본 연구는 동자개 정자의 동결보존을 위해 동결보존제의 최적 농도와 적정 희석액을 구명하여 정자를 최상의 상태로 보존하여 인공종자를 생산하는데 목적이 있다. 실험은 3종의 희석액(Ⅰ: 300 mM glycose, Ⅱ: Kurokura extender, Ⅲ: Li extender), 4종의 동결보존제(dimethyl sulfoxide, ethylene glycol, methanol and glycerol)와 4개의 동결보존제 농도(5, 10, 15, 20%)를 조합하여 대하여 조사하였다. 동결보존한 정자를 해동한 후 정자의 생존율과 정자활성지수로 동결보존 제의 효과를 평가하였다. 희석액 Ⅲ(Li extender)과 10과 15%의 methanol을 조합했을 때 동자 개 정자의 생존율과 정자활성지수는 각각 66.9 ± 8.7, 67.3 ± 13.1%과 2.6 ± 0.4, 2.6 ± 0.5로 다른 희석액과 동결보존제보다 높았다.

A literature study was conducted on the management of the pavilion of the royal tomb in the late Joseon Dynasty, focusing on “Gakneung Suri Deungnok(Records relating to the repair of royal tombs)”. This study analyzed the royal tomb management system, organized the types of damage identified in the building, and examined how the damage status was recorded by type. In the above, the records related to the 1675∼1713 repair of three JeongJaGak(Geonwonneung, Sungneung, and Mokneung), which are registered as state-designated cultural properties, are summarized in three aspects: management system, damage status, and expression words. The results of the study are as follows. First, the royal tomb pavilion was regularly inspected by Observator(觀察使) in spring and autumn, and Surunggwan(守陵官) every 5th, and Servant(守僕) regularly inspected every day and night, and also inspected and reported emergency cases of natural disasters or unexpected damage. Second, the damage status of each building was continuously observed and reported for the continuous maintenance of the buildings in the royal tomb. A total of 75 records of damage to the three royal tombs' pavilion were found to have been most frequently inspected, including 19 cases (25.3%), 14 cases (18.7%), 23 cases (30.7%) of the roof, and 19 cases (25.3%) of the roof. Third, the expression of the damage status is confirmed in various ways, such as separation, separation, burst, damage, excitation, moisture, leakage, and exfoliation. Among them, the main damage records were confirmed due to the separation of the base from the peeling, the furniture, cracks, leaks, leaks in the roof, and the collapse of the roof was able to check the damage records.

In the 16～17th centuries, the construction of T-shaped pavilion in Gyeongsangbuk-do was centered on the families of the Goseong-Lee clan and Andong-Kwon clan, who had a lot of exchanges with each other near Andong. It can be presumed that the complex structure of the T-shaped pavilion was intended to represent the technology, economic power, and social influence of the clan. After the 18th century, construction areas spread and construction subjects were diversified, but the number of new constructions decreased. It can be seen that T-shaped pavilion was erected and used for public purposes rather than personal reasons in terms of layout or flat scale. The roof of the T-shaped pavilion is very diverse depending on the wooden structure, the height of the roof and the configuration of the apex. The T-shaped pavilion, which combines two parts, has been developed in a way that strengthens not only the appearance but also the structural bonding force. The bonding strength is strengthened through the process of “roof aligning”, “roof bonding”, “structure connection”, and “structure integration”, which shows a similar tendency to the age of actual cases.

The remaining Joongjaesil(中齋室)-type pavilions in the eastern part of the Jirisan Mountain area showed more complex characteristics than those in the western part. The front number of kan(間) had two, three, and four kan (間) types, but the number of kan on the side was consistent with two regardless of the number of kan on the front. The pavillion is classified into 'central type', 'towards the backward type', and 'expanded by backwards type' according to the location of Joongjaesil. Joongjaesil's size, which characterizes the characteristics of Joongjaesil-type Pavilion, is related to the building's front number of kan. Front number of kan. When this space was increased by one space to four spaces, Joongjaesil must have two spaces. The pavilion with four bays in front showed the characteristics of being concentrated in Sancheong and Jinju, the eastern part of the Jirisan Mountain area. In the eastern region, many high pavilions remain, and the upper structure of the pavilion was insignificant, but the flat shape and public construction showed strong locality.

북방전복 Haliotis discus hannai 웅성생식세포의 분화과정과 정자의 형태를 미세구조적으로 기 재하였다. 정자의 분화과정은 정원세포기, 정모세포기, 정세포기 및 정자기의 4단계로 구분하였 다. 정원세포기에서 정모세포기로의 분화과정은 형태학적 변화가 크지 않았다. 그러나 정자변 태과정 동안 염색질 응축, 핵의 형태 변화, 첨체와 중편 및 편모 형성 등의 급격한 형태학적 변 화를 나타냈다. 북방전복의 정자는 두부, 중편 및 미부로 구성되며, 두부의 길이는 약 5.3 μm로 전자밀도가 높은 핵과 총알형의 첨체로 이루어져 있었다. 중편은 기저체와 미토콘드리아로 구 성되어 있었으며, 기저체를 중심으로 5개의 미토콘드리아가 한 층으로 배열되어 있었다. 미부 의 횡단면은 "9+2"의 미세소관 구조를 보였다. 이러한 형태 및 구조적 특징은 북방전복의 정자 는 원시형(primitive type) 정자임을 보여주는 결과이다.

We observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. According to the results from each freezing extender, the sperm membrane integrity (HOST: Hypoosmotic Swelling Test) analysis in TCGGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Glycerol 3%, Dimethylsulpoxide 3.5 M) is 59.8 ± 0.7, TCGSD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Sucrose 0.1 M, Dimethylsulpoxide 3.5 M) is 59.3 ± 0.5 were significantly higher (p < 0.05) among the experimental groups. And MMPs analysis result, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of active MMP-2 was the highest in sperms frozen in TCGSD and TCGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Dimethylsulpoxide 3.5 M), Meanwhile, sperms from the TCGGD and TCGED (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Ethylene glycol 3%, Dimethylsulpoxide 3.5 M) group showed lower level of active MMP-2 expression. Together, these results indicate that adding glycerol or sucrose to the sperm freezing buffer would not only suppress MMPs expression but also minimize DNA fragmentation, providing a mean to improve the success rate in the in vitro manipulation of rabbit sperms. Therefore, these results suggest that TCGGD or TCGSD extender method for freezing-thawing of rabbit sperm increased the viability after thawing.

This study was conducted to find out the effect that κ-Carrageenan has on the properties of dog sperm when it was added to the cryoprotectant. Extender basically was contained 1.21 g Trizma base, 0.67 g citric acid, 0.4 g glucose, 0.03 g penicillin G, 0.05 g streptomycin sulfate. Extender1 was added with 0.1%, 0.2%, 0.3%, and 0.5% carrageenan, while extender2 was supplemented with glycerol. After freezing-thawing, the motility, viability, acrosome integrity, apoptosis, and ROS (reactive oxygen specifications) of sperm were measured to analyze the effects of the supplementation of carrageenan. Total Motile (TM), Rapid Progressive Motile (RPM), Medium Progressive Motile (MPM), and Immotile were measured through the CASA system after thawing in 37 degree water. Extender with 0.2% κ-carrageenan (64.26 ± 0.49) was significantly higher than control (40.24 ± 8.27) (p < 0.05). RPMs of extender with 0.1%, 0.2% κ-carrageenan (57.64 ± 6.34, 56.47 ± 1.35) were significantly higher than the other groups (p < 0.05). Acrosome integrity was measured by dyeing to PSA-FITC with an epifluorescence microscope. Normal acrosome ratio of extender with 0.5% κ-carrageenan (61 ± 8.03) was higher than the other groups (p < 0.05). Apoptosis was measured with a FACSCalibur flow cytometer using FITC (FITC Annexin V Apoptosis Detection Kit). Treated groups of κ-carrageenan of 0.1% (0.81 ± 0.05), 0.2% (0.85 ± 0.05) were significantly higer (p < 0.05) than control. Modified SYBR/PI staining was used for determination of viability and DCF staining was used for evaluation of ROS. Viability and ROS were not significantly different from other groups. In conclusion, adding a certain concentration of carrageenan to the extender of cryopreservation, carrageenan contributes to the improvement of the sperm motility, acrosome integrity and prevention of apoptosis.

본 연구는 티모시 건초와 농후 사료 위주의 사료를 급여한 한우 씨수소 정소상체 정자 체외수정 효율 조사를 통해 정자의 활용 가능성을 조사하였다. 농후 사료는 체중의 1.8%를 급여하고 양질의 티모시 건초를 자유채식 시킨 14개월령 거세우의 정소에서 분리된 정소상체 미부의 정자를 회수하고 동결 흉해 후 체외수정을 실시한 결과는 다음과 같다. 웅성전핵과 자성전핵이 형성(2PN)된 난자는 정상수정으로, 1개의 전핵(1PN), Expanded Sperm Head (ESH), Polyspermy 형태는 비정상적인 수정의 형태로 평가하였다. 정상적으로 수정된 난자의 비율은 정소상체 정자의 경우 전체 침투율은 49.7% 그리고 정상적인 2PN을 가진 난자는 18.5%를 보였으며, 대조구 정자의 전체 침투율은 54.4%로서 정소상체 정자 보다 높은 결과를 보였으나 유의적인 차이를 보이지는 않았다. 정상적으로 2PN을 형성한 비율은 36.7%로서 정소상체 정자를 이용한 정자 보다 높았으나 유의적인 차이는 없었다. 체외수정 후 발달률 조사에서 정소 상체 정자의 분할률은 81.2%, 대조구 정자는 82.7%로 유사한 결과를 보였으나, 배반포 발달률은 정소상체 정자 24.4%와 대조구 정자 12.2%로 정소상체 정자를 사용한 난자의 발달에서는 유의적으로 높았다(p<0.05).

본 연구는 돼지의 정자와 난소내 과립막세포에서 bisphenol S(BPS)가 생존성과 활성산소 생산에 미치는 영향을 알아보고자 연구하였다. 돼지정액은 0, 5μM BPS를 처리하여 3, 6시간동안 배양하였다. 정자의 생존성은 SYBR14/PI를 이중 염색하여 분석하였으며, 활성산소의 생산을 측정하였다. 또한, BPS(0, 5, 10, 20μM)를 과립막세포에 처리하여 24, 48, 72시간동안 처리하였다. 처리 후, 세포의 생존율과 활성산소 생산(단, 5μM BPS)을 측정하였다. 그 결과, 돼지에서 정자의 생존율은 BPS에 의해 감소하였고, 활성산소의 생산은 모든 처리시간에서 증가하였다(p<0.05). 또한 과립막세포의 생존은 BPS에 의해 억제되었고, 활성산소는 유의적으로 증가하였다(p<0.05). 이상의 결과를 토대로, BPS의 노출은 정자의 활성과 번식과 관련된 세포에 나쁜 영향을 미칠 것이다.

ZAR1은 척추동물의 초기 배아 발달에 영향을 미치는 유전자로 알려져 있다. 본 연구는 두록 수퇘지 112두에서 ZAR1 유전자의 SNP을 발굴하고 ZAR1 유전자의 인트론 Single nucleotide polymorphism(SNP)과 정액의 운동학적 특성들과의 연관성을 규명하였다. 돼지의 정액 운동학적 특성을 분석하기 위해서 운동성(motility, MOT), 직선운동 속도(straight-line velocity, VSL), 곡선운동 속도(curvilinear velocity, VCL), 평균운동 속도(average path velocity, VAP), 직진성(linearity, LIN), 선형성(straightness, STR), 측두 이동거리(amplitude of lateral head displacement, ALH) 및 머리 진동수(beat cross frequency, BCF)를 측정 하였다. SNP을 탐색하기 위해서 돼지의 혈액에서 genomic DNA를 추출하여 PCR(polymerase chain reaction)을 통해 시퀀싱 분석을 하여 유전자형을 분석하였다. 그 결과 ZAR1의 non-coding 영역인 인트론에서 SNP 3개를 확인 했으며 각 각 인트론 2 영역에서 g.2435T>C, 인트론3 영역에서 g.2605G>A, g.4633A>C 변이를 보였다. g.2435T>C, g.2605G>A는 각 각 MOT(p<0.01)와 VSL(p< 0.05)에서 높은 연관성을 나타냈고 g.4633A>C는 MOT, VCL, VSL, VAP, ALH에서 높은 연관성을 나타냈다(p<0.01). 본 연구 에서는 ZAR1 유전자의 SNP이 돼지 정액의 운동학적 특성들에 영향을 미치는 것으로 나타났다. 이러한 결과는 ZAR1이 우수한 수퇘지들에서 번식능력이 높은 개체를 선발할 수 있는 후보유전자로 제시하며 다양한 돼지의 품종과 더 많은 두수를 확보하여 검증연구를 통해 우수한 능력의 수퇘지에서 활력이 좋은 개체를 선발하는 분자마커 연구의 기초자료로 사용이 가능하다.

Lonicera Caerulea(Honey berry) has been used in medical treatment in Russia, Japan, China and Korea. It has high level of vitamin C and polyphenolics. Polyphenolics can improve anti-inflammatory effect and prevent cancer, diabetes mellitus type 2. Also, Vitamin C is a representative anti- oxidant. however, It is still unknown what effect it will have on the oxidation stress of the reproductive system. In previous studies, ROS can be produced when it is exposed to heat stress and has negative effect on sperm's maturation, capacitation, hyperactivation, acrosome reaction and fusion of egg and sperm. Therefore, The purpose of this study is to investigate the antioxidant effects of L.caerulea on the sperm and egg cells of mice. At first, it conducted using ICR mouse(n=20) during 4 weeks. There are four groups of mouse(n=5 per group). Also, L.caerulea was taken by oral gavage. Group Ⅰ(control) kept at 23℃~27℃ and administer D.W(0.5ml/day), Likewise, Group Ⅱ(HB) kept at room temperature but gave HB(0.5ml/day), Group Ⅲ (HB+HS) received heat stress (40℃) using hyperthermia induction chamber and gave HB at same dose. and Group Ⅳ(HS) exposed heat stress only. Mainly, we showed degree of gene expression using Western blot in SOD, HSP 70, 17β-HSD and Real time PCR. It can find correlation between intracellular activity like steroid hormone, apoptosis under ROS and antioxidant activity of L.caerulea.

본 연구는 말 냉장정액의 정자 성상 향상을 위한 기초 연구로 국립축산과학원 난지축산연구소에서 한라마(더러브렛×제주마) 수말 정액의 보존시간대별 정자 성상을 분석하기 실시하였다. 본 연구의 공시마는 국립축산과학원 난지축산연구소에서 사육・보유하고 있는 한라마(더러브렛×제주마) 씨수마 2 두를 공시하였으며, 씨수마의 정액채취는 채취 시마다 같은 시간인 오전 9 시에서 10 시 사이에 실시하였다. 정액채취는 독성이 제거된 50ml 플라스틱 일회용 용기를 인공질(INRA model, IMV, French)에 장착하여 사용하였으며, 자외선 차단용 커버로 용기 주변을 보호하였다. 인공질은 45∼50℃의 온수를 이용하여 인공질 내부 온도가 38℃가 되도록 유지하였으며, 삽입을 원활하게 하기 위해 젤을 도포하였다. 정액은 채취 직후 10 분 이내 실험실로 운반하여 겔(gel) 제거 후 부피, 농도, 운동성 등을 조사하였다. 정장 물질 제거를 위하여 INRA96 희석제로 1:1 희석 후 50ml 튜브에 40ml 씩 분주한 후 400×g 에서 10 분 원심분리하여 seminal plasma 및 부유액을 제거하고 하층의 펠렛(정자괴)을 회수하였다(Cochran 등, 1984). 회수한 정자 펠렛은 최종 농도를 50×106/ml 로 희석하여 24h, 48, 72, 96 동안 혐기상태로 4℃냉장고에 보관하여 각각의 정자성상을 분석하였다. 말 정액은 INRA 96 희석제와 희석제에 Egg Yolk 3%가 처리 후 시간대별 정자성상을 분석하였다. 2 차 희석 후 INRA 96 과 Egg Yolk 3% 처리구에서 총 운동성은 각각 89.18%, 91.81%를 보였으며, 96 시간이 경과한 후에 정자 총 운동성은 INRA 96 이 80.71%, Egg Yolk 3% 처리구는 88.54%로 유의적인 차이를 보이지는 않았다. 본 연구의 기초 데이터를 바탕으로 정장제거 원심분리 조건, 정장제거 필터링 조건 등 정자 생존율 향상 연구 등 냉장정액의 시간대별 정자 성상분석을 통한 장기보존 방법 등의 연구가 수행되어야 할 것으로 생각된다.

This study was conducted to determine the effect of pentoxifylline levels on sperm motility, survival rate, sperm membrane integrity of frozen semen and fresh-extended equine semen in Jeju cross-bred horses. As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw, the progressive motilities were 53.25±2.87 (4mM pentoxifylline) and 50.28±2.14 (8mM pentoxifylline) and significantly higher compared to the control group(40.09±5.15) and other treatment group (16mM pentoxifylline, 41.27± 2.82). The progressive fast motility were 22.44±1.62 (4mM pentoxifylline,) and 22.74±3.07 (8mM pentoxifylline) and significantly higher compared to the control group (13.47±1.48) and other treatment group (16mM pentoxifylline, 14.66±3.68) (p<0.05). As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw were 68.96±1.64 (4mM pentoxifylline) and 67.90±6.72 (8mM pentoxifylline) and significantly higher compared to the control group (53.48±4.84) and other treatment group (16mM pentoxifylline, 58.14±2.65) (p<0.05). In conclusion, these results suggest that treatment groups with 4mM and 8mM pentoxifylline were higher compare to equine seperm mobility and the control group and treatment groups with more than 16mM pentoxifylline has a negative effect on sperm characteristics. After thawing, the total motility in post-thawed equine sperm has increased by 10 percent for 1 hour. these results suggest that pentoxifylline contributes to the improvement of the equine sperm motility and characteristics in post-thawed semen.

This study was to investigate effect of tunicamycin (TM) on sperm viability, mitochondrial activity and motility in boar semen. Collected sperm were incubated with semen extender containing 0, 1, 2, and 5 μM TM for 3, 6 and 9 h. Sperm viability was analyzed using SYBR14/PI doubling staining, and mitochondrial activity was detected using Rhodamine123/PI staining methods. Sperm viability, mitochondrial activity and motility were measured every 3 h during incubation. In results, boar sperm viability, mitochondrial activity and motility were significantly decreased in 2 and 5 μM TM groups compare to control group at all incubation time (p<0.05). In addition, mitochondrial activity and motility were significantly decreased in 1, 2, and 5 μM TM groups compare to control group at 9 h after incubation (p<0.05). These results suggest that TM can inhibit sperm viability, mitochondrial activity and motility in boar semen, and it may influence on the fertility of sperm.

In this experiment, we determined the effect of curcumin supplementation in freezing buffer for miniature pig sperm cryopreservation. Each ejaculate was diluted with modified Modena B extender and mixed with lactose-egg yolk (LEY extender, 80% v/v lactose solution [310 mM], 20% v/v egg yolk, and100 μg/mL kanamycin sulfate) and LEY-glycerol Orvus ES Paste (LEYGO, 89.5% v/v LEY, 5% v/v glycerol, 1.5% v/v Orvus ES Paste), 100 mM trehalose supplemented with 0, 10, 50, 100, and 500 μM of curcumin from turmeric, respectively. Following equilibration, the 0.5 mL French straws were frozen and plunged into LN2 tank for 7 days at least. Sperm parameter and oxidative byproducts were determined by the computer assisted sperm motility analysis (CASA) and fluorescence-activated cell sorting (FACS) as compared with each groups.Supplementation of curcumin had no effect on sperm motility, progressive motility and curvilinear velocity. However, average-path velocity and straight-line velocity were significantly higher in 10 μM curcumin group (100.9±8.8 μm/s, 61.7±2.9 μm/s, respectively) than control group (77.8±3.9 μm/s, 46.4±3.0 μm/s, respectively) (p < 0.05). In addition, the level of the O2 radical and H2O2 were comparatively decreased in curcumin groups by evaluation of ethidium and DCF fluorescence. According to the results, curcumin can improve sperm kinetic variables and alleviate ROS induced cryoinjury to pig sperm.