Effect of growth hormone on steroid content,proliferation and apoptosis in the chicken ovary during sexual maturation

The present study was undertaken to examine in vivo the effect of growth hormone (GH) on progesterone and estradiol levels and on cell proliferation and apoptosis in the chicken ovary during sexual maturation. Hy-Line chickens (10 weeks old) were injected three times a week with 200 μg recombinant chicken GH (cGH) per kilogram body weight until sexual maturity. GH treatment significantly increased ovarian weight at 16 weeks of age, i.e., ∼1 week before onset of egg laying. The progesterone content in the ovary just before and at the time of sexual maturity and the estradiol content before onset of egg laying were also elevated after cGH injections. The highest number of proliferating (positive for proliferating cell nuclear antigen) and apoptotic (positive for terminal-deoxynucleotidyl-transferase-mediated dUTP nick-end labeling) cells was found in the ovarian stroma and white follicles (>1-4 mm diameter), whereas the lowest number of these cells was detected in yellow (>8-30 mm) follicles. Administration of cGH significantly stimulated cell proliferation and inhibited cell apoptosis in the ovarian stroma and small ovarian follicles. The number of ovarian follicles and the weight of the ovary prior to the first oviposition were also higher in cGH-injected hens. Thus, prior to and after the onset of egg laying, GH participates in the growth, maturation and hormonal activity of ovarian follicles in the chicken, via the regulation of steroidogenesis, proliferation and apoptosis processes.     

Distribution of follicular growth, atresia and ovulation in the ovary of the domestic hen (Gallus domesticus) at different ages

Ovaries of laying hens of 14, 18, 30, 45 and 86 weeks of age were divided transversely into two or more distinct regions. Differences in follicular populations between these regions were observed. The central segments of the ovary in birds of 14 weeks of age produced most of the visible (greater than 0.5 mg) developing follicles (P less than 0.01). This was a transient effect during ovarian maturation. In birds of 18 weeks of age, but not yet in lay, more follicles of greater than 8 mm in diameter were observed in the anterior part of the ovary than the posterior part (P less than 0.05). Follicles of this size are almost certain to ovulate. The posterior segment of the ovary of birds of 30, 45 and 86 weeks of age contained more follicles beginning the rapid growth phase, as measured by follicles of 1.4-1.8 mm in diameter, than did the anterior segment (P less than 0.01). Higher levels of atresia in the posterior segment (P less than 0.001) resulted in fewer follicles of greater than 8 mm (P less than 0.001) and fewer post-ovulatory follicles than in the anterior segment. We conclude that most of the eggs produced during the hen's laying year must be from ovulations from the anterior part of the ovary.     

Decreased granulosa cell luteinizing hormone sensitivity and altered thecal estradiol concentration in the aged hen, Gallus domesticus

Few studies have examined the effect of age on the ovulation cycle of the hen. Our aim was to determine if changes in the ovary account for the decrease in egg production with age. Young hens (28-38 wk of age) laying at least 20 eggs per sequence and old hens (53-63 wk of age) laying 3-6 eggs per sequence were used. We determined luteinizing hormone (LH) sensitivity of the ovary of young and old hens by measuring LH stimulable adenylyl cyclase (AC) activity of the granulosa layer. We also measured theca- and granulosa-layer weights and steroid concentrations of these layers and of the serum in young and old hens. Mean basal AC activity (pg/min/mg protein) for the largest (F1) and second largest (F2) follicles from young and old hens did not differ. A significant dose-response relationship to LH was present in all groups, and AC responsiveness to increasing doses of LH was greater in the F1 and F2 follicles of young hens than in the same follicles of old hens. The F4 and F5 follicles of young hens had a significantly greater estradiol (E2) concentration (pg/mg theca protein) compared to old hens, while the E2 concentration in the F2 follicle was greater in old hens. The theca layer of the F1 follicle of old hens weighed significantly more than that of young hens, whereas the theca layer of the F3, F4 and F5 follicles from young hens weighed more than those of old hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Steroidogenic activity of chicken ovary during pause in egg laying

The present study was designed (i) to assess the changes in the activity of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and cytochrome P450 aromatase (P450arom) in the ovaries of hens which are subjected to a pause in egg laying by fasting, and (ii) relate these changes with progesterone (P(4)) and estradiol (E(2)) production in the ovary. Hy-Line Brown laying hens (n=90) were fasted for 5 days with water deprivation only on day 3 and subsequently fed every second day up to day 13 and then ad libitum. Birds were euthanized (n=18) on day 0, 3, 6, 9 and 16 of the experiment. The activities of 3beta-HSD and P450arom were evaluated in stroma with cortical follicles (<1mm) and in the wall of white non-hierarchical (1-8 mm) and yellow hierarchical follicles (>8 mm) by histochemical and immunohistochemical method, respectively. Ovarian P(4) and E(2) were measured radioimmunologically. Hens stopped egg laying on day 4 of the experiment and pause in egg laying lasted up to day 12. The hens then began to gradually resume egg laying and on day 16 all hens laid eggs. It was found that during the pause in egg laying: (i) the activity of 3beta-HSD in stroma and normal white follicles was slightly decreased while P450arom activity was significantly increased; (ii) in yellow hierarchical follicles which became atretic and regressed, activity of both enzymes were markedly decreased; (iii) ovarian P(4) production dramatically decreased, whereas ovarian E(2) production after an initial decrease significantly increased. In white atretic follicles the activity of 3beta-HSD and P450arom was very weak during the whole experiment. In conclusion, the present results indicate that during a pause in egg laying white follicles become resistant to atresia.     

Variation in the ovarian and plasma progesterone and estradiol levels of the domestic hen during a pause in laying

Little is known about the physiological events occurring in the chicken ovary during a pause in laying, therefore the aim of the present study was to examine changes in sex steroid concentration in the follicle wall and blood plasma during cessation of egg laying. The experiment was performed on laying Isa Brown hens. Control hens were fed ad libitum whereas the experimental ones were subjected to a pause in laying by complete food deprivation for 5 days and water deprivation on 3 day followed by feeding every second day up to 9 day and then ad libitum. Blood samples were taken from the wing vein each day. The hens were decapitated on day 3, 6, 9, and 16. The ovary was isolated and the following follicles were dissected: white (1-2; 2-4; 4-6; 6-8 mm) and yellow preovulatory ones (F1-F3). Progesterone and estradiol were measured in the follicle wall and blood plasma by RIA methods. The hens stopped egg laying on day 4 and began egg restoration on day 14 of the experiment. Cessation of egg laying was preceded by a decrease in estradiol and progesterone levels in the ovary as well as in the blood plasma. The plasma level of these steroids began to increase 7 days before the start of egg restoration. Autopsy of the ovary showed that the atrophy of the chicken yellow preovulatory follicles during the pause in laying was accompanied by a significant increase in the total number of white follicles.     

Dietary daidzein influences laying performance of ducks (Anas platyrhynchos) and early post-hatch growth of their hatchlings by modulating gene expression

Our previous studies demonstrated that dietary supplementation of daidzein improves egg production in duck breeders during late periods of the laying cycle. The present study was aimed to clarify whether the growth of ducklings hatched from eggs laid by daidzein-treated hens would be affected, and to elucidate the mechanisms underlying potential trans-generational effects, by determining changes of hormone levels and mRNA expression of relevant genes. Daidzein was added to the basal diet of 415-day-old duck breeders at the level of 5 mg/kg. During 9 weeks of daidzein treatment, laying rate increased by 7.70%, average egg mass tended to increase, whereas yolk/albumen ratio decreased significantly. These changes were accompanied by significantly elevated plasma T4 and E2 levels, enhanced gonadotropin releasing hormone (GnRH) mRNA, but diminished estrogen receptor (ER)-beta mRNA expression in hypothalamus of daidzein-treated hens. Ducklings hatched from daidzein-treated eggs were significantly smaller at hatching, but they caught up with their control counterparts by 4 weeks of age. Serum levels of T4, pituitary GH, hepatic GH receptor (GHR) and type-1 IGF receptor (IGF-1R) mRNA expression were all suppressed markedly in the daidzein-treated group at hatching, but this suppression proved to be temporary, as at 4 weeks of age, expression levels of all investigated genes were restored. However, it is noteworthy that at 4 weeks of age an obvious down-regulation of hypothalamic GnRH mRNA expression was detected in ducklings maternally exposed to daidzein. Our results provide evidence that maternal exposure to daidzein affects post-hatch growth in the duck with accompanying changes in the secretion of metabolic hormones and expression of growth-related genes. Although the negative effect of maternal daidzein on embryonic growth could be eliminated 4 weeks after hatching, the long-term effect of maternal daidzein on reproductive function is not to be ignored and awaits further investigation.     

Immunolocalization of MHC-II+ cells in the ovary of immature, young laying and old laying hens Gallus domesticus

The aim of this study was to localize major histocompatibility complex class II positive (MHC-II+) cells in the hen ovary, and to determine the effects of ageing and sex steroids on their frequency. Cryostat sections of ovarian tissues of immature, young laying and old laying hens and those of immature hens treated with or without diethylstilboestrol or progesterone were prepared. Sections were immunostained for MHC class II antigens using mouse anti-chicken MHC class II monoclonal antibody and observed under a light microscope. Positive cells were counted using a computer-assisted image analyser. MHC-II+ cells were localized in the ovarian stroma and theca layer of primary follicles in all birds examined. The frequency of MHC-II+ cells in the stroma and theca of primary follicles (approximately 400-600 microns in diameter) was significantly greater in young laying hens than it was in immature and old laying hens (P < 0.01). In the stroma and the theca of primary follicles of diethylstilboestrol-treated birds, the frequency of MHC-II+ cells was significantly greater than it was in the stroma and theca of control and progesterone-treated birds (P < 0.01). Progesterone had no significant effect when compared with controls. These results indicate that both the ovarian stroma and theca of follicles in the hen ovary contain MHC-II+ cells, the frequency of MHC-II+ cells increases in association with sexual maturation and decreases thereafter during ageing, and oestrogen may be one of the factors enhancing the induction of MHC-II+ cells in the ovary.     

The influence of prepubertal partial sinistral ovariectomy on the subsequent reproductive function of domestic hens (Gallus domesticus )

Each of 20 White Leghorn hens of 13 to 14 weeks were subjected to partial sinistral ovariectomy and sham-operations. In half of the hens from each group, the percentage of egg production and clutch size were noted until 50 weeks of age. The growing pattern of normal ovarian follicles was also recorded at 26 weeks of age in a rest half ofthe hens in the two groups. The percentage of egg production and the mean and variance of clutch size did not differ significantly (P / 0.05) between the partially ovariectomized and sham-operated groups. The growing yellow follicles (>8 mm) in the rapidly developing phase in these two groups did not vary, although the smaller follicles (4 to 8 mm in diameter) remained significantly (P / 0.01) more in the shamoperated control group than in the partially ovariectomized group. This observation indicates that smaller follicles (4 to 8 mm) developed in the larger (>8 mm) follicles more efficiently in partially ovariectomized hens than in the sham-operated (control) hens. In a second experiment, one group of hens had all the yellow follicles (>8 mm) removed, while a second group of hens was left untreated. On the 3rd and 6th day post treatment, the hens were examined for the presence of ovarian follicles. No significant (P / 0.05) difference in the growing pattern of subsequent follicles (2 to 4 or 4 to 8 mm) was detected due to treatment. These data demonstrate that the mechanisms that regulate follicular growth and atresia are adjust to maintain normal ovulation following partial ovariectomy.     

Ovarian follicular development, lipid peroxidation, antioxidative status and immune response in laying hens fed fish oil-supplemented diets to produce n-3-enriched eggs

The objective of the present study was to research the effect of feeding laying hens fish oil-supplemented diets to produce n-3-enriched eggs on their ovarian follicular development, serum lipid peroxidation, antioxidative status and immune response. A total of 105 white Bovens hens at 24 weeks of age were housed in cages in an open-sided building under a 16 h light : 8 h dark lighting schedule. Birds were randomly divided into five treatments and were fed, ad libitum, diets containing 0% (control), 1.25%, 2.5%, 3.5% or 5.0% fish oil from 24 to 36 weeks of age. Egg production and weight were recorded. By weeks 35 and 36 of age 15 eggs were taken at random from each treatment to determine the yolk lipid profile and cholesterol content. At the end of the experimental period, 10 females from each treatment were randomly chosen, anaesthetised and killed by decapitation. Ovary and oviduct samples were immediately weighted and ovarian follicles were classified. Serum thiobarbituric acid-reactive substance (TBARS), hepatic TBARS and hepatic glutathione peroxidase (GSH-Px) activity were measured. No clear trend was observed concerning egg production and egg yolk cholesterol. As dietary fish oil levels increased, n-3-polyunsaturated fatty acids (n-3 PUFA) increased, whereas n-6 PUFA tended to decrease in yolk lipids. No negative effects were detected in ovary and oviduct weights, expressed in both absolute terms and relative to body weight. The numbers and total weights of large yellow follicles (LYF) in the ovary were not significantly affected by fish oil supplementation. Low levels (1.25% to 2.5%) of fish oil reduced both plasma and hepatic TBARS and enhanced GSH-Px activity. It is also interesting to note that inclusion of 2.5% fish oil in laying hen diets enhanced the antibody titre in laying hens. Therefore, it could be concluded that inclusion of fish oil in laying hen diets at moderate levels increased the n-3 fatty acids content in eggs, improved antioxidative status, enhanced the antibody response and did not have a negative influence on the different reproductive morphology parameters in laying hens.     

Progesterone injection and egg production in turkey hens

An arrest in laying associated with either a polyovarian follicle (POF) or a polycystic ovarian follicle (PCOF) syndrome has been reported in turkey hens photostimulated at an early age with a constant-light photoperiod. Hens expressing the POF or PCOF syndrome had stopped laying for several weeks, but the ovary contained an increased number of mature-size and larger follicles (POF hens), which were cystic (PCOF) in some of the hens. Hens with the POF or PCOF syndrome had plasma progesterone (P(4)) concentrations that were relatively high and without surges. We hypothesized that high plasma P(4) concentrations may block ovulatory surges of LH but not the growth or maintenance of hierarchical follicles leading to development of the POF or PCOF syndrome in turkey hens. In the first six studies, hens were photostimulated with either a 14L:10D or a 24L:0D photoperiod and, after laying for 1-38 wk, were then injected daily for up to 14 days with P(4) (up to 1.50 mg kg(-1) day(-1)) and necropsied. At all ages, the oviposition rate was reduced at a P(4) dosage of 0.17 mg kg(-1) day(-1). With dosages of 0.33 mg kg(-1) day(-1) or greater, however, ovipositions stopped in most hens within approximately 2 days. For hens laying for less than 15 wk, oviductal weight and number of hierarchical follicles of P(4)-injected hens were not different from control vehicle-injected hens, but the numbers of mature, cystic, and atretic follicles were increased. For hens laying for 38 wk, when treated with P(4), oviductal weight and number of hierarchical follicles decreased, but number of atretic follicles increased. No effect of photoperiod was found on egg production, oviductal weight, or follicle number, and none of the hens developed POF or PCOF syndrome in these experiments. Two additional experiments were conducted with hens early in the reproductive period that had been photostimulated with 14L:10D or 24L:0D and injected with P(4) (0.33 mg kg(-1) day(-1)) for 10 or 12 days but not necropsied until 3 wk after the last injection. Most of the hens photostimulated with the 24L:0D photoperiod and injected with P(4), and a few of the hens photostimulated with the 14L:10D photoperiod and injected with P(4), had developed the PCOF syndrome when necropsied. The hens with the PCOF syndrome had high levels of P(4) when necropsied. From these studies, we concluded that the PCOF syndrome can be induced early in the reproduction period by photostimulating turkey hens with a 24L: 0D photoperiod, injecting them for 10 to 12 days with P(4) at a dosage of 0.33 mg kg(-1) day(-1), and then waiting 3 wk for the PCOF syndrome to develop.     

The effects of age and sex steroids on the macrophage population in the ovary of the chicken, Gallus domesticus

The role of macrophages in the function of the hen ovary has not yet been described, although these cells may be an important regulator of ovarian function in mammals. The aim of this study was to determine the changes in the frequency of macrophages during ageing and follicular atresia, and the effects of sex steroids on the macrophage population in the hen ovary. Cryostat sections of ovarian tissues of immature, young laying and old laying hens and those of immature hens treated with or without diethylstilboestrol (DES) or progesterone were immunostained for macrophage cells using mouse anti-chicken macrophage monoclonal antibody. Macrophages were observed under a light microscope and counted using a computer assisted image analyser. The frequency of macrophages in both the stroma and theca of primary follicles was significantly greater in young laying hens than in immature and old laying hens and these cells were more frequent in old laying hens than in immature hens (P < 0.01). Macrophages were more frequent in atretic follicles than in normal follicles (P < 0.01). The number of macrophages in both the stroma and theca of primary follicles of DES-treated birds was significantly greater than in those of progesterone-treated and control birds (P < 0.01). Progesterone had no significant effect on the population of macrophages. These results suggest that macrophages in the ovary increase in association with sexual maturation of birds and atresia of follicles and decrease during ageing. Oestrogen may be one of the factors that affect the population of macrophages in the hen ovary.     

Expression of follicle stimulating hormone and luteinizing hormone receptors during follicular growth in the domestic cat ovary

In order to better understand the pituitary regulation of follicular growth in the domestic cat, follicle stimulating hormone (FSH) and luteinizing hormone (LH) receptors (R) were localized and quantified in relation to follicle diameter and atresia using in situ ligand binding on ovarian sections. Expression of FSHR was homogeneous and restricted to follicle granulosa cells from the early antral stage onwards, whereas expression of LHR was heterogeneous on theca cells of all follicles from the early antral stage onward, and homogeneous on granulosa cells of healthy follicles larger than 800 microm in diameter and in corpora lutea. LHR were also widely expressed as heterogeneous aggregates in the ovarian interstitial tissue. Atretic follicles exhibited significantly reduced levels of both FSHR and LHR on granulosa cells, compared with healthy follicles whatever the follicular diameter, whereas levels of LHR on theca cells were lower only for atretic follicles larger than 1,600 microm in diameter. In healthy follicles, levels of FSHR and LHR in all follicular compartments increased significantly with diameter. Although generally comparable to that observed in other mammals, the expression pattern of gonadotropin receptors in the cat ovary is characterized by an early acquisition of LHR on granulosa cells of growing follicles and islets of LH binding sites in the ovarian interstitial tissue.     

Growth hormone (GH) receptor knockout mice reveal actions of GH in lung development

The presence of growth hormone (GH) and GH receptors (GHRs) in the lung suggests it is an autocrine/paracrine target site for pulmonary GH action and/or an endocrine site of pituitary GH action. Roles for GH in lung growth or pulmonary function are, however, uncertain. The possibility that pituitary and/or pulmonary GH have physiological roles in lung development has therefore been investigated in GHR knockout (KO or -/-) mice, using a proteomics approach to determine if an absence of GH-signaling affects the proteome of the developing lung. More than 600 proteins were detected by 2-DE in the lungs of control [GHR (+/+)] and GHR (-/-) mice at the end of the alveolarization period (at day 14 postnatally). Of these, 39 differed significantly in protein content at the p>0.05 level [6 were of higher abundance in the GHR (-/-) group, 33 were of lower abundance] and 17 differed at the p>0.02 level [5 of higher abundance in the GHR (-/-) group, 12 of lower abundance] and 7 were definitively identified by MS. Vimentin, a protein involved in cellular proliferation, was reduced in content by approximately 75% in the lungs of the GHR (-/-) mice. Three proteins involved in oxidative protection [SH3 domain-binding glutamic acid-rich-like protein, peroxiredoxin 6 (Prdx6), and isocitrate dehydrogenase 1] were also of lower content in the GHR (-/-) lungs (by approximately 88%, 81% and 70%, respectively). Prdx6 is also involved in lipid and surfactant metabolism, as is apolipoprotein A-IV, the lung content of which was reduced by approximately 73% in these mice. Proteasome 26S ATPase subunit 4, a protein involved in the non-lysosomal degradation of intracellular proteins, and electron flavoprotein alpha subunit , involved in intracellular metabolism, were also reduced in content in the lungs of the GHR (-/-) mice (by approximately 70% and 49%, respectively). These results therefore suggest that these proteins are normally dependent upon GH signaling, and that GH is normally involved in early lung growth, oxidative protection, lipid and energy metabolism and in proteasomal activity. These roles may reflect endocrine actions of pituitary GH and/or local autocrine/paracrine actions of GH produced within the lung.     

Immunohistochemical localization of aromatase during the development and atresia of ovarian follicles in prepubertal horses

Ovarian steroidogenesis from the neonatal to pubertal period in horses is poorly understood. This study was designed to immunolocalize cytochrome P450 aromatase in the ovarian follicles of slaughtered fillies ages approximately (I) 6-9 mo (<10MF); (II) 1 y (1YF); and (III) 1.5 y (1.5YF). The ovaries of adult mares were used as controls. In each age group, immunoreactivity for P450arom was observed in the mural granulosa of nonatretic follicles >5 mm in diameter. Staining intensity was dependent on the size and morphology of the follicle. In nonatretic follicles 5-10 mm in diameter, the reaction was weak and heterogeneous, while most intense staining was observed in preovulatory follicles. In follicles (diameter <20 mm) in the groups <10MF and 1YF, the reaction was less intense than in adult mare follicles of similar size. In each age group, several follicles with early or advanced signs of atresia exhibited a heterogeneous staining pattern, which subsequently disappeared in late atretic follicles. No immunoreactivity was detected in the theca interna, preantral follicle, or stroma cells. Our observations reveal that the mural granulosa of viable follicles in fillies about 6-18 mo old contains aromatase, indicating that the ovary is capable of estrogen synthesis. Immunoreactivity for P450arom was dependent on follicle size and disappeared in atretic follicles.     

Follicular growth and atresia in the ovaries of hens (Gallus domesticus) with diminished egg production rates

Diets containing 3.5, 1.0 and 0.1% calcium were fed from the age of 42 weeks to individually caged laying hens. Ovaries were examined at 46-49 and 70 weeks of age for changes in the follicular population corresponding to the lowered egg production rates of birds given calcium-deficient diets (1.0% and 0.1%) and older birds given a normal diet (3.5%). Growth rates of follicles from 3.5 mm diameter to ovulation were not changed by the level of dietary calcium in 46-49-week-old birds. The number of atretic small follicles (less than or equal to 8 mm diam.) increased in old and calcium-deprived birds, resulting in lower numbers of viable follicles in the intermediate stages of growth (3-8 mm diam.). There was also an increase in the number of small follicles (1-2 mm diam.) starting to grow in 70-week-old birds which may have partly compensated for the increased loss by atresia. Birds of all ages on all diets were able to produce large follicles up to ovulable size. The main feature of poor laying birds was a reduction in the ovulation rate due to the loss of large follicles (greater than 8 mm diam.) by atresia, an event seen rarely in the birds with good laying performance. As atresia is the normal fate of most of the small follicles, the mechanisms controlling atresia in the small follicles and the large follicles appear to be independent.     

Expression and functional role of peroxisome proliferator-activated receptor-gamma in ovarian folliculogenesis in the sheep

Peroxisome proliferator-activated receptor (PPARgamma) is a nuclear receptor that is activated by fatty acids and derivatives and the antidiabetic glitazones, which plays a role in the control of lipid and glucose homeostasis. In the present work, we tested the hypothesis that PPARgamma plays a role in reproductive tissues by studying its expression and function in the hypothalamo-pituitary-ovary axis in the sheep. PPARgamma 1 and PPARgamma 2 proteins and mRNAs were detected in whole ovine pituitary and ovary but not in hypothalamic extracts. In situ hybridization on ovarian section localized PPARgamma mRNA in the granulosa layer of follicles. Interestingly, PPARgamma expression was higher in small antral (1-3 mm diameter) than in preovulatory follicles (>5 mm diameter) (P < 0.001) and was not correlated with healthy status. To assess the biological activity of ovarian PPARgamma, ovine granulosa cells were transfected with a reporter construct driven by PPARgamma-responsive elements. Addition of rosiglitazone, a PPARgamma ligand, stimulated reporter gene expression, showing that endogenous PPARgamma is functional in ovine granulosa cells in vitro. Moreover, rosiglitazone inhibited granulosa cell proliferation (P < 0.05) and increased the secretion of progesterone in vitro (P < 0.05). This stimulation effect was stronger in granulosa cells from small than from large follicles. In contrast, rosiglitazone had no effect on LH, FSH, prolactin and growth hormone secretion by ovine pituitary cells in vitro. Overall, these data suggest that PPARgamma ligands might stimulate follicular differentiation in vivo likely through a direct action on granulosa cells rather than by modulating pituitary hormone secretion.     

Role of intrafollicular regulators and FSH in growth and development of large antral follicles in sheep

Manipulation of circulating concentrations of hormones and ovarian follicle status was carried out on Day 11-12 of the oestrous cycle in sheep. All follicles visible on the ovary were ablated by cautery and ewes were treated with oestradiol or ovine follicular fluid (oFF) to suppress FSH or with PMSG to increase circulating gonadotrophic activity. One group underwent unilateral ovariectomy which greatly increased endogenous FSH and was the only treatment which significantly affected LH pulse frequency. The size distribution of antral follicles, the extent of atresia and the mitotic index of granulosa cells of follicles on Day 15 showed that (a) treatment with oFF inhibited the growth of follicles beyond 2 mm diameter by suppressing the mitotic index of the granulosa cells and (b) the concentration of FSH in peripheral plasma was related to the ability of small antral follicles to grow during the late luteal-early follicular phase of the cycle. Subsequently, it was demonstrated that oFF inhibits, in a dose-dependent manner, folliculogenesis sustained by PMSG in ewes on Days 12-15. Inhibition of folliculogenesis was represented by a decrease in those follicles greater than 4 mm, an increase in the relative proportion of follicles less than 2 mm, and minimal change in the average number of follicles visible on the ovarian surface, and a decrease in the mitotic index of granulosa cells of follicles less than 2 mm. There was no change in the extent of atresia.(ABSTRACT TRUNCATED AT 250 WORDS)     

尼罗罗非鱼生长激素及其受体的cDNA克隆与mRNA表达的雌雄差异

尼罗罗非鱼(Oreochromis niloticus)雌雄鱼生长差异明显,为了探讨其原因,本文采用RT-PCR方法克隆了尼罗罗非鱼生长激素(Growthhormone,GH)及其受体(Growth hormone receptor,GHR)的cDNA序列,并应用半定量RT-PCR方法比较了雌、雄尼罗罗非鱼垂体GHmRNA、肝脏GHRmRNA、肌肉GHRmRNA的表达差异。序列分析表明:GH开放阅读框为615bp,共编码204个氨基酸;GHR开放阅读框为1908bp,共编码635个氨基酸。以RT-PCR方法研究了GH、GHR在各组织的分布情况,结果表明:GH仅在垂体中检测到有表达,而GHR在所检测的18种组织中均有表达,其中以肝脏、肌肉、性腺、下丘脑、胸腺表达量较高。以半定量RT-PCR方法进一步比较了雌、雄尼罗罗非鱼垂体GHmRNA、肝脏GHRmRNA、肌肉GHRmRNA的表达量,结果表明:雄鱼垂体GHmRNA和肝脏GHRmRNA的表达量均显著高于雌鱼,肌肉GHRmRNA的表达量则无显著差异,推测垂体GHmRNA和肝脏GHRmRNA表达的雌雄差异是尼罗罗非鱼雌雄生长差异的主要原因之一。     

Involvement of growth hormone (GH) and insulin-like growth factor (IGF) system in ovarian folliculogenesis

During the last decade, involvement of growth hormone (GH), insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) in ovarian folliculogenesis has been extensively studied. This review provides an update on the GH, IGF system and their role in ovarian follicular development. In vitro studies and knockout experiments demonstrated an important role of GH in preantral follicle growth and differentiation through their binding with GH receptors, which are located both in the oocyte and follicular somatic tissues. Furthermore, GH stimulates the development of small antral follicles to gonadotrophin-dependent stages, as well as maturation of oocytes. With regard to the IGF system, IGF-I has no effects on primordial follicle development, but both IGF-I and IGF-II stimulate growth of secondary follicles. Depending on the species studies and method used, these proteins have been detected in oocytes and/or somatic cells. In antral follicles, these IGFs stimulate granulosa cell proliferation and steroidogenesis in most mammals. The bioavailability of IGFs is regulated by a family of intrafollicular expressed IGF binding proteins (IGFBPs). Facilitation of IGF can be increased through the activity of specific IGFBP proteases, which degrade the IGF/IGFBP complex, resulting in the production of IGFBP fragments and release of attached IGF.