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N Dekel

Publications and source records attributed to N Dekel.

At least 55 records · Page 3Linked to original sources
PubMed1985

Mammalian fertilization as seen with the scanning electron microscope.

For several years we have been looking at mammalian gametes and their interactions with the scanning electron microscope (SEM). Examining the images produced by the SEM has given us a three-dimensional view of sperm, eggs, and egg investments. We are particularly impressed with the structural variation among gametes of different mammalian species. In this short report we examine the structure of mammalian spermatozoa, eggs, zonae pellucidae, and cumuli. Our observations and those of others have led us to believe that variation in gamete structure and function may have evolved as a mechanism for reproductive isolation of mammalian species.

Animals↗
PubMed1985

Epidermal growth factor induces maturation of rat follicle-enclosed oocytes.

Gonadotropin-induced differentiation of ovarian granulosa cells in culture is inhibited by epidermal growth factor (EGF). The present study was undertaken to test a possible inhibitory effect of EGF on LH-induced maturation of rat follicle-enclosed oocytes. We have found that EGF not only failed to affect LH action but served by itself as an inducer of maturation of follicle-enclosed oocytes. EGF action on the oocytes was dose and time dependent and could be prevented by (Bu)2 cAMP. The response of the oocytes was specific to EGF and could not be elicited by other growth factors such as nerve growth factor and insulin. The response to EGF was not limited to the large antral follicles, as oocytes enclosed by small antral follicles (less than 0.4 mm) were induced to mature by EGF as well. In addition, we have demonstrated that oocytes, induced to mature by EGF, are concomitantly uncoupled from the follicular cells. Based on these results we suggest that EGF may terminate the transfer of a follicular inhibitor to the oocyte. It is also possible, however, that EGF induces oocyte maturation by a mechanism independent of its effect on communication between the cellular components of the follicle.

1-Methyl-3-isobutylxanthine↗
PubMed1985

Characterization of the maturational changes induced by a GnRH analogue in the rat ovarian follicle.

The GnRH analogue [D-Ser(t-Bu)6]des-Gly10-GnRH-N-ethylamide (GnRHa, 2 micrograms/rat) or hCG (4 i.u./rat) was administered to hypophysectomized, PMSG-primed immature female rats. Oocyte maturation was initially detected by 2 h after GnRHa administration but the response to hCG was observed only after 4 h. Initiation of GnRHa-induced ovulation also preceded the response to hCG by 2 h. Maximal response to both these hormones was obtained at 10 and 14 h after hormone administration for oocyte maturation and ovulation respectively. The number of oocytes ovulated after GnRHa was significantly lower than that with hCG (29 +/- 4 and 50 +/- 7 per rat respectively; P less than 0.05). Expansion of the cumulus mass and secretion of mucoid material, which are characteristic responses to LH, were also observed after GnRHa administration. However, while the action of 5 micrograms ovine LH/ml on the cumulus cells was mediated by cAMP, no accumulation of the nucleotide could be detected in follicles exposed to GnRHa (10(-7) M). We conclude that even though GnRHa and LH/hCG seem to elicit similar responses in the ovarian follicle they differ in their kinetics, their efficiency and the mediator of their action.

Animals↗
PubMed1984

Gonadotropin releasing hormone: regulation of phospholipid turnover and prostaglandin production in ovarian granulosa cells.

The direct effect of gonadotropin releasing hormone (GnRH) upon ovarian function, is initiated by a rapid receptor-mediated increase in phosphatidylinositol (PI) turnover (approximately 5 min) followed by prostaglandin E (PGE, 120 min) and progesterone (120 min) formation, oocyte maturation and induction of ovulation. In contrast, luteinizing hormone (LH) stimulation of oocyte maturation and induction of ovulation is mediated by increased adenosine 3',5'-monophosphate (cAMP, 15 min), progesterone (30 min) and PGE (180 min) production. Both LH and GnRH stimulation of oocyte maturation are inhibited by dibutyryl cAMP and 3-isobutyl-1-methylxanthine, whereas induction of ovulation by the two hormones is blocked by indomethacin. GnRH and LH differ, therefore, in the mechanism leading to PGE formation, but thereafter share a common mechanism responsible for oocyte maturation and independently for induction of ovulation.

Animals↗
PubMed1984

Spontaneous maturation in vitro of cumulus-enclosed rat oocytes is inhibited by forskolin.

We have recently reported that the adenylate cyclase activator, forskolin, induces in the rat ovarian follicle both cAMP accumulation and oocyte maturation. We demonstrate here, on the other hand, that the spontaneous maturation in vitro of isolated rat cumulus-enclosed oocytes is inhibited by forskolin. The inhibitory effect of forskolin is dose dependent with an ED50 at 15 microM. Forskolin inhibition decreases gradually with time, being completely relieved by 20 h of culture. Methylisobutylxanthine significantly prolongs the duration of the inhibitory action of forskolin. In addition to its inhibitory effect on oocyte maturation, forskolin triggers the cumulus-oocyte complex to generate cAMP. Cyclic AMP accumulation is maximally stimulated by 100 microM of forskolin with an ED50 at 60 microM. The potency of the cumulus-oocyte complex to respond to forskolin in terms of cAMP accumulation decreases with time. The pattern of the decrease in the potency of the cumulus-oocyte complexes to generate cAMP corresponds with the relief of its inhibitory influence on the oocyte. These results indicate that inhibition of maturation of the cumulus-enclosed oocyte may be coupled to elevation of cAMP levels in the cumulus-oocyte complex. As isolated cumulus-free oocytes are not inhibited by forskolin, we suggest that in the cumulus-enclosed oocyte system, cAMP generated by the cumulus cells is apparently transferred to the oocyte and maintains it in a meiotically arrested state. Maturation in this system occurs upon relief of inhibition which results from cessation of cAMP generation by the cumulus cells.

Animals↗
PubMed1983

Induction of maturation in rat follicle-enclosed oocyte by forskolin.

The diterpene forskolin, which was found to be a potent and reversible activator of adenylate cyclase in intact tissues as well as in broken cell preparations, was employed to investigate the role of cAMP in the induction of oocyte maturation. We have found that forskolin can mimic the effect of LH on the ovarian follicle stimulating both cAMP accumulation and oocyte maturation. These findings suggest that LH-induced maturation in follicle-enclosed oocytes is a cAMP-mediated response.

Animals↗
PubMed1983

Dissociation between the direct stimulatory and inhibitory effects of a gonadotropin-releasing hormone analog on ovarian functions.

The paradoxical effects of gonadotropin-releasing hormone (GnRH) on the ovary have hitherto been believed to result from different regimens of administration; an acute treatment was shown to stimulate the ovary while chronic administration of the hormone inhibited LH-induced responses. In the present report we demonstrate that a single injection of a GnRH analog (D-Ala6)des-Gly10-GnRH-N-ethylamide (GnRHa, 2 micrograms/rat) is sufficient to obtain a significant inhibition (75%) of hCG-induced ovulation in PMSG-primed, either intact or hypophysectomized, immature rats. Inhibition of ovarian development, in terms of growth and ovulation, by multiple injections with GnRHa (2 micrograms/rat, twice daily for 3 days) could be obtained only upon administration of the hormone at early stages of follicular development, i.e. concomitantly with the PMSG injection. When administered after PMSG, GnRHa could not inhibit the ovary but rather induced ovulation by itself in the absence of hCG. A 12-24 h delay in initiation of GnRHa treatment triggered 65% of the rats to ovulate while a delay of 48 h resulted in 100% ovulation. Under both regimes of GnRHa administration, either the inhibitory or the stimulatory, the oocytes of the treated rats were induced to resume meiotic maturation. Since under the inhibitory regime ovulation did not occur, maturation was followed by a massive degeneration of the oocytes trapped within their follicles. These findings demonstrate that the follicular stage of development rather than the dose and/or duration of GnRHa administration determines whether GnRHa inhibits ovarian growth and ovulation, while the competence of the oocytes to respond to the GnRHa stimulus and mature is independent of hormonal priming.

Animals↗
PubMed1983

A comparative study of the mechanism of action of luteinizing hormone and a gonadotropin releasing hormone analog on the ovary.

The mechanism of action of a gonadotropin releasing hormone (GnRH) agonistic analog ([D-Ala6]GnRH) on the rat ovary has been studied in comparison to similar effects of luteinizing hormone (LH). Stimulation of meiosis resumption in vitro in follicle-enclosed oocytes by both LH and [D-Ala6] GnRH, was blocked by elevated levels of cAMP as demonstrated when either dibutyryl cAMP or the phosphodiesterase inhibitor methylisobutylxanthine was present in the culture medium. In vivo, the prostaglandin synthase inhibitor indomethacin, which blocks LH-induced ovulation, also inhibited ovulation induced by the GnRH analog in hypophysectomized rats. On the other hand, the potent GnRH-antagonist [D-pGlu1, pClPhe2, D-Trp3,6] GnRH which blocked the stimulatory effect of the agonist on oocyte maturation and ovulation had no effect on LH action. It is concluded that while a GnRH-like peptide does not seem to mediate LH action on the ovarian follicles, both LH and GnRH agonist share some common mechanistic pathways at a post-receptor locus.

Animals↗
PubMed1982

Effect of gonadotropins and prostaglandin on cumulus mucification in cultures of intact follicles.

Follicles of 28 day-old pregnant mare serum gonadotropin (PMSG)-primed rats, were cultured for up to 24 hours in the presence or absence of ovine gonadotropins, highly purified rat gonadotropins, dibutyryl cyclic AMP (dbcAMP), methylisobutylxanthine (MIX), choleratoxin (CT), or prostaglandin E2 (PGE2). The morphology of the cumulus-oocyte complexes isolated from these follicles was subsequently examined with the light microscope. Cumulus mucification was studied under the different culture conditions using scanning electron microscopy (SEM) and the hyaluronidase sensitivity test. The features of the cumulus-oocyte complexes in the control cultures did not change throughout the incubation period, while complexes from follicles incubated with LH, FSH, dbcAMP, MIX, CT, or PGE2 changed their appearance and accumulated extracellular mucoid material. Treatment of these cumuli with hyaluronidase resulted in lysis of the extracellular mucus and dispersal of the cumulus masses. The results of this study agree with our earlier observation that the maturation of the cumulus-oophorus, which occurs in vivo following the LH surge, can be induced in vitro by either gonadotropins or cAMP. Prostaglandin E2 did not affect cumulus cells, unless incubated enclosed by their follicles. This suggests that this hormone may influence the cumulus cells indirectly, probably via other components.

1-Methyl-3-isobutylxanthine↗
PubMed1980

Binding of human chorionic gonadotropin by rat cumuli oophori and granulosa cells: a comparative study.

Cumulus oocyte complexes isolated from PMS gonadotropin-primed immature rats bind [125I]hCG. Unlabeled hCG and ovine LH compete with the labeled hormone for binding sites, while rat FSH and ovine PRL are without effect. Since specific binding of [125I]hCG could not be detected in preparations of oocytes from which the cumuli had been removed, it appears that the binding exhibited by the complex can be attributed to the cumulus cells. The concentration dependence of binding is consistent with the presence of one population of high affinity (apparent Kd, 1.4 X 10(-10) M) binding sites (223 +/- 33 sites/cumulus cell). Rat granulosa cells bind hCG with equivalent apparent affinity and specificity but contain more sites (2060 +/- 180/cell) than cumulus cells.

Animals↗
PubMed1979

The effect of LH on the fertilizability and developmental capacity of rat oocytes matured in vitro.

The effect of adding LH (10 microgram NIH-LH-B8/ml) to the medium in which oocytes were undergoing maturation in vitro was studied. The fertilizability of the oocytes was evaluated in the sterile oviduct of a unilaterally ovariectomized, mated recipient. Freshly ovulated oocytes, used as a control of the method, were fertilized at a rate of 72%. Only 14% of oocytes matured in culture (without LH) were penetrated by spermatozoa, and 11% were fertilized normally. Addition of LH to the medium increased these proportions to 43 and 33% respectively. Oocytes matured in the presence of LH were able to develop into apparently normal rats. It is concluded that, although oocytes can mature in vitro spontaneously, and that these matured oocytes can be fertilized, addition of LH increases the numbers 3-fold. LH therefore has a direct maturation-promoting action on the rat oocyte-cumulus complex in vitro.

Animals↗
PubMed1978

Rat oocyte maturation in vitro: relief of cyclic AMP inhibition by gonadotropins.

The hormone-independent, spontaneous maturation that rat oocytes undergo in vitro can be inhibited by derivatives of cyclic AMP and inhibitors of cyclic nucleotide phosphodiesterase. In this study, we have shown that this inhibition of maturation can be partially relieved by preparations of ovine and rat luteinizing hormone or follicle-stimulating hormone. The ability of gonadotropins to foster the resumption of maturation in cultures of cyclic AMP-inhibited oocytes suggests that this system is suitable for studies of the hormonal control of oocyte development. The dose and time dependency of the response to gonadotropins has been examined in order to study the role of these hormones in oocyte maturation and to compare this effect to other known responses of the cumulus-oocyte complex. These studies show that highly purified preparations of rat gonadotropins are less effective inducers of maturation than the more commonly used, but considerably less purified, preparations of ovine gonadotropins. Almost complete relief of inhibition is observed, however, when the oocytes are exposed to a combination of rat luteinizing hormone and follicle-stimulating hormone. Oocyte maturation was not influenced by the sex steroids progesterone or 17beta-estradiol. Our results suggest that: (i) cyclic AMP is involved in the intrafollicular inhibition of oocyte maturation; (ii) both gonadotropins are required for maximal stimulation of the resumption of oocyte meiosis; and (iii) steroids are not involved in this response to gonadotropins.

Animals↗