성별에 따른 배양육 인지도 및 영향 요인

이경본,박길준,권희경 한국디지털정책학회 2022 디지털융복합연구 Vol.20 No.1

The Semen Property and Preservation in Shih Tzu Dogs

이경본,김민규,박병권 사단법인 한국동물생명공학회 2013 한국동물생명공학회지 Vol.28 No.2

This study was carried out to investigate the general characteristics of semen such as semen volume, pH, sperm motility and sperm concentration of the semen collected from Shih Tzu dogs (age of 24 to 48 months, weight of 4 to 8 kg) by using the method of digital manipulation of the penis. The effect of preservation temperature and time on motility of fresh semen was also investigated in the present study. Semen was collected for 16 times from 4 male Shih Tzu dogs by multiple ejaculations (four times ejaculation per dog). The average of semen volume, semen pH, sperm motility and sperm concentration of the second fraction containing small volume of the initial third fraction per ejaculation were 2.11 ± 0.31 ml, 6.25 ± 0.07, 97.59 ± 1.03% and 2.05 ± 0.14 × 108 cells/ml, respectively. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculation were 1.12 ± 0.15 ml, 5.99 ± 0.14, 16.09 ± 6.18% and 5.16 ± 2.03 × 105 cells/ml, respectively. Those of second fraction were 2.07 ± 0.29 ml, 6.36 ± 0.13, 97.31 ± 1.36% and 2.15 ± 0.30×108 cells/ml, respectively. Those of third fraction were 2.60 ± 0.29 ml, 6.63 ± 0.08, 95.72 ± 1.61% and 6.03 ± 1.83 × 107 cells/ml, respectively. Sperm motility was significantly higher at 17℃ preservation temperature than at 5℃ or 36℃ during preservation period except 1 h preservation (P<0.05). When preservation temperature was 17℃, sperm motility was 96.69 ± 1.49% at 1 h, 91.38 ± 1.90% at 6 h, 88.38 ± 2.34% at 12 h, 78.13 ± 4.58% at 18 h, 58.44 ± 8.57% at 24 h and 29.56 ± 5.06% at 30 h, respectively.

Cysteamine의 첨가배양이 돼지 난포란의 체외성숙과 배발달에 미치는 영향

이경본,한만희 한국동물생명공학회(구 한국동물번식학회) 2002 Reproductive & developmental biology Vol.26 No.1

ABSTRACT I. 서 론 II. 재료 및 방법 III. 결과 및 고찰 IV. 요 약 V. 인용문헌

자산어보(玆山魚譜)의 현대 분류학적 접근

서승희,이경본,정은영,최동욱,유형빈 全南大學校 科學敎育硏究所 2014 科學敎育硏究誌 Vol.38 No.1

This research is focused on identifying the kinds of creatures in "Jasan Eobo" and analyzing the uniqueness of what attributes they are classified. We also turned the kinds of creatures in "Jasan Eobo" into modern language through the preceding research based on Korean literature and biology in order to reclassify and analyze them in terms of modern times. "Jasan Eobo" is made up of three volumes and involves content about creatures life; a total of four types and fifty-six groups and two hundred twenty-six species, shows the taxonomic characteristics using current terminology which is not in previous atlases of fish, and we can identify these characteristics by means of taxonomic ways and nomenclature. In other words, by taking a look at creatures in "Jasan Eobo", it is possible not only discern taxonomic group of specific creatures, but also to find various nomenclature activities considering their features. In addition, there are often some unknown species when the names of some creatures in "Jasan Eobo" are changed into modern languages. That is mainly because there are miswritten letters in transcribing the "Jasan Eobo" and deficient descriptions of their features without photographs or pictures. Furthermore, in classifying "Jasan Eobo", some mutant and variant fish groups are newly categorized in the same group or a fully new group. Keyword : Jasan Eobo, modern taxonomic approach, nomenclature, mutant, variant, fish

산소농도 및 배양액이 돼지 난포란의 체외성숙과 배발달에 미치는 영향

한만희,이경본,박병권,박창식,이규승 韓國受精卵移植學會 2001 한국동물생명공학회지 Vol.16 No.3

본 연구는 돼지 난포란의 체외성숙에 적합한 배양액을 조사하고, 아울러 산소농도가 돼지 난포란을 이용한 체외수정란의 생산에 미치는 영향을 구명하고자 실시하였다. 본 연구에서 얻어진 결과를 요약하면 다음과 같다. 1. NCSU-23과 TCM-199 배양액에 10% PFF를 첨가하여 5% 및 20% 산소조건하에서 체외성숙을 유기한 결과, 배양액 및 산소조건에 따른 난핵붕괴률 및 핵성숙률에는 유의적 (P>0.05)인 차이가 없었다. 2. NCSU-23이 TCM The present study was carried out to examine the effects of concentrations and culture media on in vitro maturation and embryo development of porcine follicular oocytes. The results were summarized as fellows : 1. The rates of GVBD and nuclear maturation in NCSU-23 and TCM-199 media with 10% PFF under the conditions of 5% and/or 20% concentrations were not different among the each treatment groups(P>0.05). 2. The rates of polyspermy and mean numbers of penetrated sperm were significantly lower in NCSU-23 medium than in TCM-199 medium (P>0.05). However, the rates of polyspermy and mean numbers of penetrated sperm were not different between 5% and 20% concentrations. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were significantly higher in NCSU-23 medium under the condition of 20% concentration than in TCM-l99 medium under the condition of 5% or 20% concentrations. However, the rates of blastocyst formation and total cell numbers in blastocysts were not different between 5% and 20% concentrations. In conclusion, the use of NCSU-23 medium under the condition of 20% concentration was beneficial for porcine oocyte maturation and in vitro development.

Catalase 첨가배양이 돼지체외수정란의 배발달에 미치는 영향

한만희,이경본,천행수,박병권,이규승,서길웅 한국수정란이식학회 2002 한국수정란이식학회 학술대회 Vol.2002 No.1

Catalase(CAT)는 주요한 활성산소(ROS)의 일종인 과산화수소()가 Hydroxyl 유리기 생성에 관여하지 못하도록 를 및 로 대사시켜 주는 효소계 항산화제의 한 종류이다. 따라서 본 실험에서는 CAT가 5% 및 20% 조건하에서 1-세포기 돼지체외수정란의 배발달에 미치는 영향을 조사하였다. 돼지난포란을 10% PFF, 0.1mg/ml cysteine, 10IU/m1 PMSG, 10IU/m1 hCG 및 10ng/m1 EGF가 첨가된 NCSU23 배양액에서 22시간 동안 배양을 실시하고, 성선자극호르몬이 배제된 배양액에서 추가로 22시간을 배양하여 체외성숙을 유도하였다. 체외성숙이 유기된 난자는 난구세포를 제거하고, 2.5mM caffeine과 0.1% BSA가 첨가된 mTBM 배양액에 정자를 1.25 cells/ml의 농도로 5-6시 동안 공동배양을 실시하여 체외수정을 유도하였다. 체외수정후 1-세포기의 수정란을 0.4mg/ml BSA가 첨가된 NCSU23 배양액에 CAT를 각각 0, 100, 500 및1,000uni1 첨가하여 30 embryos/ 50u1 소적으로하여 38.8, 5% 및 5%, 5% 90% 조건하의 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT 6.03 Package를 이용하여 통계분석을 실시하였다. 체외배양 48시간에 난할률을 조사하였을 때, 대조구와 처리구간 차이가 인정되지 않았으나, 배양 7일째 배반포형성률에 있어서는 각각 22.72.7, 22.13.9, 18.74.9, 및 15.12.5%로서 처리구가 대조구보다 유의적으로 낮은 결과를 나타냈다. 그리고 산소농도에 따른 배반포형성률은 5% 조건(22.12.4%)이 20% 조건(17.22%)보다 유의적(P<0.05)으로 높은 것으로 나타났다. 총세포수에 있어서는 각각 44.44.0, 43.336, 25.42.4 및 13.41.5로서 처리구가 대조구보다 세포수가 적은 것으로 나타났으며, 산소농도에 따른 차이는 인정되지 않았다. 따라서, 체외생산된 돼지초기수정란을 배양할 때, CAT의 첨가는 효과가 없는 것으로 나타났다.다.

β-Mercaptoethanol의 첨가배양이 돼지난포란의 체외성숙과 배발달에 미치는 영향

한만희,이경본,천행수,박병권,서길웅,이규승 한국동물생명공학회(구 한국동물번식학회) 2003 Reproductive & developmental biology Vol.27 No.2

본 연구는 돼지 난포란의 체외배양액에 황화합물인 β-mercaptoethanol(β-ME)을 첨가배양함으로서 돼지난포란의 체외성숙과 체외배발달에 미치는 향을 구명하고자 실시하였다. 본 연구에서 얻어진 결과를 요약하면 다음과 같다. 1. 돼지난포란을 체외성숙배양액인 NCSU-23 배양액에 β-ME를 각각 0, 25, 50 및 100μM 첨가배양하여 성숙을 유기한 다음, 체외수정을 실시한 결과, 모든 처리구에서 핵성숙률, 정자침투율, 웅성전핵형성률, 다정자침입률 및 평균침입정 수에서 유의적인 차이가 인정되지 않았다 P>0.05). 2. 체외성숙 및 수정을 실시한 후, 배발달 배양액인 NCSU-23에 7일간 배양한 결과, 배반포 형성률은 25μM의 β-ME 처리구가 유의적(P<0.05)으로 높은 결과를 나타냈고, 총세포수에 있어서는 대조구와 처리구간 유의적인 차이가 인정되지 않았다. 3. 배발달배양액인 NCSU-23에 β-ME를 각각 0, 12.5, 25 및 5μM 첨가하고 5 및 20%산소조건 하에서 7일간 배양한 결과, 처리구별 산소농도에 따른 배발달률 및 총세포수에 있어서는 유의 적 인 차이가 없었으나, 25μM β-ME 처리구에서 유의적으로 높은 결과를 나타냈다(P<0.05). 그러나 총세포수에서는 처리구 및 대조구간 유의성이 인정되지 않았다. 결론적으로 돼지난포란의 체외성숙 및 배발달시 황화합물인 β-ME의 첨가는 25μM이 가장 적합하며, 산소농도에 따라서는 영향을 미치지 않는 것으로 조사되었다. The present study was carried out to examine the effect of β-Mercaptoethanol (β-ME) on in vitro maturation (IVM) of porcine follicular oocytes and oxygen concentration with β-ME on in vitro development (IVD) of porcine IVM/IVF embryos. The results were summarized as follows. 1. The rates of nuclear maturation, penetrated oocytes, polyspermic oocytes, pronucleus formation and mean numbers of the penetrated sperms were not significantly different using NCSU-23 maturation media for 0, 25, 50 and 100 μM β-ME (P>0.05). 2. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in oocytes matured with 25 μM β-ME (25.4±0.9%) than in those matured with 0 (14.5±1.6%), 50 (17.3±1.7%) and 100 μM (12.4±1.3%) (P<0.05). However, no differences ware found in total cell numbers of blastocyst among the treatments. 3. The rates of blastocyst formation at day 7 after in vitro fertilization were higher in the NCSU-23 Culture medium With 25 μM β-ME (23.6±2.8%) than in those Cultured With 0 (15.4±4.4%), 12.5 (17.5±2.3%) and 50 μM β-ME (18.6±2.1%) Under the 5% and 20% O₂ Concentrations (P<0.05). However, no differences was found in total cell numbers of blastocyst among the treatments. These results suggested that the addition of 25 μM β-ME in the IVM/IVD media were effective on the porcine embryo production. However, the rates of blastocyst formation and total cell numbers of blastocyst at day 7 of porcine IVM/IVF embryos were not significantly different in the NCSU-23 culture medium under 5% and 20% 02 concentrations.

말의 정소상체 정자의 동결 후 해동 온도 및 Incubation의 효과

김근중,이경본,이지혜,김은영,한길우,박강선,김민규,Kim, Keun-Jung,Lee, Kyung-Bon,Lee, Ji-Hye,Kim, Eun-Young,Han, Kil-Woo,Park, Kang-Sun,Kim, Min-Kyu 한국수정란이식학회 2013 한국동물생명공학회지 Vol.28 No.3

Cryopreservation of epididymal spermatozoa offers a potential tool for rescuing genetic material from males of genetically elite populations. Castration, catastrophic injury, sudden death or any other event that makes semen collection or mating impossible may prematurely terminate a stallion reproduction. Stallion epididymal spermatozoa vary widely in the loss of progressive motility, acrosomal integrity, and viability during freezing and thawing. The objective of this work was to investigate the effect of (1) freezing package types on cryopreservation efficiency, (2) thawing temperatures (37, 56 or $70^{\circ}C$) on Computer Assisted Sperm Analysis (CASA) parameters and (3) post-thawing incubation time (0, 1, 2 or 4h) on castrated stallion epididymis. Post-thawed sperm motility ranged between 59.69% and 64.28% ($56^{\circ}C$ and $37^{\circ}C$) in various thawing temperatures. When stallion epididymis sperm was frozen, straw was better than freezing tube on VCL (Velocity of Curvilinear Line) and VAP (Velocity of Average Path) parameter. Higher percentage of motility was observed at $37^{\circ}C$ thawing temperature even though no significant difference was observed among various temperatures. The motility, VCL, ALH (Amplitude of Lateral Head displacement), VAP, BCF (Beat-Cross Frequency) and STR (Straightness index) parameter of post-thawed sperm were significantly decreased by increasing the incubation time for all thawing temperatures. The present study showed that type of freezing package (Straw vs. Freezing tube) was not significantly different on cryopreservation efficiency. Furthermore, stallion epididymal spermatozoa frozen-thawed at $37^{\circ}C$ for 1 min resulted the highest proportion of motility and velocity movement. In addition, motility and viability of frozen-thawed stallion epididymal spermatozoa were also decreased by incubation.

마우스 수정란에 있어서 부계 DNA 손상이 부계 DNA 퇴화 및 초기 배발달에 미치는 영향

김창진,이경본 사단법인 한국동물생명공학회 2019 한국동물생명공학회지 Vol.34 No.3

This study was investigated to test whether the zygote recognized the topoisomerase II beta (TOP2B) mediated DNA fragmentation in epididymal spermatozoa or the nuclease degradation in vas deferens spermatozoa by testing for the presence of gammaH2AX (γH2AX). The γH2AX is phosphorylation of histone protein H2AX on serine 139 occurs at sites flanking DNA double-stranded breaks (DSBs). The presence of γH2AX in the pronuclei of mouse zygotes which were injected with DNA broke epididymal spermatozoa was tested by immunohistochemistry at 5 and 9 h post fertilization, respectively. Paternal pronuclei that arose from epididymal spermatozoa treated with divalent cations did not stain for γH2AX at 5 h. On the other hand, in embryos injected with vas deferences spermatozoa that had been treated with divalent cations, γH2AX was only present in paternal pronuclei, and not the maternal pronuclei at 5 h. Interestingly, both pronuclei stained positively for γH2AX for all treatments and controls at 9 h after sperm injection. In conclusion, the embryos recognize DNA that is damaged by nuclease, but not by TOP2B because H2AX in phosphorylated in paternal pronuclei resulting from spermatozoa treated with fragmented DNA from vas deferens spermatozoa treated with divalent cations, but not from epididymal spermatozoa treated the same way.

마우스 수정란에 있어서 부계 DNA 손상이 부계 DNA 퇴화 및 초기 배발달에 미치는 영향

김창진,이경본 한국동물생명공학회(구 한국동물번식학회) 2019 Journal of Animal Reproduction and Biotechnology Vol.34 No.3

This study was investigated to test whether the zygote recognized the topoisomerase II beta (TOP2B) mediated DNA fragmentation in epididymal spermatozoa or the nuclease degradation in vas deferens spermatozoa by testing for the presence of gammaH2AX (γH2AX). The γH2AX is phosphorylation of histone protein H2AX on serine 139 occurs at sites flanking DNA double-stranded breaks (DSBs). The presence of γH2AX in the pronuclei of mouse zygotes which were injected with DNA broke epididymal spermatozoa was tested by immunohistochemistry at 5 and 9 h post fertilization, respectively. Paternal pronuclei that arose from epididymal spermatozoa treated with divalent cations did not stain for γH2AX at 5 h. On the other hand, in embryos injected with vas deferences spermatozoa that had been treated with divalent cations, γH2AX was only present in paternal pronuclei, and not the maternal pronuclei at 5 h. Interestingly, both pronuclei stained positively for γH2AX for all treatments and controls at 9 h after sperm injection. In conclusion, the embryos recognize DNA that is damaged by nuclease, but not by TOP2B because H2AX in phosphorylated in paternal pronuclei resulting from spermatozoa treated with fragmented DNA from vas deferens spermatozoa treated with divalent cations, but not from epididymal spermatozoa treated the same way.