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Dax1 regulates testis cord organization during gonadal differentiation.

Mutations of the DAX1 nuclear receptor gene cause adrenal hypoplasia congenita, an X-linked disorder characterized by adrenal insufficiency and hypogonadotropic hypogonadism. Targeted deletion of Dax1 in mice also reveals primary testicular dysgenesis, which is manifest by obstruction of the rete testis by Sertoli cells and hyperplastic Leydig cells, leading to seminiferous tubule dilation and degeneration of germ cells. Because Dax1 is expressed early in gonadal development, and because Sertoli and Leydig cells are located ectopically in the adult, we hypothesized that these testis abnormalities are the result of an early defect in testis development. In Dax1(-/Y) males, the gonad develops normally until 12.5 dpc. However, by 13.5 dpc, the testis cords are disorganized and incompletely formed in Dax1-deficient mice. The number of germ and Sertoli cells is unchanged, and the expression of Sertoli-specific markers appears to be normal. However, the number of peritubular myoid cells, which normally surround the testis cords, is reduced. BrdU labeling of peritubular myoid cells is low, consistent with decreased proliferation. The basal lamina produced by peritubular myoid and Sertoli cells is disrupted, leading to open and incompletely formed testis cords. Leydig cells, which normally reside in the peritubular space and extend from the coelomic surface to the dorsal surface of the gonad, are restricted to the coelomic surface of Dax1-deficient testis. We conclude that Dax1 plays a crucial role in testis differentiation by regulating the development of peritubular myoid cells and the formation of intact testis cords. The developmental abnormalities in the Dax1-deficient testis lay the foundation for gonadal dysgenesis and infertility in adult mice and, potentially in humans with DAX1 mutations.

Animals↗

Temporal and spatial localization patterns of Gata4 during porcine gonadogenesis.

The zinc finger transcription factor Gata4, is associated with gonadal development in many species. The present study characterizes temporal and spatial localization of Gata4 throughout gonadogenesis in porcine embryos. Immunohistochemical studies illustrated that Gata4 protein is present in the coelomic epithelium prior to histological differentiation of the nascent bipotential gonad, marking the future site of both XX and XY porcine gonads. Many somatic cells of both XX and XY bipotential gonads continue to retain Gata4 immunoreactivity throughout sexual differentiation and subsequent gonadal development. Testicular cords were evident by 26 days postcoitum. Gata4 was present in Sertoli cells, identified by virtue of coexpression with Müllerian inhibiting substance and also interstitial cells including Leydig cells throughout fetal and postnatal life. Many somatic cells of the differentiating ovary including follicular cells also contained Gata4 protein throughout fetal and postnatal life. Gata4 was not present in germ cells, endothelial cells, or other undifferentiated mesenchymal cells of both XX and XY gonads. A population of Gata4-positive cells in the dorsal mesentery was continuous with the coelomic epithelium of the gonad. This localization pattern led to the hypothesis that a subpopulation of somatic cells in the dorsal mesentery moves toward the gonad. An in vitro cell migration assay demonstrated that Gata4-positive cells preferentially migrate toward explanted gonadal tissue, and morphological features of the developing gonad supported this hypothesis. This study illustrates that Gata4 is a very early marker for gonad formation, highlights species differences in temporal and spatial localization patterns, and suggests a potential role for Gata4 in the development of both XX and XY porcine gonads. Further, we suggest that mesenchymal cells of the dorsal mesentery may provide a source of somatic cells that migrate and incorporate into the gonad and contribute to various somatic cell lineages. Overall, the spatial and temporal localization patterns of Gata4 during porcine gonadogenesis implies a much earlier and wider role for Gata4 than previously reported in other species.

Animals↗

Quantification of müllerian inhibiting substance in developing chick gonads by a competitive enzyme-linked immunosorbent assay.

An immunoblotting method was used to purify a Müllerian-inhibiting substance (MIS)-specific antiserum. The serum was used to quantify the content of MIS in developing chick gonads by competitive enzyme-linked immunosorbent assay. From embryonic stages to the eleventh week after hatching, male chicken testes have a high content of MIS in the following two stages: (1) from the sixth to the eighth day and from the fourteenth to the twentieth day of incubation, and (2) from the second to the eighth week after hatching. The high content of MIS in the early embryonic stage is closely correlated with the natural pattern of Müllerian duct regression observed in the male embryo. From the sixth to the twelfth day of incubation, the female right ovary contains a higher content of MIS than that of the left ovary. Up to the fourteenth day of incubation, the content of MIS in the left ovary reaches maximum levels and then declines. The combination of MIS from right and left ovaries was found to be highest in the ninth to the fourteenth day of incubation, when the regression of the right Müllerian duct reached its highest peak. However, the question of the inability of MIS to cause regression of the female left Müllerian duct and the caudal part of the right duct is raised and discussed. The hypothesis that prenatal estrogenic hormone (diethylstilbestrol) protects the Müllerian duct has been reevaluated. It was found that estrogen does not reduce the MIS content in prenatally treated gonads.

Animals↗

Spermiogenesis in commercial poultry species: anatomy and control.

Spermatogenesis is a complicated process dependent upon several factors. Formation of a testis requires the interaction of gene-products and hormones (androgens) on pluripotent tissue. In birds, the female is the heterogametic (ZW) sex, but W chromosomal genes do not influence gonadal development in a way similar to the SRY gene on the mammalian Y chromosome. However, autosomal genes such as SRY-like HMG box gene 9 (SOX9) may influence gonadal development. Hormones affect development; male gonads subjected to estrogen form an ovotestis, whereas ovaries exposed to aromatase inhibitors form an atypical testis. Sertoli cell numbers are set early in spermiogenesis, possibly under the influence of follicle-stimulating hormone and thyroid hormone, and this may determine the number of gonial cells that can be supported. Sertoli cells make a number of substances that affect testicular development and function, particularly anti-Müllerian hormone, which inhibits female oviduct formation from the Müllerian anlage, inhibits aromatase activity to stop estrogen production, and possibly stimulates androgen production by Leydig cells. Undifferentiated primordial germ cells (PGC) migrate to the testis and are converted to spermatogonia by factors from gonadal ridge tissue and androgens. The PGC of males in the ovary form oocytes of Z genotype, whereas the female PGC in males form mostly Z sperm (with a few of W genotype). Transmission electron microscopy micrographs of turkey testis are presented, and control of spermatogenesis by hormones and cytokines is discussed. This discussion includes follicle-stimulating hormone, luteinizing hormone, inhibin, activin, follistatin, tumor necrosis factor-alpha, growth factors such as transforming growth factor-beta, interleukins, and interferon. Although information concerning paracrine and autocrine regulation of the avian testis by these substances is sparse, much can be learned from mammalian studies, in which putative roles of each of these substances have been established. How Sertoli cells cause directed apoptosis of spermatogonia using the Fas-ligand, Fas-receptor pathway is reviewed, as well as ways to circumvent this process. A possible role for ubiquitin concerning prevention of heat-induced damage to the testis is presented.

Animals↗

Effect of cyclic sound cues on sexual development in nonphotostimulated Japanese quail.

The influence of cyclic ambient sound stimuli on sexual development was studied in nonphotostimulated [6 hr light:18 hr dark (16L:18D)] Japanese quail (Coturnix coturnix japonica). The incidence of accelerated gonadal development was reduced when ambient daily sound stimuli were attenuated by the presence of a white noise mask in the animal quarters. In a second experiment nonphotostimulated (9L:15D) male quail showed a phase-dependent testicular response to a daily 3-hr radio broadcast presented at different portions of the day. The radio sound stimulus induced a higher incidence of accelerated gonadal development when presented 6 hr prior to the photophase than when presented either 3 hr prior to the photophase or when presented coincidently with the photophase onset. In a third experiment locomotion was monitored in nonphotostimulated quail (9L:15D) with a 3-hr radio sound stimulus presented 6 hr prior to photophase onset. The onset of a daily locomotor activity pattern was associated with radio sound in some individuals, but sound-induced locomotion was not consistently associated with sound-induced accelerated gonadal development.

Acoustic Stimulation↗

Expression pattern of alphavbeta3 and alphavbeta5 integrin mRNA in mouse fetal gonads.

The alphavbeta3 and alphavbeta5 integrins are known as transmembrane receptors capable of binding to the RGD amino acid peptide sequence. In mouse early gonadogenesis, some proteins containing the RGD sequence are deposited into extracellular space and participate in morphogenesis. We analyzed the expression patterns of the alphavbeta3 and alphavbeta5 integrins in mouse developing gonads (10.5-13.5 days post coitum) using whole-mount in situ hybridization. The alphav integrin mRNA was homogenously expressed in developing gonadal regions. On the other hand, the beta3 integrin mRNA was found only in large and round cells (presumptive germ cells), whereas beta5 integrin was localized in gonadal somatic cells, with the exception of coelomic epithelial cells. The beta3 integrin-expressed cells were determined to be primordial germ cells because the number of these cells was drastically reduced in busulfan-treated gonads. In this study, we demonstrated that the alphavbeta3 and alphavbeta5 integrins are widely localized in the mouse developing gonads and discussed their presumptive functions on mouse gonadogenesis.

Animals↗

Targeted disruption of the pituitary glycoprotein hormone alpha-subunit produces hypogonadal and hypothyroid mice.

Pituitary thyrotropin (TSH) and gonadotropins (LH and FSH) are thought to be critical for thyroid and gonadal development and function. Each of these pituitary hormones is a heterodimer composed of a common alpha-subunit and unique beta-subunit, and heterodimerization is required for function. No mutations in the alpha-subunit or any of the beta-subunit genes have been reported in mice. To assess directly the functional role of TSH, LH, and FSH in thyroid and gonadal development, we created a disruption of the alpha-subunit gene by homologous recombination. The homozygous mutant animals were hypogonadal and exhibited profound hypothyroidism resulting in dwarfism. Thyroid development was arrested in late gestation, but GnRH neuron migration, development of secondary sex organs, and fetal and neonatal gonadal development were normal. This establishes the importance of thyrotropin in ontogeny and reveals that fetal pituitary gonadotropins are not required for sexual differentiation or genital development in male or female fetuses. The pituitary cells that produce TSH beta-subunit exhibited dramatic hypertrophy and hyperplasia as a result of the lack of thyroid function. This proliferation response occurred at the expense of somatotrope and lactotrope cells, consistent with a derivation of these three cell types from a common precursor.

Aging↗

Exposure of juvenile roach (Rutilus rutilus) to treated sewage effluent induces dose-dependent and persistent disruption in gonadal duct development.

Wild roach (Rutilus rutilus) have been found with intersex gonads in rivers throughout the United Kingdom. The incidence of intersexuality is strongly correlated with discharges of estrogenic treated sewage effluent into those rivers, and this has led to the hypothesis that estrogenic chemicals in effluents are feminizing wild male fish. In this study, early-life stage roach (50 days post hatch, dph) were exposed for 150 days to a graded concentration (0%, 12.5%, 25%, 50%, and 100%) of treated sewage (primarily domestic) effluent to examine the effects of exposure on sexual differentiation and development. Measurement of steroid estrogens and alkylphenolic chemicals in the effluent and a resulting dose-dependent induction of vitellogenin (VTG; a female-specific, estrogen-dependent plasma protein) confirmed that the fish had been exposed and responded to "estrogen" in the effluent. Exposure to treated sewage effluent induced feminization of the reproductive ducts in "male" roach in a dose-dependent manner (in full-strength effluent, 100% of the fish had feminized ducts), indicating that the disruption of the gonad ducts seen in wild roach is the result of exposure to treated sewage effluents during early-life stages. There were no effects of treated sewage effluent exposure on germ cell development; therefore, no oocytes occurred in the testes of the feminized male roach. Subsequent, depuration of the effluent exposed fish in "clean" water for 150 days resulted in a reduction in plasma VTG but no alteration of the feminized ducts, indicating that the effect of the treated sewage effluent on reproductive duct development was permanent. The causality of oocytes in the testes of wild male roach therefore remains to be elucidated.

Animals↗

The 412 retrotransposon and the development of gonadal mesoderm in Drosophila.

We have shown that the expression of the 412 retrotransposon provides a useful early marker for the development of the gonadal mesoderm in Drosophila embryos. 412 is initially expressed in a set of parasegmentally repeated stripes from parasegments (PS) 2-14 in the mesoderm at the extended germ band stage. During germ band retraction the bulk of 412 expression declines except in dorsolateral clusters of cells in PS10, 11 and 12, where high levels of 412 expression remain. These mesodermal cell clusters are associated with germ cells and subsequently they coalesce, rounding up to form the gonads. The gonadal mesoderm thus appears to originate specifically from three abdominal parasegments, PS10, 11 and 12. We show that the maintenance of high levels of 412 expression in gonadal mesoderm is not induced by contact with germ cells, but rather depends on genetic control by the homeotic genes abdominal-A and Abdominal-B.

Animals↗

Variation in thyroid hormone action and tissue content underlies species differences in the timing of metamorphosis in desert frogs.

Hormonal control of post-embryonic morphogenesis is well established, but it is not clear how differences in developmental endocrinology between species may underlie animal diversity. We studied this issue by comparing metamorphic thyroid hormone (TH) physiology and gonad development across spadefoot toad species divergent in metamorphic rate. Tissue TH content, in vitro tail tip sensitivity to TH, and rates of TH-induced tail tip shrinkage correlated with species differences in larval period duration. Gonad differentiation occurred before metamorphosis in species with long larval periods and after metamorphosis in the species with short larval periods. These differences in TH physiology and gonad development, informed by phylogeny and ecology of spadefoot metamorphosis, provide evidence that selection for the short larval periods in spadefoot toads acted via TH physiology and led to dramatic heterochronic shifts in metamorphic climax relative to gonad development.

Animals↗

Migratory and postmigratory mouse primordial germ cells behave differently in culture.

In all vertebrate groups, the progenitors of the germ line, the primordial germ cells (PGCs) arise extragonadally and move to the developing gonad early in embryonic development. We have examined the behavior of isolated pregonadal and gonadal PGCs in vitro on feeder layers of an embryo-derived cell line. Histochemically and serologically identified pregonadal germ cells are found to be actively motile in vitro and, furthermore, show behavior characteristic of invasive cells. PGCs isolated from the developing gonad, however, show little locomotory activity and are not invasive on the same cellular substrate. These observations suggest that PGCs undergo a major change in phenotype at the time of their entry into the gonad anlagen.

Alkaline Phosphatase↗

zfh-1 is required for germ cell migration and gonadal mesoderm development in Drosophila.

In Drosophila as well as many vertebrate systems, germ cells form extraembryonically and migrate into the embryo before navigating toward gonadal mesodermal cells. How the gonadal mesoderm attracts migratory germ cells is not understood in any system. We have taken a genetic approach to identify genes required for germ cell migration in Drosophila. Here we describe the role of zfh-1 in germ cell migration to the gonadal mesoderm. In zfh-1 mutant embryos, the initial association of germ cells and gonadal mesoderm is blocked. Loss of zfh-1 activity disrupts the development of two distinct mesodermal populations: the caudal visceral mesoderm and the gonadal mesoderm. We demonstrate that the caudal visceral mesoderm facilitates the migration of germ cells from the endoderm to the mesoderm. Zfh-1 is also expressed in the gonadal mesoderm throughout the development of this tissue. Ectopic expression of Zfh-1 is sufficient to induce additional gonadal mesodermal cells and to alter the temporal course of gene expression within these cells. Finally, through analysis of a tinman zfh-1 double mutant, we show that zfh-1 acts in conjunction with tinman, another homeodomain protein, in the specification of lateral mesodermal derivatives, including the gonadal mesoderm.

Animals↗

From fertilization to adult sexual behavior.

Research has established the broad mammalian developmental plan that genes on the sex chromosomes influence gonad development which determines gonadal hormone production (or its absence) leading to modification of the genitalia and simultaneously biasing the nervous system to organize adult sexual behavior. This might be considered the "gonad to hormones to behavior" model. It is clear, however, that although this model generally works well it is incomplete. The model does not account for behavioral influences attributed to the environment or to genetic but nongonadal or hormonal factors. In this essay we probe those areas of sexual development that are neither differentiated by hormones nor activated by them. The concept of the environment used for our discussion is very broad; it incorporates considerations of both the molar and the molecular levels. The general sense of the word "environment" as something exterior to the person is retained, even if that something influences intraperson processes. In addition, we focus directly on molecular events themselves. Here the "environment" involved can be that within a DNA segment. We also expand the notion of "biologically based sex differences." Although many, and perhaps most, important sex differences arise from gonadal and hormonal development, also important are sex differences which are neither gonadal nor hormonal. All these factors affect the internal workings of the individual and intervene in structuring how the social environment might or might not modify sexual behavior. This discourse calls attention to features that are central to the so-called nature-nurture discussion.

Fertilization↗

E-cadherin in the developing mouse gonad.

Expression of the cell adhesion molecule E-cadherin in the developing mouse has been investigated by immunocytochemistry and disaggregated organ culture. The principal aims were firstly, to determine whether E-cadherin is expressed in the indifferent gonad and if so with which cell population(s) it is associated. Secondly, to investigate the pattern of expression in the mesonephros, especially in relation to ventral mesonephric tubule cells, which contribute to the somatic cell population of the gonad. Thirdly, to discover whether there are any sex differences in expression of E-cadherin in differentiating gonads. Germ cells in the indifferent gonad showed strong immunoreactivity whereas somatic cells were unstained unless their membranes were in contact with those of germ cells. Positive staining for E-cadherin was found in epithelial cells of the mesonephric duct and tubules. Staining was weak at the ventral margins of the ventral mesonephric tubules. At later stages, germ cell immunoreactivity could be correlated with stages of ovarian differentiation, being reduced or absent between germ cells at 16 days post coitum, when ovigerous cords become dissociated as a prelude to follicle formation. Stronger staining reappeared briefly at 17 days post coitum, the time of follicular cell attachment to oocytes, before waning again 2 days later. Similarly, immunoreactivity in the differentiating testis was initially restricted to the germ cell population but pre-Sertoli cells were strongly positive between 16 and 19 days post coitum. The most striking sex difference was seen in the somatic cell population, where Leydig cells of the testis became strongly positive for E-cadherin from 17 days post coitum onwards. At this time, unlike controls, dissociated cells from gonads of either sex were unable to reform their initial contacts when cultured in the presence of the antibody to E-cadherin, confirming its functional importance.

Animals↗

The genus Acipenser as a model system for vertebrate urogenital development: nephrostomial tubules and their significance for the origin of the gonad.

Early gonadal development was studied in the sterlet, Acipenser ruthenus, by means of histological and semithin serial sections and scanning electron microscopy. Special attention was given to the role of opisthonephric nephrostomial tubules and their coelomic funnels (nephrostomes, coelomostomes) in the origin of the gonad. Specimens of about 1 mm in length (about 7 days post hatching) have a continuous kidney complex (holonephros) that extends from the branchial region to the level of the cloaca and may be divided into a cranial pronephros and a caudally following opisthonephros, with no overlapping of either portion. In specimens of 10 to 25 mm in length the regression of pronephros and cranial opisthonephros can already been seen; as a consequence, these parts of the kidney complex are not involved in gonadogenesis. The initial gonadal anlage is seen in specimens of 30-40 mm in length. The somatic cells of the gonadal primordium develop from the medial lips of segmentally arranged opisthonephric nephrostomes situated in a line that extends from the level of the stomach through that of the spiral valve. The nephrostomes involved in this process belong to the first-order set of nephrons, since nephrons of higher order that arise continuously from blastema cells during further growth of the animals never send nephrostomial tubules to the coelomic surface. The cells of the medial nephrostomial lips proliferate by many mitoses. They grow over and surround the large germ cells that have accumulated on the medial side of the nephrostomes. The proliferating nephrostomial cells are elongated in shape, and their long axes are oriented in the cranio-caudal direction. By their size, shape and arrangement they replace not only the flat, polygonal, mesothelial cells of lateral plate origin on the medial side of the nephrostomes, but also those in the interstices in between. The result is the formation of a continuous gonadal crest situated medially from the nephrostomial line. At 100-130 mm in length, the gonadal crest has reached the stage of a gonadal fold that is attached to the dorsal body wall by a thin mesogonadium. Stroma cells and blood vessels start to invade the gonadal fold. At 240-290 mm in length, parts of the gonadal fold are converted into a fat body, a structure that is typically present in adult sturgeons of either sex. Nephrostomial tubules and their nephrostomes are visible only for a short period in Acipenser ruthenus. They are completely developed in specimens of 25-40 mm in length and start continuous retrogression immediately afterwards. Therefore, they cannot play an important role in excretion. Their only purpose apparently is to function as precursor tissue for other organs, such as the gonad, in establishing a route by which cells of the intermediate mesoderm can gain access to and spread out over the coelomic surface. In conclusion, the observations made in Acipenser prove that this species is a most suitable model to explain the origin of the gonad from opistho/mesonephric nephrostomial tubules in vertebrates with a less-developed or early regressing pronephros. Most mammals, including man, belong to this category.

Animals↗

Requirement of WT1 for gonad and adrenal development: insights from transgenic animals.

Development of the gonad and adrenal gland occurs in close proximity and it has been proposed that they share a common primordium. Recent knockout and transgenic studies in mice have demonstrated that besides the orphan nuclear receptor SF1, the Wilms' tumor suppressor gene WT1 is important for gonad and adrenal gland development. WT1 is a complex gene with at least 24 isoforms produced from its locus. Although the role of each isoform is not yet understood, biochemical studies and analyses of human patients suggest that they do serve different functions in the cell. Here we present our current view of the part WT1 may play in the development of the gonad and discuss its possible role in the formation of the adrenal primordium.

Adrenal Glands↗

Patterns of development of gonads, sex-drive and hormonal responses in tropical beef bulls.

The development of different traits was studied in tropical beef bulls of seven genotypes (Brahman, Africander, British and combinations of these) from approximately 500 to 910 d of age. Bulls were raised under pasture conditions without supplementation. At each examination, approximately 2 mo apart, bulls were weighed, palpated (including scrotal and testicular measurement), electroejaculated, and subjected to two libido tests with estrus-induced females. At alternate examinations, plasma luteinizing hormone (LH) was measured at 30 and 150 min post gonadotrophin releasing hormone (GnRH) injection (LH - 30 and LH - 150) and testosterone (T) was measured at 150 min (T - 150). In general, nutritional and environmental stressors appeared to impede bull reproductive development. Scrotal circumference increased nonlinearly, apparently influenced by puberty and average daily gain (ADG). Libido increased overall, albeit nonlinearly also. No apparent marked differences in development of either trait could be attributed to genotype differences, although Brahman bulls tended to display lower sexual interest. The LH-30 level was relatively high (>14 ng/ml) at 500 and 640 d of age, but then dropped markedly at 760 d followed by a slight recovery. The LH-150 level followed a similar pattern, although it was very low at 500 d of age. The T-150 level showed a reverse pattern, being lower initially and higher in the latter part of the study. No apparent genotype differences occurred. Possible contributory influences on these patterns, including the onset of puberty and sexual maturity, season and nutrition, are discussed herein.

Journal Article↗