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PubMed2002

Transcriptional activity associated with meiotic competence in fully grown mouse GV oocytes.

The involvement of cumulus cells and chromatin organisation in transcriptional activity was investigated. In addition, the relationship between transcriptional activity and meiotic competence in fully grown mouse oocytes was surveyed. Transcriptional activity was detected in fully grown oocytes in which chromatin did not surround the nucleolus in the germinal vesicle (NSN-type oocytes), but not in oocytes in which chromatin surrounded the nucleolus (SN-type oocytes). Cumulus cells seemed to downregulate transcriptional activity in NSN-type oocytes, since transcriptional activity was 3 times greater in the denuded NSN-type oocytes free of cumulus cells (DO oocytes) than in NSN-type oocytes enclosed in cumulus cells (COC oocytes). Higher transcriptional activity corresponded to lower germinal vesicle breakdown (GVB) competence of fully grown oocytes in culture. Although GVB occurred in nearly all (99%) the SN-type oocytes, it occurred in 88% of COC/NSN-type oocytes (cumulus-oocyte complex with SN-type configuration) and in 61% of DO/NSN-type oocytes (denuded oocytes with NSN-type configuration). There was a negative correlation between transcriptional activity and the capacity of a cell to complete the progression to the second metaphase (MII). In GVB oocytes, the percentage of first polar body (PBI) extrusion differed among COC/NSN-type (81%), DO/SN-type (66%), COC/NSN-type (47%) and DO/NSN-type (29%) oocytes. After activation with 10 mM Sr2+, the frequency of parthenogenetic activation was greater in SN-type oocytes (46.9%) than in transcriptionally active NSN-type oocytes (27.5%). These results suggest that transcriptional activity has a detrimental effect on the competence of meiotic maturation and subsequent activation in fully grown GV oocytes. Alternatively, active transcription in the fully grown oocytes suggests that they are still in the process of synthesising substances required for meiotic maturation and are not yet competent for these processes.

Animals↗
PubMed2006

Induced expression of pattern recognition receptors in cumulus oocyte complexes: novel evidence for innate immune-like functions during ovulation.

Ovulation is the complex, inflammatory-like process by which the cumulus oocyte complex (COC) is released from a mature, preovulatory follicle through a rupture site at the ovarian surface and requires expression of genes that generate and stabilize the expanded extracellular COC matrix. Gene profiling analyses of COCs at selected time intervals during ovulation revealed that many genes associated with immune related surveillance functions were also induced in cumulus cells. Specifically, cell surface signaling molecules known as pattern recognition receptors that act as sensors of the external environment important for the innate immune system to detect self from nonself or altered self are induced and/or expressed in cumulus cells as well as granulosa cells. These include the complement factor q1, CD14, and the Toll-like receptors (TLRs) 4, 8, and 9 as well as mediators of TLR activation, myeloid differentiation primary response gene 88 and interferon regulatory factor 3. COCs exposed to bacterial lipopolysaccharide exhibit enhanced phosphorylation of p38MAPK, ERK1/2 and nuclear factor-kappaB and increased expression of Il6 and Tnfa target genes, documenting that the TLR pathway is functional. Cumulus cells and granulosa cells also express the scavenger receptors CD36 and scavenger receptor type B1 and exhibited phagocytic uptake of fluorescently tagged bacterial particles. Collectively, these results provide novel evidence that cumulus cells as well as granulosa cells express innate immune related genes that may play critical roles in surveillance and cell survival during the ovulation process.

Amphiregulin↗
PubMed1988

Morphologic and functional studies of immature rat oocyte-cumulus complexes after cryopreservation.

The effects of freezing and thawing (F/T) on functional activity of immature rat oocyte-cumulus complexes (OCC) were studied. The OCC were divided into three groups according to the number of cumulus-cell layers surrounding them. The OCC were then frozen and thawed (F/T), with dimethyl sulfoxide (DMSO) as cryoprotectant. The survival rates after thawing increased significantly (P less than 0.001) as the number of cumulus-cell layers increased. Germinal vesicle breakdown (GVBD) was evaluated in F/T oocytes. After 2 hours, significantly (P less than 0.05) fewer oocytes demonstrated GVBD than did those in the control group. There was no difference, however, after 4 hours of culture. A significant (P less than 0.05) decrease in follicle-stimulating hormone (FSH)-dependent cyclic adenosine monophosphate (cAMP) accumulation was observed in the F/T group. However, the amount of cAMP produced was sufficient to maintain the oocyte in meiotic arrest. There was a borderline significant decrease of the coupling between cumulus cells and the oocyte in F/T OCC, as evaluated by the transport of 3H-uridine into the oocyte. It is concluded that immature rat oocytes can be successfully cryopreserved when they are surrounded by five or more layers of cumulus cells. FSH responsiveness and intercellular communication were essentially maintained. There was a slight delay in GVBD, which needs further clarification.

Animals↗
PubMed2005

Identification of a hyaluronidase, Hyal5, involved in penetration of mouse sperm through cumulus mass.

A glycosylphosphatidylinositol (GPI)-anchored hyaluronidase, PH-20, on the sperm surface has long been believed to assist sperm penetration through the cumulus mass surrounding the eggs. However, mouse sperm lacking PH-20 were still capable of penetrating the cumulus mass despite a delayed dispersal of cumulus cells. Intriguingly, a 55-kDa hyaluronan-hydrolyzing protein was abundantly present in wild-type and PH-20-deficient mouse sperm. In this study, we purified the 55-kDa mouse protein from soluble protein extracts released from epididymal sperm by acrosome reaction and identified as a hyaluronidase, Hyal5. Hyal5 was exclusively expressed in the testis and formed a 160-kbp gene cluster together with Hyalp1, Hyal4, and Ph-20 on mouse chromosome 6. Hyal5 was a single-chain hyaluronidase present on the plasma and acrosomal membranes of sperm presumably as a GPI-anchored protein. Moreover, hyaluronan zymography revealed that Hyal5 is enzymatically active in the pH range 5-7 and inactive at pH 3 and 4. Both Hyal5-enriched PH-20-free soluble protein extracts and PH-20-deficient mouse sperm were capable of dispersing cumulus cells from the cumulus mass. Cumulus cell dispersal was strongly inhibited by the presence of a hyaluronidase inhibitor, apigenin. These results suggest that in the mouse, Hyal5 may function principally as a "cumulus matrix depolymerase" in the sperm penetration through the cumulus mass and in the local hyaluronan hydrolysis near or on the surface of the egg zona pellucida to enable the proximal region of sperm tail to move freely. PH-20 may compensate in part for the functional roles of Hyal5.

Animals↗
PubMed1994

Cytoplasmic argyrophilic protein(s) in cumulus-oocyte complexes: a preliminary study.

The presence of acidic argyrophilic protein(s) in cumulus cells and oocytes was examined in bovine, ovine, porcine and murine cumulus-oocyte complexes (COCs) using a specific silver-staining method. The COCs were isolated from small (1 mm) or large (3-5 mm) antral follicles; they were fixed immediately after slaughter (first group) or after transfer to the laboratory (ca 60 min, second group) and then silver stained according to Likovskí and Smetana (1981). The argyrophilic proteins were accumulated mainly in foot processes of cumulus cells on oocytes isolated from large bovine follicles. The intensity of the reaction was less evident in small follicles. The intensity of staining gradually disappeared from cumulus cells as the maturation continued. Moreover, we were unable to detect similar strong labelling in mouse, pig and sheep COCs. Such a chronology suggests that argyrophilic protein(s) may play a role in the transition from meiotic arrest to resumption in bovine oocytes. Our results also suggest that preliminary events of maturation may occur just after slaughter, ie a relatively long time before the COCs are isolated and cultured in vitro.

Animals↗
PubMed1995

[Chronological aspects of the nuclear maturation of bovine oocytes cultivated in vitro].

In this study, the chronological changes in the meiotic progress of in vitro maturation of bovine oocytes was analysed. Oocytes (n = 1044) were obtained from a local abattoir and were classified into two groups according to the presence or absence of cumulus cells. They were incubated in microdrops (5 per drop) in TCM-199 (control), TCM-199 + 10% fetal calf serum (treatment 2) and TCM-199 + 10% estrous cow serum (treatment 3). Oocytes were fixed and stained at the end of 0, 6, 12, 18, 24 and 30 h of the beginning of in vitro culture, were evaluated according to the nuclear stage of maturation. These results demonstrated that the immature oocytes at the time of collection (0 h) were in the germinal vesicle stage (GV), and that the highest maturation rate was at 24 h of culture in all treatments. Serum treatments enhanced the maturation rates obtained (52.1 and 55.7%) compared to control (serum-free) medium (42.7%; P < 0.05) in cumulus-cell-enclosed oocytes. In denuded oocytes, the maturation rates were lower compared to cumulus-cells-enclosed oocytes in all treatments. In conclusion, meiotic progression of bovine oocytes can be influenced by the inclusion of sera in the maturation media and by the presence of the cumulus cells.

Animals↗
PubMed2007

Large-scale chromatin remodeling in germinal vesicle bovine oocytes: interplay with gap junction functionality and developmental competence.

In mammals, oocyte acquires a series of competencies sequentially during folliculogenesis that play critical roles at fertilization and early stages of embryonic development. In mouse, chromatin in germinal vesicle (GV) undergoes dynamic changes during oocyte growth and its progressive condensation has been related to the achievement of developmental potential. Cumulus cells are essential for the acquisition of meiotic competence and play a role in chromatin remodeling during oocyte growth. This study is aimed to characterize the chromatin configuration of growing and fully grown bovine oocytes, the status of communications between oocyte and cumulus cells and oocyte developmental potential. Following nuclear staining, we identified four discrete stages of GV, characterized by an increase of chromatin condensation. GV0 stage represented 82% of growing oocytes and it was absent in fully grown oocytes. GV1, GV2, and GV3 represented, respectively, 24, 31, and 45% of fully grown oocytes. Our data indicated a moderate but significant increase in oocyte diameter between GV0 and GV3 stage. By dye coupling assay the 98% of GV0 oocytes showed fully open communications while the number of oocytes with functionally closed communications with cumulus cells was significantly higher in GV3 group than GV1 and GV2. However, GV0 oocytes were unable to progress through metaphase II while GV2 and GV3 showed the highest developmental capability. We conclude that in bovine, the progressive chromatin condensation is related to the sequential achievement of meiotic and embryonic developmental competencies during oocyte growth and differentiation. Moreover, gap-junction-mediated communications between oocyte and cumulus cells could be implicated in modulating the chromatin remodeling process.

Animals↗
PubMed1983

Reproductive function following advanced ovulation induced by synthetic luteinizing hormone-releasing hormone in rats.

Reproductive function following advanced ovulation induced by a single injection of 1 microgram synthetic luteinizing hormone-releasing hormone (LH-RH) has been studied in rats. While advanced ovulation induced by an injection of the agent on the day of proestrus was followed by pregnancies occurring as frequently as those in spontaneous ovulation group, incidences of mating behavior, fertilization, and pregnancy were considerably reduced in the advanced ovulation group when induced by an injection of synthetic LH-RH during the early period of diestrus 2. The mean volume of primary oocytes of the three largest follicles at 1.00 h on the day of diestrus 2 was significantly smaller than that at 1.00 h on the day of proestrus. However, shortly before ovulation, the mean volume of secondary oocytes in the preovulatory follicles from rats treated with synthetic LH-RH at 1.00 h on the day of diestrus 2 was approximately the same as that in the preovulatory follicles from rats treated with the agent at 1.00 h on the day of proestrus. Shortly after ovulation the number of mitotic cumulus cells around the oviductal ova was significantly greater in advanced ovulation group induced by synthetic LH-RH injection during the early period of diestrus 2 than in spontaneous ovulation group. Furthermore, when examined 13 h after ovulation, the layers of cumulus cells were thicker, and cumulus cells were more tightly arranged around the ova in the advanced ovulation group induced by injection of synthetic LH-RH during the early period of diestrus 2 than those in the group with with spontaneous ovulation. These results suggest that the cumulus-oocyte complex from advanced ovulation group induced by injection of synthetic LH-RH in the early period of diestrus 2 is less potent in fertilizability than that from spontaneous ovulation group, leading to a reduction in the rate of subsequent pregnancy.

Animals↗
PubMed1984

Asynchrony between human cumulus-corona cell complex and oocyte maturation after human menopausal gonadotropin treatment for in vitro fertilization.

One hundred seventeen oocyte-cumulus-corona cell complexes (CCCs) were obtained from 15 women undergoing in vitro fertilization after human menopausal gonadotropin/human chorionic gonadotropin follicular stimulation. In each woman, five oocyte-CCCs were left intact, and the remaining one to five oocytes were freed of their CCCs by hyaluronidase (300 IU/ml) dispersal treatment. Dispersal time for individual CCCs correlated well with their degree of mucification and was significantly shorter in 20 mature CCCs as compared with 24 intermediate CCCs (1.20 +/- 0.05 versus 2.35 +/- 0.13 minutes, respectively; P less than 0.001). Oocyte maturation at collection and after 8 hours of in vitro incubation was not related to the CCC type present at harvest. Maturation of oocytes progressed regardless of CCC type; so that after 8 hours of preincubation, 73% of the oocytes attained a polar body while 20% were still at the germinal vesicle breakdown stage and 7% at the germinal vesicle stage. Overall, the 44 denuded oocytes and 72 intact oocyte-CCCs were comparable in rate of fertilization (64% versus 68%) and subsequent cleavage (75% versus 82% of all fertilized oocytes). It is concluded that in human menopausal gonadotropin-stimulated cycles, asynchrony between individual CCC mucification and oocyte maturation may occur, and that the absence of CCC does not seem to affect in vitro fertilization and cleavage rates.

Adult↗
PubMed2000

Effect of the gonadotropin-releasing hormone antagonist ganirelix on cyclic adenosine monophosphate accumulation of human granulosa-lutein cells.

OBJECTIVE: To evaluate whether the GnRH antagonist ganirelix exerts an effect on cyclic adenosine monophosphate (cAMP) production of human granulosa-lutein (GL) cells in vitro. DESIGN: In vitro cell culture study. SETTING: Research laboratory of a university hospital. PATIENT(S): Mural GL and cumulus cells were obtained from 15 patients on whom controlled ovarian hyperstimulation was being performed for intracytoplasmic sperm injection treatment. INTERVENTION(S): Mural GL and cumulus cells were cultured for 48 hours with and without 1 nM ganirelix or triptorelin. For the last 6 hours, the cells were either exposed to 1-5 IU hCG or left unstimulated. MAIN OUTCOME MEASURE(S): At the end of the culturing period, the intracellular and extracellular cAMP accumulations were measured by an (125)I-scintillation proximity assay. RESULT(S): hCG induced dose-dependent increases in total cAMP accumulation. Stimulation with 1 IU/mL hCG resulted in 9-fold and 13-fold increases, and 5 IU/mL hCG resulted in 19-fold and 14-fold increases in total cAMP release from cumulus and mural GL cells, respectively. On the other hand, treatments with 1 nM GnRH antagonist ganirelix and 1 nM GnRH agonist triptorelin did not exert any significant changes on the basal and hCG-stimulated cAMP accumulation of mural GL cells and cumulus cells as compared with controls. CONCLUSION(S): Ganirelix does not influence basal and hCG-stimulated cAMP accumulation of human GL cells in vitro. cAMP is apparently not involved in the mechanism of action of GnRH analogs in human ovary.

Adenosine Monophosphate↗
PubMed2005

Oocyte nucleus controls progression through meiotic maturation.

We analyzed progression through the meiotic maturation in oocytes manipulated to replace the prophase oocyte nucleus with the nucleus from a cumulus cell, a pachytene spermatocyte or the pronucleus from a fertilized egg. Removal of the oocyte nucleus led to a significant reduction in histone H1 kinase activity. Replacement of the oocyte nucleus by a pronucleus followed by culture resulted in premature pseudomeiotic division and occasional abnormal cytokinesis; however, histone H1 kinase activity was rescued, microtubules formed a bipolar spindle, and chromosomes were condensed. In addition to the anomalies observed after pronuclear transfer, those after transfer of the nucleus from a cumulus cell or spermatocyte included a dramatically impaired ability to form the bipolar spindle or to condense chromosomes, and histone H1 kinase activity was not rescued. Expression of a cyclin B-YFP in enucleated oocytes receiving the cumulus cell nucleus rescued histone H1 kinase activity, but spindle formation and chromosome condensation remained impaired, indicating a pleiotropic effect of oocyte nucleus removal. However, when the cumulus cell nucleus was first transformed into pronuclei (transfer into a metaphase II oocyte followed by activation), such pronuclei supported maturation after transfer into the oocyte in a manner similar to that of normal pronuclei. These results show that the oocyte nucleus contains specific components required for the control of progression through the meiotic maturation and that some of these components are also present in pronuclei.

Animals↗
PubMed1986

Ultrastructure of in-vitro oocyte maturation in cattle.

Cumulus-oocyte complexes were collected from cows at an abattoir by aspiration from small (1-6 mm) antral follicles. After different periods of culture the complexes were processed for electron microscopy. Cumulus expansion occurred at 12-18 h of culture and concomitantly enlargement of cumulus cell projections in the perivitelline space was seen. At 48 h the innermost cumulus cells flattened and adhered tightly to the zona pellucida. In the oocyte the following changes occurred: at 0-3 h of culture the perivitelline space developed; at 3-12 h disconnection of the junctions between cumulus cell projections and the oolemma, and the concomitant breakdown of the nucleus was observed; at 12-18 h the mitochondria moved from a peripheral location to a more even spatial distribution and the Golgi complexes decreased in size; at approximately 18 h the smooth endoplasmic reticulum formed large aggregates surrounded by mitochondria; at 18-21 h the first polar body was abstricted; at 24-40 h the cortical granules spread; at 30-40 h the polar body degenerated in some specimens; at 40-48 h the perivitelline space decreased in size; and at 48 h one oocyte was in the process of fragmentation. It is concluded that nuclear and cytoplasmic oocyte maturation is simulated in vitro. However, certain deviations were noticed compared to in-vivo maturation.

Animals↗
PubMed1997

Maturation, fertilization, and development of marmoset monkey oocytes in vitro.

This study was conducted to investigate 1) the capacity of in vitro-matured (IVM) marmoset oocytes to be fertilized and to support embryonic development in vitro and 2) oocyte meiotic maturation in relation to in vivo FSH administration, follicle size, and oocyte-cumulus cell status. Pairs of ovaries were collected on Day 4 of the follicular phase from adult females receiving either 1) human FSH (3 IU; n = 5) or 2) control (saline; n = 5) daily for 4 days. Antral follicles were excised from ovaries and separated into classes according to size: class 1 (660-840 microm), class 2 (> 840-1000 microm), class 3 (> 1000-1400 microm), and class 4 (> 1400 microm). A total of 823 partially naked and cumulus-enclosed oocytes (CEOs) were released from follicles and cultured in vitro. Cumulus cells remaining after 22 h were removed, metaphase II (MII) oocytes were inseminated with epididymal sperm, and resulting embryos were cultured until developmental arrest. Fluorescence microscopy was used to assess oocyte meiotic and embryo developmental progression. Oocyte germinal vesicle breakdown (GVB)- and MII-competencies increased significantly with follicular size (p < 0.01 and p < 0.0001, respectively), although they were independent of oocyte-cumulus cell associations. After 24 and 32 h in vitro, 69% and 93%, respectively, of CEOs with MII competence had completed meiotic maturation, and the rate of nuclear maturation increased progressively with follicle size (p < 0.01) and with the association of cumulus cells (p < 0.01). In vivo FSH priming slightly improved oocyte GVB- and MII-competencies (p < 0.01 and p < 0.05, respectively) and decreased the time required to achieve MII (p < 0.01). IVM oocytes from all follicle sizes fertilized (78-92%) in vitro, with 27% developing to morula- and 4% to blastocyst-stage embryos. This study demonstrates for the first time that IVM New World primate oocytes are able to support advanced preimplantation embryonic development in vitro. Oocyte meiotic competence and the time course of nuclear maturation are profoundly influenced by their follicular origin, and marginally by FSH treatment.

Animals↗
PubMed2006

Mathematical modelling of oxygen concentration in bovine and murine cumulus-oocyte complexes.

Immature oocytes benefit from nutrient modification of the follicular environment by the surrounding cumulus mass. However, the oxygen concentration that the oocyte may be exposed to could be lower than the antral follicular concentration due to the metabolism of surrounding cumulus cells. Using metabolic data previously determined, we have developed a mathematical model of O(2) diffusion across the bovine and murine cumulus-oocyte complex. From this we have determined that across a physiological range of external pO(2), less than 0.25% and 0.5% O(2) is removed by cumulus cells within the bovine and murine cumulus-oocyte complex respectively. Our model differs from others as it: incorporates a term that allows for nonlinear variation of the oxygen consumption rate with oxygen concentration; considers two regions (oocyte and cumulus) sharing a common boundary, both of which consume oxygen at different non linear rates. Cumulus cells therefore remove little O(2), thus sparing this essential gas for the oocyte, which is dependent on ATP generation via oxidative phosphorylation.

Animals↗
PubMed2006

[Granulosa cells as indicators of oocyte quality].

Granulosa-cumulus cells are located around the oocyte and may be involved in competence acquisition and in the resumption of oocyte meiosis. We assessed Bax expression in granulosa-cumulus cells and its correlation to in vitro resumption of oocyte meiosis in women presenting PCOS (polycystic ovarian syndrome), as well as the role of the chemokine SDF1-CXCL12 in human preovulatory follicle in the follow-up of granulosa cell survival and embryo quality in IVF.

Chemokine CXCL12↗
PubMed1980

Ovarian follicular and luteal physiology.

Follicular maturation and development is a complex process of interrelated intra- and extraovarian events that ultimately lead to ovulation of a mature oocyte and transformation of the ruptured follicle into a corpus luteum. The primordial follicle consists of an immature oocyte arrested in the dictyate stage of meiosis, surrounded by a single layer of relatively undifferentiated granulosa cells. The oocyte remains in the immature state because of many factors, one of which is the oocyte maturation inhibitor (OMI) secreted by granulosa cells. The oocyte subsequently increases in size, and as the antrum forms it becomes surrounded by cumulus cells. The cumulus cells may be intimately involved in the action of O,I to arrest the oocyte in the immature state within the follicle, as well as the resumption of meiosis during the LH surge. The compartments of the follicle that change most dramatically during follicular maturation are the cells lining the follicle--the granulosa and thecal cells. Under the influence of estrogen and FSH, the granulosa cells proliferate and also acquire FSH receptors. At this time, the thecal compartment differentiates and surrounds the granulosa cells, but remains separated from them by a basement membrane. Steroid secretion by the antral follicle involves the interplay of androgens, estrogens, and progestins. Both the granulosa and thecal cell compartments contribute to follicular fluid and serum levels of steroids; the interaction of both cell types may be necessary for estrogen and progesterone secretion in some species. As a consequence of the presence of an elevated number of FSH receptors, the granulosa cells of the small antral follicle are able to respond to FSH in many ways, including increased cyclic AMP accumulation, activation of the aromatase system, and induction of LH receptors, which permits the granulosa cells to later respond to LH. The mechanism by which thecal cells acquire their LH receptors is presently unknown. The granulosa cells of the follicle may indirectly control their own maturation and the number of follicles maturing through the secretion of follicular inhibin, which decreases the pituitary output of FSH. Even though the granulosa cells have acquired large numbers of LH receptors, they are prevented from luteinizing prematurely by factors in follicular fluid, including estrogen and a luteinizing inhibitor (LI). As serum LH levels increase during the preovulatory LH surge, a number of events occur: resumption of oocyte meiosis, transformation of the steroid enzyme complex from estrogen to progesterone secretion, follicular rupture, and formation of the corpus luteum. Granulosa cells form the bulk of the corpus luteum, which secretes elevated amounts of progesterone for a fixed time period depending on the species. Before ovulation the preovulatory follicle must be exposed to and respond to adequate LH and FSH levels in order for the eventual corpus luteum to secrete elevated amounts of progesterone for its normal lifespan...

Animals↗
PubMed1984

Effect of forskolin on the spontaneous maturation and cyclic AMP content of rat oocyte-cumulus complexes.

Forskolin (0-100 microM), a reversible stimulator of the catalytic subunit of adenylate cyclase, induced dose-response increases in the % germinal vesicle (GV) (ID50 = 2.68 microM, where ID50 is the dose of forskolin which maintained the meiotic arrest at the germinal vesicle stage, determined cytogenetically, of 50% cultured oocytes), and the cAMP content (determined by RIA) of cumulus-enclosed oocytes and cumulus masses. A significant positive correlation was established between the amount of cAMP within the cumulus mass and that in the corresponding enclosed oocyte (r = 0.78). In contrast, neither the % GV nor the cAMP content of cumulus-free oocytes was affected by the drug. The arresting action of forskolin upon cumulus-enclosed oocytes was dependent upon the presence of adherent cumulus cells, and was transient and fully reversible. The gradual decrease in the % GV of cumulus-enclosed oocytes cultured from 4 to 12 h in 25 microM-forskolin (84, 54 and 22% GV at 4, 8 and 12 h, respectively) was accompanied by a drastic fall in intra-oocyte cAMP at 8 h (41.2 +/- 2.5 and 1.1 +/- 0.6 fmol/oocyte at 4 and 8 h, respectively), while the cumulus cell cAMP content remained constant (135.3 +/- 14.7 and 145.9 +/- 28.7 fmol/cumulus at 4 and 8 h, respectively). Moreover, heterologous metabolic coupling, assessed by determination of the fraction of radiolabelled uridine marker that was transferred from the cumulus mass to the oocyte, significantly decreased. These results show that cumulus cell cAMP is transferred to the rat oocyte where it appears to play a pivotal role in the regulation of meiosis and that the rat oolemma does not appear to possess active catalytic subunits of adenylate cyclase in an amount adequate to stimulate measurably cAMP synthesis.

Animals↗
PubMed1998

Ovarian markers of implantation potential in assisted reproduction.

In-vitro fertilization provides evidence for the heterogeneity of human embryos; data from the UK indicating that approximately 90% of embryos selected for transfer fail to implant. The need to identify factors that promote implantation or markers of implantation is self-evident. There is an increasing awareness that the heterogeneity of follicles may have a significant impact on oocyte competence and embryo viability and that factors which contribute to the heterogeneity of follicles may provide markers of implantation in assisted conception. The markers of implantation described include the expression of 11 beta-hydroxysteroid dehydrogenase by granulosa cells in vitro; adhesion and proliferation of cumulus cells in vitro; steroidogenic activity of cumulus in vitro and perifollicular vascularity and vascular endothelial growth factors bound to granulosa and cumulus cells. These factors may provide clinically useful markers of implantation potential.

11-beta-Hydroxysteroid Dehydrogenases↗