• 한국수정란이식학회지
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• 제31권2호
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• pp.109-116
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• 2016

The recovery of epididymal sperm in animals is considered as one of the important tools to preserve high value or endangered species. However, there are no appropriate castrating indicators such as months of age in bull, sperm morphology, and motility, particularly in young Korean native bull (Hanwoo). Therefore, this study aimed to investigate sperm number, morphology, and motility of sperm in the epididymis tail of young Hanwoo bulls at 8 and 15 months of age. After castration, epididymal tails were collected and minced with blades to recover sperm. In experiments 1 and 2, sperm number, morphology, and motility were examined. Total number of sperm and percentage of normal sperm from bulls at 8 months of age was lower than that of bulls at 15 months of age after collection (P<0.05). Percentage of abnormal head, tail, proximal cytoplasmic droplet, dead and damaged acrosome of sperm from bulls at 8 months of age were higher than those of bulls at 15 months of age (P<0.05). In experiment 3, sperm motility from bulls at 8 and 15 months of age were examined before freezing and after thawing. Frozen-thawed sperm at 8 months of age showed low total motility and motile sperm with ${\geq}25{\mu}m/sec$ compared to those at 15 months of age and commercially-used sperm (P<0.05). In conclusion, sperm derived from the epididymal tail of bulls at 8 months of age showed high abnormal morphology and poor motility, which are not adequate for AI and IVF. On the other hand, sperm derived from the epididymal tail of bulls at 15 months of age showed high normal morphology and motility.

• Asian-Australasian Journal of Animal Sciences
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• 제23권4호
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• pp.430-436
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• 2010

The objective of the present studies was to develop and validate a system for isolation, purification and extended culture of pigment-producing cells in alpaca skin (melanocytes) responsible for coat color and to determine the effect of alpha melanocyte stimulating hormone treatment on mRNA expression for the melanocortin 1 receptor, a key gene involved in coat color regulation in other species. Skin punch biopsies were harvested from the dorsal region of 1-3 yr old alpacas and three different enzyme digestion methods were evaluated for effects on yield of viable cells and attachment in vitro. Greatest cell yields and attachment were obtained following dispersion with dispase II relative to trypsin and trypsin-EDTA treatment. Culture of cells in medium supplemented with basic fibroblast growth factor, bovine pituitary extract, hydrocortisone, insulin, 12-O-tetradecanolphorbol-13-acetate and cholera toxin yielded highly pure populations of melanocytes by passage 3 as confirmed by detection of tyrosinase activity and immunocytochemical localization of melanocyte markers including tyrosinase, S-100 and micropthalmia-associated transcription factor. Abundance of mRNA for tyrosinase, a key enzyme in melanocyte pigment production, was maintained through 10 passages showing preservation of melanocyte phenotypic characteristics with extended culture. To determine hormonal responsiveness of cultured melanocytes and investigate regulation of melanocortin 1 receptor expression, cultured melanocytes were treated with increasing concentrations of ${\alpha}$-melanocyte stimulating hormone. Treatment with ${\alpha}$-melanocyte stimulating hormone increased melanocortin receptor 1 mRNA in a dose dependent fashion. The results demonstrated culture of pure populations of alpaca melanocytes to 10 passages and illustrate the potential utility of such cells for studies of intrinsic and extrinsic regulation of genes controlling pigmentation and coat color in fiber-producing species.

• 한국수산과학회지
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• 제17권6호
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• pp.523-528
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• 1984

자연에서 5월중순부터 10월초기까지 산란기를 가지는 그물코쥐치, Rudarius ercodes를 대상으로 이들의 생식주기성립기구를 밝히고저 여러가지 광주기 및 온도 조건이 생식소의 활성과 퇴화 그리고 재성숙에 미치는 영향을 실험적으로 조사하였다. 이른봄에 생식소는 장일광주기(13L이상)에 의해서 활성화되어 산란기에 이르게 되며, 이때 수온상승이 보상적으로 촉진역할을 한다. 일단 임계일장($12L{\sim}13L$)이상이 작용되어 생식소 활성화가 일어난 개체들은 임계일장 이하에 되돌려도 생식소는 계속 성숙하였다. 산란종료기에는 수온에 관계없이 단일(12L)적으로 생식소 퇴화가 초래되고 있어 자연에서 일장단축이 산란기를 종료시키는 요인임을 알 수 있다. 산란기를 종결하고 퇴화중인 생식소에서도 $20^{\circ}C$ 수온의 장일광주기(13L이상)에서 재성숙하여 산란까지 이르고 있어 연중 생식주기의 인공적인 제어가 가능함을 시사하고 있다. 이때 장일광주기일지라도 $28^{\circ}C$이상의 고수온구는 성숙억제현상이 나타났다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권7호
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• pp.969-976
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• 2009

Use of oocytes from prepubertal animals for in vitro embryo production holds potential application for reducing generation intervals and increasing genetic progress through embryo transfer. The objective of these studies was to compare the effect of three sperm pretreatments (prior to in vitro fertilization) and seven embryo culture protocols on fertilization rate and (or) subsequent development of in vitro fertilized embryos derived from oocytes harvested from ovaries of 1-6 month old prepubertal Boer goats in China. Cleavage rates were highest for embryos fertilized with heparin-treated versus calcium ionophore- or caffeine-treated sperm. Similar rates of blastocyst development were observed using heparin- and ionophore-treated sperm, which were higher than obtained with caffeine-treated sperm. No differences in cleavage or blastocyst rates were observed following embryo culture in basal medias (synthetic oviductal fluid (SOF), Charles Rosenkrans 1 (CR1) or tissue culture medium-199 (TCM-199)) containing 10% fetal bovine serum (FBS). Cumulus or oviductal cell co-culture did not enhance cleavage or blastocyst rates relative to culture in SOF+10% FBS. Replacement of FBS in SOF medium with 0.3% BSA increased cleavage rates, but did not increase rates of blastocyst development. Sequential culture in SOF+0.3% BSA followed by SOF+10% FBS increased blastocyst yield versus continuous culture in SOF+10% FBS and tended to increase blastocyst yield versus continuous culture in SOF+0.3% BSA. These results demonstrate a pronounced effects of sperm pretreatments and in vitro embryo culture systems on rates of blastocyst development and provide a potential protocol (sperm pretreatment with heparin and sequential embryo culture in SOF+0.3% BSA followed by SOF+10% FBS) for generation of the significant numbers of in vitro produced blastocysts from oocytes of prepubertal Boer goats necessary for application of embryo transfer in rural regions of China for distribution of Boer goat genetics.

• Journal of Animal Science and Technology
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• 제48권5호
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• pp.651-662
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• 2006

프탈레이트 에스테르는 플라스틱 가소제로서 이용되며 또한 유제품과 같은 음식에서 미량으로 발견되고, 종종 내분비 교란물질로 의심되고 있다. 본 연구의 목적은 DBP, DINP 또는 DEHA의 주산기 노출이 랫트에 있어 성 성숙 후, 번식기능 특히 뇌의 성분화에 어떤 영향을 끼치는 지에 대해 조사하였다. 이를 수행하기 위해서, 어미에게 식물성 에스트로겐의 함유가 낮은 분말 사료에 다음과 같은 단계적 농도의 DBP (20, 200, 2000, 10000 ppm), DINP (40, 400, 4000, 20000 ppm), DEHA (480, 2400, 12000 ppm)를 혼합한 후, 임신 15일째부터 출생 후, 21일째 (이유기)까지 섭취 시켰고, 성 성숙 후, 혈청 성호르몬 및 성선자극호르몬의 레벨과 교배행동 및 성주기 회귀를 분석하였다. 그 결과, DBP, DINP 또는 DEHA의 주산기 노출에 의한 생후 20~21주째의 암수 랫트에 있어, 성호르몬 및 성선자극호르몬의 레벨뿐만 아니라 암컷의 성주기의 회귀에 대해 어떠한 영향을 주지 않았다. 이것은 시상하부－하수체－성선축의 내분비계를 제어하는 뇌의 성분화에는 이들 화학물질이 영향을 주지 않았다는 사실을 시사한다. 하지만, 수컷의 성행동 특히, 사정 (ejaculation)과 암컷의 로도시스 반응이 억제되는 것이 관찰되었다. 이러한 결과로부터 DBP, DINP 또는 DEHA의 주산기 노출은 성선자극 호르몬의 분비에는 영향을 주지 않지만, 성행동을 제어하는 시상하부의 어떤 영역에 직접적으로 작용할 가능성 즉, 뇌의 성분화에 영향을 끼쳐 성 성숙 후, 성 특이적 행동을 억제시킬 가능성을 시사한다.

• Reproductive and Developmental Biology
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• 제30권3호
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• pp.181-187
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• 2006

최근 돼지의 장기를 사람에게 이식하는 이종간 장기 이식에 관한 연구가 급속히 발전되고 있다. 그러나 돼지의 장기를 이식할 경우 가장 큰 문제점 중의 하나는 돼지 genome 내에 존재하는 내인성 레트로바이러스(porcine endogenous retrovirus; PERV)가 인간에게 그대로 전이될 수 있다는 것이다. 이에 대한 대안으로 최근 활발히 연구되고 있는 RNA 간섭을 통한 PERV RNA의 발현을 최대한 억제하는 방법이 제안되고 있는데, RNA 간섭(RNA interference)은 double-standard RNA(dsRNA)가 상보적인 표적 mRNA를 분해하여 결과적으로 표적 단백질의 발현을 특이적으로 억제하는 현상을 의미한다. 본 연구에서는 PERV에 대한 RNA 간섭 현상을 일으키는 shRNA 유전자를 레트로바이러스 벡터를 이용하여 돼지세포에 RNA)가 상보적인 표적 mRNA를 분해하여 결과적으로 표적 단백질의 발현을 특이적으로 억제하는 현상을 의미하다. 도입한 후 PERV의 발현율 감소 여부를 조사하였다. 그 결과, gag-pol 유전자와 env 유전자 발현은 각각 대조군 세포의 4%와 10% 정도로 억제되었다. 한편, virus 입자의 생산에서 gag-pol 유전자는 대조군 세포에 비해 300배 이상 억제되었으며, env 유전자에서는 20만 배 이상 억제되었다. 이상의 결과를 미루어 볼 때 형질 전환 돼지를 이용한 이종 장기 이식에 있어서 RNA 간섭 현상을 이용한 PERV의 발현을 억제하는 시도는 생물학적안전성을 크게 증가시킬 수 있을 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제44권4호
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• pp.214-223
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• 2017

Objective: In vitro fertilization (IVF) is a well-known method for the treatment of infertility. The present study aimed to compare the differences between infertile women with successful and unsuccessful IVF outcomes regarding the expression of T helper (Th) cell transcription factors and a group of related cytokines before and after exposure to their husbands' seminal plasma. Methods: This study was performed on 19 couples with unexplained infertility undergoing IVF treatment. Among the studied group, nine and 10 couples had successful and unsuccessful IVF outcomes, respectively. This study was carried out using real-time polymerase chain reaction. Results: Before seminal plasma exposure, the expression levels of T-bet (p< 0.007), $interferon-{\gamma}$ (p= 0.013), and tumor necrosis factor $(TNF)-{\alpha}$ (p= 0.017) were higher in the infertile women with IVF failure than in those with successful IVF outcomes, while those of GATA3 (p< 0.001), Foxp3 (p= 0.001), and interleukin (IL)-35 (p< 0.003) were lower. After seminal exposure, the expression of T-bet (p= 0.02), Rorc (p< 0.001), $TNF-{\alpha}$ (p= 0.001), Foxp3 (p= 0.02), and $interferon-{\gamma}$ (p= 0.001) increased in the unsuccessful IVF group, while the expression of Foxp3 (p= 0.02), Rorc (p< 0.001), IL-23 (p= 0.04), IL-17 (p= 0.02), IL-6 (p< 0.001), transforming growth $factor-{\beta}$ (p= 0.01), and IL-35 (p< 0.001) increased in the successful IVF group. Conclusion: In summary, IVF failure was associated with imbalanced Th1/Th2/Th17/Treg responses. Moreover, our results show that seminal plasma might have a positive effect on IVF outcomes via changes in peripheral blood T cell subsets.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제23권8호
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• pp.1007-1012
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• 2010

The aim of the present study was to describe whether abnormal cervical mucus discharge (A-CMD) or pathogens in cervical mucus discharge (CMD) have effects on reproductive performance of cows and heifers in estrus. Animals having clear discharges (68 cows, 38 heifers) with normal viscosity and without bad odor were grouped as normal cervical mucous discharge (N-CMD) group. The other animals (84 cows, 32 heifers) were grouped as A-CMD group. Microorganisms isolated from samples were divided into three groups as uterine pathogens (UP), potential uterine pathogens (PUP) or opportunistic uterine pathogens (OUP). Presence of PUP was associated with A-CMD for both cows (p<0.01) and heifers (p<0.02). First service conception rates (FS-CR) were lower in cows positive for PUP (p<0.01). Moreover, presence of PUP and OUP affected FS-CR in heifers (p<0.01). Although A-CMD significantly affected FS-CR in cows (p<0.04), it did not affect FS-CR in heifers. Differences in average open day for cows (p<0.02) and first service age for heifers (p<0.01) were significant between N-CMD and A-CMD groups, respectively. The current study suggested that CMD should be evaluated more carefully when there are infertility problems. In addition to the known microorganism that causes sterility and infertility in the UP group, pathogens in the PUP group should be considered for their potential to cause infertility.

• Reproductive and Developmental Biology
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• 제37권1호
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• pp.29-33
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• 2013

The objective of this work was to analyze the concentrations of progesterone (P4) and estrogen (E2) hormones changed during estrus synchronization in dairy heifers. Estrus synchronization was carried out with CIDR$^{(R)}$ (Controlled Intravaginal Drug Release) devices. Corpus luteum (CL) was classified into three grades based on its size and palpable characteristics. The concentrations of P4 and E2 were measured by enzyme-amplified chemiluminescence. Serum P4 concentration was markedly low at the estrus stage (36 hrs after removal of CIDR) compared to other stages, while E2 concentration was kept high during estrus stage. The serum P4 concentration was highest in the CL classified into grade I. These results indicate that P4 concentration could be used as a criteria for determining recipients for artificial insemination or embryo transfer in dairy cattle.