifically into the male gonad and are absolutely required for the construction of the seminiferous cords of the testis.

Animals↗
PubMed2004

Endocrine disruption and altered gonadal development in white perch (Morone americana) from the lower Great Lakes region.

High prevalences of gonadal intersex have been observed in wild fish populations in areas affected by domestic and industrial effluents. For this study, fish were collected in 1998 from the Cootes Paradise region of Hamilton Harbour in western Lake Ontario, Canada, to determine whether gonadal abnormalities, including intersex, were present in young of the year (YOY) fish. No gonadal abnormalities were observed in goldfish (Carassius auratus), common carp (Cyprinus carpio), gizzard shad (Dorosoma cepedianum), brown bullhead (Ictalurus ameiurus), pumpkinseed (Lepomis gibbosus), and bluegill (Lepomis macrochirus). However, intersex gonads were observed in 8 of 16 male white perch (Morone americana) examined in this survey. Subsequently, in 1999 and 2000 white perch estimated to be YOY to approximately 2 years of age were collected from Cootes Paradise and from two other sites in the lower Great Lakes region. Gonadal intersex was observed in male white perch collected from the Bay of Quinte (22-44%) and Lake St. Clair (45%), although the prevalence and the extent of the intersex condition were lower relative to the 83% prevalence in white perch collected in Cootes Paradise. Intersex was not observed in hatchery-reared white perch or in white perch collected from an uncontaminated reference site (i.e., Deal Lake) in the United States. An analysis of plasma collected in the spring of 2002 from male adult white perch in Cootes Paradise revealed high concentrations of vitellogenin, ranging from 49 to 1,711 microg/mL. These observations indicate that male white perch are exposed to estrogenic endocrine-disrupting substances that may be responsible for the induction of gonadal intersex.

Animals↗
PubMedDate not supplied

Gonad development in Drash and Frasier syndromes depends on WT1 mutations.

The study of the gonads of 8 cases of Drash syndrome (6 ambiguous males, 2 females) and of 2 Frasier syndrome shows that WT1 mutations gives a dysgenetic testis which is the cause of the genital ambiguity observed at birth. By contrast the same mutations have no effect on ovary development giving normal females. However intron mutations in KTS with isoforms imbalance of WT1 proteins cause streak gonads with a female phenotype in XY patients. In consequence WT1 mutations are the cause of a spectrum of male genital malformations associated with glomerulonephritis and tumors. The absence of WT1 protein detection in sertoli cells shown by immunohistochemistry for 3 cases suggests an imprinting effect of the normal WT1 allele promotor rather than a low level of protein production. A caryotype is mandatory for a correct diagnosis.

Adolescent↗
PubMed1981

Ultrastructural events during early gonadal development in Rana pipiens and Xenopus laevis.

The establishment of the undifferentiated gonad was studied in Xenopus laevis and Rana pipiens using high resolution techniques. It was found that the cells of the so-called "mesonephric blastema" had no structural resemblance to the cells of the gonadal medulla in both species. Furthermore, there was no morphological evidence that would suggest a migration of the former cells towards the incipient gonad at the time of its appearance. However, the basal lamina of the coelomic epithelium was interrupted in the region of the genital crest, and there was a definite ultrastructural similarity between the cells of this epithelium and those that first form the medulla. These observations suggest that, in amphibians, the cells of the gonadal medulla come from a cellular line arising from the coelomic epithelium and not from the "mesonephric blastema," as has been proposed.

Animals↗
PubMed2001

Drosophila homolog of the myotonic dystrophy-associated gene, SIX5, is required for muscle and gonad development.

SIX5 belongs to a family of highly conserved homeodomain transcription factors implicated in development and disease. The mammalian SIX5/SIX4 gene pair is likely to be involved in the development of mesodermal structures. Moreover, a variety of data have implicated human SIX5 dysfunction as a contributor to myotonic dystrophy type 1 (DM1), a condition characterized by a number of pathologies including muscle defects and testicular atrophy. However, this link remains controversial. Here, we investigate the Drosophila gene, D-Six4, which is the closest homolog to SIX5 of the three Drosophila Six family members. We show by mutant analysis that D-Six4 is required for the normal development of muscle and the mesodermal component of the gonad. Moreover, adult males with defective D-Six4 genes exhibit testicular reduction. We propose that D-Six4 directly or indirectly regulates genes involved in the cell recognition events required for myoblast fusion and the germline:soma interaction. While the exact phenotypic relationship between D-Six4 and SIX4/5 remains to be elucidated, the defects in D-Six4 mutant flies suggest that human SIX5 should be more strongly considered as being responsible for the muscle wasting and testicular atrophy phenotypes in DM1.

Animals↗
PubMed1998

Steroid contents of and steroidogenesis in vitro by the developing gonad and mesonephros around sexual differentiation in fetal sheep.

The aim of the present study was to establish whether the steroids, progesterone, androstenedione, testosterone and oestradiol, were present in the mesonephric-gonadal complex of female and male sheep fetuses around sexual differentiation (that is, from day 28 to day 45 of gestation, with sexual differentiation occurring at approximately day 32). A second aim was to test whether the mesonephric-gonadal complex, mesonephros (days 35-45 only) and gonad (days 35-45 only) were capable of steroid synthesis in vitro. The steroid contents in the mesonephric-gonadal complex were not detectable before sexual differentiation. However, from day 35 of gestation onwards, the mesonephric-ovarian complex contained mainly oestradiol and the mesonephric-testicular complex contained mainly testosterone: from day 35 until day 45 the increase in content of these two steroids exceeded the increase in the mass of tissue by more than fivefold. From day 40 to day 45 of gestation, the contents of the other steroids in the pathways to oestradiol increased progressively in both sexes but more in parallel with the increase in tissue mass. In contrast to the steroid contents in the tissue at recovery, the mesonephric-gonadal tissue from both sexes in tissue culture was able to synthesize most steroids before and after sexual differentiation and also to metabolise supplementary androstenedione to oestradiol. These findings suggest that many, if not all, of the steroidogenic enzymes in the pathway from cholesterol to oestradiol are present before sexual differentiation. Most of the aforementioned steroids were present in detectable amounts in isolated mesonephros and gonad of both sexes after sexual differentiation. Moreover, for both the isolated mesonephros and gonad, there were increases in the mean contents of most steroids after culture relative to the contents in the tissues at recovery. These data suggest that the mesonephros, as well as the gonad, in both sexes is capable of synthesizing steroid. It is concluded that, in the sheep fetus, the female and male gonads are steroidogenically active after sexual differentiation, that the steroidogenic enzymes develop before sexual differentiation, and that the mesonephros is a site of steroid synthesis.

Androstenedione↗
PubMed2003

Differential expression of tissue inhibitor of metalloproteinases type 1 (TIMP-1) during mouse gonad development.

In mammals, the gene Sry initiates signaling pathways triggering the differentiation of a testis from a sexually indifferent gonad. Assuming that these morphogenetic events may alter the proteolytic balance, the expression of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) was investigated in gonads from 11.5 days postcoitum (dpc) onward, when testicular organogenesis occurs. Whereas selective MMPs and TIMPs (1-3) were detected in undifferentiated gonads (11.5 dpc) and in neonatal testes, a single TIMP (TIMP-1) was expressed in a sexually dimorphic manner from 12.5 dpc onward (i.e., after overt male gonad differentiation), demonstrated by using a semiquantitative reverse transcriptase-polymerase chain reaction and a Western blot analysis. To gain insight into the role of TIMP-1, the expression of gelatinases (mRNA levels and enzyme activity) was monitored. However, no sex differences could be evidenced, indicating that TIMP-1 was not inhibiting this class of MMPs during testis organogenesis. Apart from being an inhibitor of MMPs, TIMP-1 is known to display growth promoting activities. Of interest, testicular TIMP-1 (but not TIMP-2) levels were further enhanced up to 2 weeks of age, consistent with a role in the early postnatal testicular growth. We, therefore, established an organotypic culture system in which seminiferous cords may differentiate de novo and grow, depending on culture conditions. In that system and mimicking the in vivo situation, TIMP-1 immunolocalized strongly within the male gonadal territory and weakly in female gonads, in which no organization was evident. Experiments are now under way to determine to what extent TIMP-1 is a morphogenic gene involved in seminiferous cord formation and development.

Animals↗
PubMed2004

Molecular aspects of female and male gonadal development in mammals.

In mammals, the choice between male or female development is genetically determined. Sex determination takes place when the embryonic gonad becomes either a testis or an ovary. This process is directed by genes that have been discovered by genetic analysis of sex-reversed patients and confirmed by knock-out experiments in mice. The testis-determining pathway is better known than the ovary pathway. SRY, a gene located on the Y chromosome, triggers a complex genetic cascade leading to testicular differentiation. In this cascade, two genes play a crucial role in male differentiation, SOX9 and FGF9, which contribute to testicular cord formation. However, only a minority of 46,XY sex-reversed patients can be explained by mutations in known genes such as SRY, SOX9, WT1 and SF1, suggesting that other genes influencing sex determination are yet to be discovered. In females, some rare genes that induce ovarian failure or female-to-male sex-reversal, have been found through gene-targeted inactivation in mice or positional cloning of mutations in humans and goats. In both sexes, genetic analysis of sex-reversed individuals (XX males, XX and XY hermaphrodites and XY with complete or partial dysgenesis) remains an approach of choice to isolate new genes involved in sex determination.

Animals↗
PubMed1993

Gonadal development in the opossum, Monodelphis domestica: the rete ovarii does not contribute to the steroidogenic tissues.

The gonads of 273 animals, ranging in age from newborn to adult, were examined in serial histological sections. Primordial germ cells were located in the hindgut, dorsal mesentery and gonadal primordia of neonates but were alkaline phosphatase negative. The testis differentiated between d 13 of gestation and birth, about half a day later. Testis cords, composed of pre-Sertoli cells and containing germ cells, were distributed peripherally in the gonad. Ovarian differentiation began on d 6, when an ill defined cortex and medulla became discernible. Meiosis commenced on d 14, medullary cords formed from blastema cells by d 26 and gave rise to granulosa cells around d 29. The rete ovarii was first observed in the hilar region of the gonad primordium. It penetrated maximally two thirds of the ovarian length between d 26 and d 29. It is concluded that, at least postnatally, the mesonephros does not contribute to the rete ovarii and that the granulosa cells are derived not from the rete but from the medullary cords.

Animals↗