• Journal of Aquaculture
• /
• 제9권3호
• /
• pp.205-213
• /
• 1996

Experiments were carried out to investigate the effect of GnRH-analogue (GnRH-a) on the induction of ovulation in catfish, S. asotus. Fully matured female catfish ($250\~600\;g$) received a single intraperitoneal injection of GnRH-a ($50\~200\;{\mu}g/kg{\cdot}body$ weight) showed the successful induction of ovulation. More than $86\%$ of treated females were ovulated after injection of GnRH-a ($90\;{\mu}g/kg$) at $25{\pm}1^{\circ}C$. The majority of spawning took place within 22 to 25 hours after the injection. The gonadosomatic index (GSI) and pseudo-GSI in the group treated with $120\;{\mu}g/kg$ GnRH-a were $23-30\%$ and $18-21\%$, respectively. Average fertilization and hatching rates were $94\%\;and\;81\%$, respectively. Electron microscopically, gonadotrophs of maturing female catfish were characterized by the presence of numerous small, electron-dense granules of approximately $150\~300$ nm in diameter and a few larger, less electron-dense granules of approximately $800\~1000$ nm in size in their cytoplasm. Gonadotrophs of GnRH-a treated catfish showed that their was a distinct decrease in number of small and large granules. The rough endoplasmic reticulum was composed of numerous cisternae conspicuosly dilated to various degrees.

• Journal of Embryo Transfer
• /
• 제26권4호
• /
• pp.229-235
• /
• 2011

Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Ski protein is implicated in proliferation/differentiation in a variety of cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of this study was, by means of immunohistochemical methods, to locate Ski protein in the rat ovaries during ovulation and corpora lutea (CL) formation to predict the possible involvement of Ski in luteinization. In addition, we performed to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vivo models. In order to examine the expression pattern of Ski protein along with the progress of luteinization, follicular growth was induced by administration of equine chorionic gonadtropin to immature female rat, and luteinization was induced by human chorionic gonadtropin treatment to mimic luteinizing hormone (LH) surge. While no Ski-positive granulosa cells were present in preovulatory follicle, Ski protein expression was induced in response to LH surge, and was maintained after the formation of corpus luteum (CL). These results indicate that Ski is profoundly expressed in the luteinized granulosa cells and luteal cells of CL during luteinization, and suggest that Ski may play a role in luteinization of granulosa cells.

• Journal of Aquaculture
• /
• 제21권2호
• /
• pp.96-101
• /
• 2008

To induce of maturation and ovulation, ovary with different development stage of oocytes of sevenband grouper Epinephelus septemfasciatus(n=51, TL $69.1{\pm}1.0$ cm, BW $5.8{\pm}0.3$ kg) rearing indoor-tank in mature and spawning season(June to July) were investigated by cannulation. Female with yolk globule stage oocyte($300{\sim}500{\mu}m$) was injected with human chorionic gonadotropin(HCG, 500 IU/kg BW). Oocytes developed at diameter $300{\sim}700{\mu}m$ in 24 hrs after the HCG injection, and the distribution ratio of over $800{\mu}m$ of oocytes diameter in the cannulated eggs were $91.3{\sim}98.8%(95.1{\pm}3.7%)$ in 48 hrs after the HCG injection. Ovulation was induced from 7 out of 8 female after the HCG injection. The total volume of stripped eggs was 2,480 mL, and the volume of buoyant eggs was 1,360 mL. The fertilization and hatching rates of buoyant eggs were $56.2{\sim}94.9%$ and $70.7{\sim}97.9%$, respectively. These results suggested that HCG 500 IU/kg BW effects on maturation and ovulation of female sevenband grouper with yolk globule stage of oocyte.

• Archives of Pharmacal Research
• /
• 제25권3호
• /
• pp.357-363
• /
• 2002

The effect of a new rhFSH, PG-0801, on oocyte quality, ovulation and in vitro fertilization (IVF) was examined in androgen-sterilized mice. Experimental sterility was induced by a single subcutaneous injection of testosterone propionate (TP, 1 mg/head) into 5 day old female mice. Ovulation was generated in the 10 to 13-week old TP-injected mice by a subcutaneous rhFSH injection (1, 5 or 10 IU/head) followed 48 hours later by a second rhFSH injection (1, 5 or 10 IU/head). For comparison, a subcutaneous PMSG (5 IU/head) injection was used for folliculogenesis and a hCG (5 IU/head) injection was used for ovulation. These were administered using the same protocol. The eggs were harvested from the oviducts and counted 17 to 20 hours after the second injection. IVF was performed by adding sperms ($2{\times}10^{5}/ml{\;}to{\;}2{\times}10^{6}/ml$) to determine the functional activity of the eggs, and the fertilization rate was measured. In addition, the pregnancy rate and fetal development were examined after 15-17 days of gestation. The number of oocytes recovered from the rhFSH/rhFSH group increased dose-dependently and was slightly higher than that of the PMSG/hCG group. The pregnancy rates of the group receiving 1, 5, and 10 IU of rhFSH/rhFSH were 50%, 66.7%, and 75%, respectively, which were significantly higher than that of the control (untreated) group (0%). The numbers of viable fetuses in the 1, 5, and 10 IU/head of the rhFSH/rhFSH group ($8.0{\pm}1.50$, $8.9{\pm}1.02$, and $8.9{\pm}1.12$ fetuses/dam, respectively) were comparable to that of the 5 IU/head PMSG/hCG group ($9.4{\pm}0.94$). The mice receiving rhFSH/rhFSH and PMSG/hCG showed similar fertilization rates (around 65%) via the IVF procedure. These results demonstrate that a new rhFSH, PG-0801, may be useful for inducing ovulation in functionally infertile patients and for superovulation in ovulatory patients participating in assisted reproductive technology (ART) programs.

• Clinical and Experimental Reproductive Medicine
• /
• 제26권3호
• /
• pp.483-490
• /
• 1999

Objectives: Polycystic ovary syndrome (PCOS) has the feature of excessive LH, hyperandrogenism and disturbance of folliculogenesis. Also, insulin, IGF-I and IGFBP-l are involved in the pathogenesis of PCOS. Various surgical and medical therapies have been used and the action mechanisms are related to the endocrine effect. Laparoscopic ovarian electrocautery or laser vaporization is effective in the restoration of ovulation and normal menstrual cycle with minimal invasive procedure especially in the patients resistant to medical therapy. Clomiphen citrate (CC) is used for the ovulation induction in pcas and the resistance is known to be related to insulin, IGF-I, IGFBP-l levels. This study was performed to evaluate the effect of the laparoscopic laser vaporization on the levels of LH, FSH, testosterone, IGF-I and IGFBP-l and on the ovarian response to clomiphen citrate in patients with CC-resistant PCOS. Materials and Methods: The fasting basal serum LH, FSH, testosterone, IGF-I and IGFBP-l level were measured in 10 PCOS patients with CC-resistance and 7 normal controls with regular menstrual cycle. In PCOS, after laparoscopic $CO_2$ laser vaporization, endocrine levels were measured in 1 week interval for 4 weeks and then compared with preoperative levels. Results: In PCOS group, mean serum LH/FSH ratio, testosterone, IGF-I levels were higher and IGFBP-l level was lower than control. LH/FSH ratio decreased from $2.51{\pm}0.67$ to $1.7{\pm}0.6$ (p<0.05) in 2 weeks, to $0.56{\pm}0.2$ (p<0.01) in 3 weeks and to $1.41{\pm}0.3$ (p<0.01) in 4 weeks after operation. Testosterone level decreased from $1.51{\pm}0.82ng/ml$ to $0.65{\pm}0.34ng/ml$ (p<0.05) in 2 weeks, to $0.56{\pm}0.67ng/ml $(p<0.01) in 3 weeks after operation. IGF-I level also decreased from $436{\pm}47.5{\mu}g/l$ to $187{\pm}38{\mu}g/l$ (p<0.0l) in 1 week, to $167{\pm}42{\mu}g/l$ (p<0.01) in 2 weeks, $179{\pm}55{\mu}g/l$ (p<0.01) in 3 weeks and to $120{\pm}43{\mu}g/l$ (p<0.01) in 4 weeks after operation. IGFBP-l level showed no significant change. In 8 of 10 PCOS patients, ovulation was induced with low dose clomiphen citrate. Conclusion: Laparoscopic $CO_2$ laser vaporization restores normal menstrual cycle and ovulation through endocrine effect of decreasing LH/FSH ratio, testosterone and IGF-I level and increases the response to CC. Therefore it is useful for restoration of normal menstruation and induction of ovulation in CC resistant PCOS patients.

• Korean Journal of Ichthyology
• /
• 제28권4호
• /
• pp.260-266
• /
• 2016

We determined the morphologic characteristics (body weight and abdomen inflation degree) of the mother fish of sevenband grouper, Epinephelus septemfasciatus producing the healthy eggs. Experimental fish were chosen from the reared fish in the sea cage for 6 years. The fish were divided into four size groups by body weight: 4.0~5.0, 5.0~6.0, and 6.0~7.0 kg and four stages (I~IV) by the abdomen inflation degree. After hormone treatment, we observed the ovulation amount of induced eggs, rate of buoyant, fertilization, embryonic survival, and hatching. Egg and oil globule diameter was measured. In order to observe gonadal development, we calculated gonadosomatic index (GSI) and conducted its historical analysis. The ovulation occurred from all experimental fishes over 5.0 kg. The rate of buoyant, fertilization, and embryonic survival was the highest in 6.0~7.0 kg. Hatching rate was the highest in 5.0~6.0 kg. Stage I and II did not induce ovulation. GSI was $0.31{\pm}0.10%$ in stage I, $0.74{\pm}0.25%$ in stage II, $4.68{\pm}0.40%$ in stage III and $6.86{\pm}0.12%$ in stage IV. The rate of buoyant, fertilization, embryonic survival and hatching was the highest in stage III.

• Journal of Veterinary Clinics
• /
• 제5권2호
• /
• pp.83-86
• /
• 1988

Morphometric observation on the ovarian follicles of ICR strain-female mice has been conducted to investigate the effect of induced inflammation on the follicular populations. Inflammation was induced by administering 25${\mu}\ell$ of turpentine oil intraperitoneally 2 times at the interval of 3 days. Mice showing 5-day estrous cycle about 20 days after induction of inflammation are placed in experimental group. Normal follicles of 100～399$\mu\textrm{m}$ in diameter increased at estrus but those of over 400$\mu\textrm{m}$ decreased. super-ovulation increased the number of normal follicles of over 400$\mu\textrm{m}$. The number of normal follicles of over 400$\mu\textrm{m}$ 48 hours after superovulation was not significantly different between experimental and control groups. Present results indicated that the number of preovulatory follicles 48 hours after PMSG injection did not decrease in the mice which showed regular estrous cycle even after the induction of inflammation.

• Korean Journal of Animal Reproduction
• /
• 제16권4호
• /
• pp.301-310
• /
• 1993

This experiment was performed to develop simple practical methods for short term preservation of rabbit embryos. A total of 55 cross bred does were superovulated by intramuscular injection of PMSG and HCG. Embryos were recovered at 25~30 hrs, 60~65 hrs and 80~85 hrs after mating and selected by morphological examination. Four cell stage, morulae and blastocyst embryos were stored in PBS enrich with 1, 10, 20 and 40% heat-treated FCS at 4, 20, 30 and 37$^{\circ}C$, respectively. Embryos were examined morphologically at 24, 48 and 72 hrs following storage. The result obtained in this experiment were summarized as follows: The superovulation was induced by PMSG 200 IU and HCG 100 IU. The average number of ovulation points and embryos recovered by collection time were 19.0, 15.6(25~30 hr), 17.3, 13.5(60~65 hr) and 19.2, 14.4(80~85 hr), respectively. And recovery rates of embryos recovered at 25~30 hr, 60~65 hr and 80~85 hr after mating were 62.8%(4 cell), 84.7%(morulae) and 79.6%(blastocyst), respectively. On the other hand, the average number of ovulation points collected by the no, of operations for the repeated collection was 17.3(60~65 hr), 19.2(80~85 hr) in 1st and 9.4(60~65 hr), 10.6(80~85 hr) in 2nd surgery, respectively. There was a significant decrease(P<0.05) in the number of ovulation points the 2nd surgery as compared to the 1st surgery. All of the 4-cell stage embryos stored at 4$^{\circ}C$ for 48 hrs showed the same morphology throught the storage period, on the contrary, 4-cell stage embryos stored at 2$0^{\circ}C$ and 3$0^{\circ}C$ for 48 hrs showed degeneration embryos and stored at 37$^{\circ}C$ for 24 hrs showed degeneration embryos. Morulae and blastodcyst stored at 4, 20, 30 and 37$^{\circ}C$ for 24 hrs showed degeneration embryos. All of the blastocyst stored at 37$^{\circ}C$ for 72 hrs showed degeneration embryos.

• Journal of Embryo Transfer
• /
• 제4권1호
• /
• pp.46-55
• /
• 1989

The effects of an antiprogesterone (RU 486) and an antiestrogen (tamoxifen) on ovulatory response and oocyte morphology were examined in pregnant mare serum gonadotropin (PMSG)-primed immatare female rats (28 days of age): a comparison has been made on two different regirnens primed with a "control" dose (4 IU) and a "superovulatory" dose (40 IU) of PMSG. Females for control control regimen received three consecutive injections of lmg RU486, lmg tamoxifen, or vehicle at 24, 36 and 48hr, and were killed at 72l'r after PMSG. Animals for superovalatory regimen received lmg RU486, 2.5mg tamoxifen, or vehicle fouowlag the injection schedule comparable to control regimen, and were killed at 60 and 72hr after PMSG. Compared to vehicle group, there was a significant reduction in ovulatory response as judged by the proportion of rats ovulating andi or by the mean number of oocytes per rat for each treatment of RU486 and tamoxifen in both regimens. The activity of tamoxifen in inhibiting the ovulatory response was greater in control, but less in superovulatory regimen than that of RU486 based on the dose employed for each antisteroid. In both regimens, RU 486 did not have any effect 6n the changes in the proportion of degenerate oocytes as well as ovarian weight, well tamoxifen treatment resulted in a marked promotion of oocyte degeneration as well as a great reduction in ovarian weight, compared to each parameter of vehicle group. RU486 treatment in each regimen did not alter the serum levels of any steroid hormones observed. Howerver, tamoxifen treatment was associated with significant increases in serum 17$\beta$-estradiol and decreases in progesterone in both regimens; also significant increases in androgens in superovulatory regimen. The results illustrate the relative inhibitory activity of RU486 and tamoxifen indicating major steroid hormone involved in PMSG-induced ovulation: 17$\beta$-estradiol for control and progesterone for superovulatory regimen. It also appears that tamoxifen-associated elevation of circulating 17$\beta$-estradiol andi or androgens could be in part, a contributing factor to the promotion of oocyte degeneration presumably by producing a hostile oviductal environment after ovulation.ent after ovulation.

• Korean Journal of Animal Reproduction
• /
• 제9권1호
• /
• pp.31-35
• /
• 1985

This study was carried out with 46 cows to investigate the ovarian response to the administration of PMSG, PGF2$\alpha$ and hCG, and the developmental stage of embryos recovered. Superovulation was induced by the injection of 2,000IU to 3,000IU PMSG on the days of 7-13 of the estrus cycle followed 48 hours later by the injection of 22.5mg PGF2$\alpha$. Of 29 cows treated with 3,000IU PMSG and 22.5mg PGF2$\alpha$ 18 cows were given 2,000IU hCG at the onset and 7 after artificial insemination. The results obtained were summarized as follows; 1. The number of developed follicles per cow after an injection of 2,000, 2,500, 3,000IU of PMSG and 3,000 PMSG-2,000IU hCG in combination with 22.5mg PGF2$\alpha$ were 12.6, 19.6, 21.5 and 29.3, respectively. This result indicated that the no. of developed follicles per cow was increased according to the increase of PMSG dosage and the combination with hCG injection. 2. The highest number of ovulation was 17.1 in cows treated with PMSG-hCG and the number of matured corpus luteum was increase as the dosage of PMSG was higher. 3. Ovulation rate from cows treated with 2,500IU PMSG was 71.0% and this reulst was higher than the average of ovulation rate (59.3%). 4. Average recovery rate was 36.8%(232/631), and the number of ova per cow was 5.0. 5. Of 232 recovered embryos, the number of morulae and blastocysts were 76 (32.8%) and 83 (35.8%), respectively. 6. 28.4% of total recovered embryos was abonormal morphologically.