• Laboraroty Animal Research
• /
• v.34 no.4
• /
• pp.211-215
• /
• 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.

• Journal of Animal Reproduction and Biotechnology
• /
• v.37 no.1
• /
• pp.34-41
• /
• 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
• /
• v.64 no.3
• /
• pp.432-442
• /
• 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.

• Clinical and Experimental Reproductive Medicine
• /
• v.50 no.3
• /
• pp.192-199
• /
• 2023

Objective: This study was conducted to investigate chromosomal abnormalities and their correlations with clinical and radiological findings in females with primary amenorrhea (PA). Methods: Detailed forms were recorded for 470 females, including the construction of three-generation pedigrees. Peripheral venous blood was drawn, with informed consent, for cytogenetic analysis. Results: An abnormal karyotype was found in 16.38% of participants. The incidence of structural abnormalities (6.8%) exceeded that of numerical abnormalities (6.15%). Turner syndrome represented 45% of all numerical abnormalities. Furthermore, the Y chromosome was detected in 5% of females with PA. Among the structural chromosomal abnormalities detected (n=32) were mosaicism (25%), deletions (12.5%), isochromosomes (18.75%), fragile sites (3.12%), derivatives (3.12%), marker chromosomes (3.12%), and normal variants (29.125%). An examination of secondary sexual characteristics revealed that 29.6% of females had a complete absence of breast development, 29.78% lacked pubic hair, and 36.88% exhibited no axillary hair development. Radiological findings revealed that 51.22% of females had a hypoplastic uterus and 26.66% had a completely absent uterus. Abnormal ovarian development, such as the complete absence of both ovaries, absence of one ovary, one absent and other streak, or both streak ovaries, was observed in 69.47% of females with PA. Additionally 43.1%, 36.1%, 67.4%, and 8% of females had elevated levels of serum follicle-stimulating hormone, luteinizing hormone, thyroid-stimulating hormone, and prolactin, respectively. Conclusion: This study underscores the importance of karyotyping as a fundamental diagnostic tool for assessing PA. The cytogenetic correlation with these profiles will aid in genetic counseling and further management of the condition.

• Nutrition Research and Practice
• /
• v.16 no.sup1
• /
• pp.147-159
• /
• 2022

BACKGROUND/OBJECTIVES: Complementary and alternative medicines can be used to alleviate climacteric symptoms that significantly affect the quality of life of postmenopausal women. Isoflavones are the most common plant-based therapies for postmenopausal changes, but the results of previous studies have been controversial. MATERIALS/METHODS: To investigate whether isoflavones would affect menopausal symptoms as well as ovarian hormones, we performed a systematic review and meta-analysis. The PubMed and EMBASE databases were used to perform the systematic search. Included studies were limited to randomized controlled trials (RCTs) assessing the impact of isoflavone supplementation on menopausal symptoms. RESULTS: Eleven studies were included for the final quantitative assessment. Isoflavone intervention was varied between 49.3 and 135 mg of isoflavones per day for 12 wk-2 yrs. The meta-analysis showed that supplementation of isoflavones significantly increased the estradiol levels (standardized mean difference [SMD] = 0.615, P = 0.035) and Kupperman index (SMD = 3.121, P = 0.003) but had no significant effect on hot flashes, follicle-stimulating hormone, and luteinizing hormone. However, both estradiol and the Kupperman index showed significant heterogeneity among studies (I² = 94.7%, P < 0.001 and I² = 98.1%, P < 0.001, respectively). CONCLUSIONS: Although the results showed a significant SMD in estradiol and the Kupperman index, the results should be interpreted with caution due to the high heterogeneity. Further validation with a larger RCT will be necessary. Overall, isoflavone supplementation has distinct effects on the climacteric symptoms and hormonal changes in postmenopausal women.

• Development and Reproduction
• /
• v.27 no.4
• /
• pp.185-193
• /
• 2023

Although increasing evidence of cause-and-effect relationship between BPA exposure and female reproductive disorders have been suggested through many studies, the precise biochemical and molecular mechanism(s) by which BPA interferes with steroidogenesis in the ovarian cells still remain unclear. Therefore, the purpose of this study was to discover the steroidogenic biomarker(s) associated with BPA treatment in human granulosa cell line, KGN. In this study, our results obtained via the analysis of steroidogenesis-related protein expression in KGN cells using quantitative polymerase chain reaction (qPCR) and western blot analyses revealed that the expression levels of steroidogenic acute regulatory (StAR) and aromatase decreased considerably and gradually after BPA treatment in a dose-dependent manner under BPA treatment. Further, remarkable decreases in their expression levels at the cellular levels were also confirmed via immunocytochemistry, and subsequent StAR and aromatase mRNA expression levels showed profiles similar to those observed for their proteins, i.e., both StAR and aromatase mRNA expression levels were significantly decreased under BPA treatment at concentrations ≥0.1 μM. We observed that follicle stimulating hormone upregulated StAR and aromatase protein expression levels; however, this effect was suppressed in the presence of BPA. Regarding the steroidogenic effects of BPA on KGN cells, controversies remain regarding the ultimate outcomes. Nevertheless, we believe that the results here presented imply that KGN cells have a good cellular and steroidogenic machinery for evaluating endocrine disruption. Therefore, StAR and aromatase could be stable and sensitive biomarkers in KGN cells for the cellular screening of the potential risk posed by exogenous and environmental chemicals to female reproductive (endocrine) function.

• Journal of Embryo Transfer
• /
• v.16 no.3
• /
• pp.183-191
• /
• 2001

This study was designed to establish the superior method for IVF embryos from DNA marker-proved Hanwoo cattle. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from unstimulated. immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local abesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analysed using the GLM procedure of SAS. Mean number of follicles identified on ultrasound was 5.5 $\pm$2.9 in right and 4.3 $\pm$2.8 in left ovaries, respectively. The highest follicles(16.6$\pm$2.6) were found in 5101 cow compared to others. Recovery rate of follicular oocytes in individual cow was highest in 5101 cow with 89.3% in > 2mm and 94.0% in $\leq$ 2mm follicles. Total recovery rate was significantly(P<0.01) higher in $\leq$ 2mm(85.7%, 130/154) than > 2mm follicles(74.2%, 201/271). Significantly more oocytcs of Grade IV were recovered from > 2mm follicles. Mean number follicles recovered was 4.8$\pm$3.7. 3.0$\pm$3.4 and 0.3$\pm$0.6 in $\leq$2mm, 2~6mm and $\geq$6mm follicles. respectively. Our results imply that the more fertilizablc oocytes can be recovered from invisible-immature follicles by the combination of simultaneous rectal ovarian-palpation and ultrasound-guided approach in Hanwoo cattle.

• Journal of Embryo Transfer
• /
• v.21 no.4
• /
• pp.315-322
• /
• 2006

This study was performed to investigate the physiological effects of bovine follicular fluid (bFF) or anti-inhibin serum (AI) on follicular development in Hanwoo. Saline (0.95%), bFF or AI (total of 40 ml) were administered into the jugular vein in 9 Hanwoo cows. The plasma inhibin, estradiol-17 $\beta$ (E2), and progesterone (P4) levels were measured using RIA or ELISA kit and the number of ovarian follicles was observed by ultrasonography at 72 hr after ovulation. The plasma inhibin level in bFF treatment group was significantly increased and maintained higher level from 102 hr after ovulation compared to that of saline and AI groups (p<0.05). In plasma E2 level, AI treatment group showed significantly higher level from 36 hr to 108 hr after ovulation than that of saline and bFF groups (p<0.05). After that it showed decreasing tendency. The plasma P4 level was increased in control and AI treatment groups at 68 hr after ovulation. However, it was maintained significantly lower level in bFF group from 84 hr to 180 hr compared to that of saline and AI group (p<0.05). As a result of ultrasonography at 72 hr after ovulation, higher number of follicles was shown in AI treatment group compared to bFF groups, although the difference was not statistically significant. Taken together, it can be postulated that a treatment of synthesized AI inhibits the secretion of inhibin, stimulates FSH secretion inhibited by inhibin, and induces follicular development and estrogen secretion. According to these results, a development of ovarian follicle immediately after ovulation is associated closely with inhibin in Hanwoo heifers.

• Clinical and Experimental Reproductive Medicine
• /
• v.34 no.2
• /
• pp.125-131
• /
• 2007

Objective: The aim of this study was to compare GnRH antagonist and agonist flare-up treatment in the management of poor responder patients. Methods: One hundred forty-four patients from Jan. 1, 2002 to Aug. 31, 2005 undergoing IVF/ICSI treatment who responded poorly to the previous cycle (No. of oocyte retrieved$\leq$5) and had high early follicular phase follicle stimulating hormone (FSH>12 mIU/ml were selected. Seventy-five patients received agonist flare-up protocol and 71 patients received antagonist protocol. We analyzed the number of oocytes retrieved, number of good embryos (GI, GI-1), total dose of hMG administered, implantation rate, cycle cancellation rate, pregnancy rate, live birth rate. Results: The cancellation rate was high in antagonist protocol (53.5% vs. 30.1%). The number of oocyte retrieved, the number of good embyos were high in agonist flare-up group. There was no statistical difference between GnRH agonist flare up protocol and GnRH antagonist protocol in implantation rate (14.5%, 10.1%), clinical pregnancy rate per transfer (29.4%, 21.2%) and live birth rate per transfer (21.6%, 18.2%). Although the result was not statistically significant, GnRH agonist flare up group showed a nearly doubled pregnancy rate and live birth rate per initial cycle than GnRH antagonist group. Conclusions: The agonist flare-up protocol appears to be slightly more effective than the GnRH antagonist protocol in implantation rate, pregnancy rate, live birth rate but shows statistically no significance. Agonist flare-up protocol improved the ovarian response in poor responders. However, based of the result of the study, we can expect improved ovarian response in poor responders by GnRH agonist flare up protocol.

• Clinical and Experimental Reproductive Medicine
• /
• v.34 no.1
• /
• pp.49-56
• /
• 2007

Objective: To evaluate the effect of short coasting, by withdrawing both gonadotropins and GnRH agonist (GnRHa), on the prevention in severe ovarian hyperstimulation syndrome(OHSS) without compromising pregnancy outcome. Method: Thirty-seven women who had been coasted during COH for IVF were coasted when $\geq$20 follicles > 15 mm with serum E$_2$ level of 4,000 pg/ml were detected. Coasting was initiated for one or two days depending on the status of follicle on ultrasound and serum E$_2$ level. Both gonadotropin and GnRHa were withheld for coasting. Retrospective study was carried and changes of serum E$_2$ levels, number of oocytes retrieved, fertilization rate, pregnancy rate were compared and analyzed. Results: The mean serum E$_2$ level fell from 6,993 pg/ml on the onset of coasting to 3,396 pg/ml on the day of hCG administration. The mean number of oocytes retrieved and fertilization rate were 15.7 and 70.0%, respectively. Fifteen patients were pregnant (40.6%) and implantation rate was 15.2%. Twenty-six (70.3%) patients were coasted for one day and 11 (29.7%) were coasted for two days. The mean decrease rate of serum E$_2$ level was 43% in one day coasting group and 15% (1$^{st}$ day) and 81% (2$^{nd}$ day) in two day coasting group. The pregnancy outcome was similar between the two groups. After coasting, no severe or moderate OHSS occurred in any patients and mild OHSS occurred in 3(8.1%) patients. Conclusions: Coasting for one or two days can be used successfully in the prevention of OHSS without compromising IVF cycle outcome.