• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2002.11a
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• pp.102-102
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• 2002

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.276-286
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• 1999

In the present study, to understand how gonadotropin-releasing hormone (GnRH) affects ovarian functions in superovulated rats, we examined the effects of GnRH agonist on the ovulatory response, the morphological normality and nuclear maturation of ovulated oocytes, the ovarian weight, the ovarian histology, and the circulating steroid hormone ($17{\beta}$-estradiol, progesterone and testosterone) levels in immature rats pretreated with 30IU pregnant mare serum gonadotropin (PMSG) and supplemented with 10IU human chorionic gonadotropin(hCG). GnRH agonist was intravenously injected via jugular vein catheter every 20min for 4hrs in early follicular phase (from 6hr after PMSG) of superovulated rats. In addition, GnRH antagonist, Antide, was intravenously injected in combination with GnRH agonist to verify the effects of GnRH agonist on ovarian functions. All animals were sacrificed at 72hr after PMSG administration. The administration with GnRH agonist in early follicular phase of superovulated rats caused inhibition of ovulatory response, increased the proportion of abnormal appearing oocytes(especially, in the rats of the group treated with 500ng GnRH agonist), decreased ovarian weight and promote follicular atresia, compared to those from the rats of control regimen that were not treated with GnRH agonist. In addition, the treatment with GnRH agonist in the superovulated rat distinctly decreased serum steroid hormone ($17{\beta}$-estradiol, progesterone and testosterone) levels in preovulatory phase. On the other hand, the inhibitory effects of GnRH agonist treatment in superovulation-pretreated rats on ovarian functions were totally reversed by the combination with GnRH antagonist, Antide. The nuclear maturation of oocytes recovered from the oviducts in immature rats treated with GnRH agonist and/or GnRH antagonist was characterized by prematurity and asynchronization in early follicular phase, which was similar to control group. The overall results of this study indicate that GnRH agonist disturbs directly ovarian function in early follicular phase of superovulated immature rats in terms of ovulatory response and morphological normality of ovulated oocytes. This concept has been further evidenced by the findings of a great decrease in ovarian weight, a marked increase in follicular and a distinct decrease circulating steroid hormone ($17{\beta}$-estradiol, progesterone and testosterone) levels in GnRH agonist treatment regimen in early follicular phase.

• Development and Reproduction
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• v.13 no.4
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• pp.341-351
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• 2009

The effects of serotonin (5-hydroxytryptamine, 5-HT), ecdysone (20-hydroxyecdysone, 20-E) and HCG (human chorionic gonadotropin) on the ovarian maturation of the fleshy shrimp, Fenneropenaeus chinensis were investigated. Adult female fleshy shrimps were injected with 5-HT ($5{\mu}g$/g body weight (BW) or $20{\mu}g$/g BW), 20-E ($5{\mu}g$/g BW, $10{\mu}g$/g BW or $20{\mu}g$/g BW) or HCG (5 IU/g BW or 10 IU/g BW) three times at 5-day intervals and were sacrificed 10 days after the last injection. The effects were compared against an intact control group and sham control group which received the injection of sterile saline solution as a vehicle. 20-E and HCG showed no effectiveness on ovarian maturation in fleshy shrimps. However flesh shrimp given 5-HT of $20{\mu}g$/g BW showed significant increase in gonadosomatic index (GSI) in early spring. The results suggest that treatment of 5-HT of $20{\mu}g$/g BW or more is useful approach for the nursery production of the fleshy shrimp, Fenneropenaeus chinensis aquaculture.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.3-6
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• 2003

The birth of the clone animals is influencing the frontier of research of animal biotechnology. It has effects on research of animal biotechnology itself by necessitating setting of new research subjects, modifications of the strategy of ongoing research projects, and challenges to schemes formerly considered impossible. In my talk, such topics including mass production of fertile ova and oocyte maturation will be discussed. (1) Oocytes are needed for the production of a clone by nuclear transplantation. Mitochondrial DNA inherited via the oocyte are involved also in the morphogenesis. Therefore, oocytes from the same animal must be used as recipients to produce genuine clones by nuclear transplantation. Experimenting on the assumption that selective oogenesis can be avoided, and apoptosis of oocytes can be prevented, by using ovarian angiogenic factos will be introduced. (2) It is important to clarify the factors of oocytes involving in reprogramming of somatic cells. Such factors are thought to be expressed in oocytes during oogenesis and oocyte maturation. Therefore, molecular mechanisms of oogenesis and oocyte maturation must be clarified extensively. Topics in this field including our recent advances will be discussed. (중략)

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.496-500
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• 2007

Holstein cow ovaries obtained at a slaughterhouse were used to study the influence of the oocyte collection methods (slicing, puncture, aspiration I and II) on recovery efficiency and subsequent in vitro maturation and embryonic development competence of immature oocytes recovered. In the slicing method, the whole ovarian was chopped into small pieces with a surgical blade. In the puncture method, the whole ovarian surface was punctured by 18-g needle. In other 2 aspiration methods, collected oocytes by aspirating from the visible follicles using an 18-g needle attached to a 5 ml syringe (aspiration I) or using a constant negetive pressure (-80 mmHg) with a vacuum pump (aspiration II). The oocytes were classified into 4 classes on the basis of the morphology of cumulus cells and cytoplasmic appearance of oocyte. Slicing ($9.6{\pm}0.4$) and puncture ($9.7{\pm}0.4$)yielded a larger number of oocytes per ovary than other two aspiration methods (aspiration I and II were $5.8{\pm}0.3$and $5.6{\pm}0.4$, respectively) (p<0.05). The number of the highest quality oocytes (grade A) per ovary was significantly higher in slicing ($4.2{\pm}0.2$) and puncture ($4.6{\pm}0.1$) methods than in other methods (aspiration I and II were $1.2{\pm}0.2$ and $1.4{\pm}0.2$, respectively) (p<0.05). The rate of nuclear maturation of the highest and higher quality oocytes (grade A and grade B, respectively) was not affected by the oocytes collection methods. The oocytes collection methods also did not influence subsequent embryonic developmental competence after in vitro fertilization with M II stage oocytes. It is concluded that slicing and puncture methods of the ovaries can be used as an alternative techniques to aspiration by the syringe or vacuum pump.

• Korean Journal of Animal Reproduction
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• v.13 no.2
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• pp.79-84
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• 1989

This study was carried out to investigate the effect of LH-RH and Gn-RH treatment in Holstein cows with ovarian quiescence and follicular cystic ovaries. The cows with ovarian quiescence and follicular cystic ovaries injected intramuscularly with 100$\mu\textrm{g}$, 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of LH-RH and 200$\mu\textrm{g}$ and 400$\mu\textrm{g}$ of Gn-RH respectively. The cows was diagnosed by repeated rectal palpation. The plasma progesterone and estradiol-17$\beta$ concentrations were assayed by radioimmunoassay methods. The resutls of this study were summarized as follows : 1. Ovulations were induced after treatment of LH-RH and Gn-RH. The concentrations of progesterone reached small peak level at luteal phase and estradiol-17$\beta$ reached obvious peak level with the development and maturation of the follicle during the periods of degeneration of the corpus luteum, and normal ovarian cycle activity started subsequently. 2. The cows with ovarian quiescence and follicular cystic ovaries were induced ovulation at 38.9$\pm$5.3 hrs. after treatment of LH-RH in 66.7% cows and at 52.7$\pm$7.9 hrs after treatment of Gn-RH in 60.0% cows respectively. 3. The good ovarian responses were indicated in treatment with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of LH-RH than those treated with 100$\mu\textrm{g}$ in cows with ovarian quiescence, and did not show difference of ovarian responses between treatments with 200$\mu\textrm{g}$ to 400$\mu\textrm{g}$ of Gn-RH in cows with follicular cystic ovaries.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.2
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• pp.119-125
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• 2016

Objective: The aim of this study was to report a case series of in vitro matured (IVM) oocyte freezing in gynecologic cancer patients undergoing radical surgery under time constraints as an option for fertility preservation (FP). Methods: Case series report. University-based in vitro fertilization center. Six gynecologic cancer patients who were scheduled to undergo radical surgery the next day were referred for FP. The patients had endometrial (n=2), ovarian (n=3), and double primary endometrial and ovarian (n=1) cancer. Ex vivo retrieval of immature oocytes from macroscopically normal ovarian tissue was followed by mature oocyte freezing after IVM or embryo freezing with intracytoplasmic sperm injection. Results: A total of 53 oocytes were retrieved from five patients, with a mean of 10.6 oocytes per patient. After IVM, a total of 36 mature oocytes were obtained, demonstrating a 67.9% maturation rate. With regard to the ovarian cancer patients, seven IVM oocytes were frozen from patient 3, who had stage IC cancer, whereas one IVM oocyte was frozen from patient 4, who had stage IV cancer despite being of a similar age. With regard to the endometrial cancer patients, 15 IVM oocytes from patient 1 were frozen. Five embryos were frozen after the fertilization of IVM oocytes from patient 6. Conclusion: Immature oocytes can be successfully retrieved ex vivo from macroscopically normal ovarian tissue before radical surgery. IVM oocyte freezing provides a possible FP option in patients with advanced-stage endometrial or ovarian cancer without the risk of cancer cell spillage or time delays.

• Development and Reproduction
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• v.25 no.2
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• pp.75-82
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• 2021

We studied steroid metabolites produced from red-spotted grouper ovarian follicles during maturation. Oocytes with 350-500 ㎛ diameter were in vitro incubated in the presence of [³H] 17α-hydroxyprogesterone as a precursor. Steroid metabolites were extracted from incubated media and oocytes. The extracts were separated and identified using thin layer chromatography, high performance liquid chromatography and gas chromatography-mass spectrometry. The identified metabolites were androstenedione (A₄), testosterone (T) and estrone (E₁). The metabolites of A₄ was dominant in all size of oocytes and it was the highest in 480 ㎛ diameter oocytes. The metabolites of two progestins, 17α,20β-dihydroxy-4-pregnen-3-one and 17α,20α-dihydroxy-4-pregnen-3-one were detected in the oocytes less than 480 ㎛ diameter although they were not identified definitely. In the oocytes of 480 ㎛ diameter, metabolite of progestin was the highest, and germinal vesicle (GV) was still in the middle of cytoplasm. In the oocytes of 500 ㎛ diameter, GV was began to migrate and the major metabolites were A₄ and E₁. The metabolite of E₁ was detected in all size of oocytes and it was higher than that of E₂. These results suggest that oocytes of 480 ㎛ diameter are the transitional stage involving steroidogenic shift to final oocyte maturation and potential function of E₁ during maturation process.

• Proceedings of the Zoological Society Korea Conference
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• 1996.10a
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• pp.140.2-140
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• 1996

No Abstract, See Full Text

• Clinical and Experimental Reproductive Medicine
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• v.31 no.3
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• pp.155-168
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• 2004

Objective: Melatonin, which is secreted by pineal gland play an important role in the regulation of ovarian function via seasonal rhythm and sleep in most mammals. It also has a role in the protection of cells by removing toxic oxygen free radicals brought about by metabolism. In the present study, effects of melatonin on the mouse oocyte maturation were examined using two different culture conditions provided with 5% or 21% oxygen concentration. Material and Method: Immature mouse oocytes were obtained from the ovarian follicles of $3{\sim}4$ weeks old ICR strain mice intraperitoneally injected with 5 I.U. PMSG 44 hour before. Under stereomicroscope, morphologically healthy oocytes with distinct germinal vesicle (GV) were liberated from the graafian follicles and collected using mouth-controlled micropipette. They were then cultured for 17 hour at $37^{circ}C$, 5% $CO_2$ and 21% $O_2$ (95% air) or 5% $CO_2$, 5% $O_2$ and 90% $N_2$. New modified Hank's balanced salt solution (New MHBS) was used as a culture medium throughout the experiments. Effects of melatonin were examined at a concentration of $0.0001{\mu}M$, $0.01{\mu}M$ or $1.0{\mu}M$. For the prevention of spontaneous maturation of immature oocytes during culture, dibutyryl cyclic AMP (dbcAMP) and/or hypoxanthine were included in the medium. Results: Under 21% oxygen condition, oocytes cultured in the presence of $0.01{\mu}M$ melatonin showed a significantly higher maturation rates, in terms of germinal vesicle breakdown (95.0% vs 89.0%) and polar body formation (88.1% vs 75.4%), compared to those cultured with $0.0001{\mu}M$ or $1.0{\mu}M$ melatonin. However, no difference was observed in oocytes cultured under 5% oxygen whether they were treated with melatonin or not. In the presence of $0.01{\mu}M$ melatonin, oocytes either cultured under 21% or 5% oxygen exhibited no difference in the polar body formation (85.6% vs 86.7%). However, in the absence of melatonin, oocytes cultured under 21% oxygen exhibited lower polar body formation (74.7%). When oocytes were cultured in the presence of dbcAMP alone or with varying concentrations of melatonin, those treated with both compounds always showed better maturation, i.e., germinal vesicle breakdown and polar body formation, compared to those cultured with dbcAMP alone. At the same concentration of melatonin, however, oocytes exposed to 21% oxygen showed poor maturation than those to 5% oxygen. Similar results were obtained from the experiments using hypoxanthine instead of dbcAMP. Conclusion: Based upon these results, it is suggested that melatonin could enhance the meiotic maturation of mouse oocytes under 21% oxygen concentration, and release oocytes from the meiotic arrest by dbcAMP or hypoxanthine regardless of the concentration of oxygen, probably via the removal of oxygen free radicals.