• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.33-39
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• 2017

Objective: The aim of this study was to assess the changes of follicular fluid (FF) and serum levels of cerebellin precursor protein 1 (cbln1) and betatrophin in patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) with a gonadotropin-releasing hormone (GnRH) antagonist protocol. Methods: Twenty infertile women with PCOS and 20 control women diagnosed as poor responders undergoing ovarian stimulation with a GnRH antagonist were included. Blood samples were obtained during ovum pick-up. Follicular fluid from a dominant follicle was collected from the subjects. Using enzyme-linked immunosorbent assays, FF and serum levels of cbln1 and betatrophin were measured in both groups of participants. Metabolic and hormonal parameters were also determined and correlated with each other. Results: Both groups of women had similar serum and FF betatrophin levels ($55.0{\pm}8.9ng/mL$ vs. $53.1{\pm}10.3ng/mL$, p=0.11). The serum and FF betatrophin levels of poor responders were found to be similar ($49.9{\pm}5.9ng/mL$ vs. $48.9{\pm}10.7ng/mL$, p=0.22). Conversely, the FF cbln1 levels of PCOS women were found to be significantly higher than the serum cbln1 levels ($589.1{\pm}147.6ng/L$ vs. $531.7{\pm}74.3ng/L$, p<0.02). The FF cbln1 levels of control participants without PCOS were significantly higher than their serum cbln1 levels ($599.3{\pm}211.5ng/L$ vs. $525.3{\pm}87.0ng/L$, p=0.01). Positive correlations were detected among body mass index, insulin resistance, serum insulin, total testosterone, and betatrophin levels in the PCOS group. Conclusion: Follicular fluid betatrophin and cbln1 concentrations may play a pivotal role on follicular growth in PCOS subjects undergoing IVF/ICSI with an antagonist protocol.

• 한국임상수의학회지
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• 제9권2호
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• pp.373-390
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• 1992

To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.

• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.488-500
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• 2013

The objective of this study was to compare the effectiveness of the protocols for superstimulation of follicular growth in Thai native heifers. Heifers (n = 20) were randomly divided into four groups of five heifers/group. Heifers were given a single dose by i.m. administration of 100 mg Follicle Stimulating Hormone dissolved in polyvinylpyrrolidone (FSHp) at 24 h. Ovum pick up (OPU) occurred at 72 h ($F_{24}O_{72}$ protocol; Group 1) or 96 h ($F_{24}O_{96}$ protocol; Group 2), and at 36 h and OPU at 72 h ($F_{36}O_{72}$ protocol; Group 3) or 96 h ($F_{36}O_{96}$ protocol; Group 4) after follicular ablation. The dynamics of ovarian follicular growth were monitored by twice-daily ultrasonographic examinations. Blood sample collections were performed every 12 h after initiation of treatment for assessment of FSH, E2 and P4 profiles. All heifers were subjected to eight repeated sequential sessions of OPU. The follicular deviation commenced $24{\pm}5.32$ h after follicular ablation in all groups. The circulatory FSH surged quickly from 24 to 36 h (>0.8 ng/ml) after follicular ablation and circulatory estrogen levels steadily increased from 36 h until OPU in all groups. At the end of the OPU sessions, the mean number of aspirated follicles/heifer/session in $F_{36}O_{72}$ protocol (Group 3) and $F_{36}O_{96}$ protocol (Group 4) were higher than in the two other groups (p<0.05). The number of cumulus-oocyte complexes (COCs), cleaved and day 8 blastocysts rates in the $F_{36}O_{72}$ protocol (Group 3) were higher than in the other groups (p<0.05). It can be concluded that a single dose i.m. administration of 100 mg FSHp at 36 h and OPU at 72 h after follicular ablation ($F_{36}O_{72}$ protocol; Group 3) was the most effective protocol for superstimulation of follicular growth for repeated OPU and subsequent in vitro embryo production in Thai native heifers.

• 한국수산과학회지
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• 제22권5호
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• pp.342-350
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• 1989

낙동강 하구지역에 서식하는 자주새우를 대상으로 1988년 6월부터 1989년 5월까지 성성숙과 산란에 대하여 조사한 결과는 다음과 같다. 1. 난소는 두흉부의 심장 하방에 위치하며 심장을 중심으로 전방은 촉각동맥 및 안동맥을 둘러싸고 둥글게 융합되어 있고, 후방은 1쌍의 관상으로 복부 2마디까지 뻗혀있다. 정소는 심장직하에 위치하며 심장을 중심으로 전방은 관상으로 곡정관의 형태를 취하고 있으며 후방은 1쌍의 관상의 형태를 띠고 있고, 이곳으로부터 연결된 저정낭이 제5흉지에 개구되어 있다. 2. 난자 및 정자의 배우자형성은 계절과 관계없이 단기간에 반복되고 있으며, 생식소상피의 전 구역에서 동시에 이루어지고 있다. 완숙난의 장경은 $430{\mu}m$ 전후이고, 변태된 정자는 두부 핵질이 뚜렷한 타원형의 구조를 나타낸다. 3. 생식주기는 약 40일을 간격으로 단주기성이며, 11월을 전후한 동계를 제외하고는 주년 산란종이다. 4. 포란된 난의 부화기간은 수온 $21^{\circ}C\~24^{\circ}C$의 실내사육에서 $12\~14$일이 소요되었다. 5. 포란된 개체의 평균 부화유생수는 1,602마리였고, 전장 32mm에서 1,372마리, 38.2개개에서 1,650마리였다.

• 한국가축번식학회지
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• 제24권1호
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• pp.83-88
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• 2000

본 연구는 한우에 있어서 초음파기기를 이용하여 반복적으로 생체내 난자를 회수시 난자회수에 미치는 영향과 시술후의 난소 유착과 같은 부작용을 조사하기 위하여 실시한 결과는 다음과 같다. 1. 난포란의 채란빈도에 따른 채란수에서는 주 2회 시술시 난포수는 8.7$\pm$4.2개로 주 1회 시술의 10.2$\pm$6.1개보다 다소 적었으나 유의적인 차이는 인정되지 않았으며, 채란수도 각각 4.1$\pm$3.4개 및 4.3$\pm$2.9개로 유의적인 차이가 인정이 되지 않았다(P＜0.05). 2. 반복적으로 9회 난포란을 채란한 경우 40개의 난소중 8개 (20%)가 유착이 발생하였으며 1~3 채란시 3개 (37.5%), 4~6 채란시 4개 (50%) 7~9회 채란시 1개(12.5%)가 유착이 발생하였다. 3. 반복적인 난포란 채란후 난소의 유착 여부에 따른 공란우의 발정주기를 조사한 결과 난소 유착을 보인 공란우의 8두중 7두(87.5%) 25일 이상의 장발정주기를 보였으며, 유착이 일어나지 않은 12두 중 정상 발정주기는 6두(50.0%), 단 발정주기는 1두(8.3%), 장발정주기는 5두(41.7%)였다.

• 한국수정란이식학회지
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• 제27권4호
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• pp.229-235
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• 2012

This research was investigated the relationship, in high-producing Holstein donor cows, between the number of the transferable embryos and the blood serum concentrations of Blood Urea Nitrogen (BUN), glucose and cholesterol, which affect the nutritional state of cows. CIDRs were inserted into the vaginas of twenty two heads of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_{2{\alpha}}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. The total numbers of ova collected from 3 experimental groups whose blood BUN concentrations were <10 mg/dl, 11~18 mg/dl and ${\geq}19$ mg/dl were 8.9, 12.5 and 19.0, respectively; whereas the numbers of transferable embryos were 5.8 + 1.9, 7.9 + 2.8 and 5.2 + 1.4, respectively. When glucose concentration was <60 mg/dl, the total number of collected ova was 9.9, which was smaller than when the concentration was 60~70 mg/dl or ${\geq}70$ mg/dl. When glucose concentration was 60~70 mg/dl, the number of transferable embryos was 7.1 + 2.4, which was slightly larger than the numbers 6.4 + 2.1 and 6.1 + 1.7 that were obtained when the concentrations were <60 mg/dl and ${\geq}70$ mg/dl, respectively ; however, the differences were not significant (p>0.05). When cholesterol concentrations were <150 mg/dl, 150~200 mg/dl and ${\geq}200$ mg/dl, the total numbers of collected ova were 11.2, 11.3 and 8.6, respectively. Whereas the numbers of transferable embryos were 7.1 + 2.1, 7.3 + 1.9 and 5.6 + 1.3, respectively ; however, the differences were again not significant (p>0.05). The result of this research showed no significant difference in ovum recovery rate and the number of transferable embryos according to major metabolite concentrations in high-producing Holstein donor cows. However, it is considered that the failure of maintaining proper nutritional status would cause the fall in in vivo embryo productivity.

• 한국수정란이식학회지
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• 제27권4호
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• pp.237-243
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• 2012

This research was investigated the relationship between the number of the transferable embryos and estrus expression rate, BCS (Body Condition Score), which affect the nutritional state of the cow, in Holstein donor cows. CIDRs were inserted into the vaginas of twenty two head of Holstein cows, regardless of estrous cycle. Superovulation was induced using folliclar stimulating hormone (FSH). For artificial insemination, donor cows were injected with $PGF_{2{\alpha}}$ and estrus was checked about 48 hours after the injection. Then they were treated with 4 straws of semen 3 times, with 12-hour intervals. Embryos were collected by a non-surgical method 7 days after the first artificial insemination. When BCS was $$\leq_-$$2.5, the total number of collected ova was 7.3 + 1.9, which is significantly lower (p<0.05) than the numbers 15.4 + 2.8 and 15.4 + 2.1 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively. Whereas the numbers of transferable embryos were 5.2 + 1.4 when BCS was $$\leq_-$$2.5, which was smaller than the numbers 6.0 + 2.1 and 8.5 + 1.8 that were obtained when BCSs were 2.75 and $$\geq_-$$3.0, respectively; however, the differences were not significant. As for estrus induction rate, the cow groups whose BCSs were 2.75 and $$\geq_-$$3.0 showed 100.0% and 95.0%, respectively. Whereas the cow group whose BCS was $$\leq_-$$2.5 showed 57.1%, and the differences were significant (p< 0.05). As for estrous expression rate, the cow groups whose BCSs were $$\leq_-$$2.5, 2.75 and $$\geq_-$$3.0 showed 100.0%, 100.0% and 85.7%, respectively; however, the differences were not significant. According to the result of this research, it is considered that the total number of collected ova and the number of transferable embryos will be affected by the nutritional state before and after in vivo embryo production and superovulation treatment, and that although the mechanism is not clear, poor stockbreeding management and nutritional level would cause the decrease of ovum recovery rate and the number of transferable embryos in high-producing cows. On the other hand, diverse researches on the superovulation treatment method that is suitable for high-producing Holstein donor cows would contribute to preventing ovarian cyclicity disorder, as well as to the early multiplication of cows with superior genes by increasing the utilization value of donor cows.

• 한국어류학회지
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• 제20권4호
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• pp.255-262
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• 2008

멸종위기야생동식물 1급으로 지정된 꼬치동자개, Pseudobagrus brevicorpus의 종복원을 위해 인위적인 교배를 통하여 획득한 자치어를 대상으로 성분화 과정을 조사하여 성적임성 여부를 확인하였다. 부화 후 4~5일째 자어에서 생식융기 1쌍이 장과 중신관 사이에서 관찰되었고, 그 후 유사분열이 활발히 진행되었다. 부화 후 20일째 치어에서 난원세포의 출현으로 난소 분화가 시작되었으며, 부화 후 30~40일째를 거치면서 염색인기와 주변인기의 난모세포로 성장하였다. 난황은 부화 후 80일째의 자어에서 생성되기 시작하였고, 부화 후 100일째가 되어 eosin에 염색되는 난황구가 생성되었다. 이후 난모세포는 난황이 지속적으로 축적되면서 세포의 크기와 수가 증가하기 시작하였고, 부화 후 약 10개월째에는 난막과 여포세포층에 의해 둘러싸여진 난모세포로 분화하였다. 한편 정소는 부화 후 25일경 치어에서 분화되기 시작하였고, 40일경 유사분열을 통해 정원세포를 둘러싸는 포낭을 형성하였다. 부화 후 100일째가 되면 정소는 정소엽 구조를 형성하였고 hematoxylin에 매우 진하게 염색되는 정모세포로 분화하였다. 부화 후 7개월째 치어의 정소엽 내부에 정세포가 출현한 이후, 부화 후 만 1년을 경과한 치어에서 최초로 성숙한 정자가 출현하였다. 위 결과를 바탕으로 인공적으로 부화된 꼬치동자개 자치어의 성분화는 다른 종에서 보이는 것과 같이 정상적인 발달을 보인다고 판단되었다.

• Applied Microscopy
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• 제29권2호
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• pp.241-253
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• 1999

북부 온대지방에 서식하는 박쥐는 그 대부분이 동면을하며, 그 생식유형은 특이하여 가을에 교미하여 사정된 정자는 자성생식도내에 긴 동면기간 동안 저장되어 있는 것으로 알려져 있다(예: 한국큰관박쥐). 환언하면 저장되어 있던 정자는 이듬해 봄에 배란되는 난자와 수정하여 초여름에 출산하게 된다. 본 연구는 월동하는 정자 두부 Plasma membrane의 성분과 활성 등 형태적 및 기능적 특성을 관찰 분석함으로써, 인공임신율을 높일 수 있는 보다 효과적인 한냉온도 정자저장방법을 모색하기 위하여 시도되었다. 동면하는 한국큰관박쥐와 햄스타 및 비동면동물(마우스)의 정자 Plasma membrane이, 실온과 한냉온도(박쥐-동면온도)에서, 생존율, 첨체반응을, 단백질분포, 능동수송에 관여되는 효소$(Na^+-K^+-ATPase)$의 활성 및 주사전자현미경적 조직화학상을 분석하였다. 본 연구에서 얻은 실험결과에 의거하면, 동면동물인 한국큰관박쥐와 햄스타 및 비동면동물인 마우스 정자의 생존율과 첨체반응율은, 실온과 한냉(박쥐-동면)온도간에 의의있는 차이를 발견할 수 없었고, 단백질 분포와 $Na^+-K^+-ATPase$ 효소활성 및 주사전자현미경적 조직화학적 소견에 있어서도 두 온도간에 의의있는 차이는 없었다. 이와 같이 실온과 한냉온도 간에 형태적 기능적 차이가 없다고 하는 사실은, 동면동물이든 비동면동물이든 Spermatozoa plasma membrane이 동면중 또는 동면 이후에 안정적인 수정을 보장해 주는 형태적 및 기능적 보호구조임을 시사해 준다.

• Journal of Animal Science and Technology
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• 제65권2호
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• pp.324-335
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• 2023

Korea, located in East Asia in the northern hemisphere, is experiencing severe climate changes. Specifically, the heat stress caused by global warming is negatively affecting the dairy sector, including milk production and reproductive performance, as the major dairy cattle Holstein-Friesian is particularly susceptible to heat stress. Here, we collected artificial insemination and pregnancy data of the Holstein and the Jersey cows from a dairy farm from 2014 to 2021 and analyzed the association between the conception rate and the temperature-humidity index, calculated using the data from the closest official weather station. As the temperature-humidity index threshold increased, the conception rate gradually decreased. However, this decrease was steeper in the Holstein breed than in the Jersey one at a temperature-humidity index threshold of 75. To evaluate the effects of heat stress on the oocyte quality, we examined the nuclear and cytoplasmic maturation of Holstein (n = 158, obtained from six animals) and Jersey oocytes (n = 123, obtained from six animals), obtained by ovum pick-up. There were no differences in the nuclear maturation between the different conditions (heat stress: 40.5℃, non- heat stress: 37.5℃) or breeds, although the Holstein oocytes seemed to have a lower metaphase II development (p = 0.0521) after in vitro maturation under heat stress conditions. However, we found that the Holstein metaphase II oocytes exposed to heat stress presented more reactive oxygen species and a peripheral distribution of the mitochondria, compared to those of the Jersey cattle. Here, we show that weather information from local meteorological stations can be used to calculate the temperature-humidity index threshold at which heat stress influences the conception rate, and that the Jersey cows are more tolerant to heat stress in terms of their conception rate at a temperature-humidity index over 75. The lower fertility of the Holstein cows is likely attributed to impaired cytoplasmic maturation induced by heat stress. Thus, the Jersey cows can be a good breed for the sustainability of dairy farms for addressing climate changes in South Korea, as they are more resistant to hyperthermia.