• Reproductive and Developmental Biology
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• 제30권3호
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• pp.189-194
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• 2006

Sperm-mediated gene transfer(SMGT) can be used to transfer exogenous DNA into the oocyte at fertilization. The main objective of this study was to assess efficiency of transferring mitochondrial DNA(mtDNA) fragment into boar spermatozoa in either presence or absence of liposome and quality of transfected spermatozoa. The mtDNA of chicken liver was isolated and purified by phenol and alkaline lysis extraction, and it was inserted to plasmid. The genome of transfected spermatozoa treated with DNase I was purified by alkaline lysis, and then amplified by the PCR analysis. After electrophoresis, DNA quantitation of each well was calculated by comparison of the band intensity with standard. As a result, exogenous DNA was composed of mtDNA fragment(1.2 kb) and plasmid(2.7 kb). On the other hand, efficiency of transfection by liposome($9.0{\pm}0.34ng/{\mu}l$) in SMGT was higher than that by DNA solution($6.9{\pm}0.53ng/{\mu}l$). However, there was no significant difference. Transfering exogenous DNA into spermatozoa was completed within 90 min of incubation. In another experiment, there were significant (p<0.05) differences between transfected spermatozoa using both DNA solution and DNA/liposome completes with unheated spermatozoa for viability ($70.8{\pm}1.80$ and $68.0{\pm}2.16%$ vs. $83.3{\pm}1.69%$, respectively) and motility($78.7{\pm}1.59$ and $79.3{\pm}2.14%$ vs. $86.7{\pm}1.59%$, respectively). This study indicates that exogenous mtDNA can be efficiently transferred into boar spermatozoa regardless of the presence of liposome, and transfected spermatozoa can also use insemination and in vitro fertilization to generate transgenic pig.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.110-110
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• 2003

The high incidence of polyspermic fertilization is one of the major causes lowering the overall efficiency of porcine IVF. The common procedure for IVF involves the co-culture of both gametes in the medium drop, which increases sperm concentration and incidence of polyspermy. Therefore, the present study was carried out to increase the efficiency of porcine IVF by reducing polyspermy using a modified swim-up method. This method modifies conventional swim-up washing by placing oocytes directly at the time of washing. Sperm pellet was prepared in the tube and mature oocytes were placed on cell strainer with $70 \mu m$ pore size (Falcon 2350) at the top of the tube. After insemination, the oocytes were stained for examination. Also, the developmental potential of fertilized embryos was measured to evaluate for the feasibility of this method. While having similar penetration rates in both methods ($86.67 \pm 2.36% to 83.33 \pm 1.36%$), there was a significant reduction of polyspermy in modified swim-up method ($17.50 \pm 1.60%$) compare to the control ($44.1 \pm 3.70%$ (p<0.05). Subsequent culture showed higher rate of blastocyst formation in modified swim-up method (20.44$\pm$0.99%) than the control ($15.73 \pm 3.26%$) (P<0.05), even though there was no significant difference. These results suggest that, by controlling the number of spermatozoa reaching the oocytes, porcine oocytes might be protected from polyspermy in vitro. Also, the developmental potential of the fertilized embryos using this method could be improved by increasing the pool of spermatozoa with better quality. Further optimization of the procedure required to implicate this method in routine porcine IVF.

• 한국산학기술학회논문지
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• 제22권4호
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• pp.504-512
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• 2021

TCDD로 유발할 수 있는 남성 난임 원인 중 가장 큰 비율을 차지하는 것은 정자형성 장애와 정자의 질적 저하가 있다. 이를 해결하기 위해 GYB의 한의학적 처방을 이용하여 TCDD-유도 생식독성을 유발한 랫드에서 내분비기능 및 생식독성 관련 지표에 미치는 영향을 조사하고자 본 연구를 시행하였다. 수컷 SD 랫드를 5개의 그룹으로 7마리씩 나누어 실험하였다. 정상 대조군은 vehicle 및 saline을 투여하였으며, TCDD 단독투여그룹은 TCDD(2 ㎍/kg, weeks) 및 생리식염수를 복강 내 투여하였으며, 시험 군은 6주 동안 GYB (75, 150, 300 mg/kg)를 3가지 농도로 나누어 경구로 투여하였다. TCDD를 투여한 모든 군에서 체중 감소가 나타났으며, 호르몬 변화에서 GYB 300 mg/kg 투여군에서 free testosterone의 유의한(p<0.01) 감소가 나타났다. 또한 GYB 투여군의 고환조직에서 생식세포 일부 파괴, 정세관 위축 그리고 정자 수의 감소가 농도 의존적으로 개선되는 것이 관찰되었다. 또한 Johnsen's score, serotoli cell index(SCI)에서도 농도 의존적으로 개선되었다(p<0.05). 이러한 발견은 향후 남성 난임의 해결방안으로 의학적 시술이 아닌 약물요법을 효과적으로 사용할 수 있을 것으로 기대할 수 있다.

• 대한한방부인과학회지
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• 제27권1호
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• pp.176-184
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• 2014

Objectives: The purpose of this study is to understand current situation of acupuncture and moxibustion for male infertility. Methods: We searched all relevant studies of male infertility patients treated with acupuncture and moxibustion. Electronic searches were conducted in Pubmed, the Cochrane Library, EMBASE and 4 Korean databases until October 2013. Results: A total of 10 reports were included in the review. Two case reports, one case series, three before and after studies, one cohort study, one non-randomized controlled study and two randomized controlled studies were reviewed. Although the majority of studies suggested a positive effect of acupuncture on male infertility, the quality of studies was low. Data on adverse effects were lacking. Conclusions: We found a benefit from acupuncture for poor semen quality in male infertility. These findings should be treated with caution as the evidence was low or very low quality. To further evaluate the effects of acupuncture and moxibustion in male infertility, well-designed randomized controlled studies are needed.

• Asian-Australasian Journal of Animal Sciences
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• 제3권4호
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• pp.343-346
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• 1990

A study was conducted to determine the effect of prostaglandin $F_2$ alpha ($PGF_2$ alpha) on the reaction time and seminal characteristics of buffalo bulls. Semen was collected from three Murrah bulls in three periods: pre-treatment, treatment and post-treatment. During the treatment period each bull was administered 2 ml $PGF_2$ alpha (Synchrocept, Fenprostalene) im, 1 hour prior to semen collection. In the post-treatment, semen was collected 7 days after the last injection of $PGF_2$. Semen samples were evaluated immediately after collection. Pre-collection injection of $PGF_2$ alpha has no significant effect on reaction time, semen volume, percentage motility, sperm concentration and total number of sperms per ejaculate. Fluctuations in semen color and consistency were observed. There is a significant (p<0.05) increase in the mean percentage of normal spermatozoa during the treatment and post treatment periods. Likewise, administration of PG results into a significant (p<0.05) rise on the average percentage of live sperms but this effect was not manifested in the post-treatment period. Improvement in mass activity was observed during the treatment and post-treatment periods.

• 한국발생생물학회지:발생과생식
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• 제18권4호
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• pp.187-195
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• 2014

Polycyclic aromatic hydrocarbons (PAHs), which are ubiquitous in the air, are present as volatile and particulate pollutants that result from incomplete combustion. Most PAHs have toxic, mutagenic, and/or carcinogenic properties. Among PAHs, benzo[a]pyrene (B[a]P) and dimethylbenz[a]anthracene (DMBA) are suspected endocrine disruptors. The testis is an important target for PAHs, yet effects on steroidogenesis in Leydig cells are yet to be ascertained. Particularly, disruption of testosterone production by these chemicals can result in serious defects in male reproduction. Exposure to B[a]P reduced serum and intratesticular fluid testosterone levels in rats. Of note, the testosterone level reductions were accompanied by decreased steroidogenic acute regulatory protein (StAR) and $3{\beta}$-hydroxysteroid dehydrogenase isomerase ($3{\beta}$-HSD) expression in Leydig cells. B[a]P exposure can decrease epididymal sperm quality, possibly by disturbing the testosterone level. StAR may be a key steroidogenic protein that is targeted by B[a]P or other PAHs.

• 한국동물생명공학회지
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• 제35권1호
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• pp.2-11
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• 2020

Assisted reproductive technologies (ART) merely depend on improving the oocyte maturation and their developmental competence to produce good quality embryos. Oocyte maturation passes through long and complex molecular steps starts from the early embryonic life and ends with sperm fertilization. Oocyte developmental competence can be attained by improving the nuclear and cytoplasmic mechanisms together with some epigenetic maturation. In this review, we highlight the cornerstones of oocyte maturation on both nuclear and cytoplasmic levels. Interfering or supporting these molecular mechanisms would help in the development of novel regulating agents for reproductive performance of humans and livestock species.

• Journal of Animal Science and Technology
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• 제65권4호
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• pp.683-697
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• 2023

The threat posed by increased surface temperatures worldwide has attracted the attention of researchers to the reaction of animals to heat stress. Spermatogenesis in animals such as stallions is a temperature-dependent process, ideally occurring at temperatures slightly below the core body temperature. Thus, proper thermoregulation is essential, especially because stallion spermatogenesis and the resulting spermatozoa are negatively affected by increased testicular temperature. Consequently, the failure of thermoregulation resulting in heat stress may diminish sperm quality and increase the likelihood of stallion infertility. In this review, we emphasize upon the impact of heat stress on spermatogenesis and the somatic and germ cells and describe the subsequent testicular alterations. In addition, we explore the functions and molecular responses of heat shock proteins, including HSP60, HSP70, HSP90, and HSP105, in heat-induced stress conditions. Finally, we discuss the use of various therapies to alleviate heat stress-induced reproductive harm by modulating distinct signaling pathways.

• Asian-Australasian Journal of Animal Sciences
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• 제24권10호
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• pp.1348-1357
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• 2011

The objective of this study was to compare the effect of Butylated Hydroxy Toluene (BHT), Pentoxifylline (PTX) and ${\alpha}$-tocopherol (Vit E) on semen quality parameters of Karan Fries bulls. The fortification of extender by various semen additives improves motility as well as fertility of spermatozoa. Split samples of 24 ejaculates of four Karan Fries bulls were extended in extender with or without various additives such as BHT, PTX and Vit E, and performance was evaluated at an interval of 0, 24, 48 and 72 h at refrigerated temperature (4-$7^{\circ}C$). Results of the present study revealed that addition of BHT, PTX and Vit E in extender improved sperm cell function, such as motility, viability, HOST, and acrosome integrity, as compared to the control during liquid storage up to 48 h of preservation at refrigerated temperature. There was no significant (p<0.05) difference between any of the additives up to 48 h of preservation. Overall, the results showed a significant (p<0.05) deterioration in motility after each storage interval. The results showed a significant deterioration in the acrosome integrity and plasma membrane integrity up to 48 h; subsequently, there was not much degradation of both the semen quality parameters. There was a significant increase in spermatozoal tail and total abnormality after each storage interval at refrigerator temperature (4 to $7^{\circ}C$); however, the head and mid-piece abnormalities were almost unaffected. Tail and total abnormality were least in extender fortified with BHT, PTX and Vit E at different hours of incubation as compared to the control. The addition of 1.5 mM BHT, 3.6 mM PTX and 1 mg/ml Vit E in the semen extender has more beneficial effect in terms of semen quality and preservability of spermatozoa.

• 한국수정란이식학회지
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• 제30권3호
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• pp.137-142
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• 2015

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.158T>C) of phospholipase C zeta (PLCz) gene reported to be significant association with MOT. This study was conducted to evaluate the PLCz gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.158 T>C SNP of PLCz was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.0001, p=0.0002, p<0.0001 and p<0.0001, respectively). Therefore, we suggest that the intron region of the porcine PLCz, may be used as a molecular marker for Duroc boar post-thawed semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.