• Journal of Embryo Transfer
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• v.29 no.2
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• pp.195-200
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• 2014

Endothelin 2 (EDN2) induces follicular rupture by constricting periovulatory follicles. In this study, it was investigated the mechanisms of EDN2 action on follicular rupture with respect of receptor using the conditionally granulosa cell specific EDN2 receptor type A (ETa) KO mice (gcETaKO; $ETa^{flox/-}{\cdot}Amhr2^{Cre}$). It was generated the gcETaKO mice by breeding with $ETa^{flox/-}$ mice after mono-alleic ETa knockout by $ZP3^{Cre}$ and $Amhr2^{Cre}$ mice. Fertility, ovulation and maturation rates of ovulated oocytes after super ovulation were investigated in the gcETaKO mice compared with wild-type mice ($ETa^{flox/flox}$ and $ETa^{flox/-}$) as a control group. In the gcETaKO mice, normal fertility after breeding with male mice was shown compared with wild-type mice. And, there was no significant differences in ovulation rates after super ovulation, however its maturation rates was lower than that of wild type mice. These findings show that EDN2 in follicular rupture for ovulation is related with an other ETa not in granulosa cells. Further studies are needed to investigate how EDN2 is acted in ovarian follicular rupture for ovulation.

• Journal of Embryo Transfer
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• v.18 no.2
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• pp.115-124
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• 2003

The objective of this study was to compare the effect of four different estrus induction methods in anestrus cows on the estrus induction and pregnancy following artificial insemination (AI). Sixty-five cows (3∼4 years old) were selected and divided into four different estrus induction treatment groups. Group 1, 12 cows were treated by Ovsynch program combined with GnRH and PGF$_2$a. Group 2, 12 cows were treated by "Tow plus Two" program with GnRH and PGF$_2$a. Group 3, 20 cows were treated by "Tow plus Two" program following intravaginal progesterone implantation (CIDR). Twenty one cows in Group 4 were treated by "Tow plus Two" program following follicular rupture and intravaginal progesterone implantation. Cows were then observed estrus induction and inseminated artificially at 12 h and 24 h after standing estrus. The rates of estrus induction in Group 4 (18/21, 86%) was significantly (P<0.05) higher than those in groups 1, 2 and 3 (8/12, 67%; 9/12, 75%; 14/20, 70%). In the mean time of onset of estrus after final administration of GnRH in different hormone-treated cows, the cows in Group 3 (24.2$\pm$2.2) and Group 4 (23.4$\pm$2.0) were significantly (P<0.05) shorter than that in Group 1 (28.5$\pm$4.6) and Group 2 (26.4$\pm$3.3). The rates of pregnancy diagnosed on Day 28 were significantly different between treatment groups. Significantly (P<0.05) higher rate of pregnancy was observed in Group 4 (17/20, 85.0%) than those in Groups 1, 2 and 3 (7/11, 63.6%; 8/12, 66.7%; 15/20, 75.0%, respectetively). The rate of abortion diagnosed on 49 days of gestation was significantly (P<0/05) lower in Group 4 (1/17, 5.9%) than those in Groups 1, 2 and 3 (2/7, 28.7%; 2/8, 25% and 3/15, 20%, respectively). In conclusion, combined treatments with GnRH and PGF$_2$a following follicular rupture and progesterone implant in anestrus cows was considered to be most effective in estrus induction and maintenance of pregnancy. Further studies are needed to verify the functional mechanisms of residual follicles in anestrus ovaries on retarding the response of hormonal treatments.sponse of hormonal treatments.

• Clinical and Experimental Reproductive Medicine
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• v.20 no.1
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• pp.19-29
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• 1993

Plasminogen activator (PA)-plasmin system in follicular fluid is involved in the process leading to follicular rupture at ovulation. It is well known that PA is closely associated with cellular differentiation and tissue remodeling on evidences from the study of normal and malignant tissues. This study was designed to ascertain a potential role of PA in the ovarian folliculogenesis. Immature Sprague-Dawley rats were injected with pregnant mare serum gonadotropin, followed by injection of serine protease inhibitor (SPI; mixture of 1 mol/L benzamidine and 1 mol/L amino-caproic acid) into the unilateral ovarian bursa. In the control study, mechanical effect of bursal injection and contralateral ovarian effect SPI were ruled out. Total antral follicular areas relative to total ovarian cross-sectional areas was siginificantly lower in SPI-injected ovary than in saline-injected ovary. SPI injection decreased the relative antral follicular area by 33 % respectively. Electron microscopic finding of granulosa cell in the atretic follicle showed the presence of pyknotic nucleus, blurring of neucleolemma, degeneration of mitochondria and dilation of endoplasmic reticulum. After induction of ovulation with hCG, the number of oocytes released was significantly decreased in SPI-injected oviduct than in saline-injected oviduct. From above results, author discussed that PA may play a role not only in ovulation but also in some processes of folliculogenesis.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.125-131
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• 1992

The possible effect of human follicular fluid(hFF) on the growth and development of fertilized oocytes and embryos is important because the fallopian tubes are exposed to FF after follicular rupture and the processes of fertilization and embryo cleavage occur inside the fallopian tubes. Previously, it was suggested that human FF might adversely affect on the development of early mouse embryos. In order to investigate the effect of hFF on the development of embryos, early mouse embryos were cultured in media containing various protein sources as bovine serum albumin(BSA), fetal cord serum(FCS) and FF. And we evaluated the development of early mouse embryos in terms of the morphology, cleavage rate, and cell count of blastcysts. There were no significant differences in the morula and blstocyst formation rates of 2-cell mouse embryos cultured in the media containg three different protein sources and three different concentrations of FF. The blastocyst formation rate of 1-cell mouse embryo cultured in FF group was significantly higher than that cultured in BSA group(P<0.05). The morula and blastocyst formation rates of 2-cell mouse embryos of the group cultured in the media containing FF were comparable with those of other two groups, in addition, the cell count of blastocysts of FF group in the 2-cell embryo culture was higher than those of BSA group and HCS group(P<0.01), and this finding was also noted in 1-cell embryo culture. There was no difference in the morula and blastocyst formation rates of the 2-cell mouse embryos cultured in the media containing different concentrations of FF. These results suggest that mature human follicular fluid has no inhibitory activity on the development of early mouse embryos even in high concentration and may be a good protein source which is positively associated with the development of mouse embryos in vitro especially in 1 cell embryo culture.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.1
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• pp.29-32
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• 1992

The effects of prostaglandins (PG) and indomethacin, a PG synthesis inhibitor, on ovulation in the perfused fowl ovary were studied. Laying hens were killed by cutting the jugular vein 18-19 h before expected ovulation of the second follicle of a clutch sequence. The cannulated ovary was dissected free and connected with a recycling perfusion apparatus. Agents to be studied for their effects on ovulation were added to the perfusion fluid. $PGF_{2{\alpha}}$ at 0.1 and 1 mg/l induced ovulation, with a success rate of 25% and 30%, respectively. The same doses of $PGE_2$ were effective at 60% and 63%, respectively. Indomethacin partially blocked gonadotrophin-induced ovulation. It is suggested that PGs may play a supportive role in the process of follicular rupture in the domestic fowl.

• Clinical and Experimental Reproductive Medicine
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• v.12 no.2
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• pp.25-37
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• 1985

Four points on the basal body temperature (B.B.T.) curve was correlated with the estimated time of ovulation, as determined by serial ultrasound in 50 induced menstrual cycles from 22 subjects. The time of ovulation was estimated by measuring the maximal diameter of follicles and observing the morphologic changes within the ovary from follicle to corpus luteum. The results were as following; 1. The diameter of the follicle measured at the day before disappearance was 21.1 mm on an average (S.D.: 2.14). The average follicular growth for 4 days before ovulation was measured at a rate of 2.8 mm/day, and rapid growth of follicle was observed 3.1 mm/day at the day before. 2. The changes associated with rupture of the follicles were the followings, in order of frequency; decrease in size(94%), disappearance of follicles(64%), fluid in the Cul-de-Sac(26%) and increased internal echoes(16%). 3. Only 20 of 50 cycles, exhibited a BBT dip and correlated with the estimated time of ovulation by ultrasound in 2 of which cases(10%). BBT nadir, 30 of 50 cycles, correlated in 5(16.7%). The first day of hyperthermic plateau(FDHP) and BBT coverline was exhibited in all cycles, correlated in 41(82%) and 35(70%) cases. 4. The relationship between the diameter of dominant dominant follicle, measured by ultrasound, and the basal body temperature curve were as following. During cycles in which dip was observed on the BBT curve, the follicular diameter were 10.5${\pm}$2.12 mm on 4 days prior to the point (D-4), and 12.5${\pm}$2.12 mm (D-3), 15.5${\pm$2.12 mm (D-2), 17.0${\pm}$1.41 mm (D-1) and 21.5${\pm}$2.12 mm just prior to the dip (D-0). In the nadir; 9.6${\pm}$1.67 mm (N-4), 12.8${\pm}$1.79 mm (N-3), 16.2${\pm}$1.92 mm (N-2), 18.2${\pm}$2.17 mm (N-1) and 21.4${\pm}$2.61 mm (N-0). In the First day of Hyperthemic Plateau (FDHP); 9.8${\pm}$1.36 mm (F-4), 12.4${\pm}$1.41 mm (F-3),15.1${\pm}$1.57 mm (F-2), 18.1${\pm}$1.67 mm (F-1) and 21.2${\pm}$2.25 mm (F-0). In the BBT coverline endopint; 9.9${\pm}$.39 mm (C-4), 12.5 ${\pm}$1.44 mm (C-3), 15.2${\pm}$1.64 mm (C-2), 18.0 ${\pm}$1.69 mm (C-1), and 21.2${\pm}$2.31 mm (C-0). 5. The relationship between the ultrasonographic signs of ovulation and the basal body temperature curve were as following. The BBT dip correlated with the ovulation in 2 cases, which revealed decrease in follicular diameter (100%), fluid pattem in the Cul-de-Sac (1 case, 50%) and complete disappearance of follicle (1 case, 50%). In the nadir (5 cases); the ultrasonographic signs of ovulation were decrease in follicular diameter (5 cases, 100%), fluid pattern in the Cul-de-Sac (1 case, 20%) and complete disappearance of follicle (3 cases, 60%). In the First day of Hyperthermic Plateau (41 cases); decrease in follicular diameter (40 cases, 97.6%), fluid pattern in the Cul-de-Sac (11 cases, 26.8%), appearance of internal echo and thickening of the wall (6 cases, 14.6%) and com plete disappearance of follicle (28 cases, 68.3%). In the BBT coverline endpoint (35 cases); decrease in follicular diameter (33 cases, 94.3%), fluid pattern in the Cul-de Sac (9 cases, 25.7%), appearance of internal echo and thickening of the wall (5 cases 14.3%) and complete disappearance of follicle (20 cases, 57.1%).

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.7
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• pp.917-922
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• 2008

A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif (ADAMTS1) plays a critical role in follicular rupture and represents a major advance in the proteolytic events that control ovulation. In this study, a 9,026-bp DNA sequence containing the full coding region, all 8 introns and part of the 5'and 3' untranslated region of the porcine ADAMTS1 gene was obtained. Analysis of the ADAMTS1 gene using the porcine radiation hybrid panel indicated that pig ADAMTS1 is closely linkage with microsatellite marker S0215, located on SSC13q49. The open reading frame of its cDNA covered 2,844 bp and encoded 947 amino acids. The coding region of porcine ADAMTS1 as determined by sequence alignments shared 85% and 81% identity with human and mouse cDNAs, respectively. The deduced protein contained 947 amino acids showing 85% sequence similarity both to the human and mouse proteins, respectively. Comparative sequencing of three pig breeds revealed one single nucleotide polymorphism (SNP) within exon 7 of which a G-C substitution at position 6006 changes a codon for arginine into a codon for proline. The substitution was situated within a PvuII recognition site and developed as a PCR-RFLP marker for further use in population variation investigations and association analysis with litter size. Allele frequencies of this SNP were investigated in seven pig breeds/lines. An association analysis in a new Qingping female line suggested that different ADAMTS1 genotypes have significant differences in litter size (p<0.01).