PubMed Health⌕ Search

Biomedical subjects

N Dekel

Publications and source records attributed to N Dekel.

At least 37 records · Page 2Linked to original sources
PubMed1992

Rat oocytes induced to mature by epidermal growth factor are successfully fertilized.

Epidermal growth factor (EGF), which is a known mitogen, can also induce resumption of meiosis in the rat oocyte. The present study was designed in an attempt to elucidate whether oocytes, induced to mature by EGF in a follicle-enclosed oocyte culture, are fertilizable and can further develop into two-cell embryos. For further clarification of the effect of EGF on steroidogenesis in the ovarian follicle, progesterone concentrations were determined by radioimmunoassay. We found that oocytes matured by EGF (100 ng/ml) were successfully fertilized. Even though their rate of fertilization was relatively lower as compared to that of oocytes stimulated by luteinizing hormone (LH) both in vitro and in vivo (61%, 79%, and 83% respectively), once fertilized they exhibit an equal potential for further development (EGF: 48%, LH: 45%). On the other hand, EGF-induced progesterone production was very poor. These findings strongly support the idea that both mitogenesis and meiogenesis can be mediated by common signals. The results further suggest that progesterone production and oocyte maturation, in the rat, are independent events.

Animals↗
PubMed1991

Maturation of the rat cumulus-oocyte complex: structure and function.

The cumulus cells that surround the mammalian oocyte become dispersed following the preovulatory surge of the pituitary gonadotropin, luteinizing hormone (LH). We have examined cumulus-oocyte complexes of PMSG-primed immature rats before and at 1, 2, 3, 4, 6, and 8 hr after injection of human chorionic gonadotropin (hCG), which acts on the rat ovary like the pituitary gonadotropin. Associations between projections of the cumulus cells and the oocyte were analyzed in thin sections. We observed that some cumulus projections were greatly enlarged where they associate with the oocyte. These enlarged regions were filled with numerous small vesicles. Gap junctions between cumulus cell projections and the oocytes were small. We quantitated the number and size of gap junctions between cumulus cells. The number of small gap junctions (less than 1 microM) between cumulus cells did not change significantly over the 8-hr period after hCG administration. Larger gap junctions, however, showed a general downward trend beginning after the third hour post hCG. Light microscopic observations of plastic sections revealed that dispersion of the cumulus oophorus is not observed until after 4 hr post-hCG, but between 4 and 8 hr after gonadotropin administration the cumulus becomes markedly dispersed. In the majority of the oocytes in these complexes the germinal vesicle (GV) displayed some irregularity in shape at 2 hr post-hCG, although absence of the GV was not observed until later. Our observations suggest a new means of communication in the cumulus-oocyte complex by the vesicle-filled enlargements of the cumulus cell projections at the oocyte surface. They further indicate that the decrease in metabolic coupling observed in rat cumulus-oocyte complexes soon after exposure to LH is not associated with a change in number and size of the gap junctions between the cumulus cells. We suggest that it is either the disruption of the gap junctions at the region of contact of the cumulus cell projections with the oocyte surface or the operation of a gating mechanism that blocks the junctional channels without affecting their morphological appearance that is responsible for uncoupling of the oocyte from the cumulus cells.

Animals↗
PubMed1991

Serum bioactive and immunoreactive follicle stimulating hormone during chronic treatment with gonadotropin releasing hormone agonist in elderly men.

Chronic administration of GnRH agonists "down regulates" the pituitary and decreases LH and FSH serum levels. Changes in the bioactivity of FSH have not been adequately assessed under such treatment, for lack of a proper test. We examined serum changes under GnRH agonist treatment among 12 healthy elderly men suffering only from benign prostatic hypertrophy, for up to one year, using a modification of a granulosa cell bioassay for the determination of FSH bioactivity. While radioimmunoassay-FSH decreased, we noticed a significant increase in the bioactivity of this hormone. The clinical importance of this increase is discussed.

Aged↗
PubMed1990

Rat oocyte maturation: role of calcium in hormone action.

We studied the role of extracellular calcium (Ca0) in oocyte maturation and oocyte-cumulus cells interaction in rat follicles in vitro. Luteinizing hormone (LH) or a gonadotropin-releasing hormone analog (GnRHa) induced full maturation at [Ca0] = 1.3 mM. At [Ca0] = 0.6 mM, maturation induced by LH or GnRHa was inhibited by 65%. Chelatin of [Ca0] resulted in 45% maturation and neither hormone caused a further increase of maturation. [Ca0] = 20 mM enhanced the response to suboptimal concentrations of GnRHa but inhibited that to LH. Divalent cation ionophores caused [Ca0]-dependent maturation, which was fully inhibited by dibutyryl cAMP. Changes in [Ca0] also affected oocyte-cumulus interaction. At [Ca0] = 1.3 mM, either LH or GnRHa caused partial dispersion of the cumulus. Chelation of [Ca0] also resulted in an almost complete dispersion of the cumulus. The ionophores, however, caused maturation with the oocyte-cumulus complex preserved intact. Our data suggest that GnRHa may induce maturation via cAMP-sensitive calcium mobilization into the oocyte-cumulus-granulosa complex.

Animals↗
PubMed1990

Serum bioactive and immunoreactive follicle-stimulating hormone in oligozoospermic and azoospermic men: application of a modified granulosa cell bioassay.

Serum follicle-stimulating hormone (FSH) levels measured by radioimmunoassay (RIA) usually correlate well with the rate of spermatogenesis. However, in certain cases this correlation does not exist. The purpose of this study was to establish a reliable bioassay of FSH for the andrological clinic. Follicle-stimulating hormone was measured by both standard RIA and bioassay in 98 men subgrouped into normospermic, oligospermic, and azoospermic. Bioactivity of FSH was determined using in vitro cultures of granulosa cells utilizing progesterone measurements for assessing FSH activity. Results of FSH levels obtained by both methods correlated well (r = 0.55, P less than 0.01) within themselves, and both correlated negatively and significantly with sperm concentration. The ratio between bioactivity and immunoreactivity of FSH did not correlate with sperm density. Thus, the decrease in sperm concentration and other sperm variables resulting from a germinal epithelial dysfunction was not mediated or associated with low biological activity of FSH. The application of this method can be of clinical value in cases where a discrepancy is found between serum RIA-FSH levels and sperm quality.

Animals↗
PubMed1990

Embryonic development of fertilized rat oocytes induced to mature by an analogue of gonadotrophin-releasing hormone.

A gonadotrophin-releasing hormone analogue (GnRHa) was administered to hypophysectomized immature rats. Postovulatory mature oocytes obtained under these conditions were exposed in vitro to a sperm suspension for fertilization. Developmental ability of the fertilized ova was studied by transfer of the 2-cell stage embryos to oviducts of foster mothers. The potential of oocytes, undergoing maturation in response to GnRHa, to develop into 2-cell embryos was similar to that of oocytes stimulated by hCG (76.4% and 83.1% respectively). The 2-cell stage embryos obtained from such oocytes were equally able to implant in the uteri of foster mothers (25.7% and 21.2% respectively) and subsequently develop into live embryos (15.3% and 15.2%, respectively, at Day 20 of pregnancy).

Animals↗
PubMed1988

Dissociation between the inhibitory and the stimulatory action of cAMP on maturation of rat oocytes.

The possible mediatory role of cAMP in the induction of oocyte maturation by luteinizing hormone (LH) is not yet clear since evidence for both inhibitory and stimulatory actions of the nucleotide on the oocyte has been provided. To elucidate the role of cAMP in regulation of oocyte meiosis we tried in the present study to dissociate between the inhibitory and stimulatory action of this nucleotide on oocyte maturation. To induce maturation, oocytes enclosed by their follicles were transiently exposed to either dibutyryl cAMP (dbcAMP) or to the phosphodiesterase inhibitor methylisobutylxanthine (MIX). Inhibition of maturation was obtained by the addition of the above agents to either follicle-enclosed oocytes incubated in the presence of LH or isolated cumulus-free oocytes that mature spontaneously in vitro. We found that inhibition of oocyte maturation is obtained by a relatively low dose of either dbcAMP or MIX while higher concentrations of these agents are required to induce oocyte maturation. Coupling of the oocyte to the cumulus cells, as expressed by the fraction of labeled uridine transferred from the cumulus cells to the oocyte following exposure of the follicle-enclosed cumulus-oocyte complex to MIX, was also determined. We found that uncoupling of the oocyte from the cumulus cells corresponded with the induction, but not inhibition of oocyte maturation, both by its concentration dependence and time-course. We suggest that cAMP has a dual role in regulation of oocyte maturation. Lower levels of the nucleotide act to maintain meiotic arrest, while elevated levels of cAMP mediate LH action to induce meiosis resumption.

1-Methyl-3-isobutylxanthine↗
PubMed1988

Regulation of oocyte maturation: communication in the rat cumulus-oocyte complex.

This study was undertaken to analyse the time relationships between breakdown of communication in the cumulus-oocyte complex and reinitiation of meiosis in the oocyte. In addition the possibility was examined that under conditions of established communication, cAMP is transferred from the cumulus cells to the oocyte. Coupling in the cumulus-oocyte complex and maturation of the oocyte were examined, both in vitro in follicles exposed to LH and in vivo following injection of the hormone to immature, pregnant mare's serum gonadotrophin-primed rats. Transfer analysis was performed by determination of cAMP content in cumulus-enclosed as compared to cumulus-free oocytes incubated with forskolin. It was discovered that after 1 h of culture in the presence of LH, coupling in the cumulus-oocyte complexes had decreased to 45% of its initial level, while only 10% of the oocytes had resumed meiosis by this time. The decrease in coupling in cumulus-oocyte complexes in vivo was 20% by 2 h after administration of the hormone, with all the oocytes being immature at this time. Cyclic AMP determinations revealed that cumulus-enclosed oocytes contained concentrations of cAMP three-fold higher than cumulus-free oocytes incubated under similar conditions. This study demonstrates that, in the rat, (i) a decrease in coupling precedes reinitiation of meiosis and (ii) in the presence of established communication, cAMP is transferred from the cumulus cells to the oocyte. These findings suggest that a decrease in communication can serve as the signal for oocyte maturation, possibly by preventing transfer of cAMP from the cumulus cells to the oocyte.

Animals↗
PubMed1988

Induction of maturation in follicle-enclosed oocytes: the response to gonadotropins at different stages of follicular development.

Antral follicles, isolated from either nontreated or pregnant mare's serum gonadotropin (PMSG)-primed 27-day-old rats, were incubated in the absence or the presence of either luteinizing hormone (LH), follicle-stimulating hormone (FSH), or forskolin. The effect of these agents on oocyte maturation and cyclic adenosine 3',5'-monophosphate (cAMP) accumulation was studied and compared. Both gonadotropins, LH and FSH, as well as forskolin, effectively induced maturation of oocytes enclosed by large antral follicles isolated from PMSG-primed rats. On the other hand, we found that maturation of oocytes enclosed by small antral follicles, isolated from nonprimed and PMSG-primed rats, could be induced by either FSH or forskolin but not by LH. cAMP determinations revealed that, in spite of the inability of LH to induce oocyte maturation, elevated concentrations of the nucleotide were detectable in small antral follicles exposed to this gonadotropin. Since granulosa cells isolated from the large but not the small antral follicles were stimulated by LH to generate cAMP, the elevation of cAMP concentrations in the small antral follicle apparently represented the response of the theca cells to this gonadotropin. Since it is the ability of the granulosa cells to interact with the hormone that determines whether or not oocyte maturation will occur, we suggest that the granulosa, but not the theca cells, mediate LH action to induce oocyte maturation.

Adenylyl Cyclases↗
PubMed1988

Regulation of oocyte maturation. The role of cAMP.

We suggest that cAMP is a key molecule in regulation of oocyte maturation. In the small follicle, tonic levels of cAMP are continuously transferred to the oocytes to maintain meiotic arrest. In the preovulatory follicle, in response to LH, cAMP levels are elevated. These increased concentrations of the nucleotide affect the cumulus cells and interrupt communication in the cumulus-oocyte complex. Under these conditions the flow of cAMP to the oocyte declines, inhibition is relieved, and meiosis is resumed.

Animals↗
PubMed1988

Receptors for gonadotropin releasing hormone are present in rat oocytes.

Specific receptors for gonadotropin releasing hormone (GnRH) in the rat oocyte have been identified by using two independent methods. Light microscopic autoradiography, utilizing an iodinated biologically active photoaffinity derivative of GnRH, revealed specific binding of the neurohormone to rat oocytes. Furthermore, the presence of GnRH-receptor is also evident from indirect fluorescent immunocytochemistry that shows binding of GnRH-receptor antibodies to rat oocytes which is neither detected with non immune serum nor with antiserum depleted of GnRH-receptor antibodies. These antibodies to the GnRH-receptor, also bind to both cumulus and granulosa cells but not to rat basophilic leukemia cells. The presence of specific GnRH receptors on rat oocytes provides an experimental basis for understanding the molecular events involved in GnRH-induced oocyte maturation.

Animals↗
PubMed1988

G-proteins in mammalian gametes: an immunocytochemical study.

The presence of transductory GTP(G)-regulatory proteins in mammalian gametes has been examined by indirect fluorescence immunocytochemistry. Using rabbit antisera to bovine rod beta gamma-transducin (RA beta gamma T), bovine rod holotransducin (AS-1), bovine rod alpha-transducin (RA alpha T), synthetic bovine rod alpha-transducin C-terminal decapeptide (AS-6), bovine brain alpha 39Go (RA alpha 39), and two mouse monoclonal antibodies raised against frog retinal transducin (4A), and rat brain beta-tubulin, we demonstrated the presence of corresponding immunoreactive material in both rat oocytes and bovine ejaculated sperm. Immunostaining in the oocyte was evenly distributed on the oolemma, excluding the cell cytoplasm and zona pellucida. Immunoreactive material was also present in the cumulus cells that encapsulate the oocyte. In contrast, the immunofluorescence corresponding to transductory G-proteins was confined in sperm to functionally defined regions in the head and tail, in a manner specific for each antibody. While RA beta gamma T, AS-1 and RA alpha 39 all stained the entire acrosome, AS-6 and RA alpha T stained only the acrosomal tip. Monoclonal antibody 4A stained the midpiece exclusively and anti-rat betaq-tubulin (a structural G-protein) stained the full length of the sperm tail. The existence of several G-protein types in mammalian gametes suggests their possible involvement in the regulation of various effector systems, in a manner reminiscent of somatic cells. The unique situation in sperm, where different G-proteins show distinct and specific patterns of distribution, further suggests their association with various effector systems in discrete functional domains.

Animals↗
PubMed1987

Maintenance of meiotic arrest in isolated rat oocytes by the invasive adenylate cyclase of Bordetella pertussis.

Rat oocytes resume meiosis spontaneously in vitro within 3 h after their isolation from the ovarian follicles. We report here that the spontaneous maturation of isolated rat oocytes is preceded by a drop in intracellular levels of cyclic adenosine 3',5'-monophosphate (cAMP). Further experiments were carried out to examine the possible correlation between the meiotic status and cAMP levels within the oocyte. To challenge rat cumulus-free oocytes to generate cAMP, bypassing their own adenylate cyclase, a preparation of an invasive adenylate cyclase from Bordetella pertussis was used. We found a dose-dependent elevation of cAMP levels within these oocytes that corresponded to inhibition of their spontaneous maturation. Persistent inhibition of meiosis was obtained with the continuous presence of the enzymatic preparation, whereas its removal resulted in a transient inhibition associated with a drop in cAMP. We suggest that the presence of elevated cAMP levels in the oocyte is directly responsible for the maintenance of meiotic arrest.

Adenylyl Cyclases↗
PubMed1987

Fertilization in vitro of rat oocytes undergoing maturation in response to a GnRH analogue.

Oocytes were exposed to GnRHa to induce their maturation both in vivo, by administration of the hormone to hypophysectomized rats, and in vitro, in cultures of intact ovarian follicles. Mature oocytes obtained under both these conditions were then exposed in vitro to a sperm suspension for fertilization. Fertilization of control groups of oocytes, isolated from intact or hypophysectomized PMSG-primed hCG-induced ovulators, was 88.3 +/- 3.3% (n = 331) and 90.0 +/- 2.8% (n = 427), respectively, as compared to 82.8 +/- 3.2% (n = 413) for oocytes isolated from hypophysectomized PMSG-primed GnRHa-induced ovulators. Fertilization rate in oocytes treated by GnRHa in vitro was 78.5 +/- 3.1% (n = 247) as compared to 79.3 +/- 4.1% (n = 261) in LH-treated oocytes. These results demonstrate that fertilizability of oocytes undergoing maturation in response to GnRHa is similar to that of oocytes induced to mature by LH. No differences could be detected in the proportions of abnormal oocytes (polyspermic, fragmented and dead) and the zygotes obtained after fertilization of GnRHa- or LH-treated oocytes showed similar ability to cleave.

Animals↗
PubMed1985

Activators of protein kinase C stimulate meiotic maturation of rat oocytes.

Agonistic analogs of gonadotropin releasing hormone can induce oocyte maturation in rat follicle-enclosed oocytes (1-5). Cyclic AMP does not rise following exposure of the ovarian follicle to GnRH (3) suggesting that cAMP-dependent protein kinase is not involved in the mechanism of GnRH action in this system. Protein kinase C, which is independent of cAMP, has recently been reported to mediate GnRH action in the pituitary (6-8). The possible involvement of this enzyme in the regulation of oocyte maturation has been tested in the present study. We report here that phospholipase C and direct activators of protein kinase C can mimic the response of rat oocytes to GnRH. These results suggest that GnRH-induced meiotic maturation of rat oocytes is mediated by the phospholipid-dependent protein kinase, protein kinase C.

Animals↗