• Asian-Australasian Journal of Animal Sciences
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• v.4 no.1
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• pp.73-78
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• 1991

Sixty-one 5-11 month-old California, Chinchilla and New Zealand White rabbit were employed in this investigation. Thirty-three does were superovulated by injecting FSH/HCG subcutaneously or intravenously and then sacrificed at different hours after mating. The ova were collected from the fallopian tubes with Ham's F-10 medium supplemented with 0.4% bovine serum albumin (BSA) and 1% pregnant rabbit serum (PRS). Embryos were examined under an inverted DIC microscopy for observing the stage of development. We have found that the fertilized ova formed pronuclei at 19 - 20 hr postcoitus. Approximately at 26, 64 - 78 and 84 - 88 hr after mating, the fertilized ova cleaved further to 2-cell, morulae and blastocyst stage respectively. Another 28 does were allocated to the gene transfer study. Fourteen of the 28 does were sacrificed at 19 - 20 hr to donate the pronuclear stage ova for gene injection. The other 14 does were induced to pseudopreganacy by injection of 100 IU HCG intravenous as recipients. Four hundreds and seventeen ova were injected totally and 212 gene injected zygotes were transferred into the recipient oviducts. Five recipients became pregnant and 10 fetuses were obtained. Eight of the 10 fetuses were analysed for gene incorporation, but none of them were transgenic.

• Development and Reproduction
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• v.6 no.1
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• pp.55-66
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• 2002

Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.47-52
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• 2016

The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be $6.20{\pm}2.28/donor$ from the control group and was significantly lowly recovered to be $1.57{\pm}1.72/donor$ from the group treated at a sperm concentration of $10{\times}10^6$ (p<0.05). The number of unfertilized embryo was $0.8{\pm}1.30/donor$ in control group which was significantly lower than the group treated at a sperm concentration of $4{\times}10^6$ (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.191-198
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• 2006

The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.

• Journal of Nutrition and Health
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• v.36 no.2
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• pp.154-166
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• 2003

To investigate the effects of isoflavone supplementation on both bone mineral density and hormone variation in premenopausal women who had decreased bone mass, the 24 subjects were divided into two groups: one was the underweight group, consisting of 13 subjects, and the other was the normal weight group consisting of 11 subjects. For each group, we investigated the effects of isoflavone supplementation of 90 mg/day on both bone mineral density and hormone variation during 3 menstrual cycles. Anthropometric measurements, dietary recall, and analyses of blood and urine were assessed from baseline to post-treatment. The results were as follows: The average age of the underweight group was 21.8 years old and that of the normal weight group was 23.2 years old. The comparative results for the two groups at baseline were as follows: Onset of menarche, menstrual cycle, and menstrual length were not significantly different between the groups. Serum protein, total, HDL-, LDL-cholesterol, triglyceride, Ca, P, Mg, Cu, and Zn level were not significantly different between the groups. Serum estradiol, SHBG, LH, and FSH level were also not significantly different between the groups. Lumbar spine BMD by T scores of the underweight group was significantly lower than that of the normal weight group. Serum osteocalcin, urinary DPD, and urinary pH were not significantly different between the groups. The comparative results for the two groups at post-treatment were as follows: From baseline to post-treatment, the intake of energy, nutrients and isoflavone in food did not significantly change in either group. Serum protein, total cholesterol, HDL-, LDL-cholesterol, and triglyceride levels did not significantly change in either group. Serum Ca, Cu, and Zn levels were significantly lower in both groups and serum Mg level significantly decreased only in the underweight group. Serum estradiol levels were significantly lower in both groups, but serum SHBG, LH, and FSH levels did not significantly change in either group. Lumbar spine BMD by T score of the underweight group significantly increased to 15%, but that of the normal weight group did not significantly change. Serum osteocalcin of the underweight group significantly increased to 28%, while that of the normal weight group significantly increased to 40%. Urinary DPD of the normal weight group significantly increased to 12%. The results show that the BMD of the underweight group was lower than that of the normal weight group. Therefore, the underweight group had a disadvantage in obtaining maximum bone mineral density. The results also show that isoflavone supplementation during 3 menstrual cycles was effective in increasing the bone mineral density of the lumbar spine and affected bone metabolism markers in premenopausal underweight women. Therefore, it can be concluded that sufficient intake of isoflavone could be helpful in preventing decreases in bone mass among premenopausal women, especially underweight women.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.4
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• pp.235-244
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• 2004

Objective: The effects on spermatogenesis by expression of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) were investigated. Materials and Methods: Testicular specimens were obtained from 40 infertile males due to primary testicular failure and from 10 fertile males with other urologic problems. The specimens of infertile males were devided into 4 groups according to histologic findings; Sertoli cell only syndrome (A), maturation arrest (B), hypospermatogenesis (C) and sloughing and disorganization (D). VEGF and ET-1 expression were detected with immunohistochemical stain. Results: VEGF expression on Leydig cell was detected in all cases. But, VEGF expression rates on germ cell were significantly higher in infertile group B, C, D compared to that of the control group (p<0.05). ET-1 expression rates on Leydig cell was significantly lower in all infertile group compared to that of the control group (p<0.05). But, ET-1 expression rates on Sertoli cell was significantly higher in all infertile group compared to that of the control group (p>0.05). In germ cell of infertile group, LH, FSH and prolactin were significantly decreased, and estradiol is increased in positive stain group on ET-1 immunohistochemical stain (p<0.05). VEGF and ET-1 expression were not correlated mean seminiferous tubule diameter (p>0.05). Conclusions: Abnormal spermatogenesis would be reflected in VEGF expression in germ cell.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.1049-1056
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• 2010

This study was conducted to investigate the effects of dietary glutamine (Gln) on the productive performance and egg quality of laying hens. A total of four hundred Lingnan Yellow laying hens aged 34 weeks were randomly assigned into four groups (100 laying hens/group), and fed, respectively, with diets supplemented with 0% (control group), 0.2%, 0.4%, and 0.8% Gln during the 6-week feeding period. The results were as follows. First, the productivity of laying hens fed with 0.8% Gln in diet was significantly increased (p<0.05); however, the egg quality (egg weight, yolk weight, shell weight, egg shape index, shell thickness, shell density, shell breaking strength, yolk color, yolk index, and Haugh unit) was not affected compared with that of the control group (p>0.05). Second, luteinizing hormone (LH) (p<0.01), follicle stimulating hormone (FSH) (p<0.01), triiodothyronine ($T_3$), and tetraiodothyronine ($T_4$) contents (p<0.05) in blood of laying hens fed with 0.8% Gln in diets were also significantly improved, and greater improvement in the duodenum and oviduct structure was observed in that treatment group. This study indicated for the first time that diets with 0.8% Gln were able to increase the productive performance of laying hens through stimulating hormone secretion and better development of both the duodenum and oviduct structure in laying hens.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.131-140
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• 2001

Objective: To know the effects of xenoestrogen on spermatogenesis, we investigated the expression of cytochrome P450s enzymes (CYPscc, $CYP_{17{\alpha}}$, CYP19) and $3{\beta}$-HSD genes involved in steroidogenesis. Methods: Mouse testicular cells were prepared from 15-day-old ICR mice which had only pre-meiotic germ cells by enzyme digestion using collagenase and trypsin. Testicular cells were cultured in DMEM supplemented with FSH (0.1 IU/ml) and 10% FBS or medium with estrogen ($E_2$), bisphenol-A (BPA), octylphenol (OP; $10^{-9},\;10^{-7},\;10^{-6},\;10^{-5},\;10^{-4}M$, respectively) and aroclor 1254 (A1254) known as PCBs for 48 hours. The gene expression of cytochrome P450 enzymes were examined by semi-quantitive RT-PCR. The production of estrogen and testosterone was examined by RIA. Results: As results, expression of CYPscc mRNA was not significantly decreased, but $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA were significantly dose-dependent decreased. And production of testosterone and estrogen were not different except BPA and OP group ($10^{-5}M$). Conclusion: BPA, OP and A1254 might inhibit steroidogenesis by decreasing CYPscc, $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA expression in the mouse testis. These results suggest that BPA, OP and PCBs like as an endocrine disruptors inhibit the productions of steroidogenic enzymes and decrease the production of T and E by negative feedback mechanism. Therefore, these might disrupt steroidogenesis in Leydig cells of testis and would disturb testicular function and subsequently impair spermatogenesis.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.4
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• pp.234-237
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• 2011

Objective: During stimulated IVF cycles, up to 15% of oocytes are recovered as immature. The purpose of this study was to investigate the trend of oocyte maturity in repeated ovarian stimulation for IVF. Methods: One hundred forty-eight patients were selected who underwent two consecutive IVF cycles using same stimulation protocol during 2008 to 2010. Ovarian stimulation was performed with FSH and human menopausal gonadotropin and flexible GnRH antagonist protocol in both cycles. Oocyte maturity was assessed according to presence of germinal vesicle (GV) and the first polar body. Immature oocyte was defined as GV stage or metaphase I oocyte (GV breakdown with no visible polar body) and cultured up to 48 hours. If matured, they were fertilized with ICSI. Results: Percentages of immature oocytes were 30.8% and 32.9% ($p$=0.466) and IVM rates of immature oocytes were 36.2% and 25.7% ($p$=0.077), respectively. A significant correlation was noted between percentage of immature oocytes in the two cycles (R=0.178, $p$=0.03). Women with >40% immaturity in both cycles (n=21) showed lower fertilization rate of $in$ $vivo$ matured oocytes (56.4% vs. 72.0%, $p$=0.005) and lower pregnancy rate (19.0% vs. 27.1%, $p$=0.454) after the second cycle when compared with women with <40% immaturity (n=70). In both groups, female age, number of total retrieved oocyte and embryos transferred were similar. Conclusion: In repeated ovarian stimulation cycles for IVF, the immature oocyte tended to be retrieved repetitively in consecutive IVF cycles.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.2
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• pp.169-174
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• 2010

A meta-analysis was conducted to summarize the results of studies which have described the profiles of hormones during the oestrous cycle in buffalo using a fixed effect model and a random effect model. Plasma progesterone concentrations were lowest (0.30${\pm}$0.06 ng/ml) during the peri-oestrous phase and increased (p = 0.067) through the early luteal phase to a maximum concentration (1.94${\pm}$0.03 ng/ml) during the mid-luteal phase. Circulating plasma inhibin and estradiol concentrations were lowest (0.31${\pm}$0.01 and 11.04${\pm}$0.13 ng/ml) during the mid-luteal phase, increased through the late luteal phase to maximum concentrations (0.44${\pm}$0.02 and 22.48${\pm}$0.32 ng/ml) during the peri-oestrous phase. Plasma FSH concentrations were lowest during the early luteal phase and increased through the mid-luteal phase to a maximum concentration during the peri-oestrous phase. Peripheral prolactin concentrations were lowest during the late luteal phase and increased to a maximum concentration during the peri-oestrous phase which then declined (p = 0.716) during the early luteal phase. Peripheral plasma cortisol concentrations decreased from 2.68${\pm}$0.14 ng/ml during the early luteal phase to 1.43${\pm}$0.27 ng/ml during the mid-luteal phase (p<0.001) which then increased to 2.06${\pm}$0.17 ng/ml during the late luteal phase. Plasma $T_{5}$ concentrations decreased from the late luteal phase to the peri-oestrous phase (p<0.001) which then increased during the early luteal phase. $T_{4}$ concentrations increased from the late luteal phase to the peri-oestrous phase which then decreased during the early luteal phase.