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Changes in localization of cytochrome P450 cholesterol side-chain cleavage (P450scc) in Japanese eel testis and ovary during gonadal development.

In this study, we generated and characterized a polyclonal antiserum against eel P450 cholesterol side-chain cleavage (P450scc) using a recombinant protein as the antigen. We examined the localization and abundance of P450scc by immunohistochemistry in Japanese eel testes and ovaries during artificially induced gonadal development. P450scc mRNA localization was also examined by in situ hybridization. In male eels, testicular development was induced by a single injection of human chorionic gonadotropin (HCG). In females, ovarian development was induced by weekly injections of salmon pituitary homogenate (SPH). Before HCG injection, the testis contained germ cells that were primarily type A spermatogonia. Additionally, several clusters of immunoreactive cells for P450scc were localized in the interstitial Leydig cells, but no P450scc mRNA signals were detected. This suggests that P450scc is either a relatively stable protein or it is produced by a mRNA that is present at too low a level to detect. Shortly after a single injection of HCG, expression of P450scc mRNA was stimulated and the number of immunoreactive clusters and their staining intensity were both increased. P450scc mRNA fell to an undetectable level 3 days after hormonal stimulation. Although the P450scc protein also decreased at the same time as the mRNA, it remained at a detectable level throughout this period. P450scc mRNA, but not the P450scc protein, was also detected in the spermatids and spermatozoa. The biological significance of P450scc mRNA expression at this stage is unknown. Prior to experimentation, the ovary contained oocytes that were developed to the oil-droplet stage, with several clusters of immunoreactive cells localized in the thecal layer and ovigerous lamella epithelium. Expression of P450scc mRNA was also stimulated by SPH injections in the ovary. In contrast to the testis, P450scc mRNA was continuously detected in the thecal cell layer throughout artificially induced maturation, possibly due to a repeated stimulus by the SPH injection every week. Clusters of immunoreactive cells in the thecal cell layer increased in number as ovarian development progressed. This increase in P450scc mRNA and protein may explain, at least in part, the increase in serum steroid hormones in female eels. The P450scc antiserum clearly immunostained interrenal steroidogenic cells in the head kidney of not only eel but also goldfish, indicating that this antibody could also be used in other teleost species.

Animals↗

Central nervous system regulation of gonadal development in the avian male.

Many avian species, particularly domesticated ones used for egg and meat production, are photoperiodic. Research conducted over the past several years has revealed the neural components, neurotransmitters, neuromodulators, and gonadotropins that play an important role in responding to changes in day length. An ultimate effect of the neuroendocrine response of birds to light is gonadal development and production of fertilized eggs and young for the next generation. The main purpose of this paper is to address the major neural systems that have been shown to affect reproductive function in males. Potential areas of research that would help elucidate the mechanism of neural activation of gonadal function are suggested.

Animals↗

PSM2, a novel protein similar to MCAF2, is involved in the mouse embryonic and adult male gonad development.

By screening RIKEN database, we obtained a mouse ovary and uterus derived clone (RIKEN clone ID: 5031403I04). Multiple tissues expression analysis revealed that the clone was specifically expressed in ovary and to a higher extent in testis. It is expressed in early mouse embryo, especially in the embryonic gonad from 11.5 dpc (days post coitus). Furthermore, it is also expressed continuously from newborn testis to adult. Using testis sections in situ hybridization, we found the mRNA was localized to spermatogonia, round spermatids and mainly to spermatocytes. We cloned the cDNA from mouse testis. The gene consists of 10 exons spanning approximately 48 kb on mouse chromosome 16. The cDNA encodes a putative nuclear protein of 319 amino acids containing a coiled-coil motif. The deduced protein has high similarity with human MCAF2 (MBD1-containing chromatin-associated factor 2), so we termed it as PSM2 (protein similar to MCAF2) in this article. We therefore hypothesized that PSM2 might interact with some important partners by the conserved domain and be involved in the transcription modulation during gonad development.

Amino Acid Sequence↗

Gonadal development and plasma steroid levels during pubertal development in captive-reared striped bass, Morone saxatilis.

Puberty is the period during which full sexual development occurs and the capacity to reproduce is acquired. Despite its importance, our understanding of the endocrine regulation of puberty in lower vertebrates is still limited. The objective of the present study was to describe the changes in gonadal development and plasma steroid levels in a relatively late maturing species, the striped bass, during the first four years of life. In about 65% of the females, puberty was initiated during the third year. Although gonadosomatic index (GSI) and oocyte diameter increased during this year, this first cycle was characterized by a heterogeneous population of developing oocytes, a relatively low mean maximum oocyte diameter, and an absence of yolk granules in the oocytes. Plasma 17beta-estradiol (E(2)) levels were low in all three-year-old fish, suggesting that an insufficient stimulation of vitellogenin production by E(2) may underlie the lack of vitellogenin incorporation into developing oocytes. All monitored parameters increased during the fourth year, but were still below the values attained by older females. In about 60% of the males, puberty was initiated during the first year and all males were mature by the third year. During the first two years, several immature males initiated spermatogenesis without reaching full maturity. In mature males, mean GSI, plasma testosterone, and 11-ketotestosterone levels increased simultaneously, reaching higher values each subsequent year. Our results indicate that, similar to the situation in mammals, more than one reproductive cycle is required in striped bass before complete adulthood is reached.

Animals↗

The impact of photoperiods and melatonin on gonadal development in juvenile Turkish hamsters (Mesocricetus brandti).

The reproductive response of both intact adult and juvenile Turkish hamsters has been thoroughly studied and shown to be similar, unlike the golden hamster where juveniles remain aphotoperiodic until approximately 8 weeks of age. Unstudied to date, however, is the role of the pineal and its hormone melatonin in generating the testicular response to photoperiod in juvenile Turkish hamsters. Therefore, in this study we examined the reproductive response of prepubertal male Turkish hamsters, subjected to four different photoperiods (8L:16D, 16L:8D, 20L:4D, and 24L:0D) with altered pineal gland function. At 15 days of age, long-day-born (16L:8D) hamsters were either pinealectomized, received melatonin implants, or remained untreated. Testes sizes were measured every 2 weeks. Testicular growth occurred only in untreated and beeswax implanted groups in 16L:8D. Exposure to other photoperiods inhibited testicular development in untreated and beeswax implanted animals. Removal of the pineal gland, masking of the daily melatonin rhythm with constant release subcutaneous melatonin implants, or eliminating the daily rhythm of melatonin by continuous light exposure resulted in inhibition of gonadal development. These results demonstrate that juvenile Turkish hamsters respond similarly to adults on all photoperiods and under all conditions of pineal function tested.

Animals↗

[Effects of the dietary incorporation of carotenoid pigments on gonad development and oocyte maturity in females of rainbow trout (Oncorhynchus mykiss, Walbaum, 1972)].

Four groups of rainbow trout (Onchorhynchus mykiss) each with 60 females with an average body weight of 467 g were submitted to an aleatory assay in order to compare the efficiency of four diets containing different concentrations of carotenoid pigments, and to determine if pigment concentration and source had an influence on female gonad development and oocyte maturity. The first diet was the non-pigmented control (C). The second diet contained 100 mg kg(-1) of Carophyll Pink (CR). The third and fourth diets contained 200 (RC200) and 250 (RC250) mg kg(-1) of saponified red chili oleoresin, respectively. The results indicated significant differences (p < or = 0.05) between C and RC250, with respect to the gonado-somatic index (IGS), average gonad weight (PG) and average diameter of oocyte (DO), with CR250 showing higher IGS, PG and DO values. Treatments CR and CR200 presented similar values. At the end of the experimental period, only 10% of oocytes were mature in the RC250 diet.

Adjuvants, Immunologic↗

Three daily melatonin infusions alter gonadal development but not GnRH neuron number in the medial preoptic area or diagonal band of Broca in Siberian hamsters.

Among juvenile Siberian hamsters reproductive development is associated with an increased number of unipolar GnRH-immunoreactive neurons in the diagonal band of Broca and medial preoptic area. In the present study, GnRH neuron morphology was assessed in male juvenile hamsters which were treated with three daily melatonin (MEL) infusions to initiate or delay gonadal development. Hamsters gestated in short days were transferred to constant light (LL) at day 14 of age and infused on days 18-20 with MEL for 6 h/day. This treatment stimulated testis development compared to that in saline-infused controls. By contrast, testis growth was suppressed by three 12 h MEL infusions in long-day gestated hamsters in LL from day 14. The number of unipolar and bipolar GnRH neuron subtypes was the same irrespective of MEL infusion or preweaning photoperiod. Thus, gonadal response to three MEL infusions is independent of changes in GnRH neuron number.

Animals↗

Effects of atrazine on metamorphosis, growth, laryngeal and gonadal development, aromatase activity, and sex steroid concentrations in Xenopus laevis.

African clawed frogs (Xenopus laevis) were exposed to one of eight nominal waterborne concentrations including 0, 0.1, 1.0, 10, or 25 microg/L atrazine, 0.005% ethanol (EtOH), or 0.1mg/L estradiol (E2) or dihydrotestosterone (DHT) containing 0.005% EtOH. Frogs were exposed from 72 h posthatch until 2--3 months postmetamorphosis via a 3-day static renewal exposure regimen. Atrazine at concentrations between 0.1 and 25 microg/L did not significantly affect mortality, growth, gonad development, laryngeal muscle size, or aromatase activity in juvenile X. laevis. Male frogs exposed to 1.0 microg/L atrazine had lower E2 levels compared to controls, but this response was not consistent among other concentrations of atrazine. Male and female frogs exposed to DHT had larger laryngeal dilator muscle areas compared to controls. E2-exposed female frogs had decreased gonadal aromatase activity, and E2-exposed male frogs had statistically greater plasma concentrations of E2 compared to controls.

Animals↗

Effects of environmentally relevant concentrations of atrazine on gonadal development of snapping turtles (Chelydra serpentina).

The herbicide atrazine has been suspected of affecting sexual development by inducing aromatase, resulting in the increased conversion of androgens to estrogens. We used snapping turtles (Chelydra serpentina), a species in which sex is dependent on the production of estrogen through aromatase activity in a temperature-dependent manner, to investigate if environmentally relevant exposures to atrazine affected gonadal development. Eggs were incubated in soil to which atrazine was applied at a typical field application rate (3.1 L/ha), 10-fold this rate (31 L/ha), and a control rate (no atrazine) for the duration of embryonic development. The incubation temperature (25 degrees C) was selected to produce only males. Although some males with testicular oocytes and females were produced in the atrazine-treated groups (3.3-3.7%) but not in the control group, no statistical differences were found among treatments. Furthermore, snapping turtle eggs collected from natural nests in a corn field were incubated at the pivotal temperature (27.5 degrees C) at which both males and females normally would be produced, and some males had oocytes in the testes (15.4%). The presence of low numbers of males with oocytes may be a natural phenomenon, and we have limited evidence to suggest that the presence of normal males with oocytes may represent a feminizing effect of atrazine. Histological examination of the thyroid gland revealed no effect on thyroid morphology.

Animals↗

Effects of 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT) on gonadal development in the chick embryo: a histological and histochemical study.

Fertile white Leghorn chicken eggs were exposed via intravitelline injections to dosages of 5.0, 10.0, or 20 mg 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT) in olive oil prior to incubation. Control embryos received only the olive oil vehicle. Eggs were placed in a forced-draft incubator for either 5 or 12 days. Embryos were removed and their gonadal areas prepared for histological or histochemical evaluation. Histological examination of DDT-exposed 5-day embryos revealed no significant differences in the number of primordial germ cells aggregating in the gonadal area and in the localization of acid and alkaline phosphatase activity. Embryos exposed to DDT for 12 days revealed significant alterations in both ovaries and testes. The testes of DDT-exposed embryos consisted of mostly stroma with fewer seminiferous cords than controls while ovaries of exposed embryos contained a larger number of distended medullary cords as well as a difference in the distribution of these cords when compared to controls. There was an increased alkaline phosphatase activity in the stromal cells of female gonads. Increased amounts of alkaline phosphatase activity found in the stroma at 12 days might be due to a DDT-induced stimulation of these cells to differentiate more rapidly. Acid phosphatase activity was found in the secondary sex cords of control 12-day ovaries, but was much reduced or absent in those of pesticide-exposed embryos. These results indicate that a single dosage of DDT administered to a chick embryo prior to incubation does not affect early stages of gonadal development but that effects on both ovaries and testes occur 12 days following exposure.

Acid Phosphatase↗

Anti-androgen flutamide affects gonadal development and reproduction in medaka (Oryzias latipes).

The effects of an anti-androgenic chemical on the reproduction of medaka (Oryzias latipes) were examined. Six mating pairs of medaka were exposed to flutamide (FLU) at measured concentrations of 0.101, 0.202, 0.397, 0.787, or 1.56 mg/L for 3 weeks. During the exposure period, one female died in the 1.56-mg/L treatment group, but no lesions or pathological abnormalities were observed. The fecundity and fertility of paired medaka were significantly decreased at 1.56 mg/L compared with those of the controls. Histological examination showed intersex gonads (testis-ova) in males exposed to FLU at 0.202, 0.397, and 0.787 mg/L. However, concentrations of hepatic vitellogenin (Vtg) in both sexes were not statistically different from those in the controls. These results demonstrate that FLU affected gonadal development in male medaka and reproductive capacity in paired medaka. This is the first report of testis-ova in male medaka exposed to FLU.

Androgen Antagonists↗

[Gonadal development and reproductive cycle of Lutjanus peru (Pisces: Lutjanidae) in Guerrero, Mexico].

In this paper, aspects of the reproductive biology of Lutjanus peru that can be used for the fishery management are described. Samples were taken monthly in 1993 and 1994, from commercial catches in the southern coast of Guerrero, Mexico. A global sex ratio of 1:1.37 (M:F) was found, but the sex ratio was highly variable each month. The condition factor, gonadosomatic, hepatosomatic and stomach fullness indexes were analyzed. None of the morphophysiological indexes showed any clear tendency. From 377 gonads, both male and female, five developmental stages were characterized for each sex. L. peru shows asynchronous gonad development and multiple spawning. Two spawning period were observed in 1993 (March, and August-September) and three in 1994 (April-May, July-August, and November), with a maximum in August of both years. Length at first maturity for the females of L. peru was estimated at 295 mm fork length.

Animals↗

Advanced gonadal development in male turkeys following knife cuts directed to the hypothalamus.

Turkey poults when 4 weeks of age received parasagittal knife cuts of the total hypothalamic area. Each of the two knife cuts was 1.2 mm lateral from midline and extended from the preoptic area to the mamillary hypothalamic region. Five experimentals survived only 3 to 5 days following surgery as none was able to eat or drink. Three experimentals showed prematurely developed wattles and caruncles about the neck, larger testes, and more advanced stages of spermatozoan development and tubule growth compared to sham-operated controls. When male poults were placed in a floor pen two of the three experimentals displayed mating behavior by strutting. No strutting was observed in controls. Similar to the chicken, male poults receiving parasagittal knife cuts throughout the anterior-posterior extent of the hypothalamus and thalamus showed advanced gonadal development.

Animals↗

An mRNA differential display strategy for cloning genes expressed during mouse gonad development.

The mRNA differential display technique has become a popular method for isolating novel genes in a variety of biological systems including carcinogenesis, hormone regulation, plant biology and neurobiology. We have further developed the method by optimizing different steps for the use of small amounts of material, such that differential display can be used in the study of developmental biology. Our techniques include a new assay for elimination of false positive cDNA clones and a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method for the rapid analysis of differences in gene expression. This improved mRNA differential display strategy requires less than 4 microg of total RNA. We have used it for the isolation of genes which are expressed during gonad development in the mouse. One of the cDNAs found, cDNA 4.3 which corresponds to a part of the gene encoding the steroid hydroxylase 3betaHSD I, was shown to be a valuable marker for adrenal development and for Leydig cell differentiation and organization during testis development.

3-Hydroxysteroid Dehydrogenases↗

Two Sox9 messenger RNA isoforms: isolation of cDNAs and their expression during gonadal development in the frog Rana rugosa.

Sox is a family of SRY-related testis-determining genes. We have isolated two different mRNA isoforms of the frog Sox9 gene from adult frog testis cDNAs. One form (Sox9 alpha) encodes a 482 amino acid protein containing the HMG box, whereas the other form (Sox9 beta), which completely lacks the HMG box, is a truncated 265 amino acid protein of Sox9 alpha. Sox9 alpha is 82% similar to mouse, 86% to chicken, and 77% to trout Sox9 at the amino acid level. Sox9 expression was upregulated in embryos after stage 16, and was seen in both developing testes and ovaries. The size of Sox9 transcripts was determined to be 7.8 knt by Northern blot analysis. In addition, Sox9 alpha expression was found prominently in the testis and brain among various tissues of adult frogs examined, and was considerably higher than Sox9 beta. The fact that Sox9 is expressed in both sexes suggests that this gene is involved in gonadal development of male and female frogs. This is dissimilar to the pattern in birds and mammals, in which Sox9 expression is male-specific.

Animals↗

Further examination of the kinetics of gonadal development in XY female mice.

Y-chromosomes derived from house mouse populations of Mus domesticus are able to lead to the formation of XY females (i.e. hermaphrodite individuals with both ovarian and testicular tissues) when inserted, by appropriate crosses, into the C57BL/6J genome, which usually carries the Mus musculus Y chromosome. An abnormal interaction between the testis-determining gene on the Y (of domesticus origin) and an autosomal recessive allele (of musculus origin) needed for testicular formation is the likely explanation of this phenomenon. In the present paper we analyse the histological kinetics of gonadal development in hermaphrodites obtained by introducing the Y chromosomes from wild living house mice [from Zagreb, (Croatia) and Hohenentringen (Southern Germany)] into the C57BL/6J genome. Among 88 individuals, 9 XY hermaphrodites were detected, 6 before birth and 3 afterwards. The percentage of the ovarian-type tissues in the ovotestes fell drastically between day 18 of fetal life and day 20 after birth. These findings corroborate and extend those already published for other Y domesticus chromosomes and stress the possible role of Y chromosome differentiation in speciation processes.

Aging↗

The Wilms tumor suppressor WT1 regulates early gonad development by activation of Sf1.

In mammals, several genes including the Wilms tumor suppressor gene Wt1, the Lim homeobox gene Lhx9, and the gene encoding steroidogenic factor 1 (Sf1) have been implicated in the development of the indifferent gonad prior to sexual differentiation. Interactions among these genes have not yet been elucidated. Using biochemical and genetic experiments, we demonstrate here that WT1 and LHX9 function as direct activators of the Sf1 gene. Interestingly, only the -KTS form of WT1 is able to bind to and transactivate the Sf1 promoter. This observation is consistent with differential roles for the -KTS and +KTS variants of WT1 which have been postulated on the basis of human disorders such as the Frasier syndrome. Our data suggest a pathway in which the products of the Wt1 and Lhx9 genes activate expression of Sf1 and thus mediate early gonadogenesis.

Animals↗

Sexually mature European eels (Anguilla anguilla L.) stimulate gonadal development of neighbouring males: possible involvement of chemical communication.

This study was aimed to investigate whether sexual maturation of immature male eels could be stimulated indirectly by placing them in contact with either male (Minj) or female (Finj) eels in which sexual maturation had been stimulated directly by weekly injections of human chorionic gonadotropin (hCG) or salmon pituitary extract (SPE), respectively. Untreated males were placed either in the same tank or in a separate tank that was linked to the injected fish via a recirculation system. The hormonal treatments stimulated spermatogenesis and spermiation in Minj, and ovulation in Finj as well as an increase of the ocular (Io) and gonadosomatic (GSI) indices in both sexes. Plasma levels of testosterone (T) and 11-ketotestosterone (11-KT) increased in Minj and T and 17beta-estradiol (E2) in Finj. A small peak of plasma 17,20beta-dihydroxypregn-4-en-3-one (17,20betaP) occurred during ovulation, while the plasma levels of 17alpha-hydroxypregn-4-ene-3,20-dione (17P) were undetectable in both males and females. The water conditioned by Minj and Finj induced significant, though relatively minor, increases in Io and GSI in uninjected males. In addition, uninjected fish showed small changes in plasma T and 11-KT levels, apparently related to the timing of spermiation and ovulation of Minj and Finj, respectively, as well as an activation of spermatogenesis (but not spermiation). Injected fish released free and conjugated T, 11-KT and E2 into the water, although immature eels were unable to smell (by electro-olfactogram) any of these steroids or prostaglandin F2alpha. However, immature males were highly sensitive to water extracts conditioned by spermiating Minj and pre-ovulatory and ovulated Finj. These preliminary results suggest the existence of chemical communication between maturing eels and immature males that stimulates gonad development, although the putative pheromone(s) involved has/have not yet been identified.

Anguilla↗