Evidence of new antigens in the mouse cumulus oophorus during preovulatory cumulus expansion

Amiel, Marie-Laure; Moos, Jiří; Testart, J

doi:10.1002/mrd.1080340113

Cited by 6 publications

(5 citation statements)

References 17 publications

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“…Since the expansion of the cumulus mass appears to benefit in vitro fertilization [2,3], up to now most investigators have used serum in the culture medium during IVM to obtain fully expanded porcine COCs. The results from the present study indicate that full expansion of the cumulus can be accomplished in a chemically defined medium containing IGF-1.…”

Section: Discussionmentioning

confidence: 99%

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Insulin-Like Growth Factor-1, a Component of Serum that Enables Porcine Cumulus Cells to Expand in Response to Follicle-Stimulating Hormone in Vitro1

Singh¹,

Armstrong²

1997

Biology of Reproduction

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The present study was aimed at determining whether insulin-like growth factor-1 (IGF-1) is a component of fetal bovine serum (FBS) that enables porcine cumulus cells to expand in response to FSH in vitro. Cumulus-oocyte complexes (COC) obtained from 4- to 6-mm follicles of prepubertal gilts were cultured at 39 degrees C for 24 h in media that contained human recombinant IGF-1 (50 ng/ml), FBS (15% v:v), or their combination, with or without FSH (1.5 microg/ml), and cumulus expansion was scored microscopically. Expansion was FSH dependent and was observed only when IGF-1, FBS, or both were present. The proportion of FSH-stimulated COC exhibiting full expansion in response to IGF-1 alone did not differ significantly (p > 0.05) from the proportion in those cultured with FBS or IGF-1+FBS (79 +/- 2.8% vs. 84 +/- 2.2% or 76 +/- 6.2%, respectively). In a concentration-response study, FSH-stimulated expansion was observed in a significant proportion of COC (32 +/- 2.8% vs. 0% control) at 1 ng/ml IGF-1, with the proportions increasing dose-dependently to maximal values between 10 and 75 ng/ml IGF-1, and decreasing at higher IGF-1 concentrations. Exposure of COC to an IGF-1 receptor (IGF-1R)-neutralizing antibody (Ab) for 90 min before addition of FSH and FBS dose-dependently inhibited cumulus expansion, with maximal inhibition at 10 microg/ml (1 +/- 1.0% vs. 68 +/- 1.1% control). In the absence of FBS or IGF-1, some COC had a tendency toward slight expansion when cultured with FSH, and the Ab completely inhibited that effect, suggesting that this may be due to endogenous IGF-1 production by the COC. The Ab effect was reversible and was eliminated by washing twice with fresh medium followed by culture for an additional 24 h in the presence of FSH and FBS. Expression of IGF-1R mRNA in the isolated oocyte and cumulus cells was determined by reverse tramscriptase polymerase chain reaction using sequence-specific primers. The IGF-1R message was detected in both the oocyte and cumulus cells. Collectively, these observations suggest that IGF-1 is a component of serum that enables cumulus cells to expand in response to FSH in vitro, and that the effect is receptor mediated. Since IGF-1 is present in the follicle in vivo, it may have a physiological role during gonadotropin-induced cumulus expansion.

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Section: Discussionmentioning

confidence: 99%

“…The compact layers of cumulus cells expand due to the deposition of mucoelastic material in the extracellular matrix. Optimal expansion of the cumulus mass appears to be essential both for normal ovulation [1] and for subsequent fertilization [2,3]. In vitro, cumulus exAccepted January 14, 1997.…”

Section: Introductionmentioning

confidence: 99%

Insulin-Like Growth Factor-1, a Component of Serum that Enables Porcine Cumulus Cells to Expand in Response to Follicle-Stimulating Hormone in Vitro1

Singh¹,

Armstrong²

1997

Biology of Reproduction

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“…Cells were counted using a hemocytometer and 4 3 10 4 cells were loaded in 100 ll of SFM to the top chamber. This is approximately equivalent to the number of cells in 20 COCs, with COCs estimated at 2000 cells/COC [39].…”

Section: Cell Linesmentioning

confidence: 99%

“…Fixed-endpoint assays utilized 12-lm-pore polycarbonate transwell inserts (Millipore) placed in 24-well Costar cell culture [39]. Cells were incubated at 378C/5% CO 2 for 4 h, then polycarbonate filters were removed and nonmotile cells were scraped from the upper side of the filter using a cotton swab.…”

Section: Invasion/migration Assaysmentioning

confidence: 99%

Transient Invasive Migration in Mouse Cumulus Oocyte Complexes Induced at Ovulation by Luteinizing Hormone1

Akison

Alvino

Dunning

et al. 2012

Biology of Reproduction

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Ovulation, the release of the oocyte from the ovarian follicle, is initiated by the luteinizing hormone surge. It is clear that highly controlled degradation of the follicle and ovarian wall is required for passage of the oocyte and accompanying cumulus cells from the follicle, but the mechanism has not yet been elucidated. Here we show that cumulus oocyte complexes (COCs) adopt transient adhesive, migratory, and matrix-invading capacities at the time of ovulation. We characterized cell adhesion, migration, and invasion in preovulatory and postovulatory mouse COCs collected over a time course post-human chorionic gonadotropin (hCG) administration. Adhesion of dispersed cumulus cells and intact COCs to extracellular matrix proteins present in the ovarian wall (collagens, laminin, and fibronectin) increased significantly after hCG treatment and declined immediately after ovulation. Cumulus cell migration was low in unexpanded, equine chorionic gonadotropin-only treated COCs, but increased 4, 8, and 10 h post-hCG, reaching a peak at 12 h post-hCG that coincided with ovulation. The ability of cumulus cells to migrate was rapidly diminished in COCs isolated from the oviduct within 2 h postovulation. Cell migration was cumulus cell specific and was not observed in granulosa cells. Invasion through three-dimensional collagen I and matrigel barriers by preovulatory expanded COCs was equivalent to that of a known invasive breast cancer cell line (MB-231). Cumulatively, these results demonstrate that cumulus cells in the expanded COC transition to an adhesive, motile, and invasive phenotype in the periovulatory period that may be required for successful release of the oocyte from the ovary at ovulation.

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“…Before ovulation, it controls oocyte maturation, and after ovulation it participates in the complex mechanisms controlling the access of the fertilizing spermatozoon to the oocyte [26,27]. In this light Amiel et al [1] tested the effects of an antiserum against mouse cumulus-oocyte complexes (anti COC) on in vitro fertilization of mouse oocytes and results indicated the appearence of new antigens in mouse cumulus during preovulatory expansion.…”

Section: Discussionmentioning

confidence: 99%

Localization and partial characterization of a rat ovarian granulose cell protein with a monoclonal antibody

Russinova

Vassilev

Davidoff

1993

Biology of the Cell

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Hybridoma cell lines were obtained from mouse splenocytes sensitized to granulosa cells collected from rat ovaries after gonadotropin stimulation. A monoclonal antibody (5G5) was obtained which reacted with granulosa cells and showed a positive reaction with serum-free conditioned medium containing granulosa cell secreted proteins. Immunoblotting of the conditioned medium and light- and electron-microscopic immunocytochemistry of rat ovary show that mAb 5G5 is directed against a 59-kDa protein which is located on the plasma membrane of granulosa cells. Furthermore, the immunoreactivity of the granulosa cells depends both on the degree of follicle development and on the position of the granulosa cells within the follicles. Strong immunoreactivity was observed in the innermost granulosa cell layers, close to the oocyte and the antral cavity. The results obtained show that mAb 5G5 is a useful marker of a 59-kDa granulosa cell protein which might be of importance for the development of the follicle and the oocyte maturation.

show abstract

Evidence of new antigens in the mouse cumulus oophorus during preovulatory cumulus expansion

Cited by 6 publications

References 17 publications

Insulin-Like Growth Factor-1, a Component of Serum that Enables Porcine Cumulus Cells to Expand in Response to Follicle-Stimulating Hormone in Vitro1

Insulin-Like Growth Factor-1, a Component of Serum that Enables Porcine Cumulus Cells to Expand in Response to Follicle-Stimulating Hormone in Vitro1

Transient Invasive Migration in Mouse Cumulus Oocyte Complexes Induced at Ovulation by Luteinizing Hormone1

Localization and partial characterization of a rat ovarian granulose cell protein with a monoclonal antibody

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