돼지의 난포액 내 단백질인자의 탐색과 변화에 관한 연구

지미란,정희태,양부근,이채식,박춘근,Ji, Mi-Ran,Cheong, Hee-Tae,Yang, Boo-Keun,Lee, Chae-Sik,Park, Choon-Keun 한국동물번식학회 2010 Reproductive & developmental biology Vol.34 No.3

When fully grown oocytes are removed from their follicles, they can resume meiosis and mature spontaneously under in vitro conditions. However, nuclear maturation under in vitro condition is not accompanied by complete cytoplasmic maturation, which is essential for successful fertilization and the initiation of zygotic development. This study analyzed change of proteins in follicular fluids during the porcine follicular development. Follicular fluids were collected from follicles of diameter 1~2 mm, 2~6 mm and 6~10 mm in ovary of slaughtered pigs. Total proteins were extracted from follicular fluids by M-PER Mammalian Protein Extraction Reagent. We confirmed totally 27 same spots, 1 spot from follicle fluid of 2~6 mm follicle and 5 spots from follicle fluid of 6~10 mm in diameter were analyzed by MALDI mass spectrometry and searched on NCBInr. In results, spot No. 28 from 2~6 mm follicle was Ig lambda chain C region, and spot No.32 and 33 from 6~10 mm was Apolipoprotein A-(APOA4). Spot No.29 and 31 were failed to analyze. These results indicate that the porcine oocyte during in vitro maturation depend on specific different expressed proteins may play an important roles in the sequence of molecular events in porcine oocyte maturation and follicular development.

체외에서 돼지 정자-난자의 상호작용시 투명대내 Lectin 결합

황인선,김정익,정희태,양부근,박춘근,Hwang, In-Sun,Kim, Choung-Ik,Cheong, Hee-Tae,Yang, Boo-Keun,Park, Choon-Keun 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.3

Objective: Lectins are cell-agglutinating and sugar specific proteins or glycoproteins of non-immune origin that precipitate glycoconjugates having saccharides of appropriate complementarity. Because of these properties, plant lectins have been used to help characterize the carbohydrate moieties of glycoproteins in the zona pellucida (ZP) of several mammalian species including pigs. Treatment of oocytes with various lectins blocks sperm binding to the ZP in various mammalian species. This study was undertaken to examine the distribution of sugar residues in the ZP of pig oocytes matured in vitro and the ability of spermatozoa to bind to ZP and in vitro penetration in oocytes treated with fluorescein isothiocyanate (FITC)-labelled lectins. Materials and Methods: The lectins of Banderiaea simplicifolia (BS-II, bind to $\beta$-D-N-acetylglucosamine), Canavalin ensiformis (Con A, bind to $\alpha$-D-Mannose), Lens culinaris (LCA, bind to a-D-Mannose), Ricinus communis (RCA-I, bind to $\beta$-D-Galactose) and Ulex europaeus (UEA-I, bind to $\alpha$-L-Fucose) were examined for spermatozoa penetration, binding capacity to ZP and distribution of lectins. Results: The penetration rates were significantry (p<0.05) higher in control oocytes (63%) than those treated with all lectins, but penetration rates ($40{\sim}49%$) were simililar in group treated with lectins. The incidence of monospermy was similar in oocytes untreated and UEA-I, but it was higher in oocytes treated with BS-II, Con A, RCA-I and LCA. The porcine oocytes cultured for 48 h in TC-199 medium were freed from cumulus cells and treated for 30 min with fluorescein isothiocyanate-labelled lectins. When examined under fluorescein illumination, higher (p<0.001) proportions of oocytes showed fluorescein of zona pellucida after treatment with Con A (93%), LCA (93%) and RCA-I (100%) than BS-II (37%) and UEA-I (50%). All of the oocytes treated with RCA-I exhibited strong fluorescein in the outer region of the zona pellucida while those treated with LCA exhibited strong fluorescein throughout the zona pellucida. BS-II bounded mainly to the outer region and UEA-I bounded mainly to the inner region of the zona pellucida, with either strong or weak fluorescein. At 120 min after insemination in vitro, fewer spermatozoa were bound to the zona pellucida of the oocytes treated with BS-II, Con-A and RCA-I. Of the lectins, Con A most inhibited sperm binding. Conclusions: These results suggest that $\beta$-D-Galactose residues in the porcine zona pellucida may act as primary sperm receptors and inducers of the sperm acrosome reaction and these sugar residues may be involved in the block to polyspermy.

대사에너지가 열 스트레스에 노출된 오리의 간, 십이지장 융모, 미생물, 유전자 조절에 미치는 영향

신종서(Jong Suh Shin),양부근(Boo Keun Yang),박병성(Byung Sung Park) 한국유화학회 2017 한국응용과학기술학회지 Vol.34 No.3

본 연구는 열 스트레스 하에서 오리사료 내 대사에너지(ME) 수준이 오리의 간, 십이지장 융모, 미생물, 유전자 조절에 미치는 영향을 조사하였다. 총 240마리의 육용 오리 채리밸리(Anas platyrhynchos)를 4처리구로 완전임의배치 한 후 42일 동안 사육하였다. 처리구는 ME 2900 kcal/kg, ME 3000 kcal/kg, ME 3100 kcal/kg 및 ME 3200 kcal/kg로 구분하였다. 간 조직은 처리구 사이의 차이가 없었고, 십이지장 융모 및 창자샘 길이는 ME 3000과 비교할 때 2900은 10.58% 감소하였으나 3100, 3200과의 사이에 차이는 없었다. 맹장 Latobacillus는 ME 3000과 비교할 때 2900은 9.47% 감소하였으나 3100, 3200은 각각 2.52, 3.24% 증가하였다. Total aerobic bacteria, E. coli, Coliform bacteria는 ME 3000과 비교할 때 2900은 증가하였으나 3100, 3200은 차이가 나타나지 않았다. 간에서 HSP (heat shock proteins)-mRNA 중 HSP 90-α는 ME 3000과 비교할 때 2900은 48.60% 감소하였으며 3100, 3200은 차이가 없거나 증가하였다. The object of this study was to determine the influence of dietary metabolic energy (ME) on ..... A total of 240 meat ducks Cherry valley (Anas platyrhynchos) were assigned into four treatment groups with a randomized block design for 42 days. The four treatments were: ME 2900 kcal/kg, ME 3000 kcal/kg, ME 3100 kcal/kg, and ME 3200 kcal/kg. There was no difference in liver tissue among the treatments. The duodenal villi and crypt depth length decreased by 10.58% in 2900 compared with ME 3000, but there was no difference between 3100 and 3200. Counts of caecal Latobacillus decreased by 9.47% in 2900 compared to ME 3000, but increased by 2.52 and 3.24% in 3100 and 3200, respectively. Total aerobic bacteria, E. coli and Coliform bacteria were increased by 2900 when compared to ME 3000, but there was no difference between 3100 and 3200. HSP 90-α among the heat shock proteins (HSPs)-mRNA in the liver was reduced by 48.60% in 2900 compared to ME 3000, while 3100 and 3200 showed no difference or increased.

소 체외수정란 및 체세포 복제란의 초자화 동결 후 생존성

권대진,박주희,박춘근,양부근,정희태,Kwon, Dae-Jin,Park, Joo-Hee,Park, Choon-Keun,Yang, Boo-Keun,Cheong, Hee-Tae 한국동물번식학회 2007 Reproductive & developmental biology Vol.31 No.1

본 연구는 소 체외수정란과 체세포 복제란의 초자화 동결 및 응해 후 생존능을 검토하였다. 배반포로 발육된 체외수정란 및 체세포 복제란을 초자화 동결법에 의해 동결하였다가 융해하여 생존율 및 배양 후 부화율을 검사하였다. 체외수정란 배반포를 초자화 동결 응해한 결과, 확장배반포가 배반포기 난자에 비하여 생존율(82.1%, 96/117)과 부화율(64.1%, 75/117)에서 모두 유의적으로 높았다(p<0.05). 핵이식 배반포 복제란을 초자화 동결 응해한 경우도 체외수정란과 비슷한 경향을 보여 확장 및 부화배의 생존율과 부화율이 각각 81.1%(30/37)와 78.3%(29/37)로, 배반포(각각 71.8 및 53.8%)에 비하여 다소 높게 나타났다. 본 연구의 결과는 초자화 동결 방법에 의해서 소 체외수정란과 체세포 복제란을 성공적으로 동결할 수 있으며, 특히 후기 배반포기 단계에서 초자화 동결 시 높은 생존율과 부화배 형성율을 얻을 수 있음을 보여준다. This study was conducted to examine the development of in vitro fertilized (IVF) and nuclear transfer (NT) embryos following vitrification IVF and NT embryos developed to the blastocyst stage were equilibrated by 3 steps, vitrified and thawed, and their survival and hatching rates were examined. In IVF embryos, higher survival (82.1%, 96/117) and hatching rates (64.1%, 75/117) were obtained respectively after thawing and culture in expanded blastocysts compared to blastocysts (p<0.05). High survival and hatching rates were also obtained by vitrification of NT blastocysts, especially in expanded and hatching blastocysts (81.1 and 78.3%, respectively). The result of this study shows that IVF and NT blastocysts, especially late stage blastocysts, are successfully cryopreserved by vitrification.

지렁이 분변토 발효사료가 고품질 계란생산 및 악취 저감에 미치는 영향

신종서(Jong Suh Shin),양부근(Boo Keun Yang),박병성(Byung Sung Park) 한국유화학회 2017 한국응용과학기술학회지 Vol.34 No.3

본 연구의 목표는 분변토로부터 분리한 Bacillus subtilis외 2종의 복합 생균제를 이용하여 제조한 발효사료가 산란계 생산성, 계란품질 및 양계장 악취저감에 미치는 효과를 조사하는 것이었다. 산란계 240마리를 4개의 처리구로 구분하여 완전임의 배치하였다. 실험 처리구는 대조구, 분변토발효사료 0.3, 0.5% (FECF3, FECF5), 일반생균제 0.2% (CP)로 구분하였다. 산란율은 일반사료를 섭취한 대조구와 비교할 때 FECF 처리구는 유의하게 높았으나 CP 처리구는 차이가 없었다. 호우유닛, 난각두께, 파란강도, 난황색 지수 및 농후난백 높이는 대조구와 비교할 때 FECF 처리구가 증가하였으나 CP 처리구와 차이는 없었다. 계란의 n-3 함량은 대조구와 비교할 때 FECF 처리구가 높았으나 n-6/n-3 비율은 낮았으며 CP 처리구와의 차이는 없었다. 계분을 실온에서 5일간 방치 후 암모니아 발생량은 대조구와 비교할 때 CP와 FECF 처리구가 낮았다. The aim of this study was to investigate the effects of fermented earthworm cast feed prepared with three complexed probiotics containing Bacillus subtilis separated from the earthworm casts, on egg production, egg quality and odor removal from poultry house. A total of 240 laying hens were divided into 4 treatments and completely randomized. Experimental groups were divided into control, 0.3 to 0.5% (fermented earthworm cast feed FECF3, FECF5) and 0.2% commercial probiotics (CP). The egg production rate was significantly higher in the FECF groups compared to the control group fed the normal feed, but there was no difference in the CP group. Haugh units, egg shell thickness, breaking strength, egg yolk index and albumen height were increased in FECF groups compared to control but not in CP group. The n-3 content of eggs was higher in the FECF groups than in the control group, while the n-6/n-3 ratio was lower and there was no difference from the CP group. Ammonia production was lower in the CP and FECF groups compared to the control group after 5 days storage at room temperature to poultry feces.

임신중인 생쥐에 DEHP 투여가 번식현상에 미치는 영향 Ⅱ. 임신중인 생쥐에 DEHP 투여가 자손의 번식특성과 혈액성분에 미치는 영향

박동헌,장현용,박춘근,정희태,김정익,양부근,Park, Dong-Heon,Jang, Hyun-Yong,Park, Choon-Keun,Cheong, Hee-Tae,Kim, Choung-Ik,Yang, Boo-Keun 한국발생생물학회 2004 발생과 생식 Vol.8 No.2

본 연구는 임신한 생쥐에 DEHP의 투여가 자손의 번식특성과 혈액성분에 미치는 영향을 검토하였으며 그결과는 다음과 같다. 임신기에 DEHP의 투여가 웅성 자손의 체중, 번식기관 무게 및 정액 성상에 미치는 결과는 투여구간에 커다란 차이가 없었다. 임신기에 DEHP의 투여가 웅성 자손의 혈액 성상에 미치는 영향을 조사한 결과, WBC, HB, HT, MCH 및 albumin은 투여구간에 차이가 없었으나, RBC MCV, MCHC, PLT 및 total protein은 각 투여구간에 통계적으로 유의적인 차이가 나타났다. 임신기에 DEHP의 투여가 자성 자손의 체중, 자궁 및 오른쪽 난소의 무게에 미치는 영향을 조사한 결과, 각 투여구간에 커다란 차이가 없었지만, 왼쪽 난소의 무게는 10.0mg 투여구가 대조구와 0.5mg 투여구에 비해 통계적으로 유의하게 높은 무게를 나타냈다. 임신기에 DEHP의 투여가 자성 자손의 혈액성상에 미치는 영향을 조사한 결과, RBC, HB 및 HT는 각 투여구간에 커다란 차이가 없었다. 임신기에 DEHP의 투여가 자손의 정소, 난소 및 자궁의 조직에 미치는 영향을 조사한 결과,정소에서는 투여구간에 차이가 없었으며, 난소에서는 10.0mg 투여구가 여타구보다 많은 황체를 가지고 있었으며, 자궁에서는 대조구에 비해 DEHP의 투여농도가 증가할수록 자궁내막층이 감소했으며, 1.0mg 투여구와 10.0mg 투여구는 대조구에 비해 뚜렷한 자궁내막층의 감소가 있었다. The objective of this study was to assess that the effects of DEHP administration on reproductive characteristics and blood hematological and chemical values in pups born after DEHP administration in pregnant mice. DEHP was administrated to pregnant mice by intraperitoneally injection with 0, 0.5, 1.0 and 10.0mg/kg B.W, 5 times at 3 days interval from Day 1 to Day 16 in the gestation period. The body weight and reproductive organ weight(testis, epididymis and coagulating gland) in male pups on 45 day after birth was not affected in all experimental groups, but vesicular gland in DEHP groups was significantly lower than that of control group(P<0.05). The semen characteristics of male pups were not affected in DEHP treatment groups. The WBC, HB, HT, MCH and albumin values in male pups were not affected in all experimental groups, but RBC MCV, MCHC, PLT and total protein values were significantly different among the experimental groups(P<0.05). In female pups, the effects of DEHP administration were not affected the body and uterus weight, but the left ovary in 10.0mg DEHP group was significantly heavier than in control and 0.5mg DEHP group(P<0.05). The WBC, MCV, MCH, MCHC, PLT, albumin, BUN and total protein values in female were not different in all experimental groups. The RBC, HB and HT values were significantly different among the experimental gruop(P<0.05). The historical evaluation of testis in male pups that were grown to 45 days after birth was not different in all experimental groups. The ovary in female pups had many corpus luteum in 10.0mg DEHP group. The endometriosisi of uterus was significantly decreased in DEHP group. There results suggest that low concentration of DEHP administration in pup born after DEHP administration in pregnant mice was not affered on reproductive characteristic, but was affected on blood hematological and chemical values.

Transforming Growth Factor β가 돼지 난자의 체외성숙에 미치는 영향

신명균 ( Myung Kyun Shin ),박춘근 ( Chun Keun Park ),조재원 ( Je Won Cho ),정희태 ( Hee Tae Cheung ),양부근 ( Boo Kun Yang ),김정익 ( Chung Ik Kim ) 한국가축위생학회 1999 韓國家畜衛生學會誌 Vol.21 No.3

돼지 수정란의 체외생산은 난자의 체외성숙과 체외수정에 관한 기술의 부족으로 아직까지 만족스럽지 못한 수준이다. 특히 돼지 수정란의 체외생산에는 복잡한 세포질의 성숙과정과 높은 다정자침입율 및 전핵형성의 억제등의 문제점이 있다. 본 연구에서는 돼지 난자의 체외성숙 체계를 개선하기 위하여 transforming growth factorβ(TGFβ)의 첨가가 난자 및 난구세포에 미치는 효과에 대하여 검토하였다. 체외성숙용 배지에 TGFβ를 1~10ng/ml의 농도로 첨가하여 미성숙 난자를 배양한 결과 성숙율이 높아졌다. TGFβ의 효과는 난구세포가 제거된 난자의 성숙에도 효과적이었다. TGFβ의 첨가시기에 따른 차이는 없었으나, 난구세포를 제거한 난자의 경우에는 성숙배양 전반기(59%) 또는 후반기(57%) 24시간 동안에만 TGFβ를 첨가하는 것이 48시간 동안 계속하여 첨가(27%)하는 경우 또는 비첨가(38%)에 비하여 유의적으로 높은 성숙율을 나타냈다(p<0.05). 이와 같은 결과는 난구세포가 돼지 난자의 체외성숙에 필수적이지만 TGFβ는 난구세포가 제거된 난자의 체외성숙에 어느정도 유익한 효과를 발휘하는 것으로 추측된다.

한우 후보종모우에 selenium과 vitamin E 투여가 정액성상에 미치는 영향

이성수,박노형,원유석,박동헌,김종복,양부근,Lee, Seoung-soo,Park, No-hyoung,Won, You-seog,Park, Dong-heon,Kim, Jong-bok,Yang, Boo-keun 대한수의학회 2000 大韓獸醫學會誌 Vol.40 No.2

To improve the semen production, the selenium(Se) and vitamin E(Vit. E) were administrated into Hanwoo young sire for intensification an antioxidant system and the taurine were supplemented into semen extender for improving the semen characteristics. The 16 heads ranging from twenty to thirty two months of age were randomly assigned to control group, Se-admistrated group(Se-group), Vit. E-administrated group(Vit. E-Group) and Se and Vit. E administrated group(Se and Vit. E-group). Se and Vit. E dministrated 3 times every 30 days by intramuscular injection. The administration of Se, Vit. E, and Se and Vit. E didn't affect on semen volume, sperm concentration, and total sperm number among all groups(p>0.05). Adiministration of Se improved sperm motility and viability. Motility of Se-group and control were 26.01% and 19.20%, respectively(p<0.05). Viability of Se-group(47.07%) was higher than control group(35.73%), Vit. E group(36.55%)(p<0.05). The administration of Se and Vit. E didn't affect sperm capacitation and acrosome reaction. The 100mM taurine supplement into semen extender increased the motility of frozen/thawed semen in the Vit. E-group(p<0.05) and had a beneficial effects on decreasing abnormality of frozen/thawed semen in all groups(p<0.05). These results indicate that Se administration improve sperm motility and viability, and the taurine supplement into semen extender decrease abnormality in Hanwoo young sire.

임신중인 생쥐에 DEHP 투여가 번식현상에 미치는 영향 Ⅰ. 임신중인 생쥐에 DEHP 투여가 분만 후 번식특성과 혈액성분에 미치는 영향

박동헌,장현용,박춘근,정희태,김정익,양부근,Park, Dong-Heon,Jang, Hyun-Yong,Park, Choon-Keun,Cheong, Hee-Tae,Kim, Choung-Ik,Yang, Boo-Keun 한국발생생물학회 2004 발생과 생식 Vol.8 No.2

본 연구는 임신한 생쥐에 DEHP의 투여가 분만 후 생쥐의 번식기능과 태아의 성비에 미치는 영향을 검토하였다. 임신한 자성 생쥐에 DEHP을 투여한 후 임신 19일째에 임신한 자성의 체중, 태아의 체중, 태아수 및 태아성비를 조사한 결과, 실험개시시와 실험종료시 임신한 자성의 체중은 각 투여구간에 차이가 없었으며, 자${\cdot}$웅성태아의 체중, 평균 태아수 및 태아의 자${\cdot}$웅성 성비는 투여구간에 커다란 차이가 없었다. 임신기에 DEHP의 투여가 분만한 후 어미의 번식기관무게와 혈구화학치에 미치는 효과를 검사한 결과, 각 투여구간에 커다란 차이는 없었다. 임신기에 DEHP의 투여가 분만한 후 난소와 자궁의 조직에 미치는 영향을 조사한 결과, 난소에서는 모든 투여구간에 커다란 차이는 없었으나, 자궁에서는 0.5mg 투여구가 대조구에 비해 자궁내막층과 부종이 감소했으며, 1.0mg 투여구와 10.0mg 투여구는 일부에서 자궁내막층이 감소했지만 0.5mg 투여구에서의 감소보다는 적었다. 임신중인 생쥐에 저농도의 DEHP의 투여는 임신중인 모체와 태어난 자손의 번식 특성과 혈구화학치에 영향을 미치지 않았다. This study is illustrated that 1) the effects of DEHP administration on body weight, sex ratio and litter size on 19 days in the prenatal period 2) the effects of reproductive organ weight and blood hematological values on 25 days postpartum in pregnant mice. DEHP was administrated to pregnant mice by intraperitoneal injection at 0, 0.5, 1.0 and 10.0mg/kg B.W from day 1 to day 16 in the gestation period with 5 times at 3 days interval. There were no significantly differences in the material body weight, litter size and sex ratio on 19 days in the prenatal period. The body weight of the fatal male mice was slightly lower in 1.0 and 10.0mg DEHP groups than in any other groups. The reproductive organ weight and hematological values in dam mice on 25 days postpartum were not affected by DEHP administration. The histological evaluation of ovary in dam mice on 25 days after dilivery was not different in all experimental groups, but the endometriosis and edema of uterus in dam mice were significantly decreased in 0.5mg DEHP group than that of control group.These results indicate that the administration of low concentration of DEHP was not affected on reproductive characteristic and blood hematological values in pregnant mice.

유전 및 육종 : 한우 Exostosin-1 유전자의 SNP 탐색 및 경제형질 관련성 분석

이승환 ( Seung Hwan Lee ),김범수 ( Bum Soo Kim ),조용민 ( Yong Min Cho ),양부근 ( Boo Keun Yang ),윤두학 ( Du Hak Yoon ),김남국 ( Nam Kuk Kim ),허강녕 ( Kang Nyeong Heo ),박응우 ( Eung Woo Park ) 한국동물자원과학회(구 한국축산학회) 2011 한국축산학회지 Vol.53 No.1

본 연구는 한우의 경제형질 관련 유전적 표지인자(DNA marker) 개발을 목적으로 EXT1 유전자의 다형성과 발현양상을 관찰하여 경제형질과의 관련성을 확인하고자 수행하였다. PCR-direct sequencing을 통하여 4개(T272196A, C272359T, G290964A 및 A302092G)의 SNPs를 탐색 하였으며, 탐색된 SNPs를 통하여 PCR-RFLP 기법으로 유전자형을 결정한 후 경제형질과 관련성을 분석하였다. 그 결과 T272196A 좌위에서 근내지방 육종가(p=0.014), G290964A 좌위에서 등지방두께 육종가 추정치(p=0.001), A302092G 좌위에서는 등지방두께 육종가(p<0.001) 및 등심단면적 육종가(p=0.020)에서 각각 유의적인 연관성이 인정되었다. 따라서, 본 연구를 통해 확인된 SNP를 이용하여 한우의 선발에 활용 가능할 것으로 사료된다. The aim of this study was to identify the polymorphism on exostosin-1(EXT1) gene and to associate with economic traits in Hanwoo(Korean cattle). We sequenced for detection of single nucleotide polymorphism(SNP) with 24 unrelated individuals and identified four SNPs(T272196A, C272359T, G290964A and A302092G). Relationship between the genotypes of 583 Hanwoo individuals by PCR-RFLP and economic traits were analyzed by general linear model. In EXT1 gene, there were four SNPs associated with economic traits such as eye muscle area breeding value, marbling score breeding value, backfat and thickness breeding value(p<0.05 to p<0.01). In conclusion, this study indicates an important role of EXT1 gene in determining the meat quality or economic characteristics in Hanwoo.