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.

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.

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.

임신중인 생쥐에 Bisphenol A 투여가 모체의 생식독성과 태아의 성비에 미치는 영향

박동헌(Dong-Heon Park),장현용(Hyun-Yong Jang),김정익(Choung-Ik Kim),정희태(Hee-Tae Cheong),박춘근(Choon-Keun Park),양부근(Boo-Keun Yang) 한국독성학회 2005 Toxicological Research Vol.21 No.2

Bisphenol A (BPA), a environmental endocrine disruptor, is considered to bind to estrogen receptors and to regulate the expressions of estrogen responsive genes. This study was to evaluate the effect of BPA administration on body weight, sex ratio and litter size on 18 days in prenatal periods, the effect of reproductive organ weight and blood hematological values on 24 days postpartum in pregnant mice. The female mice was administrated to low doses of BPA (0, 0.05, 0.5 and 5.0 ㎎/㎏ B.W.) by intraperitoneal injection in gestation days 0~15 with 5 times at 3 days interval. The maternal body weight, litter size and sex ratios were similar to in all experimental groups, but body weights of male and female offspring was significantly lower in 5.0 ㎎ BPA group when compared to any other groups (P<0.05). No treatment-related effects on body weight, ovary weight and blood hematological values were observed in dams on 24 days after delivery. The uterine weight in 5.0 ㎎ BPA group was slightly higher than those of any other groups, but not significantly difference. The histological evaluation of ovary in dam mice on 24 days after dilivery was not difference in all experimental groups, but the endometriosis of uterus in dam mice were significantly increased in 0.5 ㎎ BPA group when compared to control group. These results indicates that low concentration of BPA should not be considered as a selective reproductive toxicant.

임신중인 생쥐에 Bisphenol A 투여 후 태어난 차산자의 생식독성과 혈액성분에 관한 연구

박동헌(Dong-Heon Park),장현용(Hyun-Yong Jang),김정익(Choung-Ik Kim),정희태(Hee-Tae Cheong),박춘근(Choon-Keun Park),양부근(Boo-Keun Yang) 한국독성학회 2005 Toxicological Research Vol.21 No.2

Bisphenol A (BPA) is a monomer used in the manufacture of a multitude of chemical products, including epoxy resins and polycarbonate. The objective of this study was to evaluate the effects of BPA administration on reproductive characteristics and blood hematological and chemical values in offspring of pregnant dams treated with BPA. BPA was administrated to pregnant mice by intraperitoneally injection with 0, 0.05, 0.5 and 5.0 ㎎/㎏ B.W. for 5 times at 3 days interval on gestation days 1~16. There were no treatment-related effects of BPA on reproductive organ weight in male offsprings at 45 days-of-age, but body weight was the lowest in 5.0 ㎎ BPA group when compared to other groups (P<0.05). No differences in semen characteristics (sperm concentration, viability, motility and abnormality) were observed between the control and BPA treatment groups. The WBC, HB, HT, MCV, MCH, MCHC, albumin, BUN and total protein of blood hematological and chemical values in male offsprings were not difference for any treatment groups, but RBC value in BPA groups was significantly increased comparing to the control group (P<0.05). The PLT value was slightly higher in 5.0 ㎎ BPA groups than in any other group, but not significantly difference among the experimental groups. In female offsprings, the effects of BPA didn't affect to the body and ovary weight, but the uterus weight in 5.0 ㎎ BPA group was slightly heavier than that of control group (P>0.05). No statically significant difference in blood hematological values in female offsprings were observed between the control group and BPA groups, but the concentration of albumin and BUN were significantly higher in 0.5 ㎎ BPA group when compared to control and other BPA treatment groups (P<0.05). The histological evaluation of testis and ovary in growing offspring at 45 days-of-age was not difference between the control group and BPA groups, but endometriosis of the uterus in female offspring was dramatically increased in 0.5 and 5.0 ㎎ BPA groups. These founding suggest that low concentration of BPA might not have a important role on reproductive ability or blood metabolite in offspring of pregnant dams treated with BPA.