• 한국수정란이식학회지
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• 제29권2호
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• pp.183-188
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• 2014

A study was carried out on 16 indigenous ewes in Bangladesh in order to assess the reproductive physiology, the pattern of vaginal cell exfoliation and progesterone profiles during the estrous cycle period. The mean estrous cycle length and duration of estrus were $15.8{\pm}0.12$ days and $31.1{\pm}0.57$ h respectively. The exfoliated epithelial cells were categorized into parabasal, intermediate, superficial and keratinized and their relative occurrences. The percentages of parabasal, intermediate and superficial cell type during proestrus were similar. The percentage of superficial cell type during estrus was 61.7%, which was significantly (p<0.01) differ from other types of cells and stages of estrus cycle. Metoestrus was predominant with neutrophils in addition with other cell types. Dioestrus was dominated by neutrophils. On days 0 to 5 of the cycle the progesterone concentration was 0.09 to $1.6{\pm}0.07ng/ml$. The length of diestrus was 5~10 days with a range of mean progesterone level of $1.6{\pm}0.07$ to $2.8{\pm}0.11ng/ml$. Progesterone levels increased significantly (p<0.01) after Day 5 and maximum level was $2.8{\pm}0.11ng/ml$ observed on Day 10 of the estrous cycle. Thereafter it dropped rapidly to basal level of $0.11{\pm}0.04ng/ml$ on Day 0 (p<0.01). These results indicate that the pattern of exfoliation of vaginal cells along with progesterone concentration could be used to determine the reproductive stages of indigenous ewe.

• 한국수정란이식학회지
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• 제31권3호
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• pp.255-260
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• 2016

The aim of this study is to evaluate the effects of vitamin or mineral supplements on the conception rates of dairy heifers when replacing the last injection of GnRH with hCG in ovsynch protocol (experiment 1) and also to investigate whether the estrus synchronization treatment in the heifer stage affects the conception rates after $1^{st}$ parturition (experiment 2). In experiment 1, 50 heifers were randomly assigned into 3 groups: 20 heifers each in groups 1 and 2, and 10 in group 3. All three groups were treated with an intramuscular injection of GnRH on day 0 (day 0 = the day of program start), $PGF_{2{\alpha}}$ on day 7 and hCG on day 9, and were inseminated on day 10, 12~16h after hCG injection. In group 1 (vitamin group), the heifers were treated with an intramuscular injection of 5 ml of vitamin-ADE $500^{(R)}$, and group 2 (mineral group) was treated twice with an intramuscular injection of 30 ml of mineral supplement-LAPTOVET$^{(R)}$ on a one-week interval beginning on the day of hormone treatment (day 0 and day 7 respectively). Group 3 (control) was treated only with hormones. Pregnancy diagnosis was performed by ultrasonography through a rectal probe. First service conception rates (FSCR) and average services per conception (ASPC) were recorded for all subjects. Of the total 50 heifers, 6 (2 in group 1, 3 in group 2, and 1 in group 3) heifers were eliminated due to accidents during experiment 1. FSCRs were 58.8% (10/17), 66.7% (12/18) and 44.4% (4/9) in groups 1, 2 and 3, respectively. ASPCs were $1.53{\pm}0.72$, $1.27{\pm}0.59$ and $1.63{\pm}0.74$ in groups 1, 2 and 3, respectively. Although there were no significant difference between the groups, relatively good results (higher FSCR and lower ASPC) were obtained in both group 1 and 2. In experiment 2, 11 primiparous cows from group 2 of experiment 1 in heifer stage which had been treated both with the hormones for estrus synchronizing and mineral supplements (ES group), and 12 primiparous cows treated only with minerals (non-ES group) were compared to examine the effects of estrus synchronization program on conception rates after $1^{st}$ parturition. Following the examination, postpartum ASPCs were $1.55{\pm}0.82$ and $2.17{\pm}1.47$ in ES group and non-ES group, respectively. The postpartum average days open (ADO) were $116{\pm}56$ and $197{\pm}93$ in ES group and non-ES group, respectively. Although there were no significant difference between the two groups, desirable results (lower ASPC and shorter ADO) were found in ES group after $1^{st}$ parturiton. In conclusion, experiment 1 indicates that vitamin or mineral supplement with ovsynch protocol may have some positive effect on FSCR and ASPC of dairy heifers, and in experiment 2, ES program in heifer stage had a positive effect on ASPC and ADO following $1^{st}$ parturition.

• Clinical and Experimental Reproductive Medicine
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• 제31권4호
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• pp.225-234
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• 2004

Objectives: To evaluate the efficacy of GnRH antagonist cetrorelix in women undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) and to determine changes in serum hormone concentrations during cetrorelix administration. Methods: We performed a clinical trial on 30 patients undergoing COH with highly purified follicular stimulating hormone (HP-FSH) and gonadotropin releasing hormone antagonist (GnRHant), cetrorelix. FSH was administrated from day 2 or 3 of cycle with fixed dose and adjusted according to individual response. 0.25 mg of cetrorelix was injected daily subcutaneously from stimulation day 5 until the day of hCG administration. Daily ultrasound monitoring was performed for growing follicles and serum levels of luteinizing hormone (LH), estradiol ($E_2$) and progesterone were measured daily during cetrorelix administration. Up to 4 embryos were transferred. Results: Mean age of enrolled patients was $32.0{\pm}3.4$ years (mean $\pm$ S.D.). All of 30 patients underwent oocyte pick-up, and embryo transfer was done in 28 patients. The total and mean numbers of received oocytes were 196 and $6.5{\pm}4.7$, the number of fertilized eggs was 111, and the fertilization rate was 56.6%. Total duration of FSH administration was $9.2{\pm}2.2$ days and mean of $24.3{\pm}7.7$ ampules of HP-FSH was administered. Total duration of cetrorelix administration was $5.7{\pm}1.9$ days. Serum LH and progesterone levels were maintained in the range of $1.4{\sim}2.9\;mIU/mL$ and $0.3{\sim}0.6\;ng/mL$, which respectively reflected effective prevention of premature LH surge. Clinical pregnancies were achieved in 9 patients, and overall clinical pregnancy rate was 30.0% per oocyte retrieval, and 32.1% per embryo transfer. Conclusion: GnRH antagonist is safe and convenient for COH for IVF-ET and effective with optimal pregnancy rate.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.98-98
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• 2002

Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

• Reproductive and Developmental Biology
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• 제31권2호
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• pp.109-113
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• 2007

This study was conducted to evaluate the effect of cytochalasin B (CB) treatment in the activation medium on the development of somatic cell nuclear transfer (SCNT) rat embryos. Fetal fibroblast cells were isolated from a Day 14.5 fetus, and the oocytes for recipient cytoplasm were recovered from 4-week old Sprague Dawley rats. After enucleation and nuclear injection, the reconstructed oocytes were immediately exposed to activation medium consisting of 10 mM $SrCl_2$ with or without CB for 4 hr, and formation of pseudo-pronucleus (PPN) was checked at 18 hr after activation. Then, they were transferred into day 1 pseudopregnant recipients (Hooded Wistar) or cultured for 5 days to check their developmental competence in vivo or in vitro. The number of PPN was not affected by CB treatment during the activation. However, CB treatment supported pre-implantation development of rat SCNT embryos. Embryos generated by the procedures of SCNT were also capable of implanting, with 1 implantation scar found from a recipient following the transfer of 87 SCNT embryos to four foster mothers. The result of the present study shows that rat SCNT embryo can develop to post-implantation stage following treatment with CB.

• 한국수정란이식학회지
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• 제26권2호
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• pp.91-96
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• 2011

The purpose of this study was to evaluate the effects of days open on subsequent reproductive performance following to estrus synchronization in the 114 lactating dairy cows. The animals were divided into two groups according to the time of estrus synchronization; viz, ${\leq}$ 85 days, and > 85 days postpartum, respectively. The estrus synchronization protocol consisted of insertion of a controlled internal drug release (CIDR) device containing 1.9 g progesterone with an injection of 250 ${\mu}g$ gonadorelin (Day 0), an injection of $PGF_2{\alpha}$ and removal of the device on Day 7, an injection of 250 ${\mu}g$ GnRH on Day 9, and TAI 17 h later. Pregnancy diagnosis was determined at 30 to 60 days after TAI using both ultrasonography and rectal palpation. The body condition score (BCS) gradually increased over the postpartum period. In estrus synchronized cows until 85 days, conception rate on first service, number of service per conception, interval from estrus synchronization to conception, and interval from calving to conception were not significantly different among two farms (P>0.05). In estrus synchronized cows after 85 days postpartum, conception rate on first service, number of service per conception and interval from calving to conception were significantly different ($P{\leq}0.05$) between herds A and B (26.8 vs 50.0%; $2.1{\pm}1.35$ vs $1.37{\pm}0.54$ times, $237.3{\pm}97.8$ vs $164.7{\pm}69.3$ days, respectively). In estrus synchronized cows after 85 days postpartum interval from estrus synchronization to conception was greater (P<0.01) in herd B than in herd A ($63.6{\pm}57.4$ vs $26.1{\pm}24.9$). These results indicate that the time of estrus synchronization for maximized the reproductive performance is before 85 days postpartum and feeding and management is important factor for high reproductive performance.

• 한국수정란이식학회지
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• 제22권1호
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• pp.39-45
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• 2007

무발정기에 속한 Shih-tzu 견 9두를 대상으로 하여 PMSG(50IU/kg)를 10일 동안 매일 근육주사 후 마지막 10일째에 hCG(1,000IU/head)를 정맥 주사하여 인공적으로 발정을 유기 하였다. 실험견 9두 중 9두(100%)에서 발정 출혈, 회음부 반사, 외음부 종대 및 수컷 허용 등의 임상적인 발정 증상이 관찰되었으며 그중 5두(55.6%)가 임신하였고 4두는 임신되지 않았다(44.4%). 임신견 5두 중 3두(33.3%)가 자연분만하였는데 분만시 산자수는 $1.66{\pm}1.15$두 $(1{\sim}3)$이었고, 나머지 2두(40.2%) 는 PMSG 첫 투여후 38일과 41일에 조기태아사 하였다. 따라서 Shih-tzu 견에서 PMSG와 hCG 병용 투여에 의한 인공 발정 유기 효과는 발정 발현율 100%, 임신률 55.6%, 분만율이 33.3%이었다.

• 한국가축번식학회지
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• 제7권1호
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• pp.19-23
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• 1983

Present studies were conducted to investigate the developmental stage and the location of embryos in the reproductive tract at various times after ovulation, the morphologically normal after thawing of embryos preserved in liquid nitrogen, and the survival after transferring frozen-thawed embryos. The results obtained were as follows: 1. Embryo stage and location in the reproductive tract after hCG administration. For the investigation of embryo stage and location in the reproductive tract after ovulation, rabbits were laparotomized at 24, 40, 48, 72 and 120 hrs post hCG injection, simultaneously with mating. the oviducts and uteri were flushed out with PBS medium containing 50% rabbit serum, respectively. 1) Most of embryos was remained in the oviduct within 48 hrs, with the lapse of time, embryos were started to move to uterus and shifted in uterus at 72 hrs after hCG injection. 2) The representatives of embryos stage collected at 24, 40, 48, 72 and 120 hrs were 1-cell(60.4%), 8-cell to early morula (52.3, 39.3%), late blastocyst (95.5%) stages, respectively. 2. Morphological normality and survival of the frozen-thawed embryos. For the evalution of the quality and viability on the frozen-thawed embryos, immediately after thawing, embryos were assessed by morphologically normal under a dissecting microscope, and a further test of frozen-thawed embryos was made by transferring the morphologically normal embryos to the uteri of recipient rabbit induced pseudopregnancy by the injection of hCG at the time of hCG injection in donor rabbits. 1) The propotions of embryos which a, pp.ared morphologically normal was higher when 8-cell (85.7%) and morula(90.5%) were used for freezing than when 4-cell (66.7%) and blastocyst (75.8%) were used. 2) Preganacies were observed at Day 15 after transfer of frozen-thawed 8-cell (7/13), morula (19/42) and blastocyst (3/19) but not after transfer of embryos at 4-cell stage.

• 한국수정란이식학회지
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• 제13권1호
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• pp.77-86
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• 1998

A combined technology of transvaginal ovum pick-up(OPU) system with in vitro-oocyte manipulation technique can be used for improving reproductive efficiency in the cattle. The objective of this study was to establish a newly-conceived breeding program using OPU in the pregnant cows. The OPU trial was performed in pregnant cows every 10 days from 40 through 90 days of artificial insemination (Al), and number of follicles in ovary, number of retrieved oocytes and embryo development following in vitro-fertilization, were evaluated. Reduced number of follicles in the ovaries of pregnant cows was firstly detected from 70 days after A' and a significant (P<0.05) decrease in the follicle number (5.4 follicles /donor) was found at 90 days than at 40, 50, 60 and 80 days after Al (8.0~9.2). A similar pattern was also observed in the number of oocytes retrieved by OPU apparatus during experimental period. When retrieved oocytes were matured and inseminated in vitro with frozen bull semen, development of the oocytes to the blastocyst stage was not significantly affected by the retrieval time. Four embryos (morula or blastocyst stage) derived from oocytes retrieved from pregnant cows were nonsurgically transferred to four recipient cows on day 7 of estrus cycle. For the first time in Korea, three of four transferred embryos developed to live calves with normal physiological parameters. In conclusion, an effective breeding program employing pregnant cow can be developed by use of OPU trial and in vitro culture techniques of oocytes ; OPU system could be repeated in pregnant cows with no risk of abortion and viable offsprings were borne after transfer to the recipients.

• 한국수정란이식학회지
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• 제25권4호
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• pp.291-295
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• 2010

Four estrus-induced Himalayan tahrs (Hemitragus jemlahicus) were inseminated with frozen-thawed semen by laparoscopic or transcervical insemination techniques with no regard to the site of ovulation in non-breeding season. In June and July, 2009, estrus was synchronized by Eazi-Breed $CIDR^{(R)}$ (Controlled internal drug release; Pfizer Animal Health, New Zealand) insertion for 16 days and PG 600 (PMSG 400IU, hCG 200 IU; Intervet, Netherlands) injection (IM) a day before removing $CIDR^{(R)}$. Forty eight hours later, laparoscopic or transcervical insemination was done to each of two tahrs under anesthetic condition inducted by ketamine (1.5 mg/kg) and medetomidine (0.09 mg/kg). For examination of estradiol and progesterone, blood was collected right before $CIDR^{(R)}$ insertion, PG 600 injection, $CIDR^{(R)}$ removal and insemination. Estradiol levels of four tahrs (No. 1, 2, 3, 4) before $CIDR^{(R)}$ insertion and insemination were 13.3, 8.8, 14.3, 12 pg/ml and 23.5, 25.5, 21.1, 11.5 pg/ml, respectively. Progesterone levels of four tahrs (No. 1, 2, 3, 4) before $CIDR^{(R)}$ insertion and insemination were 1.8, 0.05, 0.63, 0.61 ng/ml and 1.03, 0.37, 1.48, 2.12 ng/ml. Except for No. 4 tahr, cervices showed cervical mucus and opened enough to penetrate with embryo transfer gun sheet usually used for cows. Therefore, No.4 was laparoscopically inseminated together with No. 1. In conclusion, none of four Himalayan tahrs was pregnant. However, we proved that estrus could be induced by CIDR and PG 600 injection in non-breeding season, and laparoscopic or transcervical insemination with frozen-thawed semen could be one of assisted reproductive techniques in Himalayan Tahr.