• The Korean Journal of Zoology
• /
• v.19 no.1
• /
• pp.1-6
• /
• 1976

The glycogen content of the oocytes at the various stages of meiotic division induced during culture was determined by a microspectrophotometer. The PAS intensity decreased gradually as the meiotic resumption progressed. The amount of glycongen was also decreased in the degenerated ova. Is is concluded that the glycogen consumption in necessary for the meiotic resumption and that the glycogen loss while the germinal vesicle is intact seems to lead degeneration. These results suggest that the endogenous glycogen is important to support meiosis.

• The Korean Journal of Zoology
• /
• v.31 no.2
• /
• pp.87-94
• /
• 1988

Korean frogs (R. dybowskii and R. nigromaculara) were collected from chonnam area and their oocyte maturation was induced by using in ultro follicle culture system. Follicles were isolated from the frog ovary and cultured for 24 hr in (amphibian Ringer's soluion AR) at 22 C in the presence or absence of hormones. Follicular cocytes of R. dybowskii were induced to mature (germinal vesicle breakdown, GVBD) by the presence of progesterone, 0.1 $\mu$g/2 ml and that of R. nigromaculata by 1 $\mu$g/2 ml of progesterone. Follicles of the frogs were also responded to (frog pituitary homogenate FPH) in terms of their cocyte maturation. Follicular cocytes of R. dybowskii were induced to mature by FPH at concentration of 0.01 pituitary equivalent/2 ml and that of R.nigromaculata at 0.1 pit equiv./2 ml. The culture time required for the maturation of bath frog follicles was 915 hr. The responsiveness of the follicles of korean frogs to hormones (progesterone or FPH) was nearly the same as that of R. pipiens which are most commonly used amphibians. Particularly, follicular cocytes of R. dybowskii used from February matured spontaneously without stimulation of hormones during in vitro culture. Furthermore, those cocytes were spontaneous- ly ovulted when the ovarian fragments were cultured in a flask.

• Korean Journal of Animal Reproduction
• /
• v.25 no.3
• /
• pp.243-250
• /
• 2001

The objectives of the present study were to examine the relationship between catalase (0.1 mg/$m\ell$) and xanthine (5 mM) on in vitro maturation of porcine follicular oocytes. At 48 h after maturation, the proportions of oocytes matured to metaphase-II stage were significantly higher (P＜0.05) in the medium with control (72%), catalase (73%) or catalase plus xanthine (70%) than of oocytes cultured with xanthine (54%). On the other hand, oocytes cultured in medium with catalase and/or xanthine for 30 h were not significantly different in maturation rates (6~l4%). At 36, 42 and 48 h after culture, however, the maturation rates were significantly (P＜0.05) higher in medium with (49~70%) that than without (29~50%) catalase regardless of presence of xanthine. When the oocytes were cultured with periods prolonged in medium with and without xanthine, the maturation rates did increase with high proportions at 72 h of culture. No significant differences, however, were observed in maturation rates between groups with and without catalase. On the other hand, degenerated oocytes were increased with culture periods, the proportions was significantly (P＜0.05) lower in medium with (28%) than without (47%) catalase at 120 h of culture. However, there were no significant difffrences between with and without catalase in medium added xanthine. The parthenogenetic oocytes were observed from 72 h after culture in medium with xanthine, but were no significant differences between with and without catalase. From these results, it is indicated that porcine oocytes nay respond to maturation stimulus by 72 h of culture in medium with catalase and xanthine and that parthenogenesis can be obtained with prolonged culture periods.

• Proceedings of the Korean Society of Developmental Biology Conference
• /
• 2001.08a
• /
• pp.42-44
• /
• 2001

연중 밀어 (Rhinogobiw brumeu)의 생식소 성숙을 조사하였다. 난소 및 정소의 성숙은 체장 60 mm 이상의 개체에서 뚜렷하였다. 난소는 원추형이며 정소는 쐐기모양으로 복강의 등쪽에 위치한다. 연중 성성숙지수 (gonadosomatic index)의 변화는 암컷 0.19 - 14.28 수컷 0.15-13.34로 산란시기는 5월 하순부터 6월 초순의 하계번식 어류로 사료된다. 성숙기에서 산란기에 이르는 동안 난소내 난자의 성숙정도는 불균질하며 전체 밀어 군집내에서 7월 하순까지도 성성숙지수 (GSI)가 비교적 높게 유지되는 것으로 보아 1차 산란 이후에도 소규모의 산란이 하계에 걸쳐 진행되는 것으로 사료된다. 생식소의 발달은 동계에 시작되어 난소내 난자는 춘계와 하계에 걸쳐 성장이 일어나 지름 0.5 mm의 성숙난자를 형성한다. 이 시기의 정소는 성숙한 정자가 세정관내를 가득 채우고 있다. 정자의 길이는 45 um 이며 두부의 직경은 5 um이며 미토콘드리아를 갖는 중편이 두부 기저부에 함입된 형태이며 첨체를 갖지 않는다.

• Journal of Embryo Transfer
• /
• v.12 no.2
• /
• pp.127-132
• /
• 1997

본 연구는 rat에서 PMSG도는 FSH 처리에 의한 과배란 유도가 배란율과 수정란의 질에 미치는 영향을 알아보기 위해 호르몬 처리하고 교미시킨 후 4일령에 난관과 자궁을 세척하여 정상 8-세포기 난자와 비정상 난자를 조사하였고 각 처리에서 채란된 난자 중에 정상난자를 골라 체외 배양하여 발육율을 비교 평가하였다. 미성숙 rat에서는 평균19.1개의 수정란이 채취되었으며 성숙rat에서는 14.2개가 채취되었고 미성숙 rat에서는 성숙 rat에 비해 더 많은 비율의 비정상적인 난자가 회수되었다. FSH와 LH-RH에 의한 방법이 PMSG와 HCG에 의한 방법보다 유의성 있게 많은 난자를 배란시켰으며, 비정상란의 빈도도 낮은 것으로 나타났다. 그러나 호르몬 처리에 의한 두 가지 방법은 자연배란에 의한 방법에 비해 훨씬 높은 비정상난자의 배란을 유도하였다(FSH, 20.1%;PMSG, 41.2%;자연배란 13.4%). 또한 FSH처리에 의해 회수된 난자보다 체외 발육율이 높은 것으로 나타났다. 그러므로 rat에서 PMSG와 FSH를 이용하여 과배란을 유도할 수 있으나 배란된 난자의 비정상율은 자연배란에 비해 훨씬 높았고, 과배란 유도시 호르몬의 종류에 따라 체외 배양율에도 영향을 미치는 것으로 나타났다.

• The Korean Journal of Zoology
• /
• v.28 no.2
• /
• pp.99-107
• /
• 1985

In order to investigate the synergistic effect of dbcAMP and progesterone which are known as the agents to inhibit maturation of mammalian oocytes in vitro, the present studies were done and the results were obtained as follow: 1) if 10 $\\mug$/ml of dbcAMP or 2 $\\mug$/ml of progesterone was given into the medium, each of the agents at the concentration above did not give any inhibitory effect on the maturation of the mouse oocytes in vitro; 2) however, when the two agents at the concentration shown above were given together into the medium, the mouse oocytes were arrested at GV stage; and 3) the oocytes, precultured in the medium containing two agnts at the same concentration as above for four hours, resumed their maturation division upon transfer to the plain medium for the extended culture. Thus, it was found that dbcAMP and progesterone were capable to suppress the maturation of mouse oocytes at the suboptimal concentration when they were given together, and such inhibitory effect was reversible.

• The Korean Journal of Zoology
• /
• v.18 no.4
• /
• pp.181-196
• /
• 1975

Rabbit follicular oocytes were cultured in a basic medium containing 0.4% bovine serum albumin (BSA), carbohydrates and amino acids in various combinations. Osmolarity of the medium was maintained at 308 mOsm. The carbohydrates, pyruvate, lactate and glucose were all about equally beneficial, but not essential for rabbit oocyte maturation. Glutamine and proline, but not methionine or phenylalanine stimulated oocyte develoment. Glutamine stimulated more follicular oocytes to develop to prophase and metaphase II than did any of the three carbohydrates tested alone or in combination. Ammonia production after 24 hours of culture was highest in medium containing glutamine(15.2$\\mu$g/ml) but this was not inhibitory to maturation. Negligible amounts of ammonia were found with the other amino acids added. The optimum level of osmolarity for rabbit oocyte maturation appears to be ranged from 250$\\sim$310 mOsm with the maximum level of 270 mOsm. With 0, 0.08, 0.4, 2, 10 and 50 mM of glutamine in the medium, plus BSA but without carbohydrates, 30, 73, 70, 71, 59, 45% of the oocytes developed to prophase or metaphase II respectively. This indicates that no carbohydrate is required of the maturation of rabbit oocytes when 0.08$\\sim$2 mM of glutamine is included, which are the optimum range. Follicular oocytes could develop in the medium containing $^14 C$-glutamine and BSA but without carbohydrates or other organic compound. From the $^14 CO_2$ produced and TCA precipitable material isolated, it is suggested that glutamine probably is utilized by oocytes and cumulus cells as a source of energy as well as for protein synthesis.

• Journal of Embryo Transfer
• /
• v.26 no.3
• /
• pp.147-152
• /
• 2011

This study was conducted to examine whether efficiency of oocyte production from superovulated prepubertal goats. Fifteen prepubertal and twenty adult goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotrophin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes was activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at 38.5$^{\circ}C$ in an atmosphere of 5% CO$_2$, 5% O$_2$, 90% N$_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I + II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytcs. The cleavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes.

• Journal of Embryo Transfer
• /
• v.18 no.2
• /
• pp.85-90
• /
• 2003

This study examined the viability of canine oocytes following storage at 4 or 38$^{\circ}C$ for 5 h. Cumulus intact oocytes were collected from domestic dog following ovariohysterectomy at local veterinary clinics. In Exp I, the oocytes that collected from ovary transport different temperatures (4 or 38$^{\circ}C$) for 5 h, were cultured for (24 or 48 h). Survivability of oocytes judged by morphological appearance and PI (propidium iodide) staining. The survival rates at 4$^{\circ}C$ ovary transport group showed significantly lower than control group (0%; 0/129 vs. 72.9%; 129/177) 48 h after culture (P<0.05). In Exp II, to assess nuclear maturation of control group oocytes (ovary transported at 38$^{\circ}C$) after in vitro cultured for 24, 48 or 96 h. After 24 h and 48 h of culture, the metaphase I to metaphase II stages (MI to MII) was 8.3% (6/72) and 8.9% (9/101), and which was not increased at 96h (9.5%; 8/84). These results show that canine oocytes remarkably sensitive to low temperature and the percentage of oocytes reaching MI to MII did not increase 96 h after culture.

• Journal of Embryo Transfer
• /
• v.18 no.3
• /
• pp.263-267
• /
• 2003

This study was carried out to test the efficacy of pharmacological inhibitors of the cell cycle transition in keeping bovine oocytes at the germinal vesicle(GV) stage and the reversibility of this inhibition. Bovine oocytes were incubated for 22∼24 hrs in the presence of various inhibitors : cycloheximide (2$\mu\textrm{g}$/$m\ell$), 6-DMAP (2 mM), and roscovitine (50$\mu$M). Bovine oocytes cultured with any of the inhibitors were significantly blocked at the GV stage. Reversibility of pharmacological inhibitors was assessed by culturing oocytes an additional 22∼24 hours in inhibitor-free medium. Examination of oocytes revealed that the inhibitory effect was fully reversible and effect of resuming meiotic progression on nuclear maturation varied according to the various inhibitors. This study suggests that cycloheximide, 6-DMAP and roscovitine can be applied to control meiotic arrest and resumption in maturation culture of bovine oocytes in vitro. More investigations are needed to better understand how the cell cycle of oocyte is blocked without problems to future developmental competence.