The synthesis and fate of glycodelin in human ovary during folliculogenesis
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The ontogeny of glycodelin in human ovarian follicles during folliculogenesis was studied. Glycodelin immunoreactivity began to be detected in the granulosa cells and thecal cells of late secondary follicles. Immunoreactivity was also found in both the luteinized granulosa cells and cumulus cells obtained from women undergoing the assisted reproduction treatment. However, only the luteinized granulosa cells, and not the cumulus cells, expressed glycodelin mRNA. Results also showed that the cumulus cells took up radiolabelled glycodelin and partially deglycosylated some of it. Glycodelin (and a partially deglycolsylated form of glycoldelin) appeared to complex with two cytoplasmic or membrane components of the cumulus cells. The data also demonstrated that ZIF-1, a glycoprotein isolated from human follicular fluid, was immunologically similar to glycodelin. In conclusion, we suggest that glycodelin is synthesized in the granulosa cells of ovarian follicles at late secondary follicle stage. It then may be released into the follicular fluid from where it is taken up and partially modified by the cumulus cells.Keywords:
Follicular fluid
Granulosa cell
The ontogeny of glycodelin in human ovarian follicles during folliculogenesis was studied. Glycodelin immunoreactivity began to be detected in the granulosa cells and thecal cells of late secondary follicles. Immunoreactivity was also found in both the luteinized granulosa cells and cumulus cells obtained from women undergoing the assisted reproduction treatment. However, only the luteinized granulosa cells, and not the cumulus cells, expressed glycodelin mRNA. Results also showed that the cumulus cells took up radiolabelled glycodelin and partially deglycosylated some of it. Glycodelin (and a partially deglycolsylated form of glycoldelin) appeared to complex with two cytoplasmic or membrane components of the cumulus cells. The data also demonstrated that ZIF-1, a glycoprotein isolated from human follicular fluid, was immunologically similar to glycodelin. In conclusion, we suggest that glycodelin is synthesized in the granulosa cells of ovarian follicles at late secondary follicle stage. It then may be released into the follicular fluid from where it is taken up and partially modified by the cumulus cells.
Follicular fluid
Granulosa cell
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Antral follicle
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Follicular fluid
Atrial natriuretic peptide
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The mammalian cumulus-oocyte complex (COCs) promotes oocyte growth and development during long stages of folliculogenesis and oogenesis. Before ovulation, the follicle is formed by a variety of fully differentiated cell populations; cumulus cells (CCs) that tightly surround the female gamete, granulosa cells (GCs) and theca cells (TCs) which build the internal and external mass of the follicular wall. It is well documented that CCs surrounding the oocyte are necessary for resumption of meiosis and full maturation of the gamete. However, the role of the granulosa cells in acquisition of MII stage and/or full fertilization ability is not yet entirely known. In this article, we present an overview of mammalian oocytes and their relationship to the surrounding cumulus and granulosa cells. We also describe the processes of GCs differentiation and developmental capacity. Finally, we describe several markers of mammalian GCs, which could be used for positive identification of isolated cells. The developmental capacity of oocytes and surrounding somatic cells a fingerprint of folliculogenesis and oogenesis.
Granulosa cell
Gamete
Growth differentiation factor-9
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Abstract We have examined the proteins associated with the mucous matrix of the rat cumulus oophorus and compared them to the composition of rat serum, follicular fluid, ampullary fluid, and oocyte–cumulus cell extract. The cumulus matrix was dispersed using Streptomyces hyaluronidase, and the proteins were analyzed by highresolution two‐dimensional polyacrylamide gel electrophoresis and compared with proteins of the serum, proestrous follicular fluid, and postvulatory ampullary fluid and extracts of oocytes and cumulus cells. In addition to albumin and transferrin, which were common to all the fluids analyzed, the cumulus material contained many proteins in common with the follicular fluid and the ampullary fluid. However, the protein extract of the cumulus matrix also contained four major proteins not present in the other fluids analyzed. Two of these proteins were acidic and heterogenous in charge and size (MW ∼81,000 and 100,000). The other two proteins were more basic and occurred at MW ∼90,000 and 150,000. Our results show that the extracellular matrix of the cumulus contains proteins that are not present in the fluids that surround the oocyte.
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Contents Remarkably little is known about folliculogenesis in the dog. Objectives were to characterise (1) changes in follicle/oocyte diameter and granulosa cell number and (2) localisation of fibroblast growth factor (FGF)‐2 and FGF‐7 during dog ovarian follicle development. Fourteen ovarian pairs were excised and processed for histological evaluation. Two to four serial sections/bitch were stained with hematoxylin, and follicle/oocyte diameters and granulosa cell number were determined at each developmental stage. Mean follicle and oocyte size were compared among stages by one‐way analysis of variance. Relationships between follicle and oocyte size and granulosa cell number were determined using correlation and regression analysis, respectively. Another eight serial sections/bitch were processed for immunostaining to determine FGF‐2 and FGF‐7 localisation. Primordial and primary follicles were similar in size, but smaller than the progressively increasing (p < 0.05) diameter of the later stages. Oocyte diameter gradually increased (p < 0.05) among oocytes derived from primordial, primary, secondary and early antral follicles with no difference (p > 0.05) thereafter. Oocyte size and granulosa cell number increased (p < 0.01) with follicular diameter. Except during anoestrus, FGF‐2 occurred in oocytes and granulosa cells of primordial to secondary follicles. In adult bitches, FGF‐7 was localised in granulosa cells of primary and secondary follicles and also occurred in the theca layer of antral follicles during prooestrus and oestrus. In summary, folliculogenesis in the domestic dog occurs in two phases: pre‐antral phase characterised by increasing follicle size in association with oocyte growth and granulosa cell proliferation and antral phase linked with marked granulosa cell proliferation and accumulation of antral cavity fluid. Finally, the temporal localisation pattern of FGF‐2 implies its role in follicular activation, whereas FGF‐7 activities appear related to later folliculogenesis.
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We tested the current popular hypothesis that sulfated glycosaminoglycans (GAGS) in murine follicular fluid may inhibit the precocious expansion of the cumulus oophorus in the antral follicle before the LH surge. Using porcine follicular oocytes, the effect of GAGS as a form of follicular fluid was examined in the cumulus exapansion. Degree of expansion of the cumulus-oocyte complexes cultured in porcine follicular fluid was greater then that in medium supplemented with 10% fBS and 0.02 Au/㎖ FSH. However, the cumulus oophorus of native oocytes tightly connected with mural granulosa cells in hemi-sec ted follicles remained at intact cumulus oophorus after incubation in the above conditions. In contrast to the results in mice, heparin could not inhibit cumulus expansion in pig, despite of higher concentrations (range: 100㎍∼10㎎/㎖) than those of total GAGS in porcine follicular fluid. The results indicate that GAGS may not be a main factor in regulation of cumulus oophotus during follicular development in pig, and the maintenance of tight cumulus oophorus may be due to a factors) from mural granulosa cells and contact between membrana granulosa and cumulus cells.
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Abstract : Transforming growth factor beta (TGFB) signaling regulates key reproductive events via TGFBR1/TGFBR2. To determine potential effect of overactivation of TGFB signaling in the oocyte, we generated a mouse model containing a constitutively active TGFBR1 using growth differentiation factor 9 (Gdf9)-Cre (i.e., TGFBR1-gCA). Follicle counting demonstrated that the number of primordial, primary, and secondary follicles was reduced in TGFBR1-gCA ovaries compared with controls at P7. Concomitantly, abnormal follicle structures were detected in TGFBR1-gCA ovaries, evidenced by INHA staining. These results suggest that sustained activation of TGFBR1 using Gdf9-Cre disrupts folliculogenesis via affecting ovarian reserve and follicle growth/development. Apoptosis analysis using ovaries at critical timepoints during follicular development did not reveal alteration of oocyte apoptosis in TGFBR1-gCA ovaries. Immunostaining was performed to determine the molecular identify of the tumors. The results showed that ovarian tumor tissues from TGFBR1-gCA mice were positive for granulosa cell markers FOXL2, INHA, and FOXO1, supporting the formation of granulosa cell tumors in these mice. In the ovary culture system, SB-505124 seemed to improve follicle development in TGFBR1-gCA ovaries. Therefore, sustained activation of TGFBR1 using Gdf9-Cre leads to the development of ovarian neoplasms reminiscent of granulosa cell tumors.
Growth differentiation factor-9
Granulosa cell
Immunostaining
Anti-Müllerian hormone
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The stage of folliculogenesis at which the human zona pellucida (ZP) is initiated and the cells responsible for the origin of the ZP continue to be controversial. This study characterizes the development of the ZP during human folliculogenesis using ovarian samples donated from patients requesting ovarian storage.Follicles (from n = 18 patients, 14-40 years old) within fresh tissue and following development in a xenograft system were stained, using immunohistochemical techniques, for the presence of the three human ZP proteins, ZP1, ZP2 and ZP3. Over 500 primordial follicles and >20 follicles at each developmental stage were examined.All three ZP proteins were detected within the oocyte of the primordial follicle. Presence of ZP1 and ZP3 was observed in the majority of primordial oocytes (93% and 95%, respectively), whereas ZP2 was detected in only 32% of these follicles. The three ZP proteins were detected in the cytoplasm of cuboidal granulosa cells and their distribution correlates with developmental stages throughout folliculogenesis.ZP proteins were detected in both the oocyte and the granulosa cells as early as the primordial follicle stage in the human. The detection of ZP proteins in the quiescent primordial follicle suggests that these proteins have been present since oogenesis.
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As the human ovarian follicle enlarges in the course of a regular cycle or following controlled ovarian stimulation, the changes in its structure reveal the oocyte environment composed of cumulus oophorus cells and the follicular fluid (FF). In contrast to the dynamic nature of cells, the fluid compartment appears as a reservoir rich in biomolecules. In some aspects, it is similar to the plasma, but it also exhibits differences that likely relate to its specific localization around the oocyte. The chemical composition indicates that the follicular fluid is able to detect and buffer excessive amounts of reactive oxygen species, employing a variety of antioxidants, some of them components of the intracellular milieu. An important part is played by albumin through specific cysteine residues. But the fluid contains other molecules whose cysteine residues may be involved in sensing and buffering the local oxidative conditions. How these molecules are recruited and regulated to intervene such process is unknown but it is a critical issue in reproduction. In fact, important proteins in the FF, that regulate follicle growth and oocyte quality, exhibit cysteine residues at specific points, whose untoward oxidation would result in functional loss. Therefore, preservation of controlled oxidative conditions in the FF is a requirement for the fine-tuned oocyte maturation process. In contrast, its disturbance enhances the susceptibility to the establishment of reproductive disorders that would require the intervention of reproductive medicine technology.
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