The present study was carried out to evaluate the effect of superovulation treatments on ovarian responses, oocyte recovery rates and grades of collected oocytes using an ultrasound-guided transvaginal approach in Korean native cows. Superovulation in cows was induced with two different regimenes: 1) FSH-decreasing dose(n=8): the cows were received twice per day for three days of the total dose of 400 mg of FSH-p, 2) FSH-single dose(n=9): the cows were administrated a single dose of 400 mg of FSH-p in 25% PVP. The Observation of visible follicles and collection of oocytes were performed 12 hours following the last FSH in FSH-decreasing dose group and 48 hours after the FSH-single dose injection. All visible follicles larger than 6 mm were punctured and aspirated with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use. The mean number of visible follicles(> 6 mm) was significantly(P<0.05) higher in the FSH-decreasing dose treatment (22.811.9) and FSH-single dose treatment (20.612.0) groups than the non-treatment group(7.0$\pm$8). The mean recovery rate of oocytes was not significantly(P<0.05) different between the treatment and control groups, but the mean number of collected oocytes was significantly(P<0.05) higher in the FSH-decreasing dose treatment( 12.611.5) and FSH-single dose treatment (11.813.6) groups than the non-treatment group(3.7$\pm$0.5). In conclusion, the FSH-single dose treatment at superovulation in cows for ultrasound-guided aspiration might increase the number of aspiratable follicles and the recovery rate of follicular oocytes as the FSH-decreasing dose treatment.
Kim, Hak-Soon;Shin, Chang-Jae;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Chang, Yoon-Seok
Clinical and Experimental Reproductive Medicine
/
v.15
no.1
/
pp.25-34
/
1988
The intrafo1licular echoes of cumulus oophoruses within ovarian follicles were assessed with the use of ultrasound in 86 women taking part in an in vitro fertilization(IVF) or gamete intrafallopian transfer(GIFT) program, stimulated with pure follicle-stimulating hormone(FSH)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin (hCG). When intrafo1licular echoes were clearly separated from the follicular wall or relatively dispersed within the follicle, they were considered to be a dissociated cumulus, and when they were only slightly prominent from the follicular wall, they were suspected to be a nondissociated cumulus. A cumulus was seen in 62.1% of the follicles larger than 10 mm diameter and 75.1% of them were dissociated. The larger the follicles in size, the more the cumuluses in number and dissociation. The number of follicles and intrafollicular echoes per woman was not different whether or not she would be pregnant, but the number of dissociated cumuluses was significantly more in pregnant women. The number of observed dissociated cumuluses correlated significantly with the number of recovered mature oocytes. When an intrafollicular echo is seen, it can be taken as evidence of a sign of maturity of that particular follicle and oocyte. Ultrasonographic monitoring of intrafollicular echoes and follicular size is very helpful to predict follicular maturation in ovulation stimulation cycles.
In the mammalian ovary, follicular development and atresia continuously occur during the reproductive cycles. Follicular atresia occurs through granulosa cell apoptosis. Apoptosis is known as the physiological cell death, which is regulated by bcl-2 gene family. In the bcl-2 gene family, bcl-2 and bcl-xLong are known as inhibitors of apoptosis, whereas bax and bcl-xShort are known as inducer of apoptosis. We thought that bcl-2 protein is associated with follicular development and atresia. But it is not known that the distribution of cells containing bcl-2 protein during follicular development and atresia. Therefore, to examine the distribution of cells with bcl-2 protein during ovarian follicular development and atresia, the immunohistochemistry was used in the rat ovary. Bcl-2 immunoreactivity was localized in the interstitial cells, theca externa cells and granulosa cells around of antrum. All positive signals were observed in the cytoplasm of these cells. Positive signals were strongly observed in the interstitial and theca externa cells of growing antral follicles. While, positive signals were weakly observed in these cells from atretic antral follicles. Positive signals were very weakly observed in the granulosa cells of growing and atretic antral follicles. According to these data, we suggested that bcl-2 proteins which were strongly expressed in the interstitial cells and theca externa cells of growing antral follicles inhibit follicular atresia. And we purposed that bcl-2 proteins regulated follicular development and atresia through the action of bcl-2 gene family.
Kim, Kyeoung-Hwa;Park, Chang-Eun;Yoon, Se-Jin;Lee, Kyung-Ah
Clinical and Experimental Reproductive Medicine
/
v.32
no.3
/
pp.269-277
/
2005
Objectives: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. Methods: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (${\underline{c}}onnective$ tissue growth factor/${\underline{c}}ysteine$-rich 61/${\underline{n}}ephroblastoma$-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. Results: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. Conclusions: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.
To investigate the effects of ovarian cysts on the controlled ovarian hyper-stimulation cycles, 16 patients with 16 paired cycles for IVF-ET were analyzed. These patients had taken both type of cycles, i.e., with cyst(cyst group) and without cyst(control group). Mean diameter of ovarian cysts in cyst group was 18.2mm. There were no significant differences in hormone levels in early follicular phase between two groups. No significant differences were found in total dosage of hMG(IU) administered during the ovarian stimulation $843.8{\pm}123.0$ vs $891.0{\pm}129.8$, serum estradiol level (pg/ml) on the day of hCG administration($1542.8{\pm}1100.6$ vs $1567.5{\pm}1193.0$), the number of aspirated follicles $10.0{\pm}3.4$ vs $11.2{\pm}4.3$ and oocytes $5.3{\pm}3.3$ vs $6.2{\pm}3.1$, the fertilization rate(51.2 % vs 57.2 %) and the cleavage rate(40.5 % vs 52.0 %). Serum estradiol terminal patterns during COH in one group tended to be repeated in the other group. In conclusion, this study suggests that small ovarian cysts do not adversely impact on the controlled ovarian hyperstimulation parameters in IVF - ET program and the presence of small ovarian cyst without concomitant high basal serum estradiol level is not an indication of the cancellation of the controlled ovarian hyperstimulation for IVF-ET.
This study was carried out to investigate the changes in ovary in repeatedly superovulated rabbits. A total of 57 New Zealand White and Californian, 25 mature virgin and 32 immature does were used in this study. For induction of repeated superovulation, PMSG and HCG were injected at 17-day and 30-day intervals for mature does and 17-day intervals for immature ones. The repeatedly superovulated does at 17-day intervals were induced luteolysis of pseudopregnant corpus luteum with PGF2${\alpha}$ on Day 8 to 9 p.c. The effect of repeated superovulation on reproductive organs was investigated on Day 3 p.c. in mature does and on Day 3 and 6 p.c. in immature ones, respectively. 1. In mature virgin does, the number of ovulation points in the 2nd and 3rd superovulation period averaged 7.0 and 5.0 at 17-day intervals and 13.4 and 6.0 at 30-day intervals, respectively. These numbers were statistically similar to 9.5 ovulation points in the control. However, there were less (p<0.05) ovulation points in those periods compared with 22.1 ovulation points in the 1st superovulation period. 2. In immature does, the number of ovulation points in the 2nd and 3rd superovulation period averaged 5.3 and 2.3, respectively. These numbers were significantly (p<0.05) decreased than 17.1 ovulation points in the 1st periods. The number of ovulation points in the 2nd superovulation period was similar to that in the control, but there was a significant (p<0.05) decrease in the number of ovulation points in the 3rd period as compared to the control. 3. In mature virgin does, the number of visible normal and hemorrhagic follicles (>1.0mm diameter) on day 3 p.c. averaged 19.1 and 8.9 in the 1st superovulation period, respectively. In the 2nd 3rd superovulation period, the number of normal follicles averaged 8.3 and 15.5 at 17-day intervals and 17.8 and 14.5 at 30-day intervals. The number of hemorrhagic follicles in the 2nd and 3rd superovulation period averaged 6.3 and 2.0 at 17-day intervals and 5.2 and 7.8 at 30-day intervals, respectively. There was a slight decrease, although not significant, in the number of normal and hemorrhagic follicles in the 2nd and 3rd period at 17-day intervals compared to that in the 1st period. 4. In immature does, the number of visible normal follicles on day 3 and day 6 p.c. in the 1st superovulation period averaged 27:3 and 26.1, respectively. The follicles on day 3 p.c. tended to increase slightly more than that in the cortrol, but the average number of normal follicles on day 6 p.c. did not differ from that in the control. The number of visible hemorrhagic follicles on day 3 and day 6 p.c. in the 1st of follicles in the 1st superovulation period average 10.2 and 9.9, respectively. There was a slight increase in the number of follicles in the 1st period compared to that in the control. In the 2nd and 3rd superovulation period, the number of normal follicles revealed a slight decrease in the 3rd period, but the number of hemorrhagic follicles was not different between periods. 5. The number of growing follicles with incipient intral formation on day 3 p.c. in mature does of the 1st superovulaton period average 29.7 and the average number of growing follicles in the 3rd period was 26.7 at 17-day intervals and 31.0 at 30-day intervals, respectively. These numbers did not differ from that in the control. In immature does, the number of growing follicles averaged 57.7, 45.0 and 59.3 in the 1st, 2nd and 3rd superovulation period, respectively. There was a slight but not significant decrease in the number of growing follicles in the 3rd period compared to that in the control.
Purpose: In this rearch, we made a plan to investigate the effects of Changbuyeukgun-Tang(CET) on the polycystic ovary(PCOS) induced by estradiol valerate(EV) in rats. PCO was induced by single intramuscular injection with EV(4㎎) in female rats. Rats(n=8) in normal group were injected with sesame oil and orally administrated distilled water for 8weeks Rats in PCO control group(n=8) were injected with EV and orally administrated distilled water for 8weeks. Rats in CET treated group(n=8) were injected with EV and orally administrated CET for 8weeks. Methods: We measured the weights of the body, the ovary, the uterus and the adrenal gland. And also, we analyzed ovarian histopathology, NGF and CRF immunohistochemistry. Results: The results were as follows1. the weights(㎎) of the ovaries in CET treated group were significantly increased(p<0.001) compared with PCO control group. 2. the number of the mature follicles in CET treated group were significantly increased(p<0.01) compared with PCO control. 3. the number of the atresia follicles in CET treated group were significantly decreased(p<0.01) compared with PCO control. 4. the number of the corpora in CET treated group were significantly increased(p<0.05) compared with PCO control. 5. the expressions of NGF-immunoreactive cells in the ovarian granulosa cells of rats in CET treated group were less observed than PCO control group. Conclusion: According to these results, we finally concluded that Changbuyeukgun-Tang(CET) has the inhibitory effect on the development of EV-induced polycystic ovary. And we deduced that the effect of it may be related to the decreased NGF activities in the ovary.
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.
Objective: Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. This report examined the expression of Prx isotype I in the rat ovary after hormone treatment. Methods: Immature rats were injected with 10 IU of pregnant mare's serum gonadotropin (PMSG) to induce the growth of multiple preovulatory follicles and 10 IU of human chorionic gonadotropin (hCG) to induce ovulation. Immature rats were also treated with diethylstilbestrol (DES), an estrogen analogue, to induce the growth of multiple immature follicles. Northern blot analysis was performed to detect gene expression. Cell-type specific localization of Prx I mRNA were detected by in situ hybridization analysis. Results: During follicle development, ovarian Prx I gene expression was detected in 3-day-old rats and had increased in 21-day-old rats. The levels of Prx I mRNA slightly declined one to two days following treatment with DES. A gradual increase in Prx I gene expression was observed in ovaries obtained from PMSG-treated immature rats. Furthermore, hCG treatment of PMSG-primed rats resulted in a gradual stimulation of Prx I mRNA levels by 24 hours (2.1-fold increase) following treatment, which remained high until 72 hours following treatment. In situ hybridization analysis revealed the expression of the Prx I gene in the granulosa cells of PMSG-primed ovaries and in the corpora lutea of ovaries stimulated with hCG for 72 hours. Conclusion: These results demonstrate the gonadotropin and granulosa cell-specific stimulation of Prx I gene expression, suggesting its role as a local regulator of follicle development.
Kim, Jong-Dae;Seo, Il-Bock;Lee, Dong-Nyung;Kim, Hyung-Jun
The Journal of Korean Obstetrics and Gynecology
/
v.24
no.1
/
pp.27-41
/
2011
Objectives: This study was designed to investigate the effect of Ojeok-san(OJS) on the estradiol valerate(EV)-induced polycystic ovaries(PCO) in rats. Methods: PCO was induced by single intramuscular injection with estradiol valerate(EV)(4mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for sixty days. PCO control group (n=8) were injected with EV and orally administrated distilled water for sixty days. OJS treated group(n=8) were injected with EV and orally administrated OJS for sixty days. Then, we measured weights of body, ovaries, adrenal glands and uterus of rats. The histomorphometrical changes of ovaries were also evaluated. And we observed the NGF and CRF expression by immunohistochemistry. Results: The results were as follows - The weights(mg) of ovaries in OJS treated group($57.4{\pm}9.4$) were significantly increased(p < 0.05) compared with PCO control group($42.3{\pm}8.5$). - The numbers of mature follicles in OJS treated group($9.3{\pm}2.5$) were significantly increased(p < 0.05) compared with PCO control group($6.1{\pm}2.1$). - The expressions of NGF-immunoreactive cells in the ovarian granulosa cells in OJS treated group were weaker than PCO control group. Conclusion: From the these results, we concluded that Ojeok-san(OJS) has inhibitory effect on the development of EV-induced polycystic ovary. And it's effect maybe related with decreased NGF activities in the ovary.
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