• 농업과학연구
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• 제35권2호
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• pp.167-176
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• 2008

Supplementation of serum and estrogen in in vitro maturation(IVM) medium was shown to improve embryo development and quality in several species. This study investigates the effect of ovarian estrus stage on canine oocyte quality and supplementation of medium with canine serum or estrogen on IVM of canine oocytes. As results, in experimental 1, IVM oocytes collected from follicular stage ovaries to MII stages($10.2{\pm}1.5%$) was higher (p<0.05) with 10% canine estrus stage serum than control($1.3{\pm}1.6%$), anoestrus stage serum($4.0{\pm}1.6%$), luteal stage serum($2.7{\pm}1.7%$) and 10% FBS($1.3{\pm}1.6$). In experimental 2, 10% canine estrus stage serum supplementation has highest maturation rate to MII stages($10.0{\pm}1.8%$) and there were significant differences(P<0.05) with another treatment in follicular stages group. In order to investigate the synergic effect of estrous serum and estrogen supplementation, different estrous stage groups of oocytes were cultured with 2 ug/ml estrogen plus various concentrations of different reproductive stage serum and FBS(experimental 3). As results, the rate of maturation to metaphase II(MII) stage was significantly higher(p<0.05) in oocytes from the follicular stage supplemented with estrogen and 10% canine estrus stage serum(11.5%) compared to the other groups(6.0 - 8.8%). The present study was demonstrated that canine serum and the estrus cycle of the bitch affect the meiotic competence of oocytes. Hormonal influences within the follicle may be one of the factors responsible for the greater proportion of maturation of oocyte to MII from bitches at the follicular phase.

• 한국수정란이식학회지
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• 제18권3호
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• pp.187-193
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• 2003

본 연구는 한우에 GnRH ＋ PGF$_2$$\alpha$＋GnRH (Ov-synch)를 처리하여 배란동기화를 시켰으며, 2차 GnRH 투여후 배란시간, 수태율, 계절별 수태율, 산차별 수태율을 조사하였으며, 시험축은 2산 이상의 개체를 무작위로 선발하여 실험에 공시하였으며, 배란동기화 처리후 1회 인공수정을 실시하고 수태율을 환산하였다. 호르몬 처리방법으로는 GnRH ＋ PGF$_2$$\alpha$ ＋ Gn-RH(Ov-synch)법을 이용하였으며, 배란시간의 조사는 초음파를 이용하여 2차 GnRH 투여 후 24시간부터 31시간까지 2시간 간격으로 난소를 촬영하여 배란 여부를 조사하였다. 1. 배란동기화 처리후 24시간부터 31시간까지 2시간 간격으로 난소의 상태를 확인한 결과 28∼30시간 사이에 80%(20/25)로 가장 많이 배란된 것으로 나타났다. 2. 수태율에 있어서는 계통별 1회 수정 수태율은 고급육 계통이 48.1%(38/79)로 다소 높게 나타났다. 3. 산차에 따른 수태율은 1∼2, 3∼4산차에서 각각 44.3, 55%로 나타났다. 4. 계절별로는 여름보다는 봄과 가을에서 47.3%로 다소 높은 경향이였다.

• 한국발생생물학회지:발생과생식
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• 제24권3호
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• pp.159-165
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• 2020

Previously, we reported adverse effects of low-dose nonylphenol (NP) exposure on the reproductive parameters of F1 female mice. In the present study we further investigated the pathohistological effect of NP exposure on the reproductive organs in F1 female mice. NP exposures were continuously conducted from parental pre-mating period until the postnatal day (PND) 33 of F1 offspring for vaginal examination. Mice were sacrificed on PND 30 and the reproductive tissue weights were measured. The initial (at PND 21) body weights of the NP-50 group animals were significantly lower than those of control group animals, and the weight deficit were recovered when the terminal (PND 33) body weights were measured. Early vaginal opening was found in NP group animals (p<0.05). Pathohistological studies revealed that NP-treated F1 animals showed prominent increase in the ovarian follicle numbers (p<0.01), and decrease in the diameter of uterine myometrium (p<0.01), and increase in the diameter of luminal epithelium (p<0.05). The present study demonstrated that the subchronic low-dose NP exposure induced early beginning of puberty and pathohistological abnormalities in ovary and uterus of F1 mice. Further studies are needed to achieve a better understanding on the action mechanism of NP in pubertal onset and to find a way to avoid a hazardous situation provoked by NP exposure.

• 한국패류학회지
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• 제26권3호
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• pp.245-254
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• 2010

For the studies of germ cell development and maturation in the ovary, the gametogenic cycle and the number of spawning seasons per year in female Ruditapes philippinarum were investigated by quantitative statistical analysis using an Image Analyzer System. Compared with the results by qualitative and quantitative analyses, monthly variations in female gonad indice by qualitative histological analysis showed a pattern similar to that of the female gametogenic cycle calculated by quantitative statistical analysis. The number of spawning seasons occurred once per year, from June to October. In quantitative statistical analysis using an image analyzer system, monthly changes in the portions (%) of the ovary area to total tissue areas in females increased in March and reached a maximum in May, and then showed a rapid decrease from June to October when spawning occurred. And also monthly changes in portions (%) of follicle areas to the ovary area and in portions of oocyte areas to ovarian tissue areas in females began to increase in March and reached a maximum in May, and then. rapidly dropped from June to October when spawnig occurred. From these data, it is apparent that the number of spawning seasons occurred once per year, from June to October. Monthly changes in the number of the oocyte per $mm^2$ and in mean diameter of the oocyte in captured image which were calculated for each female slide showed a maximum in May and reached the minimum from December to February. Therefore, female R. philippinarum showed a unimodal gametogenic cycle during the year.

• Clinical and Experimental Reproductive Medicine
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• 제48권3호
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• pp.245-254
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• 2021

Objective: In humans, polycystic ovary syndrome (PCOS) is an androgen-dependent ovarian disorder. Aberrant gene expression in folliculogenesis can arrest the transition of preantral to antral follicles, leading to PCOS. We explored the possible role of altered gene expression in preantral follicles of estradiol valerate (EV) induced polycystic ovaries (PCO) in a mouse model. Methods: Twenty female balb/c mice (8 weeks, 20.0±1.5 g) were grouped into control and PCO groups. PCO was induced by intramuscular EV injection. After 8 weeks, the animals were killed by cervical dislocation. Blood serum (for hormonal assessments using the enzyme-linked immunosorbent assay technique) was aspirated, and ovaries (the right ovary for histological examinations and the left for quantitative real-time polymerase) were dissected. Results: Compared to the control group, the PCO group showed significantly lower values for the mean body weight, number of preantral and antral follicles, serum levels of estradiol, luteinizing hormone, testosterone, and follicle-stimulating hormone, and gene expression of TGFB1, GDF9 and BMPR2 (p<0.05). Serum progesterone levels were significantly higher in the PCO animals than in the control group (p<0.05). No significant between-group differences (p>0.05) were found in BMP6 or BMP15 expression. Conclusion: In animals with EV-induced PCO, the preantral follicles did not develop into antral follicles. In this mouse model, the gene expression of TGFB1, GDF9, and BMPR2 was lower in preantral follicles, which is probably related to the pathologic conditions of PCO. Hypoandrogenism was also detected in this EV-induced murine PCO model.

• Clinical and Experimental Reproductive Medicine
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• 제46권2호
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• pp.76-86
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• 2019

Objective: This study was performed to explore the possibility that each oocyte and its surrounding cumulus cells might have different genetic expression patterns that could affect human reproduction. Methods: Differential gene expression analysis was performed for 10 clusters of cumulus cells obtained from 10 cumulus-oocyte complexes from 10 patients. Same procedures related to oocyte maturation, microinjection, and microarray analyses were performed for each group of cumulus cells. Two differential gene expression analyses were performed: one for the outcome of clinical pregnancy and one for the outcome of live birth. Results: Significant genes resulting from these analyses were selected and the top 20 affected pathways in each group were analyzed. Circadian entrainment is determined to be the most affected pathway for clinical pregnancy, and proteoglycans in cancer pathway is the most affected pathway for live birth. Circadian entrainment is also amongst the 12 pathways that are found to be in top 20 affected pathways for both outcomes, and has both lowest p-value and highest number of times found count. Conclusion: Although further confirmatory studies are necessary, findings of this study suggest that these pathways, especially circadian entrainment in cumulus cells, may be essential for embryo development and pregnancy.

• Clinical and Experimental Reproductive Medicine
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• 제46권2호
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• pp.87-94
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• 2019

Objective: The primary objective of this study was to compare clinical pregnancy rates in intrauterine insemination (IUI) treatment cycles with transabdominal ultrasound guidance during intrauterine catheter insemination (US-IUI) versus the "blind method" IUI without ultrasound guidance (BM-IUI). The secondary objective was to compare whether US-IUI had better patient tolerability and whether US-IUI made the insemination procedure easier for the clinician to perform compared to BM-IUI. Methods: This was a randomized controlled trial done at the Reproductive Medicine Unit of General Hospital Kuala Lumpur, Malaysia. We included women aged between 25 and 40 years who underwent an IUI treatment cycle with follicle-stimulating hormone injections for controlled ovarian stimulation. Results: A total of 130 patients were recruited for our study. The US-IUI group had 70 patients and the BM-IUI group had 60 patients. The clinical pregnancy rate was 10% in both groups (p> 0.995) and there were no significant difference between the groups for patient tolerability assessed by scores on a pain visual analog scale (p= 0.175) or level of difficulty for the clinician (p> 0.995). The multivariate analysis further showed no significant increase in the clinical pregnancy rate (adjusted odds ratio, 1.07; 95% confidence interval, 0.85-1.34; p= 0.558) in the US-IUI group compared to the BM-IUI group even after adjusting for potential covariates. Conclusion: The conventional blind method for intrauterine catheter insemination is recommended for patients undergoing IUI treatment. The use of ultrasound during the insemination procedure increased the need for trained personnel to perform ultrasonography and increased the cost, but added no extra benefits for patients or clinicians.

• Laboraroty Animal Research
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• 제34권4호
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• pp.211-215
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• 2018

The following experiments were designed to examine the effect of serum of spayed dogs on superovulation response in mice and rats. In Experiment 1, female mice at diestrus (n=30) were divided into three equal groups and superovulated with either administration of 5 IU pregnant mare serum gonadotropin (PMSG) or recombinant follicle stimulating hormone (rFSH) (reducing dose from 2.5 to 0.5 IU) and 5 IU human chorionic gonadotropin (hCG) administered 48h later. Serum of spayed dogs was administered intraperitoneally at a reduced dose from 0.1 to 0.025 mL in a 48 h period. In Experiment 2, female rats (n=30) at diestrus stage were divided into three equal groups. Superovulation was induced using either 30 IU PMSG, or a dose reduced from 5 to 1 IU rFSH and 25 IU hCG administered 48h later. Serum of spayed dogs was administered in a reduced dose from 0.6 to 0.1 mL in a 48 hour period. Female mice and rats were mated 24 h following hCG administration. On day 14 after mating, animals were euthanized and ovarian sections were fixed for histopathological evaluation and corpus luteum (CL) counting. No significant difference observed in mean (${\pm}SEM$) number of CLs between the PMSG group and the mice that received serum of spayed dog ($10.4{\pm}1.3$ vs $9.2{\pm}1.0$). Mean (${\pm}SEM$) number of CLs tended to be lower in rats that received serum of spayed dog than those of rats which received either PMSG or rFSH ($15.1{\pm}1.9$ vs $23.6{\pm}3.1$ and $23.1{\pm}2.9$, P=0.06, respectively). In conclusion, serum of spayed dogs is able to induce a superovulatory response in mice and rats.

• 한국동물생명공학회지
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• 제37권1호
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• pp.34-41
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• 2022

In vivo oocytes grow and mature in ovarian follicles whereas oocytes are matured in vitro in plastic culture dishes with a hard surface. In vivo oocytes show a superior developmental ability to in vitro counterparts, indicating suboptimal environments of in vitro culture. This study aimed to evaluate the influence of an agarose matrix as a culture substrate during in vitro maturation (IVM) on the development of pig oocytes derived from small antral follicles (SAFs). Cumulus-oocyte complexes (COCs) retrieved from SAFs were grown in a plastic culture dish without an agarose matrix and then cultured for maturation in a plastic dish coated without (control) or with a 1% or 2% (w/v) agarose hydrogel. Then, the effect of the soft agarose matrix on oocyte maturation and embryonic development was assessed by analyzing intra-oocyte contents of glutathione (GSH) and reactive oxygen species (ROS), expression of VEGFA, HIF1A, and PFKP genes, and blastocyst formation after parthenogenesis. IVM of pig COCs on a 1% (w/v) agarose matrix showed a significantly higher blastocyst formation, intra-oocyte GSH contents, and transcript abundance of VEGFA. Moreover, a significantly lower intra-oocyte ROS content was detected in oocytes matured on the 1% and 2% (w/v) agarose matrices than in control. Our results demonstrated that IVM of SAFs-derived pig oocytes on a soft agarose matrix enhanced developmental ability by improving the cytoplasmic maturation of oocytes through redox balancing and regulation of gene expression.

• Journal of Animal Science and Technology
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• 제64권3호
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• pp.432-442
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• 2022

The purpose of this study was to analyse the effects of light colour on rabbit reproductive performance and the expression of key follicular development genes. Rabbits (n = 1,068, 5 months old, 3.6-4.4 kg live body weight) were divided randomly into four groups, housed individually in wire mesh cages and exposed to red, green, blue, and white light-emitting diode (LED) light (control). The lighting schedule was 16 L : 8 D-15 d / 150 lx / 6:00 am-22:00 pm (3 d preartificial insemination to 12 d postartificial insemination). Red light and white light affected the conception rate and kindling rate and increased the total litter size at birth (p < 0.05). The effects of red light on litter size at weaning, litter weight at weaning, and individual weight at weaning increased compared with the green and blue groups. The effects of red light on live litter size at birth were increased compared with those in the blue group (p < 0.05). Compared to white light, green and blue light reduced the number of secondary follicles (p < 0.05). Compared to red light, green and blue light reduced the number of tertiary follicles (p < 0.05). Compared with white light, red LED light resulted in greater ovarian follicle stimulating hormone receptor and luteinizing hormone receptor mRNA expression (p < 0.05). Compared with green and blue LED light, red LED light resulted in greater B-cell lymphom-2 mRNA expression (p < 0.05). Compared with green LED light, red LED light inhibited FOXO1 mRNA expression in rabbit ovaries (p < 0.05). Red light can affect the reproductive performance of female rabbits and the expression of key genes for follicular development.