• Journal of Veterinary Clinics
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• v.9 no.1
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• pp.323-332
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• 1992

The present study was carried out to examine the effect of oviduct epithelium and its conditioned medium on e development of early bovine embryos in vitro. Oocytes obtained from ovarian follicles of slaughtered cows were cultured in TCM199 with 10% fetal calf serum for 22-24hrs and then fertillzed in vitro using frozen-thawed semen treated with BO-caffein, BO-BSA(20mM heparin added). Oviduct epithelium was collected in each stage of the estrus cycle and conditioned medium was the medium in which oviduct epithelium in early luteal stage was cultured. In vitro fertilized bovine embryos of 1～2 cell were co-cultured with oviduct epithelium from different estrus cycles, cultured in conditioned medium, and cultured in rabbit oviduct. The cleavage rates of in vitro fertilized early bovine embryos co-cultured with oviduct epithelial cell from early luteal, luteal and follicular phase of estrus cycle(67.2～70.8%) and cultured in conditioned medium(56.7%) were significantly(p<0.05) higher than that of the control(44.2%) The rate of development to morula or blastocyst stage in oviduct epithelial cell co-culture(15.3～32.5%) from three phase of estrus cycles and conditioned medium(14.5%) were significantly(p<0.05) higher than that of the control(5.2%). The oviduct epithelial cell from early luteal phase gave a significantly( p<0.05) higher rate of development to morula or blastocyst stage than both luteal and follicular phase. The results of in vivo culture in rabbit oviduct of early bovine embryos were 52.1% for the cleavage rate and 26.7% for the rate of development to morula or blastocyst stage.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.2
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• pp.96-105
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• 2022

The ovary undergoes substantial physiological changes along with estrus phase to mediate negative/positive feedback to the upstream reproductive tissues and to play a role in producing a fertilizable oocyte in the developing follicles. However, the disorder of estrus cycle in female can lead to diseases, such as cystic ovary which is directly associated with decline of overall reproductive performance. In gene expression studies of ovaries, quantitative reverse transcription polymerase chain reaction (qPCR) assay has been widely applied. During this assay, although normalization of target genes against reference genes (RGs) has been indispensably conducted, the expression of RGs is also variable in each experimental condition which can result in false conclusion. Because the understanding for stable RG in porcine ovaries was still limited, we attempted to assess the stability of RGs from the pool of ten commonly used RGs (18S, B2M, PPIA, RPL4, SDHA, ACTB, GAPDH, HPRT1, YWHAZ, and TBP) in the porcine ovaries under different estrus phase (follicular and luteal phase) and cystic condition, using stable RG-finding programs (geNorm, Normfinder, and BestKeeper). The significant (p < 0.01) differences in Ct values of RGs in the porcine ovaries under different conditions were identified. In assessing the stability of RGs, three programs comprehensively agreed that TBP and YWHAZ were suitable RGs to study porcine ovaries under different conditions but ACTB and GAPDH were inappropriate RGs in this experimental condition. We hope that these results contribute to plan the experiment design in the field of reproductive physiology in pigs as reference data.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.35-41
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• 2008

This study was undertaken to investigate the distribution of major histocompatibility complex class II positive (MHC II+) (HLA-DP-like) cells in the cow uterus (cervix, corpus and cornu uteri) and to compare these cells between the estrus and diestrus phases of the estrous cycle. Twenty-nine multiparous cows were used. Tissue samples from the middle of the cervix, the corpus and the right cornu were taken immediately after slaughter at the estrus or diestrus phase. Streptavidin-biotin peroxidase complex staining was used to detect MHC II+ cells. The number of MHC II+ cells per unit area of tissue was counted using image analysis software under a light microscope. Numerous MHC II+ cells were found in the endometrium (cervix, corpus and cornu uteri) in both estrus and diestrus. MHC II+ cells were found in the surface epithelium of the cervix uteri in diestrus, but in the corpus uteri in both estrus and diestrus and in the cornu uteri in estrus. MHC II+ cells were also found freely in the lumen of the glands and between the gland epithelia of the corpus and cornu uteri in both estrus and diestrus. There were also MHC II+ cells in the connective tissue of the myometrium and perimetrium (outside the endometrium) and around the blood vessels. Endothelial cells were frequently positive for MHC II staining. More MHC II+ cells were found in the endometrium than outside the endometrium in both estrus and diestrus (p<0.001). However, there was no difference in the numbers of positive cells between estrus and diestrus either in the endometrium or outside it. These results are the first evidence for HLA-DP-like MHC II+ cells in the bovine uterus. They indicate that antigen presentation by HLA-DP-like MHC II+ cells of the uterus is not influenced by hormonal status.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.92-92
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• 2002

Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to evaluate semen quality after cryopreservation and to evaluate the Pregnancy rate after insemination (AI). Fifty infertilie dogs (age 2∼3 years) were selected for the study and divided into three different estrus induction treatment groups. Group 1: dogs (n=15) were given clomifene (0.1 mg/kg) orally for five days at 12 hr intervals. Group 2: dogs (n=15) were given bromocriptine (50 $\mu\textrm{g}$/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Group 3, n=20) when pro-estrus occurred. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. The ejaculated semen to freeze was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM: TES, 209 mM: citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 cm above the surface of liquid nitrogen (LN$_2$) for 23 min. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, the pregnancy rate of dogs treated with a combination of GnRH and bromocriptine was more effective than use of clomifene or bromocriptine only. In addition, frozen-thawed semen can be used successfully far artificial insemination in dog.

• Korean Journal of Agricultural Science
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• v.35 no.2
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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.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.253-262
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• 1989

This study was performed to investigate the patterns of progesterone secretion after induction of estrus in premature, metestrous and anestrous bitches. A total of 22 bitches were used. Of them 18 bitches were treated with hormone to induced estrus and 4 bitches were untreated and served as controls. Estrus was induced with $PGF_{2{\alpha}}$, estrone, estradiol-$17{\beta}$, PMSG and HCG(Treatment A), and with PMSG and HCG(Treatment B). Blood samples were collected via the cephalic vein at 2 to 5 days interval. Blood samples were centrifuged (1,200g, 10min.) within 30 minutes after collection and plasma was stored at $-20^{\circ}C$ until analyzed for the progesterone concentrations. Plamsa progesterone concentrations were measured by radioimmunoassay. The results of estrous induction were determined by estrous signs, ovarian response, egg recovery and progesterone patterns. The results obtained were as follows; 1. All bitches in treatment A showed estrous signs, however the ovarian response and egg recovery were not detectable and the levels of progesterone were nearly same as before. 2. In the treatment B, premature and metestrous bitches showed only estrous signs, however 5 of 7 anestrous bitches (71.4%) showed estrous signs, ovarian response and changes of progesterone levels. In conclusion, clinical estrous behavior can be induced during any phase of the estrous cycle, but ovulation should be induced only if induction occur approximately 4 months or more after the previous estrus.

• Journal of Veterinary Clinics
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• v.37 no.6
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• pp.301-304
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• 2020

The gestation period and parturition days were accurately predicted by measuring progesterone concentration in the plasma from 40 pregnant companion bitches. The mean length of the estrous cycle based on plasma progesterone concentrations were 8.14 ± 1.39 (Mean ± SD) days for proestrus, 9.19 ± 2.01 days for estrus, and 55.38 ± 1.96 days for diestrus phase, respectively. The gestation length from each based on the days was 65.61 ± 2.47 days from the first day of estrus after the first vaginal discharge, 63.21 ± 0.99 days from the day when plasma progesterone concentrations increase above 4.0 ng/ml, and 54.51 ± 3.51 days from the first day of diestrus, respectively. Therefore, the parturition day was estimated 65 days from the first day of estrus after the first vaginal discharge, 63 days from the day when plasma progesterone concentrations increase above 4.0 ng/ml, and 54 days from the first day of diestrus, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.2
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• pp.188-196
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• 2010

The objective of this study was to characterize the litter sizes in Huanghuai goats with high prolificacy (HP, five or more kids born per litter on at least one occasion), and to compare their peripheral blood concentrations of progesterone and estradiol with those of goats with poor prolificacy (PP, up to three kids born per litter on any occasion). The circulating concentrations of progesterone and estradiol were measured by radioimmunoassay from daily blood samples taken during natural estrus cycles and at 1-5 days after ovariectomy. Estrus was synchronized using two doses of a prostaglandin analog. Litter size for the HP goats increased up to a parity of five and decreased thereafter. The percentage of goats with litter sizes of $\geq$4 from parities 3 to 6 ranged from 44.5% to 58.3%. Although small differences in litter size were obtained for goats of parities three, four and six relative to five, parity five does had the highest mean litter size. Progesterone concentrations began to rise earlier and were higher in the HP than in the PP goats on most days of the luteal phase, but not during the follicular phase of the cycle or after ovariectomy. There was a significant difference between the two groups (p<0.05) in the magnitude of the progesterone plateau. Mean estradiol concentrations in the HP group remained significantly higher than in the PP group (p<0.05) during the estrus cycle. There were two estradiol peaks in the HP goats during the early luteal phase, but only one in the PP group. Measurements of individual corpora lutea (CL) in vitro showed that there was a greater prevalence of small CL (<6 mm in diameter) in the HP group than in the PP group (p<0.05). After ovariectomy, the estradiol level on day 1 was significantly higher than at the nadir during the estrus cycle in both the HP (p<0.01) and PP (p<0.05) goats, while levels decreased by 12.3% and 26.2% respectively compared with the mid-luteal period in HP and PP goats (p<0.05). The overall mean estradiol concentrations in HP goats were lower than in the PP group, but no significant differences were found between groups at 1-5 days after ovariectomy.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.180-185
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• 2020

Objective: Tyrosine phosphorylation is an essential process in many biological systems, including the male reproductive system. The presence of tyrosine-phosphorylated (TyrPho) proteins has been well documented in male reproductive organs, but research in fertile females is still limited. Methods: The ovary, oviduct, and uterus of adult female Sprague-Dawley rats in the estrus phase were used to localize TyrPho proteins using an immunohistochemical technique. These proteins were separated and their expression patterns were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis, respectively. Results: TyrPho proteins were localized in the cytoplasm of the oocyte except the antral fluid; in the granulosa cells, theca cells, and stromal cells of the ovary; at the apical surface of oviductal epithelial cells; and in the basal epithelium and submucosa of the uterine wall. Moreover, we found that 72-, 43-, and 28-kDa TyrPho proteins were localized in the ovary, while 170-, 55-, and 43-kDa proteins were localized in the oviduct. In the uterus, we detected four major bands, corresponding to 61-, 55-, 54-, and 43-kDa TyrPho proteins. Conclusion: Given that these TyrPho proteins were found in major reproductive organs in the estrus phase, these proteins may play important roles in female fertility.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.897-900
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• 2000

The experiment was conducted to study the effect of GnRH administration at induced estrus on pituitary and ovarian response in buffalo heifers. Eight Murrah river buffaloes of 12 to 13 months of age were treated with $PGF_{{2}{\alpha}}$ and eCG combination. GnRH (Fertagyl) 200 ug was injected (iv) at estrus in four heifers (treated group) while saline (2 ml, iv) was injected in remaining four heifers (control group). Blood was collected through jugular catheter to estimate plasma FSH, LH and estradiol level. The pretreatment plasma FSH, LH and estradiol values ranged from $8.46{\pm}1.97ng/ml$ to $12.31{\pm}1.30ng/ml$, $0.87{\pm}0.21ng/ml$ to $1.19{\pm}0.29ng/ml$ and $19.09{\pm}2.38pg/ml$ to $20.24{\pm}1.00pg/ml$ respectively. The plasma estradiol concentration elevated significantly (p<0.05) within 24 hr after eCG administration and reached its peak levels of $154.09{\pm}17.28pg/ml$ and $181.95{\pm}31.82pg/ml$ at estrus in respectively treatment and control groups. The plasma FSH and LH concentrations did not increase during follicular development after eCG administration while initial significant (p<0.05) increases in both plasma FSH and LH concentrations occured within 5 and 10 min, reaching peak levels of respectively $110.06{\pm}23.56ng/ml$ and $13.15{\pm}3.13ng/ml$ within 90 min after GnRH injection was detected. A sharp and significant decline in plasma estradiol concentration ($59.27{\pm}8.78pg/ml$) associated with synchronized ovulation within 24 hours after GnRH injection was recorded. The observation suggest that the hypophysis of prepubertal buffaloes treated with eCG have gonadotrophins awaiting the releasing factor to evoke release of gonadotrophin during the follicular phase to induce synchronized ovulation.