• 한국수정란이식학회지
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• 제31권3호
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• pp.287-291
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• 2016

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. Estrogen receptors 2(ESR2) is involved in estrogen related apoptosis in cell cycle spermatogenesis, but their functions have not been confirmed in pig until now. Therefore, this study was conducted to analyze their association with sperm motility and kinematic characteristics. DNA samples from 105 Duroc pigs with records of semen motility and kinematic characteristics [Total motile spermatozoa (MOT), Curvilinear velocity(VCL), Straight-line velocity(VSL), the ratio between VSL and VCL(LIN), Amplitude of Lateral Head displacement(ALH)] were analyzed. A SNP in coding region of ESR2 g.35547A > G in exon 5 was associated with MOT (p < 0.05) in Duroc population. Therefore, we suggest that the porcine ESR2 gene may be used as a molecular marker for Duroc boar semen quality, although its functional effects were not defined yet. These results might shed new light on the roles of ESR2 in spermatogenesis as candidate gene for boar fertility, but still the lack of association across populations should be considered.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.241-246
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• 2010

The objective of this study was to investigate the effect of storage time on fresh boar semen in Androhep and Beltsville Thawing Solution (BTS). Boar semen samples extended in each extender were stored at $17^{\circ}C$ up to 4 days. Sperm motility kinematics was evaluated by computer assisted sperm analyzer (CASA) and capacitation status by chlortetracycline (CTC)/Hoechst 33258 staining. Sperm motility (%) was not decreased during storage in BTS and Androhep. No significant difference between extenders was observed. Only significant differences in kinematic parameters on linearity during storage were found. The percentage of dead sperm significantly decreased during storage (p<0.05). Also the percentage of noncapacitated, capacitated, and acrosome-reacted sperm significantly modified during storage (p<0.05). However, there was no significant difference between extenders except proportion of capacitated sperm. This finding supported that modification in these parameters was not significantly different between extenders during this short-term storage. Our finding strongly indicated that both Androhep and BIS maintained favorable conditions for motility, motility kinematics, and capacitation status during short-term storage. Despite modifications in some parameters were apparent during sperm storage in extenders, these may not affect the fertilizing capacity of boar semen.

• 한국양식학회지
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• 제11권1호
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• pp.67-75
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• 1998

감성돔 (Acanthopagrus schlegeli) 정자의 냉동 보존시 정자의 생리활성과 보존효과를 비교하기 위하여, 순환여과 사육시스템에서 사육한 전장 $25.9{\pm}1.7$ cm, 체중$292.8{\pm}53.7$ g의 성어로부터 얻은 정액을 재료로 하여 희석액별, 동해방지제별 해동후 정자의 활성, 생존율 및 알에 대한 수정능력을 평가하였다. 희석액별 냉동보존에서 해동정자의 생존율은 3% sodium citrate에서 $80{\pm}1.4$%로 가장 높았으나, 수정률은 5.4% glucose에서 $63.4{\pm}4.4$%로 가장 높았다. 동해방지제별 냉동보존에서 해동정자의 수정률은 5~15% glycerol을 동해방지제로 사용하였을 때, 50.1~69.4%로 DMSO 보다 나은 효과를 보였다. 감성돔 정자의 냉동보존을 위한 적정 희석액과 동해방지제는 5% glucose와 5% glycerol이었다. 냉동하지 않은 감성돔 정자의 머리는 구형으로 직경 $1.5{\pm}0.1$ ${\mu}$m, 길이 $1.3{\pm}0.1$${\mu}$m였으며, 치밀한 핵질로 채워져 있었다. 편모는 전형적인 9+2 구조를 나타냈다. 그러나 냉동후 해동시킨 정자중에서는 염색질의 과립화, 세포막의 이탈에 의한 그 용적이 커지는 구조변화를 보였다.

• 한국임상수의학회지
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• 제26권6호
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• pp.647-649
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• 2009

양측성 잠복고환을 가진 18개월 연령의 잉글리쉬 불독에서 고환하강고정술을 실시하였고 수술 1달 후 GnRH를 투여하였다. 수술 전후 정액검사를 실시하였으며 인공수정을 통해 생식능력을 검증하였다. 수술 전과 수술 2달 후 정액검사 결과, 무정자증으로 판단되었으나 수술 4개월 경과 후 6마리의 정자가 관찰되었다. 수술 7개월 후 정자수를 제외한 나머지 정자 소견에서 정상을 나타내었으며 수술 8개월 후 채취된 정액을 암컷 불독에게 인공수정한 결과 6마리의 건강한 산자를 생산하였다. 양측성 잠복고환견에서 성성숙 이후라 할지라도 고환하강고정술과 GnRH 투여를 통해 정자형성과 수정능력이 회복 가능함을 알 수 있었다.

• Clinical and Experimental Reproductive Medicine
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• 제15권2호
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• pp.149-155
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• 1988

To increase fertilization rate in vitro, separation of viable spermatozoa from the seminal plasma and its other components may be a useful procedure. Ejaculates from healthy men, whose semen analysis findings were normal in 19, and abnormal in 10, were filtered using the glass-wool filtration technique to yield a concentrated, viable sperm samples for IVF, and the usefulness and safety of this method were evaluated. The recovery rate of motile sperm in abnormal groups was 46.2% and 54.5% in normal group. The % motility was increased significantly compared with original sample after filtration, and the grade motility was improved, too. The sperm population with normal morphology was also increased significantly in both group. Using transmission electron microscopy, the ultrastructural integrity of acrosomal segment was examined in order to evaluate the potentially hazardous effect of glass-wool filtration to sperm head, however, sperm population with normal ultrastructure was increased compared with that of original ejaculate after separation. The filtered sperm was then processed for IVF, as the fertilizing capacity is the ultimate parameter of the sperm function. In abnormal group, the fertilization rate(41.5 %) and the ET rate per stimulated cycle were much lower than that of mormal group(69.6%). However, the cleavage rate and the number of embryos transfered per ET cycle were comparable with those of nomal group. The results suggest that the glass-wool filtration of sperm, particularly in oligo-asthenozoospsrmia, may be useful and safe method in the preparation of sperm for IVF.

• 한국수정란이식학회지
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• 제30권3호
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• pp.143-147
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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. 35756 T>C) of Estrogen Receptor 1 (ESR1) gene reported to be significant association with MOT. This study was conducted to evaluate the ESR1 gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.35756 T>C SNP of ESR1 was significantly associated with frozen semen motility and kinematic characteristics. The g.35756 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.001). The SNP was also significantly associated with ALH (P<0.05). Therefore, we suggest that the g. 35756 T>C polymorphism in the intron 1 region of the porcine ESR1 gene could potentially be applied in frozen semen programs to improve MOT trait, but only after validation in other populations.

• 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.

• Journal of Animal Science and Technology
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• 제63권2호
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• pp.262-271
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• 2021

Communication among epididymal epithelial cells creates the best luminal condition where spermatozoa mature, transport and are stored. Vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) have been used as signal indicators for clear and basal cells of the epididymal epithelium, respectively, in mice, rats, bats, and pigs; however, these two markers have not yet been described in the epididymis of bulls. Here, we examined the presence and distribution of the B1 subunit of V-ATPase (B1-VATPase) and KRT5 in the distinct regions of adult bovine epididymides, specifically, the caput, corpus, and cauda. Immunofluorescence staining and confocal microscopy showed that narrow shaped-clear cells were placed in the caput and corpus regions of the bovine epididymis; however, they were absent in the cauda epididymis. In addition, B1-VATPase was highly expressed in the cauda spermatozoa; however, it was rarely detected in the caput spermatozoa. On the other hand, KRT5-positive cells, basal cells, were maintained beneath the basal lamina and they had the traditional form with a dome-shaped morphology from the caput to cauda region of the bovine epididymis. The co-expression of B1-VATPase and KRT5 was confined to basal cells placed in the basal region of the epithelium. In summary, 1) clear cells were present with region-specific localization, 2) B1-VATPase was present in the corpus and cauda spermatozoa but absent in the caput, 3) co-expressed cells with B1-VATPase and KRT5 were present in the adult bovine epididymis, and 4) B1-VATPase was not a specific marker for clear cells in the bovine epididymis. Therefore, the perfect epididymal luminal condition created by the specific expression and localization patterns of B1-VATPase might be necessary to obtain fertilizing capacity of spermatozoa in the bovine epididymis.

• Animal Bioscience
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• 제35권5호
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• pp.670-676
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• 2022

Objective: This study was conducted to examine influence of skimmed milk-based extender (SM), INRA 96 extender and BotuSemen Gold extender on parameters of stallions' ejaculate during storage. Methods: In this study, 14 stallions between 4 and 20 years of age were monitored. Total and progressive motility, viability and morphology of sperm were evaluated at time intervals of 24, 48, and 72 hours after collection. Results: The total motility, progressive motility, and values of sperm with normal morphology were significantly higher in the INRA 96 and BotuSemen Gold extenders than in the SM (p<0.01). The sperm viability differed significantly in all extenders (p<0.01). The highest value of sperm viability was in INRA 96 (64.69%±0.67%) and lowest in SM (59.70%±0.81%). The highest differences occurred at 72 hours of storage. Values of total motility, progressive motility and sperm viability decreased over time (p<0.01). In case of sperm morphology there was no statistically significant decrease between 48- and 72-hour time intervals. Conclusion: It can be concluded that the extenders with a chemically defined composition have shown better indicators of insemination capabilities in ejaculates than the SM. The BotuSemen Gold extender is a suitable alternative to the INRA 96, when used within 48 hours; after 72 hours of storage, however, the INRA 96 showed a higher share of viable spermatozoa.

• 한국가축번식학회지
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• 제24권2호
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• pp.209-216
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• 2000

본 연구는 1996년부터 1999년까지 다비 AI 센타에서 사육되는 종모돈 199두와 종빈돈 4,181두를 가지고 종모돈의 정액생산과 번식성적에 미치는 품종, 연령, 계절, 산차 및 교배방법의 영향을 구명하고자 실시한 바, 그 얻어진 결과는 다음과 같았다. 1. 정액량은 Landrace (L) 종이 266.8 $m\ell$로 제일 많았고, Yorkshire (Y) 종이 그 다음으로 많았으며, Duroc (D) 종이 제일 적었다. 정자 운동성은 품종간에 차이가 없었다. 정자농도는 Landrace 종이 4.7$\times$109 sperm/$m\ell$로 제일 낮았고, 정액량이 제일적은 Duroc 종이 5.7$\times$$10^{9}$ sperm/$m\ell$ 로 제일 높았다. 종모돈의 연령에 따른 정액량은 2세 때가 213.3 $m\ell$로 제일 많았고, 4, 3 그리고 l 세의 순서로 정액량이 감소하였다. 2. 종모돈의 계절에 따른 정액량은 겨울철이 228.6 $m\ell$로 제 일 많았고, 가을, 여름, 봄의 순서로 정액량이 감소하였다. 정자농도는 봄철이 5.9$\times$109 sperm/$m\ell$로 제일 높았으며, 여름, 겨울, 그리고 가을의 순서로 낮았다. 3. 교배방법에 따른 복당 생시체중은 품종간 처리구간에 차이점을 인정할 수 없었으나 복당 출생시 생존수에 있어서는 자연종부후 인공수정을 실시한 처리구에서 9.5두로 제일 많았으며, 인공수정을 실시한 처리구에서는 9.2두, 그리고 자연종부를 실시한 처리구에서는 8.9두로 처리구간에 유의성 (P＜0.01)이 인정되었다. 4. 교배그룹간에서는 L (우)$\times$Y (♂) 교배조합과 L (♀)$\times$L (♂) 교배조합이 Y (♀)$\times$ L (♂) 교배조합 Y (♀)$\times$Y (♂) 교배조합보다 복당 생시체중이 무거웠으며, 복당 출생시 생존 자돈수에 있어서도 비슷한 경향을 나타내었다. 자연종부와 인공수정간에 복당 생시체중과 복당 출생시 생존자돈수는 유의차가 없었다. 5. 산차간에 있어서는 2~6 산차의 복당 생시체중과 복당 출생시 생존 자돈수가 각각 13.5 kg 과 9.5 두로 1산차와 7~9 산차에 비하여 우수하였다.