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Expression profiles of Dax1, Dmrt1, and Sox9 during temperature sex determination in gonads of the sea turtle Lepidochelys olivacea.

Sex determination is controlled either by genetic or environmental factors. In mammals Sry initiates determination but no homologue of this gene exists in non-mammalian species. Other genes of the mammalian sex-determining pathway have been identified in gonads of different vertebrates. Sox9, Dax1, and Dmrt1 are expressed at the onset of gonadal development in birds and reptiles. In the sea turtle Lepidochelys olivacea, a species with temperature sex determination (TSD), Sox9 is expressed in undifferentiated gonads at male- (MPT) or female-promoting temperatures (FPT). At MPT, Sox9 remains expressed in male gonads, but at FPT it is downregulated coinciding with the onset of the ovarian morphologic differentiation and female sex determination. At MPT however, male sex is determined early than at FPT in still undifferentiated gonads suggesting that other genes maintain Sox9 expression in testis. Here we used RT-PCR to study the expression profiles of Dax1, Dmrt1, and Sox9 in gonads of embryos of L. olivacea incubated at MPT or at FPT. The profiles were correlated with sex determination during and after the temperature-sensitive period (TSP). Dax1 maintained similar levels at both temperatures during the TSP. The Dax1 expression level increased significantly in ovaries compared to testes at stage 27, once they were morphologically distinct. The expression levels of Dmrt1 were higher at MPT than at FPT at all stages, in contrast with Sox9 levels which were similar at both temperatures at stages 23-25. Together, current results suggest that, whereas Dax1 is not involved in TSD in L. olivacea, upregulation of Dmrt1 and downregulation of Sox9 may play a role in male and female sex determination, respectively.

Amino Acid Sequence↗

Naturally occurring antihormones: secretion of FSH antagonists by women treated with a GnRH analog.

Follicle-stimulating hormone (FSH) is a glycoprotein essential for gonadal development and steroidogenesis. Recent studies suggest that deglycosylation of FSH results in the formation of antagonistic proteins that are capable of binding to gonadal receptors but that are devoid of bioactivity. Treatment of hypogonadal women with an antagonist of gonadotropin-releasing hormone substantially decreased serum FSH bioactivity with minimal changes in immunoreactivity. Chromatofocusing and size fractionation of the serum samples indicated the secretion of immunoreactive FSH isoforms that are devoid of bioactivity but that are capable of blocking FSH action in ovarian granulosa cells. These findings provide the first demonstration of naturally occurring circulating antihormones. These FSH antagonists may play an important role in the physiology and pathophysiology of the gonads.

Biological Assay↗

Zinc transport activity of Fear of Intimacy is essential for proper gonad morphogenesis and DE-cadherin expression.

Embryonic gonad formation involves intimate contact between germ cells and specialized somatic cells along with the complex morphogenetic movements necessary to create proper gonad architecture. Previously, we have shown that gonad formation in Drosophila requires the homophilic cell-adhesion molecule Drosophila E-cadherin (DE-cadherin), and also Fear of Intimacy (FOI), which is required for stable accumulation of DE-cadherin protein in the gonad. Here, we present an in vivo structure-function analysis of FOI that strongly indicates that zinc transport activity of FOI is essential for gonad development. Mutant forms of FOI that are defective for zinc transport also fail to rescue morphogenesis and DE-cadherin expression in the gonad. We further show that expression of DE-cadherin in the gonad is regulated post-transcriptionally and that foi affects this post-transcriptional control. Expression of DE-cadherin from a ubiquitous (tubulin) promoter still results in gonad-specific accumulation of DE-cadherin, which is strongly reduced in foi mutants. This work indicates that zinc is a crucial regulator of developmental processes and can affect DE-cadherin expression on multiple levels.

Amino Acid Sequence↗

Reproductive cycle of hepatus pudibundus (Herbst, 1785) (Crustacea, Decapoda, Calappidae) in Ubatuba, SP, Brazil.

This study aimed to analyze the reproductive cycle of the crab Hepatus pudibundus (Herbst, 1785). The crabs were collected in Ubatuba, SP (23 degrees 26'S and 45 degrees 02'W) monthly from the July, 1992 to June, 1993. Samplings were performed with an "otter trawl". In the laboratory the crabs were numbered, sexed and measured for their maximum carapace width. The gonad developmental stage and the moult stage were also macroscopically registered. In all sampled months ovigerous females and crabs in both sexes with developed gonad were observed. Moult activity in all size classes was verified. Such fact indicates that this species can suffer moult process after their maturity. In despite of temperature being pointed a one of the most important ecological factor in the reproductive biology of aquatic animals, it was verified any relation between water temperature and ovigerous ratio. The oscillations of the environmental factors in the Ubatuba region sea probably were not enough to alter the continuity of the physiological processes related to H. pudibundus reproduction.

Animals↗

Termination of photorefractoriness in the brahminy myna, Sturnus pagodarum: role of photoperiods and gonadal hormones.

In brahminy myna a photosensitive species, long days caused full gonadal development followed by rapid regression, whereas short days inhibited these responses. Experiments were performed to investigate the effects of duration of photoperiod and gonadal hormones on the recovery of photosensitivity to long photoperiods in male birds. Groups of photorefractory birds were subjected to 8-, 9- or 11-hr daily photoperiods for 45 (6.5 weeks) or 63 (9 weeks) days and then transferred to 15 h daily photoperiods for 60 days to check for the regaining of photoresponsivity. A control group was held under 15L:9D throughout the period of study. Another experiment included three groups of photorefractory males, which were maintained on 9L:15D for 9 weeks and administered with, birth-1day-1 alternately for first 30 days olive oil or different doses (10 or 100 micrograms) of testosterone propionate (TP)/bird/day alternately for first 30 days, and then transferred to 15L:9D for another 30 days to test the recovery of photosensitivity. The results indicated that (i) a period of exposure to short daylengths is required to dissipate photorefractoriness, (ii) termination of photorefractoriness is dependent on the length and duration of photoperiods and (iii) TP inhibits the recovery of photosensitivity in a dose dependent manner.

Animals↗

Expression of androgen receptors in upper human fetal reproductive tract.

BACKGROUND: Androgens play a key role in human fetal development. All androgens act through a single intracellular androgen receptor (AR), which is encoded by a single copy gene on the X chromosome. ARs are expressed as early as 9 weeks in the epithelium and mesenchyme of the urogenital sinus, paramesonephric (Müllerian) and mesonephric (Wolffian) ducts. METHODS: Using immunohistochemistry, we investigated the distribution of ARs in the gonads and lower genital tracts of 54 human fetuses at 8-11 weeks of gestation. Gender was determined by PCR. RESULTS: The AR was expressed in a similar pattern in both male and female fetuses. There appears to be no difference in expression in the mesonephros or the mesonephric ducts when male and female pelvises were compared. Expression in the female paramesonephric duct was within the epithelium, whereas, in the male pelvises, expression was in the mesenchyme of the paramesonephric duct. When AR expression was compared in the ovary and testes, both gonads seem to express AR at 9 weeks, but this expression was extended into the 10th week of gestation in the male. CONCLUSION: The specific pattern of AR expression implies a key role in gonadal development. However, the pattern of staining was similar in the gonads at 8 and 9 weeks in both sexes, although staining persisted longer in the testis until the 10th week. AR expression, therefore, is not a key determinant of human gonadal differentiation.

Embryo, Mammalian↗

A case of gonadal dysgenesis, breast development, Graves' disease, and low bone mass.

OBJECTIVE: To describe a case of XY gonadal dysgenesis with Tanner stage 4 breast development in the absence of a hormone-producing gonadal neoplasm and with Graves' disease and low bone mass. METHODS: The clinical features, laboratory results, and cytogenetic findings in the patient are presented, and the potential mechanisms of breast development are discussed. A MEDLINE search was performed, and related articles in the English-language literature published between 1955 and 2001 were reviewed. RESULTS: A 23-year-old African American woman was referred to the University of Louisville Hospital for evaluation of hyperthyroidism. About 4 months before this referral, hyperthyroidism was diagnosed, and treatment with methimazole was initiated. She continued to have thyrotoxicosis. Additionally, systemic review disclosed a history of primary amenorrhea. Physical examination revealed a tall phenotypic female patient with Tanner stage 4 breast development. Pelvic examination showed normal findings except for sparse pubic hair. Laboratory evaluation confirmed the diagnosis of Graves' disease as well as primary gonadal failure. Pelvic ultrasonography revealed a small uterus and bilateral adnexal masses (0.9 by 0.6 cm). On chromosomal analysis, a 46,XY karyotype was found. Further analysis of Y-DNA by polymerase chain reaction confirmed the presence of an intact Y chromosome, and no microdeletions were identified. Dual-energy x-ray absorptiometry demonstrated a Z-score of -4.7 and -4.2 at the lumbar spine and right hip, respectively. Graves' disease was successfully treated with (131)I. Laparoscopy was performed to resect streak gonads. On histologic examination, no typical ovarian, testicular, or neoplastic tissue was identified. The breast development in this patient remains unexplained. CONCLUSION: To the best of our knowledge, this is the first case report of a tall XY female patient with breast development in the absence of a hormone-producing gonadal neoplasm and without clearly identifiable gonads. Breast development was most likely related to estrogens, possibly produced by either streak gonads at the time of puberty or peripheral conversion of androgens, or to increased sensitivity of breast tissue to estrogens. Graves' disease is likely coincidental and could contribute to bone loss in such subjects.

Absorptiometry, Photon↗

Sex difference in the action of activin-A on cell proliferation of differentiating rat gonad.

The effect of recombinant bovine activin-A on DNA synthesis in fetal rat gonads and mesonephroi was studied in vitro by using [3H]thymidine autoradiography to evaluate the role of activin in the regulation of gonadal development. mRNA levels of inhibin-beta A and -beta B, activin receptor II and IIB (ActRII and ActRIIB), and follistatin were studied by Northern hybridization in the fetal rat gonads and mesonephroi. Activin stimulated thymidine incorporation in ovaries and female mesonephroi on days 14 and 15 postcoitum (pc) in a dose-dependent manner, as assessed by autoradiography. In contrast, activin inhibited thymidine incorporation in testes and male mesonephroi on day 14 pc in a dose-dependent way. On day 18 pc, the stimulatory effect of activin in ovary was nearly lost, whereas that in mesonephros was still pronounced. Activin had no effect on thymidine incorporation in testes and male mesonephroi on day 15 or 18 pc. Inhibin-beta A mRNA was seen in testes and mesonephroi in the male and in mesonephroi in the female of 15 and 18 day pc embryos. Inhibin-beta A mRNA expression was also seen in the testes and epididymes of newborn male rats and in the ovaries of 11-day-old postnatal rats. Inhibin-beta B mRNA was present in both testes and ovaries from 15 day pc onward. ActRIIB mRNA was most abundant in the testes, ovaries, and mesonephroi of 15 day pc embryos. The expression of ActRIIB message diminished during aging. ActRII mRNA was ubiquitously expressed during development. Follistatin mRNA was seen in the ovaries from 15 day pc onward and in the oviducts at 11 days postnatally. The present novel findings of activin subunit and receptor mRNA expression and activin action on gonadal and mesonephric cell proliferation suggest an important, sexually dimorphic role for this substance in fetal gonadal differentiation.

Activin Receptors↗

Perinatal influences of melatonin on testicular development and photoperiodic memory in Siberian hamsters.

We assessed the influence of perinatal melatonin on reproductive development and adult responsiveness to melatonin. Testicular growth in an intermediate day length (14 : 10 h light/dark cycle) was substantially reduced in Siberian hamsters gestated by pinealectomised compared to pineal-intact females; gonadal development was normalised in offspring of pinealectomised dams that were pinealectomised at 3-4 days of age. Hamsters deprived of melatonin only during gestation, or both pre- and postnatally, underwent testicular involution during treatment with melatonin in adulthood. Photoperiodic histories acquired prenatally did not endure as long as those acquired by adult hamsters. Hamsters first exposed to melatonin in adulthood were not more proficient in acquiring photoperiodic histories than were normal males. These findings indicate that pre- versus postnatal differences in melatonin signal duration determine rates of testicular development. Exposure to melatonin perinatally does not appear to organise the neuroendocrine substrate that mediates effects of day length and melatonin on the gonads of adult hamsters.

Age Factors↗

The endocrine control of reproduction and molt in male and female emperor (Aptenodytes forsteri) and adelie (Pygoscelis adeliae) penguins. I. Annual changes in plasma levels of gonadal steroids and LH.

Changes in plasma LH, testosterone, and estrogens were investigated throughout reproduction and molt in free-living male and female emperor (Aptenodytes forsteri) and adelie (Pygoscelis adeliae) penguins. In both sexes and species, plasma LH and gonadal steroids were severalfold above basal level at the time of arrival on the breeding grounds, suggesting that environmental cues (especially decreasing daylength in emperors) rather than mating and courting primarily stimulate gonadal development and reproduction. In both species a marked increase in plasma LH (both sexes), testosterone (males), and estrogens (females) corresponded with the time of maximum gonadal size, and peak values were obtained at the time of copulation, i.e., in emperors about 10-15 days prior to egg laying. In females, plasma LH and estrogens dropped to low levels between copulation and egg laying. Similarly, following copulation in males plasma testosterone fell to lower levels that in emperors were maintained during incubation and brooding of the non-thermally emancipated chick. Plasma LH levels followed the same trend as testosterone, falling after copulation and falling further prior to molt. Though lower than at copulation, plasma LH was higher in incubating (males) and brooding (males and females) emperors than during rearing of the thermally emancipated chicks, suggesting that plasma LH might be related to incubating, brooding, and territorial behavior. In male and female emperors and in male adelies, plasma gonadal steroids and LH were at basal levels throughout molt.

Animals↗

Development of the subfornical organ and area postrema of the male albino mouse. Karyometric effect of neonatal and prepuberal castration.

We have studied the karyometric development of the nuclei of the ependymal cells and neurons of the subfornical organ and the area postrema in the male albino mouse from the 5th to the 190th postnatal day. We have found similar patterns of development in both although the area postrema showed more significant postnatal oscillations than those of the subfornical organ, suggesting a more intimate chronological relationship to gonadal development. We have furthermore analyzed the development in two experimental groups: in the one animals were castrated at birth, in the other, castration was made on the 20th postnatal day. We have found that neonatal castration produced a significant decrease of nuclear sizes; this was more evident in the subfornical organ than in the area postrema in earlier stages of development while the response was similar in both at peripuberal ages. The response to prepuberal castration was similar in both organs.

Aging↗

Expression and role of PDGF-BB and PDGFR-beta during testis morphogenesis in the mouse embryo.

The role played by PDGF in testis morphogenesis is still incompletely understood. The present study investigates the expression and potential role of platelet-derived growth factor-BB (PDGF-BB) and its receptor, PDGF receptor beta (PDGFR-beta), during mouse testis cord formation, and the possibility that the growth factor may be involved in the migration to the gonad of mesenchymal cells of mesonephric origin. Studies from this laboratory have previously shown that mesenchymal cells that migrate from the mesonephros into the gonad, to form peritubular myoid cells and most of the intertubular cells, can be identified by the presence on their surface of the p75 neurotrophin receptor (p75NTR), and can be isolated to near-purity by immunomagnetic selection with anti-p75NTR antibody. We show here that mesonephric p75NTR(+) cells also bear the PDGFR-beta, and are able to migrate and proliferate in vitro in response to PDGF-BB. PDGF-BB is expressed at higher levels in male than female developing gonads, suggesting a role for this factor in testis development. Such a role is further supported by the observation that addition of PDGF-BB to serum-free medium is sufficient to allow organ-cultured male 11.5 days post-coitum urogenital ridges to form testis cords. Finally, we show that mesonephric cell motility and growth induced by exposure to PDGF-BB involve mitogen-activated protein kinases (MAPK) and phosphatidylinositol-3 kinase (PI3-K) pathways, as MAPK inhibitor U0126 and PI3K inhibitor Ly294002 inhibit migration and proliferation in vitro assays. The present findings support the hypothesis that the PDGF/PDGFR system plays a key role in testis morphogenesis in the mouse embryo.

Animals↗

Development of the gonad in normal and anencephalic human fetuses.

The development of the germinal and somatic cells in the gonads of normal and anencephalic fetuses was investigated. Interstitial cells were reduced in number in both sexes and in males the population of gonocytes in profiles of seminiferous tubules was reduced when Leydig cells were seeminaly absent. Ovarian differentiation up to about 34 weeks after conception was apparently normal, although precise counts of the numbers of germ cells were not carried out. Nevertheless, oogonia increased in number by mitotic divisions, oocytes progressed through meiotic prophase to the diplotene stage, and primordial follicles formed normally. By contrast, 'growing' and small antral follicles, which are a constant feature of control ovaries, were not observed in the anencephalic specimens.

Anencephaly↗

Guidance of primordial germ cell migration.

Primordial germ cells (PGCs), the progenitors of the gametes, migrate from the position where they are specified towards the region where the gonad develops. To reach their target, the PGCs obtain directional cues from cells positioned along their migration path. One such cue, the chemokine SDF-1, has recently been found to be critical for proper PGC migration in zebrafish and in mice. In Drosophila, too, a molecule that is structurally related to chemokine receptors and is important for PGC migration has been identified. The ability to visualize chemokine-guided migration at a high resolution in vivo in these model organisms provides a unique opportunity to study this process, which is relevant for many events in normal development and disease.

Animals↗

Life-cycle toxicity of 4-nonylphenol to medaka (Oryzias latipes).

We studied the chronic effects of 4-nonylphenol (4-NP) on reproductive status of medaka (Oryzias latipes) over two generations of continuous exposure. The exposure study of the parental (F0) medaka was begun on embryos within 24 h postfertilization and continued with monitoring through embryological development, hatching, posthatch survival, growth, sexual differentiation, and reproduction under flow-through exposures to mean measured 4-NP concentrations of 4.2, 8.2, 17.7, 51.5, and 183 microg/L for up to 104 d. Eggs spawned from the F0 fish at 102 and 103 d posthatch were also examined for hatchability, survival after hatching, growth, and sexual differentiation until 60 d posthatch. The 183-microg/L treatment significantly reduced the embryo survival and swim-up success of the F0 fish. The cumulative mortality after swim-up of the F0 fish exposed to 17.7 and 51.5 microg/L were significantly higher than the control mortality. No concentration-related effect of 4-NP was observed on the growth of surviving F0 fish at 60 d posthatch. However, the sex ratio estimated from the appearance of their secondary sex characteristics was skewed toward female in the 51.5-microg/L treatment. Additionally, gonadal histology showed that 20% of the fish in the 17.7-microg/L treatment and 40% in the 51.5-microg/L treatment had testis-ova, indicating that 4-NP affects the gonadal development and survival of medaka at similar concentrations in juveniles. The sex ratio of the F0 fish in the 51.5-microg/L treatment was completely skewed toward female; subsequently, the effects on fecundity and fertility in this generation were monitored at mean measured concentrations of 4.2, 8.2, and 17.7 microg/L from 71 to 103 d posthatch. Fecundity was unaffected by any of the treatments examined. The mean fertility in the 17.7-microg/L treatment was reduced to 76% of that in the controls, although no statistically significant differences were determined. Overall, these results indicate that the lowest-observed-effect concentration (LOEC) and no-observed-effect concentration (NOEC) of 4-NP through the life cycle of the F0 medaka were 17.7 and 8.2 microg/L, respectively. In the F1 medaka, no significant effects were observed on hatching success, posthatch mortality, or growth, but sexual differentiation at 60 d posthatch was affected. Induction of testis-ova in the gonads of the F1 fish was observed in both the 8.2- and the 17.7-microg/L concentrations. The results indicate that 4-NP can have significant effects on reproductive potential of medaka at concentrations as low as 17.7 microg/L.

Animals↗

Selectivity in mobilisation of stored fatty acids by maturing cod, Gadus morrhua L.

Lipids, proteins and carbohydrates diminish in the liver and muscle of cod when (a) food is inadequate or (b) the gonads are maturing. The results reported in this paper appear to show that the mobilisation of the lipids differs according to whether the need is just for energy (simple starvation) or for building up gonads. Two groups of fish from the same catch were starved. In one, the gonads were developing, while in the other the gonads had been surgically removed. Significantly more of the fatty acid C22:6 was mobilised from the liver lipids in the group with developing gonads, this fatty acid being present in the greatest amount in the gonads of either sex. The fatty acid C18:1 is also important in the gonads, and this was also preferentially removed from all the livers of maturing-starving fish when compared with gonadectomised-starving, though here the effect was not statistically significant. Some discrimination therefore appears to be exercised when hepatic lipids are removed for gonad development. No selectivity was observed in the mobilisation of fatty acids from the muscle.

Aging↗

Seasonal adaptations of Siberian hamsters. II. Pattern of change in daylength controls annual testicular and body weight rhythms.

Studies of rodent photoperiodism almost without exception have employed fixed day lengths (DLs) and abrupt transitions from long to short DLs. Because the natural progression of changes in DL carries predictive information and may have physiological consequences, we determined seasonal fluctuations in testis size and body weight in Siberian hamsters maintained in a year-long pattern of increasing and decreasing DLs. A cycle of gonadal development, regression, and recrudescence, and corresponding changes in body weight, were observed in hamsters maintained in separate simulated natural photoperiods (SNPs) in which DLs neither fell below nor exceeded the putative critical DL of 13 h. Gradually decreasing DLs as long as 15.3 h induced gonadal regression, and DLs as short as 12.3 h supported ponderal growth, depending on the hamster's prior photoperiodic history. DLs experienced by hamsters during development influenced the adult incidence of responsiveness to short DLs. Photorefractoriness to short DLs occurred earlier in hamsters kept in static 10-h than in 12-h DLs. Increasing DLs in winter had little impact on the rate of gonadal recrudescence and weight gain. These data extend earlier investigations in showing the photoperiodic history determines gonadal responses over a broad range of DLs and influences gonadal responsiveness to short DLs and the triggering of the interval timer underlying recrudescence.

Adaptation, Physiological↗

Comparative biochemical composition in gonad and adductor muscle of triploid and diploid catarina scallop (Argopecten ventricosus Sowerby II, 1842).

Biochemical components of gonad and adductor muscle for diploid and triploid catarina scallop, Argopecten ventricosus, were evaluated and compared at four periods in 1 year (January, April, June, and October). Two comparisons were done. The first one compared an untreated control (diploid) vs. a triploidy-treated group for which the percentage of triploids was 57%. The second comparison was done on a group derived from within the triploidy-treated group, separating diploids (internal control) from triploids ('putative triploids'). Regardless of which comparison, in the gonad diploid scallops had larger concentrations of proteins, carbohydrates, lipids, and acylglycerols than triploid scallops. This reflects the maturation processes in diploid scallops vs. the sterility seen in most triploid scallops, and it is particularly supported by the consistently larger concentration of acylglycerols in gonads of diploids than in triploids. The gonad index of the internal control (diploid) group was significantly larger than that seen in the putative triploid group at all sampling periods but October, when none of the gonad biochemical components were different between ploidy groups.Triploid scallops had a significantly larger muscle index than diploids from April to October. This can be caused by a larger gain in muscle tissue in triploids than diploids from January to June. However, there were no consistent differences in any of the biochemical components evaluated in adductor muscle of diploids and triploids. The use of freshly ingested food rather than reserve mobilization from muscle in diploids is suggested by these results. Nutrients derived from ingested food are apparently used for muscle growth in triploids, whereas in diploids those nutrients serve primarily for gonad development. The importance of freshly ingested food for maintenance and growth is suggested because the decline in biochemical components seen in October in both muscle and gonad was paired with a decline in weight of those two organs, especially when the control groups are considered, but a decrease was also evident for the triploid groups. This may have been caused by the presence of El Niño, with its characteristic high water temperatures and low productivity.

Journal Article↗