• Title/Summary/Keyword: Embryonic effects

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Hsp90 Inhibitor, 17-AAG, Affects Early Embryonic Development and Apoptosis of Bovine Embryos (Hsp90의 저해제인 17-AAG의 처리에 따른 소 수정란의 배발달 및 세포사멸 양상)

  • Hong, Joo-Hee;Min, Sung-Hun;Lee, E-Nok;Son, Hyeong-Hoon;Park, Hum-Dai;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.307-311
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    • 2011
  • Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG ($33.1{\pm}9.6$ vs $21.7{\pm}8.3%$). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP-treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The mRNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.

IN VITRO DEVELOPMENT OF BOVINE ONE-CELL EMBRYOS FERTILIZED IN VITRO IN SERUM- AND FEEDER CELL-FREE CULTURE SYSTEMS

  • Ohboshi, S.;Hanada, K.;Zhao, J.;Hattori, M.;Fujihara, N.;Umetsu, R.;Yoshida, T.;Tomogane, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.5
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    • pp.583-590
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    • 1996
  • The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium (mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera (CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage (8.6-21.4%), dependent upon their product lots, although the development might not be affected at least by serum progesterone levels. ${\beta}$-Mercaptoethanol (${\beta}$-ME) supplemented into mSOFM was effective to the embryonic development (27.8%), as well as the co-culture system with cumulus cells (19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and ${\beta}$-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin (BSA, fraction V), but not polyvinyl alcohol (PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture (CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA (7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium (final 2.0 mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with ${\beta}$-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos.

Embryonic Stem Cell-Preconditioned Microenvironment Effects on Epidermoid Carcinoma

  • Ryoo, Zae Young;Kim, Myoung Ok
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.275-281
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    • 2012
  • Embryonic stem cell-preconditioned microenvironment is important for cancer cells properitities by change cell morphology and proliferation. This microenvironment induces cancer cell reprogramming and results in a change in cancer cell properties such as differentiation and migration. The cancer microenvironment affects cancer cell proliferation and growth. However, the mechanism has not been clarified yet. Using the ES-preconditioned 3-D microenvironment model, we provide evidence showing that the ES microenvironment inhibits proliferation and reduces oncogenic gene expression. But ES microenvironment has no effect on telomerase activity, cell viability, cellular senescence, and methylation on Oct4 promoter region. Furthermore, methylation of Nanog was increase on ES-preconditioned microenvironment and supports results that no difference on RNA expression levels. Taken together, these results demonstrated that in the ES-preconditioned 3-D microenvironment is a crucial role for cancer cell proliferation not senescence.

The Effect of Dammarane Glycosides of Panax ginseng on Primary Cultured Chicken Brain Cells (인삼 Dammarane Glycoside류 분획물이 일차배양한 계배의 뇌세포에 미치는 영향)

  • Park, Mi-Jung;Song, Jin-Ho;Kim, Young-Choong
    • YAKHAK HOEJI
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    • v.33 no.1
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    • pp.39-45
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    • 1989
  • Effects of dammarane glycosides of Panax ginseng on primary cultured chicken embryonic brain cells were studied by microscopic observation and determination of the activity of pyruvate dehydrogenase complex (PDHC). Brain cells were prepared from the brain of 10-day-old chicken embryo and cultured with either a standard medium consisted of 85% Dulbecco's Modified Eagle Medium (DMEM), 10% horse serum and 5% chicken embryonic extracts or a deficient medium consisted of 90% DMEM and 10% horse serum. It was observed that dammarane glycosides of Panax ginseng seemed to show the tendency to stimulate the neurite outgrowth of brain cells which were cultured with a deficient medium under microscopic observation. The activity of PDHC in brain cells cultured with a deficient medium was increased by dammarane glycosides of Panax ginseng.

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New role of E3 ubiquitin ligase in the regulation of necroptosis

  • Seo, Jinho;Lee, Eun-Woo;Song, Jaewhan
    • BMB Reports
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    • v.49 no.5
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    • pp.247-248
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    • 2016
  • Necroptosis is a well-known form of caspase-independent cell death. Necroptosis can be triggered by various extrinsic stimuli, including death ligands in the presence of receptorinteracting protein kinase 3 (RIPK3), a key mediator of necroptosis induction. Our recent studies have revealed that C-terminus HSC-70 interacting protein (CHIP), an E3 ligase, can function as an inhibitor of necroptosis. CHIP−/− mouse embryonic fibroblast showed higher sensitivity to necrotic stimuli than wild-type mouse embryonic fibroblast cells. Deleterious effects of CHIP knockout MEFs were retrieved by RIPK3 depletion. We found that CHIP negatively regulated RIPK3 and RIPK1 by ubiquitylation- and lysosome- dependent degradation. In addition, CHIP−/− mice showed postnatal lethality with intestinal defects that could be rescued by crossing with RIPK3−/− mice. These results suggest that CHIP is a negative regulator of RIPK1 and RIPK3, thus inhibiting necroptosis.

Effects of Ginseng and Its Saponins on Experimental Amnesia in Mice and on Cell Cultures of Neurons (인삼 및 인삼 사포닌이 쥐의 건망증 및 신경세포배양에 미치는 영향)

  • Saito Hiroshi;Nishiyama Nobuyoshi;Iwai Akihiko;Kawajiri Shinichi;Himi Toshiyuki;Sakai Toshimi;Fukunaka Chizu
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.92-98
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    • 1988
  • The present study was performed to find the effects of ginseng and its saponins. which is written in Chung Yao Ta Tsu Tien as anti-amnesia in its chief indication. on experimental amnesia in mice. In the step through test. ginsenoside $Rb_1\;(GRb_1)\;and\;GRg_1$ facilitated the registration of memory and antagonized the electroconvulsive shock (ECS)-induced inhibition of the retention of memory. Moreover. $GRg_1$ antagonized the EtOH-induced inhibition of the retrieval of memory. In the step down test. $GRb_1\;GRb_2\;and\;GRg_1$ antagonized the ECS-induced inhibition of the retention of memory. Moreover. $GRg_1$ antagonized the EtOH-induced inhibition of the retrieval of memory and facilitated the acquisition of short term memory. In the shuttle hox and lever press tests. they have no effects on acquisition and retrieval of memory. except $GRb_1\;GRb_1$ depressed the retrieval of conditioned avoidance response in the shuttle box test. After the end of four tests. the effects of these orally administered drugs on sedative. analgesic. antipyretic and anticonvulsant actions. and on spontaneous and exploratory movements were tested in doses of less than 500mg/kg. but they had none of these effects. Present study may indicate that $GRg_1$ had effects on the retrieval of memory and on the acquisition process of learning response. The recent research on the role of NGF for the survival. regeneration and regulation of brain in adult animals. indicated the importance of NGF on dementia and amnesia. During our research on the specificity of the neurite out growth induced by NGF. we found that the effect of NGF was potentiated by $GRb_1$ in organ cultures of chick embryonic dorsal root ganglia. Then. the effect of $GRb_1$ on neuronal cell survivalin cell culture system was studied. $GRb_1$ potentiated the NGF-mediated increase of neurofilaments in cell cultures of chick embryonic sensory and sympathetic neurons. NGF with $GRb_1$ also showed a tendency to increase the number of surviving neurons of rat embryonic cerebral cortex. NGF increased choline acetyl transferase activity in cell cultures of rat embryonic septum area neurons. but $GRb_1$ did not potentiate NGF activity in cell cultures of rat embryonic septum area neurons. Present study may indicate that $GRb_1$ plays an important role for the survival or regeneration of neurons in the brain.

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Effects of Enzyme Inducers and Glutathione on the Embryotoxicity of Cyclophosphamide in Cultured Rat Embryos (효소유도제 및 glutathione이 전배자배양된 랫드태자에서 cyclophosphamide의 독성에 미치는 영향)

  • 한순영;신재호;권석철;강명옥;이유미;김판기;양미라;박귀례;장성재
    • Toxicological Research
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    • v.11 no.1
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    • pp.31-36
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    • 1995
  • Cyclophosphamide (CP) must be enzymatically activated by cytochrome P450(CYP)-linked mixed-function oxidation pathway to be either mutagenic or teratogenic. Influences of alterations in hepatic mixed-function oxidase acitivity and glutathione (GSH) content on the embryotoxicity of CP were studied in rat whole embryo culture system. The embryotoxicity of CP was compared using rat S-9 fraction (S-9) pretreated with chemicals inducing different CYP isozymes, acetone (ACE), Aroclor 1254 (ARO), $\beta$-naphthoflavone (NAF) and phenobarbital (PHE). When 10.5 day embryos were cultured in the immediately centrifuged rat serum for 48 hrs using general gas char{ging schedule, CP$(40{\mu}g/ml)$ with S-9 induced by either NAF or PHE increased the incidence of realformations and significantly decreased embryonic growth compared with the non-induced S-9 group. ACE or ARO induced S-9 group showed no significant difference in embryonic growth. These data suggest that PB and/or NAF inducible CYP isoenzymes are mainly involved in the activation of CP. To examine the effect of GSH on the embryotoxicity of CP, 10.5 day embryos were exposed to CP and S-9 after preincubation with 10 mM of GSH for 3 hrs. In the GSH pretreated group the growth of embryos increased significantly compared with that of the untreated group, suggesting that GSH may protect embryos in culture from some toxic effects of CP.

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Developmental Abnormalities in Zebrafish Angiogenesis with Chronic Exposure to Crude Oil and Dispersant

  • Lee, Suyeon;Kim, Kyoohyun;Kim, Hyunjin;Yeo, Sang-Yeob
    • Korean Journal of Environmental Biology
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    • v.31 no.1
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    • pp.10-18
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    • 2013
  • Oil spills have occurred throughout the years of industrialization and represent a global challenge as they affect vast areas of the ocean. The toxicity of crude oil to aquatic organisms has been extensively investigated, but the potential impacts of crude oil on vertebrate development remain largely unknown. Here, we investigated the effects of dispersants used in treating a recent oil spill, as well as that of crude oil, on vertebrates by using the zebrafish (Danio rerio) model species, which has been widely used in empirical studies of both early embryonic development and adult physiology. Chronic exposure to crude oil resulted in marked developmental abnormalities, including pericardial edema, abnormal trunk vessel development, retardation of axonal branching, and abnormal jaw development. Embryonic development was affected more severely by exposure to the oil-dispersant combination than to the oil alone. Thus, the zebrafish in vivo model system suggests that dispersant treatment can have detrimental developmental effects on vertebrates and its potential impact on marine life, as well as humans, should be carefully considered in clean-up efforts at the site of an oil spill.