Gonadotropin-releasing hormone: effects on the in vitro perfused rabbit ovary

Gonadotropin-releasing hormone (GnRH) has been shown to inhibit ovulation in gonadotropin-primed hypophysectomized rats and steroid production in cultured rat granulosa cells. To determine if similar effects of GnRH can be observed in another species, the extracorporeal perfused rabbit ovary was utilized. Two groups of rabbit ovaries were exposed to GnRH in a pulsatile fashion at two dose levels (Group I, 2.56 X 10(-8) M; Group II, 2.56 X 10(-7) M). Contralateral ovaries were not perfused with GnRH. Human chorionic gonadotropin (hCG) was added to the perfusate of all ovaries 30 min after the onset of perfusion. Ovulation occurred in all ovaries exposed to hCG in the presence or absence of GnRH. Ovulatory efficiency was similar in both the experimental and control groups. No statistical difference could be determined in the time of ovulation, stage of maturity of oocytes, or percent of degeneration of ovulated or follicular oocytes. Progesterone production was not inhibited in the GnRH-treated ovaries. In contrast to observations in the rat, GnRH does not exhibit a direct inhibitory effect on ovulation or steroid production in the rabbit.     

Effect of histamine and histamine blockers on the ovulatory process in the vitro perfused rabbit ovary

An increase in the content of histamine in the ovary following luteinizing hormone (LH) release and the inhibition of ovulation in the rabbit by antihistamines suggest that histamine may be involved in the ovulatory process. The effects of various doses of histamine and antihistamines on ovulation were investigated using the in vitro perfused rabbit ovary system. Histamine (100 ng/ml) added to the perfusate at hourly intervals induced ovulation, although at a rate below that observed following human chorionic gonadotropin (hCG) administration. Cimetidine (10 micrograms/ml), an H2 blocker, inhibited histamine-induced ovulation, while the H1 blocker, chlorpheniramine (66.7 micrograms/ml), failed to do so. Neither cimetidine nor chlorpheniramine was able to block ovulation following hCG (50 IU). In all experimental groups in which histamine was used to induce ovulation, both extruded ova and follicular oocytes remained in an immature stage and displayed little evidence of degeneration. In contrast, a high percentage of ova exposed to hCG were mature. Ovarian edema was increased in ovaries in which ovulation occurred, regardless of treatment. A linear correlation was noted between ovulatory efficiency and degree of ovarian edema. Histamine may be an intermediary in the mechanism of follicular rupture, but does not support ovum maturation. However, the inability of H1 and H2 antagonists to block hCG-induced ovulation raises questions regarding the role of histamine in the physiologic process of ovulation.     

Effects of high and low preovulatory concentrations of progesterone on ovulation from the isolated perfused rabbit ovary

Rabbit ovaries were isolated surgically before the ovulatory gonadotrophin stimulation and perfused in vitro. Untreated, control ovaries never ovulated. Ovaries treated in vitro with ovine LH ovulated 10-14 h later and the oocytes had undergone germinal vesicle breakdown (GVB). LH induced increases in progesterone secretion from the treated ovaries. A 3 beta-hydroxysteroid dehydrogenase blocker ('Compound A') effectively reduced progesterone secretion into the perfusate and follicular fluid to very low levels but had no effect on ovulation rate or on oocyte maturation. Excessively high progesterone levels were obtained artificially in perfusates by addition of exogenous steroid; the number of ovaries ovulating was markedly reduced but there was no effect on oocyte maturation. It is concluded that the rise in progesterone that normally occurs immediately after the LH surge is not a prerequisite for ovulation in the rabbit. However, progesterone may have a modifying effect on LH-induced follicle rupture when at a pharmacologically high level.     

Influence of calcium and magnesium deprivation on ovulation and ovum maturation in the perfused rabbit ovary

The role of calcium (Ca++) and magnesium (Mg++) in the ovulation process was studied using in vitro perfused rabbit ovaries. Ovaries were perfused with or without human chorionic gonadotropin (hCG) in Ca++/Mg++-free medium (M199) alone or combined with standard M199 to yield varying concentrations of Ca++ and/or Mg++. In all ovaries perfused with hCG, ovulatory efficiency was similar regardless of the concentration of Ca++ and/or Mg++. In ovaries perfused in Ca++/Mg++-free medium without hCG, ovulatory efficiency was similar to that in ovaries perfused with hCG. As Ca++/Mg++ levels were increased without hCG, ovulatory efficiency declined. Ovulation time was significantly accelerated in ovaries perfused in Ca++/Mg++-free medium with or without hCG. Most ovulated ova from ovaries perfused without hCG were immature. With hCG, degree of ovum maturity was directly related to ovulation time. Ovarian smooth muscle contractions were undetectable in 3 ovaries perfused in Ca++/Mg++-free M199 despite occurrence of ovulation. Smooth muscle contractions were recorded in 2 of 3 ovaries perfused in standard M199 with hCG. These results indicate: 1) Ca++/Mg++ exclusion results in rapid follicle rupture and immature ova; 2) oocyte maturation appears to be gonadotropin-dependent; 3) ovulation occurs in the absence of ovarian smooth muscle contractions during perfusion with Ca++/Mg++-free medium.     

Effect of inhibition of oxygen free radical on ovulation and progesterone production by the in-vitro perfused rabbit ovary

The potential role of oxygen free radicals in hCG-induced ovulation was investigated using the free radical scavenging enzymes superoxide dismutase (SOD) and/or catalase with the in-vitro perfused rabbit ovary preparation. SOD (25 micrograms/ml) and SOD + catalase (25 micrograms/ml) significantly reduced the % of large follicles that ovulated during perfusion (P less than 0.005). Neither maturity nor degeneration of ovulated ova and follicular oocytes was affected by SOD and/or catalase. Progesterone concentration in the perfusate was significantly increased in the SOD + catalase treatment group (P less than 0.01). These results indicate a significant role for oxygen free radicals in the process of ovulation.     

Induction of meiotic maturation of follicle-enclosed oocytes of rabbits by a transient increase followed by an abrupt decrease in cyclic AMP concentration.

The involvement of cyclic adenosine monophosphate (cAMP) in mammalian oocyte maturation was assessed using cultures of rabbit cumulus-oocyte complexes and perfused rabbit ovaries. Rabbit cumulus-oocyte complexes were cultured in Brackett's medium with or without forskolin at 10(-4), 10(-5) or 10(-6) mol l-1 for 3-6 h. At 3 or 4 h spontaneous meiotic maturation was significantly (P < 0.05) inhibited by forskolin at 10(-4) mol l-1. With prolonged incubation, spontaneous maturation progressed despite exposure to forskolin. In the second experiment ovaries were perfused for 12 h with forskolin (10(-4), 10(-5) or 10(-6) mol l-1) or medium alone. Neither ovulation nor degeneration of follicular oocytes occurred in any perfused ovary. The percentage of follicular oocytes achieving germinal vesicle breakdown was significantly (P < 0.001) increased in response to forskolin in a dose-related manner. In an additional experiment, ovaries were perfused with forskolin at 10(-4) mol l-1. A significant increase in the cAMP content in the follicle was observed within 30 min, but the ability to produce cAMP in response to forskolin decreased as the duration of perfusion was increased. Intraoocyte cAMP increased significantly within 30 min and reached its maximum 2 h after exposure to forskolin. Thereafter, cAMP levels in the oocytes decreased abruptly. This drop in intraoocyte cAMP concentration was followed by the resumption of meiosis. The alterations of intraoocyte cAMP contents following exposure to hCG in vivo paralleled those observed in the ovaries perfused with forskolin. These data suggest that a transient, but not continuous, increase in cAMP concentration after the gonadotrophin surge may be required to initiate oocyte maturation.     

Lectin binding sites of the mouse ovary, intraovarian and ovulated ova

Fluorescein isothiocyanate (FITC) labeled lectins were used to study the distribution of specific binding sites in histologic sections of mouse ovaries as well as ovulated ova. Four distinct patterns of reactivity of the components of the follicle (exclusive of the ovum) and the surrounding ovarian stroma were recognized: uniform staining of granulosa cells, theca cells and surrounding stroma; weak to moderate staining of the granulosa cells and strong staining of the theca cells and stromal cells; no reactivity of the granulosa cells coupled with strong reactivity of the theca and stromal cells; no reactivity with any component of the cumulus oophorus. Three lectins (from Triticum vulgare, Arachis hypogaea and Maclura pomifera) distinctly accentuated the basal lamina of the cumulus oophorus. The reaction of lectins with oocytes and zona pellucida revealed six distinct patterns: no reactivity with either structure; weak reactivity with the cytoplasm of the oocyte and no reactivity with the zona pellucida; very strong reactivity with the cytoplasm of the oocyte and no reactivity with the zona pellucida; moderate reactivity with both the oocyte and the zona pellucida; moderate reactivity with the oocyte and very strong reactivity with the zona pellucida; no reactivity with the oocyte and moderate reactivity with the zona pellucida. The same pattern of reactivity was seen in the ovulated ova in the oviduct except for the lectin from Arachis hypogaea, the reactivity of which changed upon ovulation and/or fertilization of the ovum. These data provide a list of lectin markers for distinct components of the mouse ovary.     

Studies of ovulation in the perfused ovary of the fowl (Gallus domesticus)

A system was developed for the in-vitro perfusion of the fowl ovary. The ovaries were isolated 16-18 h before expected ovulation of the first follicle of a clutch sequence and perfused at 41 degrees C with Eagle's culture medium containing L-thyroxine and insulin. The efferent perfusion pressure was maintained at 30-40 mmHg. This model was used to investigate the mechanism of ovulation. Addition of LH (1 U) to the perfusate induced ovulation (46%) but LH (1 U) + FSH (1 U) was more effective (88%; P less than 0.05). Progesterone at 100 micrograms alone also induced ovulation (80%). Clomiphene prevented gonadotrophin-induced ovulation. These results suggest that progesterone may act directly on the ovary as a final hormone to induce ovulation in the domestic fowl.     

Leukocyte supplementation increases the luteinizing hormone-induced ovulation rate in the in vitro-perfused rat ovary

Ovaries from eCG-primed (20 IU s.c. on Day 28) rats were perfused from the morning of Day 30 of age in a recirculating system initially containing a buffered blood cell-free medium (M199 + 4% BSA) for periods of up to 21 h. The addition of ovine LH (0.1 micrograms/ml) at 0 h of perfusion resulted in ovulations in all 6 ovaries perfused (3.2 +/- 0.7 ovulations per treated ovary; mean +/- SEM), whereas none of the 6 control ovaries ovulated. Rat leukocytes (50 x 10(6)), added at 7 h of perfusion significantly increased the number of LH-induced ovulations (7.8 +/- 0.5; p less than 0.05). All ovulated oocytes showed resumption of meiosis as judged from the presence of germinal vesicle breakdown. Ovaries perfused with leukocytes but without LH did not ovulate. Histological examination of ovaries 14 h after leukocyte administration showed a considerable number of perifollicular extravasated white blood cells. These findings indicate that leukocytes participate in the normal ovulatory process as part of an inflammation-like reaction.     

The pattern of reproduction in female Columbian black-tailed deer, Odocoileus hemionus columbianus.

Ovarian cycles and the pattern of reproduction in female black-tailed deer in British Columbia were ascertained largely through examination of the ovaries from 444 females. Cyclic development and degeneration of single follicles of ovulatory size occurred several weeks before first ovulation. As the breeding season approached, a second or third large follicle developed in each cycle but in 48% of adult females the follicles were at different stages of maturation. Those failing to rupture at first ovulation luteinized 1 to 2 days thereafter. The first ovulation of the season, in November, never resulted in a lasting pregnancy even though some ova were penetrated by spermatozoa and began to cleave. First ovulation was apparently 'silent' in five of seven females for their ova lacked spermatozoa. Of sixty-one pregnant females, fifty-nine conceived at second ovulation; the other two conceived at subsequent ovulations more widely spaced than the 8- to 9-day interval between first and second ovulations. The synchrony of ovulatory cycles among adult females was such that half of them ovulated for the second time in a span of 7 or 8 days. Primary CL that formed after first ovulation grew to an average maximum volume of only about 45 mm3, whereas those originating at second ovulation grew to twice that size within 5 to 8 days. First generation CL shrank from 35 mm3 to 10 mm3 within 2 days. They disappeared within 18 months but corpora albicantia persisted for the life of the female. The possible ecological significance of the reproductive pattern is discussed.     

Lectin binding sites of the mouse ovary,intraovarian and ovulated ova

Summary Fluorescein isothiocyanate (FITC) labeled lectins were used to study the distribution of specific binding sites in histologic sections of mouse ovaries as well as ovulated ova. Four distinct patterns of reactivity of the components of the follicle (exclusive of the ovum) and the surrounding ovarian stroma were recognized: 1. uniform staining of granulosa cells, theca cells and surrounding stroma; 2. weak to moderate staining of the granulosa cells and strong staining of the theca cells and stromal cells; 3. no reactivity of the granulosa cells coupled with strong reactivity of the theca and stromal cells; 4. no reactivity with any component of thecumulus oophorus. Three lectins (from Triticum vulgare, Arachis hypogaea and Maclura pomifera) distinctly accentuated the basal lamina of thecumulus oophorus. The reaction of lectins with oocytes and zona pellucida revealed six distinct patterns: 1. no reactivity with either structure; 2. weak reactivity with the cytoplasm of the oocyte and no reactivity with the zona pellucida; 3. very strong reactivity with the cytoplasm of the oocyte and no reactivity with the zona pellucida; 4. moderate reactivity with both the oocyte and the zona pellucida; 5. moderate reactivity with the oocyte and very strong reactivity with the zona pellucida; 6. no reactivity with the oocyte and moderate reactivity with the zona pellucida. The same pattern of reactivity was seen in the ovulated ova in the oviduct except for the lectin from Arachis hypogaea, the reactivity of which changed upon ovulation and/or fertilization of the ovum. These data provide a list of lectin markers for distinct components of the mouse ovary.     

Effects of prolactin on ovarian plasmin generation in the process of ovulation.

The present study was undertaken to assess the effects of prolactin (PRL) on gonadotropin-induced plasmin generation in the in vitro-perfused rabbit ovary. The ovarian plasmin activity was determined by measuring plasmin bound to its major inhibitor, alpha 2-plasmin inhibitor (alpha 2 PI-Plm). In the first experiment, exposure to hCG enhanced ovarian alpha 2 PI-Plm generation from 1.2 +/- 0.3 ng/min/ovary in unstimulated ovaries to 2.9 +/- 0.3 ng within 2 h. The concentration of alpha 2 PI-Plm reached a maximum at 4 h and then declined. A second peak occurred 8 h after hCG administration; however, the ovarian alpha 2 PI-Plm generation without hCG was very low throughout the entire perfusion period. In the subsequent experiment, the addition of PRL(10-10(3) ng/ml) to the perfusate inhibited hCG-induced ovulation in a dose-dependent manner. Exposure to PRL at 10(3) ng/ml significantly (p less than 0.05) inhibited hCG-induced alpha 2 PI-Plm generation in ovaries throughout the entire perfusion period. Furthermore, PRL inhibited hCG-stimulated alpha 2 PI-Plm generation at 4 h after hCG administration in the perfused rabbit ovaries in a dose-dependent manner. In conclusion, PRL directly inhibits hCG-induced ovulation in rabbit ovary, at least in part, by a mechanism depending upon inhibition of the plasmin-generating system in the preovulatory follicles.     

The involvement of oxytocin in ovulation and in the outputs of cyclo-oxygenase and 5-lipoxygenase products from isolated rat ovaries

The effects on ovulation of a specific anti-oxytocin rabbit serum (anti-OT) (50.0 microliters) given by intrabursal injection into the right ovaries of etherized adult female rats at proestrus, were explored by counting the number of ovulated ova present within the right oviducts. Left ovaries were not treated and served as control ovaries. Control rats were treated with male normal rabbit serum (NRS) (50.0 microliters) given by intrabursal injections into the right ovaries of animals at proestrus. Ovulation was induced by injection of human chorionic gonadotrophin (hCG). Anti-OT administered into the right ovarian bursae of proestrous rat ovaries evoked a significant 51% inhibition of ovulation in comparison with that observed in control non-injected left ovaries (p less than 0.01). Also, when the ovulation of right ovaries injected with anti-OT was compared with that of left ovaries injected with NRS, the number of ovulated ova in the right side was significantly smaller (30%) than on the contralateral side (p less than 0.02). However, in rats pre-treated with hCG the intrabursal injection of oxytocin (OT) (50.0 mU/ml) into right and left ovaries failed to alter the number of ovulated ova compared with that of rats receiving intrabursal injections of saline. The basal control and the OT-evoked synthesis and release of endogenous prostaglandin E2 (PGE2) and PGF2 alpha were explored in ovaries isolated from prepuberal rats injected with pregnant mare's serum gonadotrophin (PMSG), two days prior to sacrifice. OT augmented the basal release of PGF2 alpha but did not influence that of PGE2. Moreover, the conversion of exogenous 14C-arachidonic acid (14C-AA) into different prostanoids and into 5-HETE, in the presence and in the absence of added OT (50.0 mU/ml), was studied in rat ovaries isolated in proestrus. The challenge with OT augmented the basal synthesis and release of PGF2 alpha and of 5-HETE from 14C-AA, but failed to influence the formation of products generated via the cyclo-oxygenase pathway, namely 6-keto-PGF1 alpha, PGE2 and thromboxane B2 (TXB2). Therefore, the present results suggest that ovarian OT may play a role in the ovulatory process, via generation of PGF2 alpha to enhance contractions of ovarian smooth muscle and of 5-HETE to promote follicular collagenolysis.     

A prostacyclin analogue, iloprost, augments the ovulatory response of the LH-stimulated in vitro perfused rat ovary.

Ovaries from immature rats, primed with pregnant mare's serum gonadotropin (PMSG; 20 IU, on day 28), were perfused in vitro in a recirculating system for 21 h from the morning of day 30 of age. Stimulation with luteinizing hormone (LH; 0.1 micrograms/ml) in vitro at 0 h of perfusion resulted in 2.4 +/- 0.75 (mean +/- SEM) ovulations per treated ovary, whereas no ovulations occurred in the unstimulated group. When the addition of LH was supplemented hourly for 10 h with a stable prostacyclin analogue, Iloprost, at concentrations of 0.01 microM or 0.1 microM, the ovulation rate increased significantly (p less than 0.05) to 6.6 +/- 1.3 and 10.2 +/- 2.4 ovulations per treated ovary, respectively. Iloprost (0.1 microM) did not cause any follicular ruptures when added by itself at every hour up to 10 h. The addition of Iloprost did not affect the release of cyclic adenosine 3',5'-monophosphate (cAMP), progesterone or estradiol from unstimulated or LH-stimulated ovaries. All ovulated oocytes had resumed meiosis as judged from the absence of a germinal vesicle. These data indicate a positive modulatory role of prostacyclin in the LH-induced ovulatory process for the rat.     

Prostaglandin F2a, an ovulatory intermediate in the in vitro perfused rabbit ovary model

PGF_2a has been proposed as a mediator of mammalian ovulation. To elucidate further the role of PGF_2a in the process of ovulation, PGF and PGF_2a metabolite were measured by radioimmunoassay in the perfusate of an perfused rabbit ovary preparation.Perfusion medium samples were collected over a 10 to 12 hour period from ovaries perfused with tissue culture M199 (total volume 150 ml, sample volume 3 ml) to which varying amounts of hCG had been added. [The PGF_2a antisera a 40% cross reaction with PGF_1a, hence total PGF was measured with this antisera.] Both PGF and PGF_2a metabolite showed a linear increase with time and numbers of ovulations.PGF media accumulation was 575 pg/ovary/ovulation/hr and PGF_2a metabolite accumulation was 367 pg/ovary/ ovulation/hr. Medium prostaglandin content could be correlated with numbers of ovulations, ovulatory efficiency (number of ovulations/total follicles) but total follicles. These data best fit a model of independent ovulatory units producing PFG_2a without recruitment or interaction between them. We infer the PGF and PGF_2a metabolites in this system can be used as a direct index of the ovulation process.     

Hysterectomy in the rat: surgical disruption of the vascular channels reduces number of ova shed.

Cycling rats were hysterectomized and/or unilaterally ovariectomized (ULO) on day 2 (metestrus). Collateral blood supply to the remaining ovary via the uterine artery was left intact or disrupted. Animals were killed in metestrus after one complete estrous cycle. Control rats were also killed at this time. Counts of tubal ova revealed that intact rats ovulated an average of 4.4 +/- 0.4 eggs per ovary (N = 8). Following ULO, rats (N = 8) ovulated 9.6 +/- 0.2 EGGS. Ligation of the uterine artery decreased the number of eggs ovulated in ULO rats (N = 8) to 5.4 +/- 1.1. Hysterectomized rats (N = 8) ovulated 4.8 +/- 0.5 eggs per ovary. If the blood supply was disrupted, a reduction to 2.7 +/- 0.2 eggs per ovary occurred (N = 8). Hysterectomized and ULO rats (N = 8) ovulated 10.3 +/- 0.4 eggs from the remaining ovary but only 5.0 +/- 1.0 eggs if the collateral blood supply of the uterine artery was not intact (N = 10). The results demonstrate that disruption of the vascular channels during the surgical procedures of hysterectomy and/or ULO results in a reduction of the expected ovulation number.     

Changes in the cytologic and electrophysiologic parameters of rat oocytes in response to prostaglandins F2 alpha and E2

Influence of prostaglandins F-2 alpha and E-2 on ovulation character and on the state of the ovulated gametes has been studied in rats at the preovulatory period (the preparations have been injected for 3 days, beginning from the 1st day of early proestrus). Under the effect of prostaglandins the amount of postovulated ova decreases, especially in the animals given prostaglandin E-2. Inhibition of ovulation in these animals is accompanied with an increased part of hyperpolarized ova, appearance of some signs of degeneration in the group of depolarized oocytes and absence of oocytes with polarized membrane at the metaphase I stage. These facts can be interpreted as desynchronization of maturation processes and ova ovulation. Mechanisms of the changes revealed in the ovulatory process and the state of gametes under the prostaglandins effect are discussed.     

Estimation and manipulation of glutathione levels in prepuberal mouse ovaries and ova:Relevance to sperm nucleus transformation in the fertilized egg

Glutathione (GSH), the major low-molecular-weight thiol in mammalian cells, is believed to be a necessary factor for the transformation of the disulfide-stabilized sperm nucleus into the male pronucleus after fertilization. Its concentration in mouse ova, isolated from the ampulla of the oviduct after hormone-induced superovulation of 3–4-week-old mice, has been determined by an enzymic cycling microassay. The level found was 1.80 pmol per ovum. Mean ovum diameter was estimated as 71–72 μm, indicating a GSH concentration of 9–10 mM in the mouse egg. Administration of L-buthionine S, R-sulfoximine, an inhibitor of GSH biosynthesis, during the 2 days preceding ovulation, reduced ovum GSH content below 0.20 pmol (<1.0 mM). The mean GSH concentration of the hormone-stimulated ovaries was reduced from 3.2 mM to 0.2 mM under these conditions. It has also been demonstrated that measurement and manipulation of ovum and ovarian levels of GSH can aid in studying its function in ovaries, ova, and early embryos. Hormone-induced superovulation was achieved in BSO-treated prepuberal C57B1/6 X SJL mice whose ovaries contained less than 10% of control levels of GSH. Over 50% of the isolated ova were fertilized in vitro. However, abnormal one-cell embryos resulted in which the maternally derived pronucleus coexisted with an unchanged sperm nucleus, thus confirming that adequate levels of GSH are necessary for initiating transformation of the fertilizing sperm nucleus.     

Effects of dibutyryl cyclic AMP on oocyte maturation and ovulation in the perfused rabbit ovary

The involvement of cyclic AMP (cAMP) in mammalian oocyte maturation was assessed using cultures of rabbit cumulus-oocyte complexes and in-vitro perfused rabbit ovaries. Rabbit cumulus-oocyte complexes were cultured in Brackett's medium with or without dibutyryl cyclic AMP [Bu)2cAMP) at 10(-3), 10(-4) or 10(-5) M for 4-12 h. At 4 h spontaneous meiotic maturation was significantly inhibited by (Bu)2cAMP (P less than 0.025). With prolonged incubation, spontaneous maturation progressed despite exposure to (Bu)2cAMP. When ovaries were continuously perfused in vitro for 12 h with (Bu)2cAMP (10(-3) M) or medium alone, (Bu)2cAMP stimulated ovarian progesterone production, but did not affect ovulation or maturation of follicular oocytes. When ovaries were perfused in vitro with or without (Bu)2cAMP (10(-3), 10(-4) or 10(-5) M) for the first 2 h and then transferred to medium without (Bu)2cAMP for an additional 10 h, ovulation did not occur, but transient exposure to (Bu)2cAMP stimulated a dose-related increase in maturation of follicular oocytes. Degeneration of follicle-enclosed oocytes and cumulus-oocyte complexes was not affected by exposure to (Bu)2cAMP. These results suggest that transient, but not continuous, elevation of cAMP after the gonadotrophin surge may be required for the initiation of oocyte maturation.     

Cytogenetic analysis of mouse oocytes after experimental induction of follicular overripening

Graafian follicle overripening was induced in (1) adult mice by inhibiting the ovulatory discharge of gonadotrophins with antibodies to LH-RH and (2) immature mice by injection of PMSG to promote follicular maturation before the neuroendocrine system was competent to produce an ovulatory stimulus. The numbers of follicles capable of meiotic maturation after exogenous LH were sharply reduced during the period of overripening and there was a corresponding increase in the proportion of cystically enlarged follicles, many of which were undergoing atresia. Freshly ovulated ova were collected after delaying ovulation for 2 days and prepared for cytogenetic study of metaphase chromosomes. The incidence of non-disjunction and other errors was indistinguishable from that of ova collected after spontaneous ovulation during 4- or 5-day cycles.