Endocrine regulation of ovarian antral follicle development in cattle

Antral follicle growth in cattle occurs in two distinct phases; the first 'slow' growth phase spans the time from antrum acquisition to a size of approximately 3 mm detectable by transrectal ultrasound, and the second 'fast' phase is gondadotrophin-dependent and includes cohort growth, dominant follicle (DF) selection, and DF growth. This review summarises current concepts of the relative roles FSH and LH, ovarian and metabolic hormones play mainly in the second phase of antral follicle growth in animals of different reproductive and nutritional states. It is proposed that differential FSH response may enable one cohort follicle to become selected, and that follicular secretions, particularly inhibin, suppress FSH and thus are responsible for DF selection and dominance. Acute dependence of the DF on LH pulses will determine DF lifespan, and the LH pulse profile can be influenced by metabolic hormones such as leptin, providing one possible link for nutritional state and reproduction. Direct ovarian effects of acute and chronic changes in growth hormone, insulin and insulin-like growth factor (IGF)-I have been described on cohort follicles, DF oestrogen activity and on DF growth. Influences of metabolic hormones on early antral follicles undergoing their first 'slow' growth phase are less well described, yet metabolic hormones appear to enhance growth into the cohort available for FSH-induced emergence, and may influence subsequent developmental competence of oocytes.     

缝隙连接在小鼠卵泡发育过程中的作用

缝隙连接广泛分布于各组织细胞中,由其构成的胞间通道允许小分子在胞间直接传递,在胞间通讯方面起着重要的作用。缝隙连接由连接蛋白(Cx)组成,已发现Cx家族有20多个成员。在哺乳动物卵泡发育过程中,卵母细胞与周围的颗粒细胞之间形成的缝隙连接,介导胞间通讯,对原始生殖细胞迁移、卵母细胞减数分裂能力恢复、颗粒细胞分层、卵泡成腔、黄体形成、促性腺激素信号传递均有非常重要的调节作用。本文根据近年来相关的研究报道,总结了不同的缝隙连接在小鼠卵泡发育过程中的调节作用。     

CPEB3 deficiency in mice affect ovarian follicle development and causes premature ovarian insufficiency

Premature ovarian insufficiency (POI) is a heterogeneous and multifactorial disorder. In recent years, there has been an increasing interest in research on the pathogenesis and treatment of POI, owing to the implementation of the second-child policy in China. Cytoplasmic polyadenylation element-binding protein 3 (CPEB3) is an RNA-binding protein that can bind to specific RNA sequences. CPEB3 can bind to and affect the expression, cellular location, and stability of target RNAs. Cpeb3 is highly expressed in the ovary; however, its functions remain unknown. In this study, Cpeb3-mutant mice were used to characterize the physiological functions of CPEB3. Cpeb3-mutant female mice manifested signs of gradual loss of ovarian follicles, ovarian follicle development arrest, increased follicle atresia, and subfertility with a phenotype analogous to POI in women. Further analysis showed that granulosa cell proliferation was inhibited and apoptosis was markedly increased in Cpeb3-mutant ovaries. In addition, the expression of Gdf9, a potential target of CPEB3, was decreased in Cpeb3-mutant ovaries and oocytes. Altogether, these results reveal that CPEB3 is essential for ovarian follicle development and female fertility as it regulates the expression of Gdf9 in oocytes, disruption of which leads to impaired ovarian follicle development and POI.Subject terms: RNA-binding proteins, Infertility     

Bone morphogenetic protein-4 acts as an ovarian follicle survival factor and promotes primordial follicle development

The growth and development of follicles within the ovary are highly dependent on autocrine and paracrine signaling involving growth factors from granulosa cells, theca cells, stromal interstitial cells, and the oocytes. The growth factor bone morphogenetic protein-4 (BMP-4) and its receptor (BMPR-IB) have been detected in ovaries, and a mutation in BMPR-IB has been associated with abnormal ovulation rate. The objective of the current study was to examine the role that BMP-4 plays in the early stages of primordial follicle development. Ovaries from 4-day-old rats were placed into a whole-ovary organ culture system for 2 wk to investigate the effect that treatment with exogenous BMP-4 has on early follicle development. BMP-4-treated ovaries had a significantly higher proportion of developing primary follicles and fewer arrested primordial follicles than did untreated controls. This indicates that BMP-4 promotes primordial follicle development and the primordial-to-primary follicle transition. Ovaries were also treated with neutralizing antibody against BMP-4 to determine effects of removing endogenously produced BMP-4. Interestingly, ovaries treated with BMP-4 antibody were markedly smaller than controls. This was associated with a progressive loss of oocytes and primordial follicles, a progressive increase in cellular apoptosis, and an accompanying loss of normal ovarian tissue morphology over time. Immunocytochemistry localized BMP-4 protein to isolated stromal cell populations, selected stromal cells (i.e., pretheca cells) associated with developing primordial follicles, and the basement membrane of follicles. Ovaries were treated with BMP-4 and RNA collected after organ culture to determine whether BMP-4 signaling affects expression of other growth factors. Kit ligand and basic fibroblast growth factor expression was unchanged, but TGFalpha expression was decreased in whole ovaries. Taken together, these data suggest that BMP-4 plays an important role in promoting the survival and development of primordial follicles in the neonatal ovary.     

促卵泡成熟激素受体与卵巢癌发生发展的关系研究进展

近年来促性腺激素及其受体与卵巢癌的关系研究受到人们的广泛关注,目前已在基础性研究和流行病学调查方面取得了巨大的进展.在体内和体外模型中都已证实,作为促性腺激素成分之一的促卵泡成熟激素通过其受体对卵巢上皮细胞的细胞增殖、凋亡、细胞粘附、侵袭和转移等生物学功能发挥了重要影响.这些研究证明了促卵泡成熟激素受体和卵巢癌的发生发展密切相关,它不但增强卵巢肿瘤细胞的增殖活性,还激活多种信号通路,促进肿瘤细胞的侵袭和转移能力;促卵泡成熟激素受体的脱敏化可能是卵巢癌发生的重要分子机制;从基因水平上,其多态性还与卵巢癌的易感性密切相关;由于它介导上皮性卵巢癌的多种恶性生物功能,因此,它是一个潜在的抗卵巢癌治疗的重要靶标.虽然如此,但是卵巢癌的发生是一个相当复杂的过程,促卵泡成熟激素及其受体在卵巢癌发病中的作用机制还不清楚.     

Low density lipoprotein receptor relatives in chicken ovarian follicle and oocyte development

The normal development of the chicken oocyte within the ovarian follicle depends on the coordinated expression and function of several members of the low density lipoprotein receptor gene family. The human low density lipoprotein receptor is the prototype of the gene family; since its discovery and the elucidation of the medical significance of mutations in the LDLR gene, many additional family members have been discovered and characterized, and some important advances have resulted from studies in the chicken. I describe the analogies as well as the differences that exist between the molecular genetics of the mammalian and avian members of this important gene family, with emphasis on receptor-mediated oocyte growth. Recent progress in the molecular characterization of the chicken genes whose products mediate oocyte growth, follicle development, and accessory pathways is described in detail, and emerging information of preliminary nature is included. As the availability of chicken genome sequence data has enhanced the rate of progress in the field, our understanding of the physiological roles of members of this receptor family in general has already gained from studies in the avian model system.     

Prostaglandin signaling and ovarian follicle development in the silkmoth, Bombyx mori

Previous work on in vitro culturing of silkmoth (Bombyx mori) ovarian follicles has shown that starting from middle vitellogenesis, follicles develop according to an endogenous developmental program that does not require the presence of extra-ovarian factors. In this paper, we are reporting on our investigation for a possible involvement of autocrine/paracrine signaling by prostaglandins in the control of silkmoth ovarian follicle development. Using an initial rapid test that evaluates the formation of a protective eggshell around the oocyte, we are showing that aspirin and indomethacin, potent inhibitors of prostaglandin biosynthesis, block the transition of cultured vitellogenic follicles into choriogenesis. More detailed studies involving analyses of temporal expression patterns of genes known to be expressed in follicular epithelium cells at specific stages of ovarian development revealed that inhibition of prostaglandin biosynthesis arrests stages of follicle development from middle vitellogenesis to late choriogenesis. The arrest could be reversed by the addition of exogenous prostaglandins or cAMP into the culture media leading to the conclusion that the production of prostaglandins triggers cAMP-mediated intracellular signaling that allows the developmental progression of the follicles. Finally, because neither prostaglandins nor cAMP is capable of rescuing a developmental block effected at mid-vitellogenesis by the ecdysone agonist tebufenozide, we are proposing that prostaglandins have a role in the maintenance of normal physiological homeostasis in the ovarian follicles rather than a more specific role in developmental decision-making at distinct stages of follicle development.     

The ovarian follicle and fertility

The precise roles of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in the control of preovulatory follicle growth has been re-examined. Suppression of both pulsatile LH secretion and FSH or specific suppression of FSH results in an inhibition of preovulatory follicle growth beyond 2.5 mm dia. Infusion of sheep FSH alone in physiological amounts in the presence of basal, non-pulsatile LH results in the growth of preovulatory follicles. Co-infusion of large amplitude pulses of LH reduced or abolished this effect of FSH. It is suggested that: (1) FSH controls the number of follicles which develop; (2) selection of the large follicle destined to ovulate is directly related to the decline in the plasma concentration of FSH occurring during the period of follicle selection--thus, only the follicle(s) which can withstand this withdrawal of FSH will continue to develop; and (3) pulses of LH may directly affect the action of FSH on the follicle and play an important, hitherto unrecognized role in the selection of the ovulatory follicle by actively inducing atresia.     

Local roles of TGF-beta superfamily members in the control of ovarian follicle development

Members of the transforming growth factor-beta (TGF-beta) superfamily have wide-ranging influences on many tissue and organ systems including the ovary. Two recently discovered TGF-beta superfamily members, growth/differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15; also designated as GDF-9B) are expressed in an oocyte-specific manner from a very early stage and play a key role in promoting follicle growth beyond the primary stage. Follicle growth to the small antral stage does not require gonadotrophins but appears to be driven by local autocrine/paracrine signals from both somatic cell types (granulosa and theca) and from the oocyte. TGF-beta superfamily members expressed by follicular cells and implicated in this phase of follicle development include TGF-beta, activin, GDF-9/9B and several BMPs. Acquisition of follicle-stimulating hormone (FSH) responsiveness is a pre-requisite for growth beyond the small antral stage and evidence indicates an autocrine role for granulosa-derived activin in promoting granulosa cell proliferation, FSH receptor expression and aromatase activity. Indeed, some of the effects of FSH on granulosa cells may be mediated by endogenous activin. At the same time, activin may act on theca cells to attenuate luteinizing hormone (LH)-dependent androgen production in small to medium-size antral follicles. Dominant follicle selection appears to depend on differential FSH sensitivity amongst a growing cohort of small antral follicles. Activin may contribute to this selection process by sensitizing those follicles with the highest "activin tone" to FSH. Production of inhibin, like oestradiol, increases in selected dominant follicles, in an FSH- and insulin-like growth factor-dependent manner and may exert a paracrine action on theca cells to upregulate LH-induced secretion of androgen, an essential requirement for further oestradiol secretion by the pre-ovulatory follicle. Like activin, BMP-4 and -7 (mostly from theca), and BMP-6 (mostly from oocyte), can enhance oestradiol and inhibin secretion by bovine granulosa cells while suppressing progesterone secretion; this suggests a functional role in delaying follicle luteinization and/or atresia. Follistatin, on the other hand, may favor luteinization and/or atresia by bio-neutralizing intrafollicular activin and BMPs. Activin receptors are expressed by the oocyte and activin may have a further intrafollicular role in the terminal stages of follicle differentiation to promote oocyte maturation and developmental competence. In a reciprocal manner, oocyte-derived GDF-9/9B may act on the surrounding cumulus granulosa cells to attenuate oestradiol output and promote progesterone and hyaluronic acid production, mucification and cumulus expansion.     

Magnetic resonance image attributes of the ovarian follicle wall during development and regression

We analyzed image characteristics in T(1)-, T(2)-, and diffusion-weighted in vitro magnetic resonance (MR) images acquired at predefined stages of the ovarian cycle in 36 heifers to test the hypothesis that MR image attributes of the follicle wall reflect the physiologic status of ovarian follicles (viable, atretic, dominant, subordinate). Numerical pixel values (NPV), standard deviation of pixel values (heterogeneity), and area under the curve were used to assess images of follicle walls. Pixel values of the wall were used to calculate a regression line from which intercept, slope, and coefficient of determination were calculated. In T(1) images, NPV of dominant follicles were less likely to fit a regression line at the preovulatory phase than at any other phase (P < 0.1). Preovulatory dominant follicles had lower area under the curve in diffusion-weighted images than early and late static dominant follicles of the anovulatory wave (P < 0.02). Subordinate follicles in the presence of a preovulatory dominant follicle had lower mean NPV in T(1)- and T(2)-weighted images and lower intercepts in T(1)-weighted images than subordinate follicles of the anovulatory wave (P < 0.02). Early atresia of dominant follicles was identified at the late static phase by greater area, mean NPV, and slope in T(2)-weighted images (P < 0.02). Preovulatory dominant follicles had poor fit of NPV to a regression line in T(1)-weighted images and lower area under the curve in diffusion images. Atretic follicles had brighter walls with more acute transitions from follicular fluid to stroma in T(2)-weighted images and more heterogeneous walls in diffusion images. The MR image attributes of the follicle wall reflected the physiologic status of dominant and largest subordinate follicles.     

Aspects of follicle and oocyte stage that affect in vitro maturation and development of bovine oocytes

Success of in vitro maturation (IVM) and production of bovine embryos as related to aspects of follicle source and oocyte size were evaluated. First, it was determined that bovine oocytes continue growing in all follicular sizes studied, including >1- to 15-mm follicles. Populations of oocytes were collected from surface visible (peripheral) and cortical follicles from the same ovaries. When the number of oocytes from both peripheral and cortical follicles was combined, the yield of oocytes was approximately double that collected from 1 ovarian site alone. Oocytes from cortical follicles were smaller than those from the surface population, and the smaller cortical oocytes had a lower potential for both meiotic maturation and embryo development Only cortical oocytes with the largest diameters underwent IVM and subsequently developed to blastocysts at rates comparable to oocytes from peripheral follicles. As the diameter of the oocytes recovered from peripheral follicles increased, so did their developmental potential. When the stage of the estrous cycle was observed, it was found to have no effect on developmental potential. Finally, oocytes which extruded polar bodies at an earlier time during maturation were, on average, larger than those which extruded polar bodies later. The results serve a practical purpose in assisting selection of oocytes capable of developing into blastocysts and they give useful correlates of oocyte competencies based on knowledge of follicle source and oocyte stage.     

Magnetic resonance image attributes of the bovine ovarian follicle antrum during development and regression

The magnetic resonance images and maps of bovine ovaries acquired at defined phases of follicular development and regression were studied to determine whether magnetic resonance image attributes of the follicular antrum reflect the physiological status of dominant and subordinate ovarian follicles. Ovariectomies were performed at day 3 of wave one, day 6 of wave one, day 1 of wave two and at >/= day 17 after ovulation. The timings of ovariectomies were selected to acquire growing, early static, late static and regressing follicles of the first wave and preovulatory follicles of the ovulatory wave. Pre-selection and subordinate follicles were also available for analysis. Serum samples were taken on the day of ovariectomy and follicular fluid samples were taken after imaging. Numerical pixel value and pixel heterogeneity in a spot representing approximately 95% of the follicular antrum were quantified in T(1)- and T(2)-weighted images. T(1) and T(2) relaxation rates (T(1) and T(2)), proton density, apparent diffusion coefficients and their heterogeneities were determined from the computed magnetic resonance maps. The antra of early atretic dominant follicles showed higher T(2)-weighted mean pixel value (P < 0.008) and heterogeneity (P < 0. 01) and lower T(2) heterogeneity (P < 0.008) than growing follicles. Subordinate follicles in the presence of a preovulatory dominant follicle had higher T(1), T(1) heterogeneity, proton density, proton density heterogeneity, and lower mean pixel value in T(1)-weighted images than subordinate follicles of the anovulatory wave (P < 0.04). T(1) relaxation rate heterogeneity and proton density heterogeneity were positively correlated with follicular fluid oestradiol concentration (r = 0.4 and 0.3; P < 0.04). T(2) relaxation rate heterogeneity was positively correlated with follicular fluid progesterone concentration (r = 0.4; P < 0.008). Quantitative differences in magnetic resonance image attributes of the antrum observed among phases of follicular development and regression coincided with changes in the ability of the dominant follicle to produce steroid hormones and ovulate, and thus were indicative of physiological status and follicular health.     

Association of the prion protein and its expression with ovarian follicle development in cattle

The cellular form of the prion protein (PrP(C)) has been detected in many tissues including reproductive tissues. While its function is unclear, it has been suggested to act as a receptor for an unidentified ligand and/or as an antioxidant agent. We tested the hypothesis that PrP(C) is differentially expressed in dominant, growing, compared to subordinate bovine ovarian follicles. Using both microarray analysis and quantitative real-time PCR, the level of prion protein mRNA (Prnp) in both theca and granulosa cells was measured. We found that levels of Prnp were significantly higher in the theca cells of dominant compared to subordinate follicles but similar among granulosa cells from different follicles. This difference was apparent immediately after selection of the dominant follicle and continued to the dominance stage of the follicle wave. Levels of the protein for PrP(C) were also higher (P < 0.05) in theca cells of dominant compared to subordinate follicles. In conclusion, elevated PrP(C) was associated with ovarian follicle growth and development and we suggest that it may play a role in the success of follicle development.