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PubMed2002

Ontogeny, tissue distribution, and hormonal regulation of insulin-like growth factor binding protein-2 (IGFBP-2) in a marine fish, Sparus aurata.

In this study, we have cloned insulin-like growth factor binding protein (IGFBP)-2 from a marine hermaphroditic fish species, the gilthead sea bream (Sparus aurata), and determined its structure, ontogeny, tissue distribution, and hormonal regulation. The sea bream IGFBP-2 precursor consists of 286 amino acids, including a putative signal peptide of 22 residues and a mature protein of 264 residues. The overall sequence of sea bream preIGFBP-2 is 52-39% identical to that of zebrafish, chick, mouse, rat, pig, sheep, bovine, and human preIGFBP-2. The cysteine-rich N- and C-terminal domains, which are believed to be important for IGF binding, show a greater degree of conservation with a sequence identity of 73-56% and 53-50%, respectively. Sea bream IGFBP-2 contains 18 cysteine residues and their alignment is identical to those of other vertebrate IGFBP-2s. The Arg-Gly-Asp (RGD) sequence, present in the C-terminal domain of all known IGFBP-2, is also present in the sea bream. Northern blot analysis of RNA samples extracted from adult liver and developing larvae, revealed two transcripts of about 1.3 and 2.5 kb. RT-PCR analysis showed that IGFBP-2 was expressed in all adult tissues studied, with the highest levels found in liver and skin. IGFBP-2 transcripts were detected in gonad during reproductive cycle of S. aurata. Highest levels of IGFBP-2 mRNA were found in bisexual young gonads, and the levels decreased with gonad development and relatively high levels of IGFBP-2 mRNA were found in the ovary during spawning. By contrast, testicular IGFBP-2 mRNA levels were very low. RT-PCR detected IGFBP-2 mRNA throughout development in unfertilized eggs, embryos, and larvae, with highest levels observed after day 3 post-hatching, suggesting that this mRNA is the product of both the maternal and embryonic genomes. High variability in steady-state levels of hepatic IGFBP-2 mRNA was noted in adult fish, resulting in a statistically insignificant response to growth hormone treatment. These results suggest that the structure of IGFBP-2 is conserved in sea bream and that IGFBP-2 mRNA is expressed during early development and in gonad during the reproductive cycle, suggesting that it may play a role in gilthead sea bream development and reproduction.

Amino Acid Sequence↗
PubMed2000

The significance of rudimentary nephrostomial tubules for the origin of the vertebrate gonad.

Initial gonadal development was studied in 30- to 40-day-old bovine embryos. The results were interpreted in conjunction with findings on pro- and mesonephric organization in larval forms of Ichthyophis kohtaoensis (Gymnophiona, Amphibia). In bovine embryos vestigial nephrostomial tubules are the immediate precursors of the blastemas for adrenocortical, rete, gonadal and Mullerian infundibular development. From the study of Ichthyophis it can be concluded that the vestigial nephrostomial tubules seen in the bovine embryo are pronephric and not mesonephric in nature. As a consequence, the indifferent mammalian gonad is defined as a modified homologue of the pronephros situated in the zone of pro-/mesonephric overlapping. Such an overlapping of the two kidney generations in the fully developed state is clearly seen in Ichthyophis. Overlapping of the mesonephros with the modified pronephros (gonad) is necessary to allow intercalation of mesonephric tubules (efferent ductules in mammals) into the male seminal excurrent duct system.

Alkaline Phosphatase↗
PubMed1980

A comparative study of the development of the fetal testis and ovary in the monkey (Macaca fascicularis).

The gonadal development of the Macaca fascicularis fetus was studied between 37 and 118 days on serial semi-thin and thin sections. The testis and the ovary began to differentiate at the same age (37 days); the definitive architecture of the testis was acquired at 43 days, while a cortex and a medulla did not form in the ovary until 55 to 60 days. In spite of the time-lag and the divergent development, the testis and the ovary evidenced three comparable stages; the main event of these stages was the centrifugal role of the mesonephros. The first stage (37-43 days) included the centrifugal and antero-posterior differentiation of the sex cord anlages from the mesonephric mesenchyme in contact with the proximal loops of the anterior tubules (for a detailed study see Dang and Fouquet, 1979). From 43 days (second stage), a remainder of the mesonephric mesenchymal blastema of the gonad supplied the rete system. The mesonephric tubules fused secondarily with that system which was connected to the sex cords. Whereas in the testis, the rete blastema did not play a direct role in organizing testicular structures, but only in forming excretory pathways, in the ovary, it invaded the medulla (whose initial sex cords degenerated) and penetrated to the ovigerous cords of the cortex. The rete ovarii blastema was probably the major source of periovocyte cells. The third stage included the differentiation of a steroidogenic interstitial tissue (from 50 days in the testis; at about 60 days in the ovary) and is further involution; these processes were similar in both sexes. Observation of the fine structure showed the development of the male and female gonocytes to be the same; the prespermatogonia and the oogonia could be characterized by the formation of nuclear vacuoles. The Sertoli cells and the periovogonial cells showed the same features.

Animals↗
PubMed2003

The role of GATA in mammalian reproduction.

GATA transcription factors are emerging as critical players in mammalian reproductive development and function. GATA-4 contributes to fetal male gonadal development by regulating genes mediating Müllerian duct regression and the onset of testosterone production. GATA-2 expression appears to be sexually dimorphic being transiently expressed in the germ cell lineage of the fetal ovary but not the fetal testis. In the reproductive system, GATA-1 is exclusively expressed in Sertoli cells at specific seminiferous tubule stages. In addition, GATA-4 and GATA-6 are localized primary to ovarian and testicular somatic cells. The majority of cell transfection studies demonstrate that GATA-1 and GATA-4 can stimulate inhibin subunit gene promoter constructs. Other studies provide strong evidence that GATA-4 and GATA-6 can activate genes mediating gonadal cell steroidogenesis. GATA-2 and GATA-3 are found in pituitary and placental cells and can regulate alpha-glycoprotein subunit gene expression. Gonadal expression and activation of GATAs appear to be regulated in part by gonadotropin signaling via the cyclic AMP-protein kinase A pathway. This review will cover the current knowledge regarding GATA expression and function at all levels of the reproductive axis.

Animals↗
PubMed2006

Differential expression of IGF-I mRNA and peptide in the male and female gonad during early development of a bony fish, the tilapia Oreochromis niloticus.

Insulin-like growth factor I (IGF-I) plays a key role in the complex system that regulates bony fish growth, differentiation, and reproduction. The major source of circulating IGF-I is liver, but IGF-I-producing cells also occur in other organs, including the gonads. Because no data are available on the potential production sites of IGF-I in gonad development, developmental stages of monosex breedings of male and female tilapia from 0 day postfertilization (DPF) to 90 DPF were investigated for the production sites of IGF-I at the peptide (immunohistochemistry) and mRNA (in situ hybridization) level. IGF-I mRNA first appeared in somatic cells of the male and female gonad anlage at 7 DPF followed by IGF-I peptide around 9-10 DPF. Gonad anlagen were detected from 7 DPF. Starting at 7 DPF, IGF-I peptide but no IGF-I mRNA was observed in male and female primordial germ cells (PGCs) provided that IGF-I mRNA was not under the detection level, this observation may suggest that IGF-I originates from the somatic cells and is transferred to the PGCs or is of maternal origin. While in female germ cells IGF-I mRNA and peptide appeared at 29 DPF, in male germ cells both were detected as late as at 51-53 DPF. It is assumed that the production of IGF-I in the germ cells is linked to the onset of meiosis that in tilapia ovary starts at around 28 DPF and in testes at around 52-53 DPF. In adult testis, IGF-I mRNA and peptide occurred in the majority of spermatogonia and spermatocytes as well as in Leydig cells, the latter indicating a role of IGF-I in the synthesis of male sex steroids. In adult ovary, IGF-I mRNA and IGF-I peptide were always present in small and previtellogenic oocytes but only IGF-I peptide infrequently occurred in oocytes at the later stages. IGF-I expression appeared in numerous granulosa and some theca cells of follicles at the lipid stage and persisted in follicles with mature oocytes. The results suggest a crucial role of local IGF-I in the formation, differentiation and function of tilapia gonads.

Animals↗
PubMed2003

Maternal transfer of photoperiodic information in Siberian hamsters. vi. effects of time-dependent 1-hr melatonin infusions in the mother on photoperiod-induced testicular development of her offspring.

We tested in Siberian hamsters the nature of the maternal signal that relays photoperiodic information to the developing fetuses. As previous investigations have identified maternal hormonal and circadian components in this process, the specific goal of this presentation is to determine quality of the signal that connotes daylength when it is imparted to the fetus. Does the function of the signal received by the fetus best support the coincidence or duration hypotheses of photoperiodic induction? Pregnant hamsters received 1 or 8 hr melatonin or vehicle infusions everyday. Juveniles of intact mothers gestated on 16 hr of light per day (16L) experienced maximal suppression of testicular development when reared on 14L. However, when intact mothers gestated on 10L received a 1-hr melatonin infusion daily at 20:00-21:00 hr, their young responded to 14L with greatly accelerated testicular development. In the absence of the maternal pineal gland (and, therefore, the maternal melatonin signal), the effects of maternal melatonin infusions were reversed. Here, only the juveniles of 16L-gestated females infused at 20:00-21:00 hr daily responded to 14L with enhanced testicular development. All other groups showed the same extent of gonadal development, independent of the time or type of infusion their mothers received. Testicular development on 14L of all juveniles from pinealectomized mothers gestated on 10L was of the same magnitude, regardless of the type and time of infusion their mothers received during pregnancy. The results suggest that the maternal signal transferred to the fetuses during gestation consists not only of the daily melatonin signal, but also some circadian-based component that greatly affects the effect of the former. The timing, and not the duration, of the maternal melatonin signal with respect to the animals' (mother and fetus) circadian day is of crucial importance in the transfer of photoperiodic information from mother to fetus.

Animals↗
PubMed2002

Vertebrate sex determination: many means to an end.

The differentiation of a testis or ovary from a bipotential gonadal primordium is a developmental process common to mammals, birds and reptiles. Since the discovery of SRY, the Y-linked testis-determining gene in mammals, extensive efforts have failed to find its orthologue in other vertebrates, indicating evolutionary plasticity in the switch that triggers sex determination. Several other genes are known to be important for sex determination in mammals, such as SOX9, AMH, WT1, SF1, DAX1 and DMRT1. Analyses of these genes in humans with gonadal dysgenesis, mouse models and using in vitro cell culture assays have revealed that sex determination results from a complex interplay between the genes in this network. All of these genes are conserved in other vertebrates, such as chickens and alligators, and show gonad-specific expression in these species during the period of sex determination. Intriguingly, the sequence, sex specificity and timing of expression of some of these genes during sex determination differ among species. This finding indicates that the interplay between genes in the regulatory network leading to gonad development differs between vertebrates. However, despite this, the development of a testis or ovary from a bipotential gonad is remarkably similar across vertebrates.

Alligators and Crocodiles↗
PubMed1989

Two gene members of the murine HOX-5 complex show regional and cell-type specific expression in developing limbs and gonads.

This study reports the expression domains of two murine HOX gene members of the HOX-5 complex (Hox-5.2, Hox-5.3). These two genes have very similar homeodomain sequences, as well as temporal and spatial specificities of expression. They are both expressed at very posterior levels in the central nervous system, in sclerotome derivatives and in a few internal organs. In addition to these expression domains which are shared with other HOX genes, transcripts from both Hox-5.2 and Hox-5.3 are present at high levels in developing limbs. After an early homogeneous expression in mesodermal limb bud cells, transcription becomes restricted to cartilage-differentiating cells. In addition, Hox-5.2 is a marker for gonadal development. The possible involvement of such genes during inductive processes or organogenesis is discussed.

Amino Acid Sequence↗
PubMed2001

Follicle-stimulating hormone and its alpha and beta subunits in rainbow trout (Oncorhynchus mykiss): purification, characterization, development of specific radioimmunoassays, and their seasonal plasma and pituitary concentrations in females.

Gonad development in fish, as in mammals, is regulated by two gonadotropins (GTHs), FSH and LH. The function of LH in fish has been clearly established; however, the function(s) of FSH is less certain. The lack of specific and sensitive assays to quantify FSH and its alpha and beta subunits has hindered studies to assess physiological function. In this study, gel filtration chromatography, ion exchange chromatography, and HPLC were employed to purify FSH and its subunits from pituitary glands of rainbow trout (Oncorhynchus mykiss), and the identities of the isolates were confirmed by amino acid analysis. Polyclonal antibodies were raised against the free GTHalpha2 and free FSHbeta subunits to develop specific RIAs. The sensitivities of the intact FSH, GTHalpha2, and FSHbeta assays were 1 ng/ml, 0.2 ng/ml, and 0.1 ng/ml, respectively, and the cross-reaction of these molecules with each other and with intact LH in the heterologous assays was <10.4% throughout. Pituitary and plasma samples diluted in parallel with the standards in all three assays and spiked sample recoveries were >90% throughout. Measurement of plasma and pituitary concentrations of intact FSH in female rainbow trout confirmed the established seasonal profiles. Concentrations of free GTHalpha2 subunit were elevated both in the plasma and in the pituitary in females at ovulation (maximum concentrations: 34.93 +/- 6.3 ng/ml in plasma; 37.63 +/- 5.79 microg/pituitary). In both the plasma and the pituitary, free FSHbeta subunit was present throughout the reproductive cycle but at very low concentrations when compared with both free GTHalpha2 and intact FSH. The presence of free GTHalpha2 subunit in the plasma similarly occurs in mammals, but its functional significance in fish has yet to be established.

Animals↗
PubMed1990

Effects of corticosterone, metapyrone, and ACTH on testicular function at different stages of the breeding cycle in migratory redheaded bunting, Emberiza bruniceps.

To study the seasonal reproductive responses, corticosterone (0.2 and 2 micrograms/day), metapyrone (0.6 mg/day), and ACTH (0.25 IU/day) were administered in male redheaded bunting, Emberiza bruniceps, during different phases of the annual gonadal cycle. All of the three treatments inhibited gonadal growth during the preparatory phase, suggesting that during the early stage of annual gonadal growth, an optimum level of the hormones of pituitary-adrenal axis is essential. In the progressive and breeding phase, while corticosterone administration had no effect, metapyrone inhibited the annual gonadal development, suggesting the necessity of increasing or increased adrenal steroid hormone during this phase. In the regressive phase, while corticosterone enhanced the rate of gonadal regression, metapyrone treatment did not allow the gonad to regress. Further, ACTH administration induced full breeding condition in the regressing gonad. It is suggested that while a higher level of adrenal steroids and a low ACTH (by exogenous corticosterone) may enhance the testicular regression, moderate levels of ACTH (caused by exogenous metapyrone) did not allow regression and increased ACTH levels (by exogenous ACTH) caused full development of the regressing gonad. Thus the sensitivity of the neuroendocrine-gonadal axis not only varies in response to photoperiod, but the reproductive system also responds differently to pharmacological administration of the above three hormones and drugs during different phases of the breeding cycle.

Adrenocorticotropic Hormone↗
PubMed1996

[The histophysiological characteristics of ovarian development with an altered level of thymic peptides at infancy in mice].

Thymic peptides effect on the female gonad development was studied: consequences of these substances dysbalance in infancy (T-activine administration of 0.1 microgram/g of body weight in dose since 8 to 21 postnatal day) was analysed. Morphofunctional diagnostics of the ovarian state is based on the spontaneous organism reaction to the drug administration (to 21-days-old animals) and on the distant consequences of its activity (in 1-, 2-, 3-months female animals). Thymic peptides dysbalance in infancy promotes only transitory disorders of the ovarian histophysiology, expressing in transitory inhibition of the follicle genesis with the simultaneous atretic process activation, which results in puberty delay with the following complete restoration of reproductive function. Taking into account that in the early neonatal period gonad reacts to T-activine administration with the development of strong ovarian pathology, the results obtain allow to conclude that the reactivity to thymic peptides content oscillations reduces so as the significance of the factors cited in the ovarian morphogenesis regulation in proportion with the animals' maturity.

Adjuvants, Immunologic↗
PubMed1989

Origin of somatic cells and histogenesis in the primordial gonad of the Japanese tree frog Rhacophorus arboreus.

Gonadal development in Rhacophorus arboreus, a sexually semidifferentiated type of tree frog, was observed by means of the electron microscope, and cell proliferation kinetics were examined autoradiographically. The genital ridge consisted of coelomic epithelial cells and primordial germ cells. The gonadal medulla was formed by the segregation of epithelial cells within the primordial gonad. Thereafter, the medullary cell mass was well developed and oogenesis began in the gonadal cortex, irrespective of genetic sex. During metamorphosis, the ovarian cavity was formed in the medullary mass. This ovarian structure developed further in females. In males, on the other hand, a layer of medullary cells comprising the epithelium of the ovarian cavity proliferated rapidly and reformed a large cell mass. The degeneration of ovarian follicles and the formation of cell cords (rudimentary seminiferous tubules) were seen in the cortex. These cell cords were separated from the superficial epithelium and continued to the medullary mass (rudimentary testicular rete). These results clearly indicate that both the cortical and medullary cells are derived from the coelomic epithelium and that the development of the cortex and medulla is not always antagonistic in the course of sexual differentiation.

Animals↗
PubMed1992

Gonad formation and development requires the abd-A domain of the bithorax complex in Drosophila melanogaster.

The abdominal-A (abd-A) gene, a member of the bithorax complex, is required for the correct identity of parasegments (PS) 7 through 13. Mutations in iab-4, one of the cis-regulatory regions of abd-A, transform epidermal structures of PS 9 and also cause loss of gonads in adult flies. Here, we describe a developmental and molecular analysis of the role of iab-4 functions in gonadal development. In flies homozygous for a strong iab-4 allele, gonadogenesis is not initiated in the embryo because the mesodermal cells fail to encapsulate the pole cells. Flies homozygous for weaker iab-4 mutations sometimes form ovaries. The ovary-oviduct junctions are abnormal, however, and egg transfer from the ovary to the uterus is blocked in the adult. To localize the sites that require iab-4 function, we have analyzed animals chimeric for the mutant and wild-type cells. These chimeras were generated by three kinds of transplantation experiments: pole cells, embryonic somatic nuclei or larval ovaries. Our results suggest that iab-4 is required in the somatic cells of the gonadal primordia, but not the germ line. In addition, the formation of functional ovary-oviduct junctions and egg transfer also requires iab-4 functions in the somatic cells of the ovary and in at least one additional somatic tissue.

Animals↗
PubMed2001

Sexual differentiation of germ cell deficient gonads in the medaka, Oryzias latipes.

To determine whether germ cells perform any function in gonadal sexual differentiation, development of gonads in the medaka, Oryzias latipes, after exposure to busulfan was investigated. Busulfan suppressed proliferation of early germ cells, thus significantly reducing the number of germ cells and generating regions without germ cells in the developing gonads. Globular structures were observed in the parenchyma in these regions. The structure was male specific, developed at the same time as acinus (seminiferous tubule precursor), surrounded by the basal lamina, and contained characteristic desmosomes. These results strongly suggest that these globular structures are the precursors of seminiferous tubules devoid of germ cells. In the ovary, no follicles were observed but a well-developed ovarian cavity was evident. From these results we conclude that differentiation of gonadal parenchyma cells, except for follicular ones, is not germ cell dependent, though morphological differentiation of the somatic cells seems to follow the differentiation of germ cells.

Alkylating Agents↗
PubMed1996

Inhibin and activin have antagonistic paracrine effects on gonadal steroidogenesis during the development of the chicken embryo.

Primary cultures of gonadal cells from chicken embryos were used to explore the paracrine role of inhibin and activin during gonadal development. Fetal testicular and ovarian cells secreted high amounts of immunoactive inhibin. FSH caused a dose-related increase of cAMP and immunoactive inhibin concentrations in testicular cell cultures. Postreceptor signalling through the protein kinase A (PKA) pathway was confirmed by additional experiments with 8-bromo-cAMP, 3-isobutyl-1-methyl-xanthine (MIX), prostaglandins, forskolin, and choleratoxin. The relative ability of these agonists to stimulate cAMP production did not always correlate with their ability to stimulate inhibin secretion. Experiments with phorbol 12-myristate 13-acetate suggested that the regulation of immunoactive inhibin secretion also involves the protein kinase C (PKC) pathway. In addition, it was shown that recombinant human (rh)-inhibin increases the conversion of pregnenolone to androgens whereas rh-activin has the opposite effect. Recombinant human follistatin, an activin-binding protein, antagonized the actions of rh-activin and to a lesser extent those of rh-inhibin. In conclusion, these results show that during the development of the chicken embryo, gonadal inhibin secretion may be regulated by hormones and by local factors such as prostaglandins. Cross talk between the PKA and PKC pathways may be involved in this regulation. Recombinant human inhibin and rh-activin may have antagonistic roles in the paracrine control of gonadal steroidogenesis.

Activins↗
PubMed2006

Development of chronic tests for endocrine active chemicals. Part 2: an extended fish early-life stage test with an androgenic chemical in the fathead minnow (Pimephales promelas).

The Endocrine Modulators Study Group (EMSG) of the European Chemical Industry has proposed an extended fish early-life stage (ELS) test based on OECD test guideline 210 in combination with a fish pair-breeding reproduction study as a possible alternative for fish full life cycle testing. In this paper the androgen methyldihydrotestosterone (MDHT) was tested in an extended ELS test with fathead minnow supplementary to such a test with the weak estrogen 4-tert-pentylphenol (4TPP). Main endpoints were secondary sexual characteristics (SSC), plasma vitellogenin (VTG) induction and gonadal development. Early blastula embryos were exposed to 0, 0.10, 0.32 and 1.0 microgMDHTl(-1) for up to 114 days post-hatch (dph). A batch of fish exposed to 1.0 microg l(-1) was transferred to clean water after 30 or 63 dph for the remainder of the study. Ethinylestradiol (EE2) was included as estrogenic reference substance at 0.01 microg l(-1). Exposure to MDHT had no significant effect on hatching success or survival, but significantly increased the condition factor of fish exposed for 63 and 114 dph (up to 150% of the control). At 63 dph MDHT exposure induced appearance of tubercles on the snout (a male SSC) of more than 80% of fish. Compared to the controls, plasma VTG was not detectable or significantly lower in fish exposed to MDHT at 0.10 microg/l, but not significantly affected at higher MDHT concentrations. Both lower levels of MDHT significantly inhibited the development of female gonads as of 30 dph. Fish exposed to MDHT at 0.32 and 1.0 microg l(-1) showed higher incidences of mixed sex gonads (10-25%) and smaller testes or dysplasia of gonadal tissue. Dysplasia was present in 80% of the fish continuously exposed to 1.0 microg l(-1) up to 114 dph, but reversible when fish were transferred to dilution water. Results indicate that suppression of ovarian development was the most sensitive endpoint for MDHT exposure after 30 dph. Other endpoints (e.g., growth and SSC) required exposure during at least up to 63 dph to yield a significant effect. Androgenic effects on VTG production required even longer exposure, i.e., until sufficient number of females had matured.

Androgens↗
PubMed1984

Gonadal tumors in patients with gonadal dysgenesis and sex chromosomal rings and fragments.

Patients with female phenotypes and dysgenetic gonads harboring testicular tissue have a markedly increased risk of developing gonadal tumors. Cytogenetic demonstration of Y chromatin is the currently accepted criterion for performing prophylactic gonadectomies in these women. We studied four patients with dysgenetic gonads containing either testicular tissue or germ cell tumors. All had small sex chromosomal fragments which could not be characterized by conventional cytogenetic studies. Clinical features, DNA replication studies, and immunologic assays of Xga and H-Y antigens failed to correlate consistently with the gonadal histology. We recommend prophylactic gonadectomies and subsequent hormone replacement in all patients with female phenotypes, gonadal dysgenesis, and cytogenetically indeterminate sex chromosomal fragments.

Adolescent↗
PubMed2005

Hystological analysis of ovarian development of the characiform Oligosarcus hepsetus (Cuvier, 1829) in a Brazilian reservoir.

A histological analysis was performed of the ovarian development of Oligosarcus hepsetus (Cuvier, 1829), a medium-sized carnivorous Characiform species in the Lajes reservoir, Brazil. A total of 125 fishes, collected monthly between April 2001 and June 2002 were examined by routine macroscopic and histological techniques. Eight phases of the oocyte development were described, and 4 stages and 4 substages of gonadal development were proposed. Spawning in reservoirs, similarly to that in rivers, is in batches, which favors juvenile survival and suggests that the impoundment of this area did not influence the reproductive strategy of this very successful species in a dammed environment.

Animals↗