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Effect of photoperiod and temperature on gonadal activity and plasma steroid levels in a reared strain of the mummichog (Fundulus heteroclitus) during different phases of its annual reproductive cycle.

Mummichog, a spring and summer spawning teleost, were exposed to various photoperiod and temperature conditions to investigate the environmental regulation of the annual reproductive cycle. In early spring, latter phases of gonadal development (vitellogenesis in females and active spermatogenesis in males) were effectively accelerated by warm temperature (16 degrees C) regardless of the photoperiod (11L or 16L), although internal factor(s) may be concerned with triggering the initiation of the development. In late summer, intense gonadal regression which leads to the termination of the spawning period was accelerated by a short day length (</=13L) in both yearlings and underyearlings. In underyearlings, however high water temperature and internal factor(s) may also be concerned. In early autumn, early phases of gonadal development (growth of cortical alveolus stage oocytes in females and basal spermatogenesis in males) were induced by moderate or lower temperatures (</=22 degrees C) even under a short day length (11L). From the middle of autumn to early winter, this fish is probably in the "refractory period," and did not progress to the latter phases even under adequate temperature and long day length conditions (22 degrees C-16L). Mummichog showed a probable circa-annual rhythm of gonadal activity under constant temperature and photoperiod conditions (22 degrees C-16L): this rhythm may be the basis of the trigger of the gonadal development prior to the spawning period, the termination of the spawning period in underyearlings, and the occurrence of the "refractory period" during autumn. Plasma concentrations of estradiol-17beta in females and testosterone in males correlated well with the gonadal development during early spring and the regression during late summer. However, there was no correlation between plasma steroid levels and the degree of progress during the early phases of gonadal development in autumn, suggesting other factor such as direct action of GtH(s) or mediating substance other than sex steroids for these phases.

Animals↗

Gonadal tumors developed from consecutively transplanted spleens bearing gonad implants.

This study was conducted by consecutively transplanting spleens, which had gonads implanted previously. A total of 84 cases for infantile testicles and 106 cases for ovarian follicles were performed. In the case of ovarian implants, the results were determined by the total number of follicle implants. A modified spleen transplantation technique called double implantation of ovarian follicles was applied to increase the amount of the implants. In this technique, an extra spleen is implanted into the potential donor so that the ovarian follicles can be implanted to two different spleens, doubling the amount of implants. Through consecutive spleen transplantation, we observed the results beyond a typical rat's life span. In many of these cases, we found more aggressive forms of malignant tumor, seminoma and dysgerminoma. We present the results and discuss possible pathogenic mechanisms of tumor formation.

Animals↗

The development of the sexually indifferent gonad in the prosimian, Galago crassicaudatus crassicaudatus.

The morphogenesis of the sexually indifferent gonad of the primate Galago crassicaudatus crassicaudatus was studied by high-resolution light microscopy and electron microscopy in 15 embryos aged 26 to 33 days. Onset of gonadal development follows the morphogenesis of the mesonephros by a conspicuous interval and is identified as the time when the first primordial germinal cells arrive in the region ventral to the central third of the mesonephros; this is followed by intense proliferation of the coelomic mesothelial cells lining the area. They become organized into short piles that deepen in the underlying mesenchyme, enclosing the germinal cells in the process. Rapidly, the piles become confluent forming a compact mass, the gonadal blastema, which is soon cleaved into gonadal cords by stroma and vascular lacunae. The mesonephros becomes involved in the morphogenesis of the gonad only in late stages of development when anatomic continuities become established between the capsules of its regressing glomeruli and the elongating gonadal rete cords. These observations show that in the Galago the somatic cells of the gonadal blastema, i.e., the precursors of the definitive testicular and ovarian sustentacular cells, derive from the coelomic mesothelium in contrast to other mammals, e.g., ruminants and rodents, where they are of mesonephric derivation. This important point is discussed in light of the differences that exist among species with regard to the structural complexity, functionality, and stages of differentiation/involution of their mesonephroi on the one hand, and the time of gonadal development on the other.

Animals↗

Development and regulation of growth and differentiated function in human and subhuman primate fetal gonads.

We have attempted to summarize the research on primate fetal gonadal development that has occurred over the past three decades. Many similarities exist between fetal gonadal development in human and subhuman primates; therefore, comparisons and analogies between these species can be made. Fetal gonadal development is a complex process dependent on timely maturation and differentiation of several cell types with different functions. Adequate development is important for normal sexual development and intact adult fertility potential as well as for intrauterine priming of neural centers in the central nervous system. While the fetal primate testis is active in steroidogenesis, the fetal ovary seems to be quiescent throughout most of gestation, although some ovarian steroidogenic enzymes have been demonstrated. Growth and development of both gonads are controlled during late gestation at least in part by pituitary hormones, while earlier in gestation other yet undefined regulators (placental, intragonadal) likely also are active. The main goal of this review was to demonstrate that gonadal growth and differentiation, both in males and females, is regulated by endocrine factors as well as by intragonadal, autocrine/paracrine agents. Although many parts of the puzzle are still missing it is probable that, similar to fetal development of other endocrine tissues and to events in postnatal gonads, these local regulators have important functions. Currently, primate fetal gonadal research is lacking in at least two key aspects: 1) the definition of paracrine and autocrine nonsteroidal factors that are involved in the regulation of gonadal growth and differentiation in vitro; and 2) in vivo studies in subhuman primates that might better help to clarify the biological roles of the multiple extra- and intragonadal hormones and their complex interactions. To date, the regulation of gonadal steroidogenesis has been investigated more thoroughly than the regulation of gonadal growth. Most of our knowledge stems from observations of gonadal development in anencephalics or subhuman primates after pituitary ablation. Because of the constraints of small organ size and limitation of material, studies of fetal primate gonadal development have been limited. Given such limitations, new molecular biological techniques, including polymerase chain reaction and in situ hybridization, may provide the means of addressing these questions. Further, because of these limitations, sensitive cell separation techniques need to be developed to achieve enriched primary gonadal cell cultures from individual gonads.

Animals↗

Lectin-binding carbohydrates in sexual differentiation of rat male and female gonads.

Development and sexual differentiation of the mammalian gonad involve changes in the type and distribution of different proteins and glycoproteins in and around the epithelial gonadal cords, the future seminiferous tubules in the testis, and follicles in the ovary. To study changes in cellular carbohydrate-containing compounds in the sex-specific morphogenesis of rat gonads, sections from embryonic, fetal and early postnatal gonads were labelled with seven different fluorescein isothiocyanate (FITC)-conjugated plant lectins of various carbohydrate-binding specificities. Double labelling of laminin with tetramethylrhodamine isothiocyanate (TRITC)-conjugated antibodies was used to outline the epithelial tissues. From the results we conclude that the abundance and similar distribution of carbohydrates in the early gonads of both sexes supports their sexually indifferent nature. Furthermore, the basement membranes of the differentiating gonadal cords in both sexes have common features, which differ, however, from those of the other developing urogenital organs. Changes in carbohydrate composition appear with the sexual differentiation of the gonads; the similarity of the changes in lectin binding to the gonadal cords of embryonic and fetal male, and to postnatal female, suggests similar mechanisms of cell-cell interactions in both sexes although activated at different developmental stages. These carbohydrate specificities at the tissue level should be taken into account together with cell-type specific changes, e.g. in the formation of the zona pellucida, when the phenomenon of polymorphic expression of different compounds is integrated into theories of epithelial differentiation.

Animals↗

The differentiation of germ cells and gonads during development of carp (Cyprinus carpio L.). A study with anti-carp sperm monoclonal antibodies.

Gonadal development, germ cell differentiation and the appearance of membrane antigenic determinants, specific for male and female germ cells during gonadogenesis, was studied in larval and juvenile carp (Cyprinus carpio L.) until 25 weeks after fertilization. Indirect immunofluorescence studies with four monoclonal antibodies raised against carp spermatozoa revealed that monoclonal antibody WCS 29 stained the outer membranes of primordial germ cells in larvae from 3 days after fertilization. The monoclonal antibodies WCS 3 and 17 reacted with the outer membranes of germ cells from 7 weeks after fertilization onwards, simultaneously with the onset of germ cell proliferation. With monoclonal antibody WCS 28 germ cell membranes were clearly stained from 18 weeks after fertilization. Similar reactions were observed in both sexes, however, female germ cells reacted at an earlier developmental stage with the monoclonal antibody WCS 28 than male germ cells. In the developing testis the monoclonal antibodies stained all types of spermatogenic cells. In the ovary, however, only oogonia and early prophase oocytes showed a positive reaction with the four monoclonal antibodies. The results indicate that germline-specific antigens are present on the outer membranes of primordial germ cells and their male and female descendants, with the exception of elderly oögenic stages. It is assumed that the appearance and disappearance of these membrane antigens reflect differentiation steps of germ cells during gonadogenesis.

Animals↗

Effects of the synthetic estrogen ethinylestradiol on early life stages of mink frogs and green frogs in the wild and in situ.

Estrogenic contaminants are known to disrupt growth and development in amphibians. Field-based research is needed to elucidate their potential impacts on wild populations. Hatch success, larval growth and development rates, and gonad development were examined in native amphibians exposed to low ng/L concentrations of 17alpha-ethinylestradiol (EE2) in a whole-lake addition experiment at the Experimental Lakes Area, northwestern Ontario, Canada. Egg masses were reared in situ in the EE2-amended lake and in two reference lakes in 2001 and 2002. Hatching success was reduced significantly in green frogs (Rana clamitans) but not in mink frogs (Rana septentrionalis) exposed to EE2. Ethinylestradiol had no consistent effect on mass or development stage of hatchlings in the early larval stages of the caging study. Ethinylestradiol had no effect on sex ratios of either species in situ, and no intersex gonads were observed in exposed or reference green frog tadpoles or in reference mink frog tadpoles. However, 5.6% (total n = 18) and 12.5% (total n = 56) of EE2-exposed mink frog tadpoles were intersex in the 2001 and 2002 caging studies, respectively. Wild mink frog tadpoles also were examined, and EE2 had no effect on sex ratios. No intersex gonads were observed in reference lake tadpoles or in tadpoles from the experimental lake prior to EE2 additions; however, 2.4, 0, and 28.6% of wild EE2-exposed first-year tadpoles had intersex gonads (2001, 2002, and 2003, respectively). These results indicate that exposure to EE2 in the wild and in situ at concentrations comparable to those detected in effluents and, occasionally, in surface waters can impact gonad development and hatch success in native amphibians.

Animals↗

tcl-2 encodes a novel protein that acts synergistically with Wnt signaling pathways in C. elegans.

Mutations in tcl-2 cause defects in the specification of the fates of the descendants of the TL and TR blast cells, whose polarity is regulated by lin-44/Wnt and lin-17/frizzled, during Caenorhabditis elegans development. In wild-type animals, POP-1/TCF/LEF, is asymmetrically distributed to the T cell daughters, resulting in a higher level of POP-1 in the nucleus of the anterior daughter. The POP-1 asymmetric distribution is controlled by lin-44 and lin-17. However, in tcl-2 mutants, POP-1 is equally distributed to T cell daughters as is observed in lin-17 mutants, indicating that, like lin-17, tcl-2 functions upstream of pop-1. In addition, tcl-2 mutations cause defects in the development of the gonad and the specification of fate of the posterior daughter of the P12 cell, both of which are controlled by the Wnt pathway. Double mutant analyses indicate that tcl-2 can act synergistically with the Wnt pathway to control gonad development as well as P12 descendant cell fate specification. tcl-2 encodes a novel protein. A functional tcl-2::gfp construct was weakly expressed in the nuclei of the T cell and its descendants. Our results suggest that tcl-2 functions with Wnt pathways to control T cell fate specification, gonad development, and P12 cell fate specification.

Amino Acid Sequence↗

Embryonic development of the porcine indifferent gonad and testis.

The early gonadal development in the pig from day 18 to day 36 post conception, when distinct testes were present in male embryos, has been studied. The development of the porcine gonad followed the general mammalian pattern. During testicular differentiation, the germ cells and the relevant somatic cells, the Sertoli cells, became enclosed in testicular cords, thus creating an intracordal germ cell compartment and an extracordal compartment. The development of conspicuous cell junctions and the production of a basal lamina were evident in the Sertoli cells during the early testis development. Their origin in this species remains unclear.

Animals↗

Expression of wilms' tumor gene and protein localization during ovarian formation and follicular development in sheep.

Wilms' tumor protein (WT1) is a transcriptional repressor essential for the development of mammalian kidneys and gonads. To gain insight into possible roles of WT1 in ovarian formation and follicular function, we studied patterns of mRNA and protein localization throughout fetal gonadal development and in ovaries of 4-wk-old and adult sheep. At Day 24 after conception, strong expression of WT1 mRNA and protein was observed in the coelomic epithelial region of the mesonephros where the gonad was forming. By Day 30, expression was observed in the surface epithelium and in many mesenchymal and endothelial cells of the gonad. Epithelial cells continued to express WT1 throughout gonadal development, as did pregranulosa cells during the process of follicular formation. However, WT1 expression was not observed in germ cells. During follicular growth, granulosa cells expressed WT1 from the type 1 (primordial) to the type 4 stages, but thereafter expression was reduced in type 5 (antral) follicles, consistent with the differentiation of granulosa cells into steroid-producing cells. The possible progenitor cells for the theca interna (i.e., the cell streams in the ovarian interstitium) expressed WT1 heterogeneously. However, differentiated theca cells in antral follicles did not express WT1. Strong expression of WT1 was observed during gonadal development, which is consistent with a role for WT1 in ovarian and follicular formation in the ewe. WT1 was identified in many cells of the neonatal and adult ovaries, including granulosa cells, suggesting that this factor is important for preantral follicular growth. However, the decline in WT1 expression in antral follicles suggests that WT1 may prevent premature differentiation of somatic cells of the follicle during early follicular growth.

Animals↗

Effects of waterborne exposure of octylphenol and oestrogen on pregnant viviparous eelpout (Zoarces viviparus) and her embryos in ovario.

Exposure to oestrogenic chemicals (xeno-oestrogens) may have severe effects on embryonic development. The present study investigates whether the oestrogenic endocrine disruptor 4-tert-octylphenol (4-tOP) or 17beta-oestradiol (E(2)) is accumulated in the viviparous fish the eelpout (Zoarces viviparus) and transferred to the embryos in ovario and subsequently affects embryonic development, including gonadal differentiation. Pregnant eelpouts were exposed to nominal concentrations of 25 micro gl(-1) or 100 micro gl(-1) 4-tOP (OP25 or OP100, respectively) or 0.5 micro gl(-1) E(2) in water. During 4-tOP exposure, the compound accumulated in both plasma and ovarian fluid in a concentration-dependent manner. In the mother fish, the oestrogenic biomarkers, vitellogenin (Vtg) in plasma, Vtg mRNA in liver and oestrogen-binding activity in liver, were all induced by 4-tOP (and by E(2)) at an actual concentration of 14 micro gl(-1). E(2) and 4-tOP were examined for their potency to disturb the maternal-foetal trophic relationship by disturbing the physiology of the ovary and by changing the distribution of essential nutrients normally transported to embryos during pregnancy. After exposure to E(2) or 4-tOP, calcium was depleted from the ovarian fluid and the level of free amino acids available in maternal plasma was decreased. A marked overall effect on ovarian components, including the ovarian sac, ovarian fluid and embryonic mass, was evident. Embryonic growth was significantly decreased, which might in part be attributed to disturbances of the maternal-foetal trophic relationship. Marked inductions of Vtg mRNA and Vtg protein, determined by RT-PCR and immunohistochemistry, respectively, were found in embryos from the OP100 group - the only group to show considerable accumulation of an oestrogenic compound in the ovarian fluid. A different pattern of gonadal development was found in embryos from the OP100 group compared with embryos from the control, OP25 or E(2) groups, in which approximately 50% had normal ovaries and 50% had normal presumptive male gonads. In the OP100 group, 46% had normal ovaries but, in contrast to controls, only 22% had normal presumptive male gonads, whereas the remaining 32% had abnormal male gonads with structures resembling the endo-ovarian cavity of a female gonad. As oestrogen receptor (ER) expression was detected by in situ hybridisation in early differentiating gonads, these effects could be mediated by direct interaction of the xeno-oestrogens with gonadal ER. In conclusion, this study indicates that the xeno-oestrogen 4-tOP can be transferred from the water via the mother fish to the ovarian fluid and can subsequently disturb the maternal-foetal trophic relationship and cause severe effects on embryonic development, including gonadal differentiation in ovario.

Animals↗

Effects of temperature on photoperiodically induced reproductive development, circulating plasma luteinizing hormone and thyroid hormones, body mass, fat deposition and molt in mountain white-crowned sparrows, Zonotrichia leucophrys oriantha.

The mountain white-crowned sparrow, Zonotrichia leucophrys oriantha, breeds in subalpine meadows throughout many mountainous regions of western North America. Mathematical analysis of 20 years of egg-laying dates at Tioga Pass, California (3030m elevation) indicated a highly predictable breeding season suggesting that precise environmental cues such as the annual change in day length were important for regulating reproductive function. Additionally, it appeared that there was sufficient yearly variation in the timing of breeding to suggest that other environmental cues may also be important for regulating adjustments in reproductive development and regression. Captive populations of Z. l. oriantha showed strong responses in gonadal development following transfer to longs days (15L 9D) and low temperature (5 degrees C) slowed down photoperiodically induced gonadal growth and subsequent regression, in both males and females. High temperature of 30 degrees C tended to accelerate gonadal development and regression whereas gonadal development was intermediate in a group exposed to 20 degrees C. Prior exposure to these temperature regimes while on short days (9L 15D) had no effect on body mass, fat, or plasma levels of luteinizing hormone (LH) and thyroid hormones. Curiously there was no effect of temperature on photoperiodically induced rises in LH in either sex despite marked effects on gonadal growth. Brood patch development was also enhanced in females exposed to 30 degrees C. Corticosterone levels measured in a subset of plasma samples from this experiment indicated no effect of temperature suggesting that the retarded gonadal development at 5 degrees C was not a result of thermal stress. Although there was a robust effect of photostimulation on thyroid hormone levels in blood of both sexes, temperature treatment had no effect on tri-iodothyronine (T3) concentrations. However, plasma levels of thyroxine (T4) were lower initially at 5 degrees C versus 20 and 30 degrees C treatments. This may be related to the protracted gonadal cycle at 5 degrees C versus the truncated gonadal cycle at 30 degrees C. Molt score, an indication of post-reproductive state and onset of photorefractoriness, was delayed in birds exposed to 5 degrees C. Body mass, and to a lesser extent fat score, tended to be lowest in birds exposed to 5 degrees C compared with those at 20 and 30 degrees C. These results demonstrate that ambient temperature significantly affected photoperiodically induced gonadal development and regression in these birds. The endocrine mechanisms underlying these effects require further study.

Adipose Tissue↗

Sex differentiation and pubertal development of gonads in the viviparous mosquitofish, Gambusia affinis.

The ontogenetic development of gonads from embryo to adult was observed histologically in the viviparous teleost, Gambusia affinis. Primordial germ cells (PGCs) appeared in the subendodermal space of the embryo 14 days before birth, and then transferred to the dorsal mesentery to form paired genital ridges 12 days before birth. The PGCs proliferated in the genital ridge, forming gonadal primordia 10 days before birth. All gonadal primordia differentiated to the ovary containing oocytes 2 days before birth, but then redifferentiated to the ovary and testis just after birth. This indicates that the mosquitofish is a juvenile hermaphroditic species. The characteristics of gonadal sex differentiation just after birth were enlargement of the oocytes in females, and invasion of somatic cells from the hilar region to an inner portion of the gonad in males. The paired ovary fused at the basal area 5 days after birth, then on the ventral and dorsal portions, developing into a single ovary 10 days after birth. During this time a single ovarian cavity was formed on the dorsal portion of the ovary. The paired testes fused only at the basal area and became a single testis having two main lobes 10 days after birth. The oocytes gradually developed and began vitellogenesis 100 days after birth, but did not reach maturation until 110 days after birth. Spermatogenic cells formed cysts at 20 days, began meiosis at 70 days, and matured to form sperm balls 90 days after birth. The male fish sexually matured earlier than the female.

Animals↗

Influence of different photoperiods on development of gonad, cloacal gland and circulating thyroid hormones in male Japanese quail Coturnix coturnix japonica.

One day old chicks of Japanese quail were exposed to different photoperiods (LD, 8:16, 13.5:10.5, 16:8 and LL) and observations (testes weight, cloacal gland size, body weight and circulating thyroxine and triiodothyronine) were taken at the age of 3, 5, 7, 9 and 16 weeks. Results indicate that immediate reproductive development occurred in birds exposed to long photoperiods (greater than 12 hr). Growth under LD 8:16, was not apparent till 7th week and by 16 weeks, degree of gonadal development was similar in all the birds, irrespective of photoperiodic treatment. Whereas body weight of the intermediate and long day (LD 13.5:10.5, 16:8 and LL) treated birds increased upto 5th week and remained constant thereafter. But the chicks maintained under short day length (LD 8:16), showed spontaneous increase till the end of the study and birds were much heavier compared to all other groups. Plasma T4 concentration increased with increasing age till 9th week and remained unaltered thereafter. On the other hand T3 level did not change till 7th week followed by a decline. It is suggested that the initiation and degree of gonadal growth in quail depends on the availability of daily photoperiod, until the achievement of full breeding condition. Peak level of T4 observed in 9 week old birds may be involved in the development of photorefractoriness at that age.

Aging↗

Immunohistochemical detection of gonadotropin-like material in the pituitary of brown hagfish (Paramyxine atami) correlated with their gonadal functions and effect of estrogen treatment.

Since hagfish are members of the most primitive group of living vertebrates, studies on their reproduction are indispensable for understanding phylogenetic aspects of vertebrate reproductive system. Nevertheless, our knowledge of the reproductive physiology of the hagfish, especially of the pituitary-gonadal axis, is almost completely lacking. In the present study, the relationship between the amount of immunoreactive gonadotropin (GTH)-like material in the pituitary gland and gonadal conditions was examined in the brown hagfish, Paramyxine atami. First, pituitary sections were stained immunohistochemically with anti-ovine LHbeta, and the degrees of the accumulation of GTH-like material were compared among three different groups of gonadal conditions; juveniles and adults with and without developing gonads. Immunoreactive GTH-like material was heavily accumulated in adults with developing gonads, whereas it was not or only weakly accumulated in juveniles or adults without developing gonads. Thus, there was a strong positive correlation between the amount of GTH-like material and gonadal conditions. Second, effect of estradiol benzoate on GTH-like material was examined using three groups of juvenile hagfish: initial control, sham control, and experimental animals. Experimental animals received estradiol benzoate resolved in sesame oil intraperitoneally every third day for 1 month, whereas sham control animals received the same doses of sesame oil. GTH-like material was heavily or moderately accumulated in most estrogen-treated animals, whereas it was not or weakly accumulated in initial or sham control animals. Thus, estrogen treatment in juvenile hagfish resulted in the large increase in the amount of GTH-like material. From these results, it is suggested the presence of not only GTH but also the hypophysial-gonadal feedback system in the hagfish.

Animals↗

Matrix metalloproteinases regulate mesonephric cell migration in developing XY gonads which correlates with the inhibition of tissue inhibitor of metalloproteinase-3 by Sry.

In the mouse, the sex determining gene Sry, on the Y chromosome, controls testis differentiation during embryogenesis. Following Sry expression, indifferent XY gonads increase their size relative to XX gonads and form cord-like structures with the adjacent mesonephros, providing XY gonad somatic cells. This mesonephric cell migration is known to depend on Sry, but the molecular mechanism of mesonephric cell migration remains unknown. In this study, it was shown that cells expressing Sry induced proliferation of mesonephric cells migrating into male gonads, and inhibited expression of the tissue inhibitor of metalloproteinases (TIMP)-3 gene, which is the endogenous inhibitor of matrix metalloproteinases (MMP). In addition, the mesonephric cell migration was blocked by a chemically synthesized inhibitor of MMP in a gonad/mesonephros organ co-culture system with enhanced green fluorescent protein transgenic embryos. The findings indicate that MMP may play a critical role in mesonephric cell migration, and the function of MMP may be regulated by a Sry-TIMP-3 cascade. These findings are an important clue for the elucidation of testicular formation in developing gonads.

Animals↗