• Title/Summary/Keyword: rat embryo

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Effects of Glucose and Inorganic Phosphate on the Development of Rat 8-Cell Embryos In Vitro (Glucose와 Inorganic Phosphate가 Rat 8-세포기 난자의 체외배양에 미치는 영향)

  • 이홍미;진동일
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.251-258
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    • 1996
  • This study was designed to evaluate the potential inhibitory effects of glucose (5.56 mM vs. 0 mM) and/or phosphate (potassium phosphate, 1.19 mM vs. 0 mM) on the in vitro devel-opment of rat 8-cell embryos (n=345 embryos from 36 mature rats). Evaluation of embryos at 48 h for developmental stage (STG) indicated that 37% (31/84), 70% (64/91), 69% (59/85), and 77% (67/85) developed to the blastocyst stage in media with glucose+phosphate, glucose only, phosphate only, and no glucose or phosphate, respectively. Embryo development (2.90${\pm}$0.097 for STG) in medium with glucose + phosphate was significantly reduced (P<0.001), while no significant differences were observed between all other media (3.4~3.5${\pm}$0.093-0.097 for STG). Evaluation of embryos for final cell number (FCN) indicated that the greatest number of cells (nuclei) resulted in medium with glucose alone (29.3${\pm}$0.97 cells, P<0.001). No significant differences were observed for FCN for the remaining three media (l7.5${\pm}$1, 04 cells, 18.6${\pm}$1.Ol cells, and 19.8${\pm}$1.01 cells for glucose+phosphate, phosphate only, and no glucose or phosphate, respectively). Our results suggest that glucose and phosphate together exert an inhibitory effect on 8-cell rat embryo development, while glucose alone was beneficial, yielding greater numbers of cells per embryo.

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Effects of the Oocyte and Developmental Stages of the Rat Embryos after the Vitrified Freezing on the Survival Rate(FDA-test) (Rat 난포란과 수정란의 발육단계별 유리화 동결 융해후 생존성(FDA-test)에 미치는 영향)

  • 고혁진;김중계
    • Journal of Embryo Transfer
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    • v.11 no.1
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    • pp.41-50
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    • 1996
  • This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.

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A Study on the Activity of Alkaline Phosphatase of Rat Oviduct During Early Embryonic Development (흰쥐 초기배아 발생기간 중 수란관조직의 알카리성 Phosphatase활성도에 관한 연구)

  • Kim, Sung-Rye;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.18 no.1
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    • pp.41-48
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    • 1991
  • The present investigation has been undertaken to elucidate the functional role of ovarian steroids on the mechanism of oviduct differentiation during early embryonic development in rat. The activity of alkaline phosphatase (ALPase) was measured in the oviduct tissue under different steroids treatment regime on day 9 pregnancy. The ALPase activity of the oviduct of pseudopregnant rat was compared with that of normal pregnant rat. The results of day 9 pregnancy rat oviduct clearly demonstrated that $17{\beta}-estradiol$ and progesterone were effective in pseudopregnant rat oviduct. In the ovary intact group the ALPase activity was similar in both of normal and pseudopregnant oviduct, but in the $17{\beta}-estradiol$ treated group the ALPase activity in normal pregnancy was significantly higher than that in pseudopregnancy. The effect of estradiol on the normal pregnant rat oviduct was apparently found on day 3 and day 9 pregnancy. This study, therefore, clearly demonstrates that $17{\beta}-estradiol$ is much potent in oviduct tissue differentiation. It is suggested that absence of $17{\beta}-estradiol$ effect on pseudopregnant rat oviduct is due to there is no embryo passing througth the oviduct.

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Sex Differences in the Glucose-6-Phosphate Dehydrogenase Activity of the Rat Livers at Various Stages of Development (출생전후 및 성별로 본 쥐의 간세포에서의 G-6-PD 활성)

  • Hahn, Sahsook
    • The Korean Journal of Zoology
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    • v.17 no.1
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    • pp.17-22
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    • 1974
  • The G-6-PD (glucose-6-phosphate dehydrogenase) activity, the first step in the pentose phosphate shunt, of rat livers during prenatal and pstnatal development in different sexes was studied. The enzyme activity is very high (54.2 units) at 16 days of embryo ad then decreases to a low level (17.6 units) at 13days after birth. There are significant increase between 13 and 15 days of age and continuously increases to the level of 53.2 units at 19days of age. The G-6-PD activity in female rat livers was slightly higher than in males.

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Effects of Rice Embryo and Embryo Jelly with Black Rice Bran Pigment on Lipid Metabolism and Antioxidant Enzyme Activity in High Cholesterol-Fed Rats (쌀 배아와 흑미 미강 색소 첨가 배아젤리가 고콜레스테를 식이 흰쥐의 지질대사와 항산화효소 활성에 미치는 영향)

  • Cho, Min-Kyung;Kim, Mi-Hyun;Kang, Mi-Young
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.200-206
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    • 2008
  • We investigated the effect of rice embryo and embryo jelly with black rice bran pigment on lipid metabolism and antioxidant activity. Thirty 4-week-old male Sprague-Dawley rats were fed high cholesterol diets supplemented with 15% rice embryo and 25% embryo jelly added black rice bran pigment, respectively, for 6 weeks. Plasma and hepatic lipid profile, lipid peroxidation, and the activity of antioxidant scavenger enzymes in liver were examined. Supplementation with rice embryo and embryo jelly had no effect on food intakes in high cholesterol-fed rats. The plasma triglyceride concentration was not significantly different among the groups. Supplementation with rice embryo and embryo jelly resulted in lower plasma and hepatic total cholesterol (TC) concentration and high-density lipoprotein-cholesterol (HDL-C)/TC ratio and atherogenic index compared to the control group, while the plasma HDL-C concentration tended to elevated. Rice embryo and embryo jelly tended to lower plasma and hepatic levels of thiobarbituric acid reactive substances than the control group. Moreover, hepatic antioxidant enzyme activities, including superoxide dismutase and glutathione peroxidase, were significantly higher in the rice embryo and embryo jelly groups. In conclusion, rice embryo and embryo jelly was very effective in improving the lipid metabolism and reducing oxidative stress by up-regulating the hepatic antioxidant enzymes in high cholesterol-fed rats.

Inhibitor of DNA Binding Protein (Id)1 and Id2 mRNA Expression on Folliculogenesis in Rat Ovary (랫드 난소에서 난포 발달에 따른 DNA 결합 단백질 억제인자 (Inhibitor of DNA Binding Protein) Id1 and Id2 mRNA 발현)

  • Hwang, Seong-Soo;Lee, Pyung-Hee;Ko, Yeoung-Gyu;Yang, Byoung-Chul;Seong, Hwan-Hoo;Min, Kwan-Sik;Yoon, Jong-Taek
    • Journal of Embryo Transfer
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    • v.23 no.3
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    • pp.183-187
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    • 2008
  • This study was conducted to analyze the expression pattern of inhibitor of DNA binding proteins (Id)1 and Id2 mRNA on folliculogenesis in rat ovary. The ovaries were obtained from 27 days old Sprague-Dawley rat, fixed, dehydrated, and paraffin embedded. For in situ hybridization, anti-sense and sense Idl and Id2 cRNA probes were prepared and applied to the ovarian section. The ovarian sections were coated with NTB-2 emulsion. After that, the slides were developed and counterstained with hematoxylin and eosin staining. In oocytes, the hybridizational signals of Id1 mRNA were strong in primordial and primary follicles, however, there were no signals in that of atretic or preovulatory follicles. The Id2 mRNA signals were also strong in the oocytes of primordial, primary and secondary follicles. Interestingly, the Id2 mRNA was expressed specifically granulosa cells, but nor in oocyte or theca cells in dominant and preovulatory follicles. Based on these results, Id1 and Id2 mRNA was expressed specifically at follicle stages and follicular tissue and might be closely related with follicle development.

Gene Expression Patterns of the Endogenous Antioxidant Enzymes in Linuron-Treated Rat Ventral Prostates after Castration

  • Yon, Jung-Min;Lin, Chunmei;Lee, Yoon-Bok;Lee, Beom-Jun;Yun, Young-Won;Nam, Sang-Yoon
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.101-105
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    • 2012
  • Linuron is a pesticide with a weak anti-androgenic property, which impacts male reproductive organs. In this study, to clarify whether linuron affects the cellular antioxidant system of ventral prostate, gene expression patterns of the representative antioxidant enzymes such as glutathione peroxidase (GPx), selenoprotein P (SePP), and superoxide dismutase (SOD) were investigated in the rat ventral prostates exposed to linuron using real-time RT-PCR analyses. Sprague-Dawley rats castrated at 6 weeks old were treated with linuron (25, 50, or 100 mg/kg per oral) daily for 10 days after testosterone propionate administration (0.4 mg/kg) subcutaneously. As compared to normal control animals, mRNA levels of phospholipid hydroperoxide GPx (PHGPx), SePP, and Mn SOD significantly increased in the prostates exposed to linuron (25, 50, and 100 mg/kg). However, cytosolic GPx (100 mg/kg) and Cu/Zn SOD (25, 50, and 100 mg/kg) mRNA levels significantly decreased in the ventral prostates. These results indicate that linuron upregulates the expressions of PHGPx, SePP, and Mn SOD mRNAs, but down-regulates the expressions of cytosolic GPx and Cu/Zn SOD in rat prostates, suggesting that linuron may have dual effects in the cellular antioxidant system of prostate.

Embryotoxicity of Ochratoxin A in Cultured Rat Embryonic Midbrain Cells and Whole Embryos (흰쥐 배양 전배자 및 중뇌세포에서 Ochratoxin A의 독성)

  • Hong, Jin-Tae;Park, Kui-Lea;Han, Soon-Young;Park, Ki-Sook;Kim, Hyung-SIk;Oh, Se-Dong;Park, Hee-Jung;Lee, Rhee-Da;Jang, Seung-Jae
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.336-344
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    • 1998
  • Effects of ochratoxin A (OTA) on embryo development were studied in cultured whole embryos from 9.5 day gestation rat for 48 h. OTA (more than $0.5{\mu}g/ml$) induced microcephaly in the cultured rat whole embryos. Protein and DNA content, and DNA synthesis were significantly inhibited by OTA. We next examined whether the microcephaly seen in cultured whole embryo partially results from inhibition of differentiation of embryonic midbrain cells. Embryonic midbrain cells were extracted from 12 day gestation rat embryos, and cultured for 96 hr. OTA ibhibited cell differentiation about 50% over control. We also tested whether OTA-induced embryotoxicity would be associated with oxidative damages. We measured the ${\gamma}$-glutamyltranspeptidase (${\gamma}$-GT) and glutathione peroxidase (GPX) activities, and glutathione (GSH) content in both cultured whole embryos and embryonic midbrain cells. OTA decreased GSH content, whereas slightly increased ${\gamma}$-GT activity, but GPX activity was not significantly changed. These results show that OTA caused the microcephaly and its effect may be partially due to the inhibition of cell differentiation of embryonic midbrain cells, but the role of oxidative damages is not clear in embryotoxicity.

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Teratogenic Effects of Phenytoin on Rat Embryos in Culture (랫드에 있어서 배양배자에 대한 Phenytoin의 최기형성 효과)

  • Kim, Jong-Choon;Lim, Kwang-Hyeon;Chung, Moon-Koo;Roh, Jung-Koo
    • Toxicological Research
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    • v.14 no.3
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    • pp.357-363
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    • 1998
  • The teratogenic potential of the anticonvulsant drug phenytoin (PHT) has been well documented both in the human and in the experimental animals. However there are few reports on the effects of PHT on embryonic development in rats in vitro. The present study was performed to evaluate the teratogenic effects of PHT using whole-embryo culture system in rats. Sprague-Dawley rat embryos were explanted on gestational day (GD) 9.5 and cultured for 48 hrs in the immediately centrifuged and heat-inactivated rat serum containing 0,25,50, or $100{\mu}g$ PHT/mL. At the end of culture period the embryos were scored for morphological development according to the procedure of Van Maele-Fabry, and their total protein contents were determined. At 100 ${\mu}$g/mL of culture medium. PHT caused significant reduction in developmental score and protein content of embryos and a high incidence morphological abnormalities (100%). Characteristic malformations included altered yolk and embryonic circulation, craniofacial hypoplasia, neural tube schisis, branchial arch defects, abnormal ratation, and limb bud hypoplasia, among others. There were no adverse effects on embryonic growth and development at concentrations of 25 and 50 ${\mu}$g /mL of culture medium. The results indicated that the dysmorphogenic effect of PHT on cultured embryos is due to a direct interference with embryonic development.

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Cell Surface Interaction with Expression of Fas Ligand Mediates Prolactin-Induced Apoptosis In Rat Luteal Cell Culture (Rat 황체세포 배양에 있어서 Prolactin에 의한 황체퇴행 및 Fas Ligand의 발현)

  • 장규태;박미령;선동수;윤창현
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.179-190
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    • 1998
  • Prolactin (PRL) surge in cycling rats at proestrous afternoon has previously been reported as an inducer of apoptotic cell death of luteal cells. This death-inducing action of PRL seeins unusual, because PRL can he categorized as a cell-survival factor, if other known physiological functions of PRL are taken into account. In this study, the apoptotic action of PRL was assessed in cultured cells prepared from rat luteal tissue and underlying molecular /cellular mechanism of PRL-induced luteolysis was analyzed. The latest crop of corpora lutea (CLs) were enucleated from rat ovaries at 18:00 h on the proestrous day before the next ovulation. Donor rats were pretreated with CB154, a dopamine agonist, in order to he exempted from the endogenous PRL surge. The harvested GLs were dispersed and cultured with or without PRL (2$\mu$g /ml) for 24 or 48 h. An addition of PRL to the culture medium changed the parameters indicative of cell death via apoptosis: a decrease in cell viability (MTT) and an increase in chromatin condensation. Most of the DNA breakdown in nuclei induced by PRL occurred in steroidogenic cells which were identified by 3$\beta$-HSD activity staining, and the number of 3$\beta$-HSD-positivecells were significantly decreased. Interestingly, most of the cells with an apoptotic nucleus adhered to one or more intact and seemingly non-steroidogenic cells. Because the expression of Fas has heen shown to be abundant in murine ovary, and Fas is known to have an exact physiological role in occurrence of apoptotic cell death, the membrane form-Fas ligand (rnFasL) was quantified in the cell lysate. An addition of PRL increased expression of mFasL. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, in place of PRL, enhanced the apoptotic parameters. Cumulatively, the apoptotic PRL action was addressed to cells unknown than steroidogenic lute~ cells. The most prohable candidate for the direct target cells is Tcells in the luteal tissue that can express mFasL in response to PRL.

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