• Journal of Veterinary Science
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• 제21권3호
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• pp.48.1-48.18
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• 2020

Background: Mature oocytes at the metaphase II status (MII-stage oocytes) played an important role in assisted reproductive technology in non-human primates. Objectives: In order to improve the proportion of MII-stage oocytes retrieval, three different superovulation protocols were performed on 24 female cynomolgus monkeys. Methods: All the monkeys received once-daily injection of follicle-stimulating hormone (25 international unit [IU]) on day 3 of the menstruation, 3-day intervals, twice daily for 8-12 days until the time of human chorionic gonadotropin (1,500 IU) injection, on the 14-17th day of menstruation collecting oocytes. The difference between protocol I and protocol II was that 0.1 mg the gonadotropin-releasing hormone agonist was injected on day 1 of the menstruation, while the difference between personalized superovulation protocol and protocol II was that oocytes could be collected on the 14-17th day of menstrual cycle according to the length of each monkey. Results: The total number of oocytes harvested using the personalized superovulation protocol was much higher than that using protocol I (p < 0.05), and the proportion of MII-stage oocytes was significantly greater than that from either superovulation protocol I or II (p < 0.001 and p < 0.01 respectively), while the proportion of immature oocytes at the germinal vesicle was less than that from superovulation protocol I (p < 0.05). Conclusions: The personalized superovulation protocol could increase the rate of MII-stage oocytes acquired, and successfully develop into embryos after intracytoplasmic sperm injection, and eventually generated fetus.

• Clinical and Experimental Reproductive Medicine
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• 제38권1호
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• pp.31-36
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• 2011

Objective: To determine whether the serum ${\beta}$-human chorionic gonadotropin (hCG) profile following preimplantation genetic diagnosis (PGD) is lower than that of intracytoplasmic sperm injection (ICSI) cycles. Methods: A total of 129 PGD cycles and 1,161 age-matched ICSI cycles, which resulted in pregnancy (serum ${\beta}-hCG{\geq}5$ mIU/mL) on post-ovulation day (POD) 12 were included. We compared the mean serum ${\beta}$-hCG levels on POD 12, 14, 21, and 28, doubling time of serum hCG, and created a cut-off value for predicting a singleton pregnancy in each group. Results: The mean serum ${\beta}$-hCG concentration of the PGD group was significantly lower than that of the control group on POD 12, 14, and 21. The doubling time of serum ${\beta}$-hCG at each time interval showed no significant difference. The cut-off-value of serum ${\beta}$-hCG for predicting a single viable pregnancy was 32.5 mIU/mL on POD 12 and 113.5 mIU/mL on POD 14 for the PGD group, which was lower than that for the control group. Conclusion: Blastomere biopsy may decrease the ${\beta}$-hCG-producing activity of the trophoblasts, especially in early pregnancy. Setting a lower cut-off value of serum ${\beta}$-hCG for predicting pregnancy outcomes in PGD may be needed.

• 보건교육건강증진학회지
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• 제1권1호
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• pp.57-71
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• 1983

Sex education is necessary for the youth that they should have an adequate sexual knowledge, attitudes and behaviors in their adolescent period. Four major objectives of this study are as follows; 1. To know the level of sexual knowledge of commercial girl's high school students in Seoul. 2. To know their actual state for the attitudes and behaviors toward sex. 3. To compare the sexual knowledge, attitudes and behaviors of day time school students with those of night time school students. 4. To compare the factors associated with their sexual knowledge, attitudes and behaviors with the individual level. Data were collected on 986 students in Seoul from April 1 to April 10, 1983. As the result of this survey, the following conclusions were obtained. 1. The level of sexual knowledge. (1) The level of knowledge of day time high school students about the physiology of female and pregnancy is shown higher than those of night time students. 64.2% of the respondents are aware of the organ producing ovum. 56.4% the ovulation period. 95.6% the cause of pregnancy. 74.5% the pregnantable period and 12.7% the place of fetilization. (2) Out of 986 respondents. 71.8% knew about contraceptive method correctly, and day time school students knew litter better than night time school students, by showing 73.9% and 69.7% respectively. They knew about contraceptive method with 'oral pills'. 'menstrual cycles', 'condoms'. and 'loops' in the same order. 3) Kinds of veneral disease was correctly known by 37.9%. Day time students knew much better than night time school students. by showing 67.8% and 7.9%. respectively. Transmission method of veneral disease was correctly known by 28.3%. Day time students knew much better than night time students, by showing 51.2% and 5.3%, respectively. (4) The major information source of sexual knowledge was 'book and magazine' (39.9%) and 'friends' (27.4%). 2. Actual state of attitudes and behaviors toward sex. (1) Out of 986 respondents, 84.0% answered that premarital purity should be kept. (2) Out of 986 respondents, 60.8% had an acquaintance of the opposite sex. 45.2% of students with opposite sex reported introduction of their friends as the main channel of making an acquaintance of the opposite sex. (3) Of those who responded to this study 13.8% reported having masturbation, 21.5% kissing, 6.2% petting and 3.7% sex intercourse. (4) 64.8% had sexual problems, which was mensturance (27.2%), aquaintance of the opposite sex (25.4%). The main method to solve the problems were consultation with 'friends' and 'books and magazine' percentage being 39.1% and 30.8%, respectively, whereas very small students discussed with 'teachers' and 'parents' percentages being 0.3% and 5.9%, respectively. (5) Out of 986 respondents, 62.2% had experience in educating about sex, the percentage of day time school was higher than that of night time school. (6) 88.2% of students wanted sex education in school, which were 'general information about sex' (35.4%), 'sexual morality and solution of sexual desire' (18.5%), 'aquaintance of the opposite sex' (13.3%) and 'marriage and role of man and woman' (12.4%) in the same order. They wanted to be instructed about sexes in the lecture of physical education, home economics, biology, military training (59.9%), regular curriculum (17.5%), special lecture (16.9%) and by the counselor or school nurse (5.7%). 3. Analyse concerning the factors about the knowledge, attitudes and behaviors. (1) The better school record was, the higher the level of sexual knowledge. (2) Those who have the religion considered the permarital purity more important than those who have not, the percentage showing 86.7% and 80.7%, respectively. (3) The result of dyad analysis of making acquaintance of the opposite sex in the friendship network showed that a high index of the acquaintance of the opposite sex tends to be a high adoption of making one at the individual level in the group, while the low index tends to be a low adoption of making one in the group.

• Asian-Australasian Journal of Animal Sciences
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• 제11권5호
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• pp.480-485
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• 1998

Application of trans rectal ultrasonography to buffaloes (n=6) to follow the growth of large antral follicles individually, on each day of one interovulatory interval revealed that follicular turnover during oestrous cycle occured in waves. There was a predominance of a two-wave pattern (5/6 animals) compared to that of a three-wave pattern (1/6 animals). For two-wave pattern, the first wave emerged at Day $0.20{\pm}0.19$ (Day 0 = day of ovulation) and was marked by development of a dominant anovulatory follicle which grew in size from $5.40{\pm}0.24mm$ at the day of detection to a maximum diameter of $12.40{\pm}0.81mm$ on Day $8.60{\pm}1.57$, with a growth rate of $0.88{\pm}0.17mm/day$ and then regressed, with a mean persistence of $19.40{\pm}1.54$ days. The second wave emerged at Day $9.20{\pm}1.06$ and was marked by development of a dominant ovulatory follicle which grew in size from $4.20{\pm}0.37mm$ at the day of detection to a maximum diameter of $13.80{\pm}0.37mm$ on Day $21.00{\pm}1.38$, with a growth rate of $0.66{\pm}0.12mm/day$ and then ovulated on Day $21.60{\pm}1.25$, with a mean persistence of $11.80{\pm}1.39$ days. The maximum diameters attained and the growth rates of dominant anovulatory and dominant ovulatory follicles, and the mean number of follicles ${\geq}3mm$ diameter detected at the time of emergence of first and second waves ($11.80{\pm}1.74$ and $9.00{\pm}2.81$, respectively) were not significantly different. In the animal which showed a three-wave pattern, the first, second and third waves emerged on Days 1, 10 and 19, respectively. All animals, except one had at least one subordinate follicle in the first or second or both waves. The subordinate follicles increased in diameter over a few days and then regressed. The results indicate that in buffaloes, the follicular turnover during oestrous cycle occurs predominantly in a two-wave pattern.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.47-55
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• 2003

Objectives: Bok, Bcl-2-related ovarian killer, is a proapoptotic Bcl-2 family protein identified in the ovary based on its dimerization with the antiapoptotic protein Mcl-1. The present study examined the hormonal regulation and localization of Bok messenger RNA levels in the mouse ovary during the follicle development. Methods: The animals were implanted subcutaneously with Silastic brand capsules containing the synthetic estrogen, DES at $21{\sim}23$ days of age. Ovaries were collected $1{\sim}3$ days after implantation for RNA analysis and in situ hybridization. Some mice were removed capsule for $1{\sim}2$ days to induce ovarian follicle apoptosis. Ovaries were also collected from 26 day-old immature mice at various times after treatment with 10 IU PMSG. Some mice received a single intraperitoneal injection of 10 IU hCG to induce ovulation, and ovaries were obtained at different time intervals for Northern blot and in situ hybridization analysis, respectively. Results: Treatment of immature mice with diethylstilbestrol (DES) for $24{\sim}48$ h increased ovarian Bok mRNA levels. Bok mRNA was remained the same levels in mice removed DES for $24{\sim}48$ h to induce apoptosis. High signals of Bok mRNA after DES treatment were detected in granulosa cells of early antral follicles. Treatment of immature mice with PMSG for 12 h increased markedly ovarian Bok mRNA expression which was detected mainly in preantral and atretic follicles. Interestingly, low levels of Bok mRNA were also expressed in granulosa cells of preovulatory follicles. Treatment of PMSGprimed mice with hCG stimulated strongly ovarian Bok mRNA expression at $6{\sim}9$ h. At that time, Bok mRNA was expressed in granulosa cells of atretic and small growing follicles. Conclusion: These results demonstrate that Bok is one of proapoptotic Bcl-2 members expressed in early growing and atretic follicles during the ovarian follicular development. Gonadotropins induce a transient increase of Bok gene expression in granulosa cells of preantral and preovulatory follicles indicating some role in the ovulatory process.

• Clinical and Experimental Reproductive Medicine
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• 제15권2호
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• pp.103-117
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• 1988

Ovulation induction was done with 3 different regimens as clomid combined with HMG, HMG only, and FSH combined with HMG in 28 menstrual cycles for IVF-ET and GIFT program. The appearance of endogenous LH surge, estradiol plateau, atypical LH surge, and time from initiation to peak of LH surge in serum and urine were observed and compared in 3 groups. 1. The estradiol concentration of serum LH surge day was similar in three groups but 1st group (Clomiphene Citrate+Sequential HMG) was slightly higher at $1924.0{\pm}865.1\;pg/ml$. In regards to the existence of serum estradiol plateau, 3rd group (FSH+Sequential HMG) was highest at 60%, and 1st group and 2nd group (HMG only) were similar at 33% and 44% respectively. 2. The number of ovarian of ovarian follicle which was more than 18mm in diameter was $4.1{\pm}2.0$, $4.2{\pm}2.1$ respectibely for 2nd group and 3rd group. Although the numbers were slightly higher thean 1st group for each ovarian follicle, serum estradiol value per follicle was higher for 1st group at $583.0{\pm}261.2pg/ml$. 3. When measuring the urine LH surge according to Hi-Gonavi and according to the standard set by three different types of surge, simultameous satisfaction for 1st group, 2nd group, 3rd group was two cases, five cases, four cases respectively at 40%, and the remained cases were composed of numorous type combination which satisfied the two definition, simultaneously in this study, the LH surge starting time was determined only in the cases tow combination were satisfied simultaneously at first, but there are something to study more. In one case of the 3rd group. 4. The concentration of LH surge start in urine and serum of 2nd group was highest at306. $0{\pm}287.2IU/l$ and $34.0{\pm}9.9mIU/ml$ and 1st group was low at $116.6{\pm}66.1IU/l$ and 7.4mIU/ml. The urine and serum value of LH was highest at $1644.4{\pm}988.8IU/l$, $65.9{\pm}15.0mIU/ml$ for 2nd group, 1st group was low at urine, and 3rd group was low of serum. With pregnancy established, the LH concentration of urine was relatively high but on the contrary the LH concentration of serum was low compared to urine concentration. 5. Time from LH surge start to the maximun of urine and serum value was highest at 15. $7{\pm}9.1$ hrs and $10.8{\pm}4.9$ hrs for 1st group and 3rd group. With pregnancy established, time was shortened for urine but on the contrary serum showed an increase in time. 6. The concentration of LH which increases with time on urine was highest at 2nd group ($194.6{\pm}76.8\;IU/hour$). The lowest increase for serum was at 3rd group (2.1mIU/hour). With pregnancy established, urine showed more increase than control group ($266.5{\pm}47.4\;IU/hour$) and for serum there was similar increase ($3.4{\pm}0.8\;mIU/hour$). 7. There were two examples of non-typical surge from 1st group and 3rd group each. Among these three cases showed a reestablishment of LH surge after 10-24 hours. 8. Endogenous spontaneous Lh surge occurs once for each 2nd group and 3rd group.

• 한국수정란이식학회지
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• 제24권3호
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• pp.169-176
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• 2009

Domestic bitches are non-seasonally monoestrus; spontaneously ovulate only once or twice occurs at anytime of the year. Estrus induction has been applied infrequent estrus, misleading ovulation, mating difficulties, failure to conceive after normal mating, pregnancy failure and biological research. Protocol of estrus induction which included variable hormones such as FSH, GnRH, and PMSG have been applied for the last decades. Recently, Bromocriptine, one of anti-prolatin/dopamine agonist has been occasionally applied for estrus induction. The study was carried out to investigate the effective method for the induction of estrus in bitches using different hormone treatments, and the initiation time of estrus from hormone treatment by assessments of cytological observation and blood plasma progesterone concentration. A total of 54 bitches on anestrus were selected for the study and divided randomly into 8 treatment groups as follow. Control, natural estrus; FSH (L), FSH (1.5 mg/kg, twice a day, $Falltrophin^{(R)}$, Vetrepharm); FSH (H), FSH (3.0 mg/kg, twice a day); GnRH+FSH, GnRH (5 ug/kg, once first day, $GNADON^{(R)}$, Dongbang)+FSH (3 mg/kg, SID); PMSG, PMSG (50 IU/kg, every third day $FOLLIGON^{(R)}$, Intevet); GnRH+PMSG, GnRH (5 ug/kg, only first day)+PMSG (50 IU/kg, every third day); GnRH, GnRH (5 ug/kg, only first day); Bromocriptine, bromocriptine (0.3 mg/kg, SID, $Parlodel^{(R)}$, Novartis). The bitches were evaluated clinical sign, cytological exam and $OVUCHECK^{(R)}$ Premate for assessment of estrus induction. Estrus induction rates were significantly (P<0.05) higher in GnRH+PMSG (100%) compared to others. PMSG and GnRH+PMGS (87.5 and 100%) and Bromocriptine (77.8%) were higher than others except GnRH+PMSG. Analysis of vaginal smear has proved to be effective a correct assessment of estrus induction with assay of progesterone concentration by $OVUCHECK^{(R)}$ Premate. Proestrus initiated by the $6^{th}$ after induction in most case. In conclusion, bromocriptine is an effective drug for estrus induction in bitches and assay of progesterone concentration by $OVUCHECK^{(R)}$ Premate with examination of vaginal smear that should be useful to detection of estrus induction of estrus induced bitches.

• 한국동물학회지
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• 제29권1호
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• pp.13-22
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• 1986

특정 계통의 생쥐 초기 배아의 체외 배양때에 나타나는 "In Vitro 2-Cell Block" 현상을 규명할 것을 목적으로 하고 본 실험이 행해졌다. 먼저 이 현상이 발생하는 ICR 계통의 생쥐의 수정란 또는 2세포기의 배아를 일정시간 대 (배란을 유도하기 위한 hCG주사시간을 기준)를 두고 수란관으로부터 회수한 뒤 이를 3-4일간 배양하면서 배낭으로까지의 발생능력을 알아보았다. 그 결과 hCG주사 후 약 30시간이 지난 뒤 수란관에서 회수한 수정란이나 2세포기의 배아의 일부가 배낭으로까지 발생하였으며 만일 48시간이 지나면 수란관 내에서 회수된 배아는 대부분이 2세포기 배아이며 이것들은 거의 배낭으로 발생하였다. HCG 주사 후 27시간이 지난 수란관으로부터 회수한 수정란을 2시간에서 24시간을 배양한 뒤 이들을 다시 수란관에 이식하여 기관배양법에 의해 72시간 배양하고 다시 수란관 밖에서 배아를 24시간 배양해 본 결과, 배아들이 수란관 밖의 환경에서 배양된 시간이 길수록 이것들을 다시 수란관내에 되돌려 준다 하더라도 배낭으로까지 발생할 능력을 크게 상실하였다. 이같은 실험 결과로 보아 생쥐의 수정란이 "2-Cell Block" 현상을 극복하기 위해서는 수정후 일정시간 이상을 수란관이라는 환경내에 머물러 있어야 한다는 것을 알게 되었다. 즉, 배아는 수란관에 오래 머물러 있을수록, 그리고 수란관으로부터 축출되더라도 다시 수란관으로 돌려보내질 때까지 밖에 머물러 있는 시간이 짧을수록, 수정란 혹은 2세포기 배아의 배낭으로의 발생능력은 정상에 가깝게 유지되는 것이다. "2-Cell Block"에 걸려있는 2세포기의 배아는 배양 후 24시간까지에는 광학현미경적인 관찰 결과 정상적인 형태를 보여주고 있었으나 48시간이 되면 핵의 이상응축이 나타나며 72시간이 경과하면 세포질 내에 비정상적인 공포들이 나타나고 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제13권9호
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• pp.1205-1209
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• 2000

The present study was undertaken to investigate the effect of stimulation of follicular development with eCG on the peripheral levels of inhibin and FSH in Murrah buffaloes. Estrus was synchronized in five normally cycling females by insertion of Crestar (Intervet, Boxmeer, Holland) implants for nine days. Estradiol valerate was administered i.m. on the day of implant insertion. On the 10th day of the induced estrous cycle a single dose of 3000 IU eCG (Folligon, Intervet, Boxmeer, Holland) was given, followed by treatment with 25 mg of $PGF_2$ alpha (Lutalyse, Upjohn, Belgium) 48 h later. Blood samples were obtained during the induced estrus, on cycle day 10 (luteal phase), at the superovulatory estrus (43 h after PGF) and during the periovulatory period (64 h after PGF). Ultrasonography was done daily to monitor follicular development. Plasma concentrations of inhibin and FSH were determined by specific radioimmunoassays. Differences between $mean{\pm}SEM$ values of different phases of the cycle were compared by ANOVA. The mean number of small (2-5 mm), medium (6-9 mm) and large (>10 mm) follicles observed two days after eCG treatment and on the day of superovulatory estrus was $2.8{\pm}0.31$, $5.2{\pm}0.30$ and $1.4{\pm}0.09$ and $1.9{\pm}0.21$, $2.8{\pm}0.40$ and $5.0{\pm}0.83$, respectively. The mean number of ovulations was $3.6{\pm}0.37$ and the mean number of unovulated follicles was $6.1{\pm}0.47$. Most of the follicles >10 mm in diameter had ovulated (72%). The mean ${\pm}SEM $ of plasma inhibin concentration $(2584.15{\pm}17.92pg/ml)$ during the superovulatory estrus was significantly higher $(p{\leq}0.05)$ than during the induced estrus $(749.87{\pm}17.29pg/ml)$, the luteal phase $(1099.54{\pm}24.98pg/ml)$ and periovulatory period $(1682.71{\pm}29.88pg/ml)$, respectively. $Mean{\pm}SEM$ plasma FSH concentration during the induced estrus $(10.35{\pm}0.41ng/ml)$ was not different from that during the superovulatory estrus $(8.52{\pm}0.39ng/ml)$, but was significantly higher $(p{\leq}0.05)$ than during the luteal phase $(2.81{\pm}0.42ng/ml)$ and periovulatory period $(5.7{\pm}0.28ng/ml)$. These data indicate that treatment with eCG in buffaloes for inducing superovulation results in a significant elevation in plasma inhibin levels and a decrease in plasma FSH levels during the superovulatory estrus. Thus, we suggest that the elevated plasma inhibin coming from fully developed follicles continued for a long time which results in inhibition of FSH leading to poor ovulation in the remaining follicles, which may be the cause of suboptimal superovulatory response.

• 한국수정란이식학회지
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• 제27권3호
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• pp.141-147
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• 2012

This study was operated to establish induction using ultrasonography by estimating the relation of follicle size and estrus manifestation. Clinical estrus symptoms were observed 97.4% in cows and 87.5% in heifers when overall 55 cows were induced to estrus in a single dose of $PGF_{2{\alpha}}$ after verifying CL through ultrasonography, which means estrus hours among those 52 cows showing the clinical estrus symptoms were estimated 2.39 days on cows and for 2.37 days on heifers which showed no differences (p>0.05). The estrus manifestation hours according to the follicle size in cows didn't have any significance each other (p>0.05), though estrus hours was 54 hours (the shortest) with follicle size bigger than 10 mm and were made up within 69 hours. The estrus manifestation hours according to the follicle size in heifers didn't have any significance each other (p>0.05) and took around 42 hours (the shortest) with follicle size of 5mm (the smallest) and were made up within 66 hours. Follicles after $PGF_{2{\alpha}}$ injection were ovulated and assigned to many phases as follows; Group 1 (growing phase) - continuously growing into ovulation, Group 2 (growing and static phase) - delaying in growth after the growth of follicles, Group 3 (static and growing phase) - growing after growth delay, Group 4 (regressing and new growing phase) - the follicle is closed and a new follicle grows. In addition, the process of follicle development and estrus hours had no significance each other (p>0.05), though estrus manifestation hours in Group 1 and 2 was relatively short, and in Group 3 and 4 for a relatively long time. In the result of all above, the estrus manifestation hours after $PGF_{2{\alpha}}$ injection has no differences accoring to the follicle size in cows and heifers. Therefore, High pregnancy rate is obtained when practicing artificial insemination within 3 days in estrus or TAI in 72 to 80 hours after adminitrating $PGF_{2{\alpha}}$.