A novel structural biological assessment for reproductive and genetic toxicity in farm animals using nuclear magnetic resonance technique
A novel structural biological assessment for reproductive and genetic toxicity in farm animals using nuclear magnetic resonance technique
批准号:
09356008
负责人:
MIYAMOTO Hajime
金额:
$20.54万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (A).
财政年份:
1997
资助国家:
日本
项目状态:
已结题
起止时间:
1997 至 2000
中文摘要
本研究利用结构生物学技术,即三维和功能性活体核磁共振(NMR)观察技术,建立了一种评价环境内分泌干扰物(EDs)对家畜生殖、遗传毒性和致畸性的新方法。(1)体内评估:应用新型体内13 P<31>和131 H-NMR光谱学以及13 <31>P和131 H-NMR成像技术,准确和无创地评估ED诱导的致畸性。以极低剂量(0.001- 1,000 ng/kg体重)对妊娠小鼠施用ED(作为天然雌激素的雌二醇17 β、作为阳性对照试剂的己烯雌酚、合成雌激素、双酚A、植物雌激素等),然后<31>无创地和定量地获取胚胎的体内13 P-NMR光谱以评估能量代谢(13 <31>P-ATP)。胚胎ATP水平显着下降,但没有显着的毒理学变化检测到常规的组织学和生化方法。因此,体内NMR技术是高度敏感的(比常规方法敏感至少1,000倍),并且可用于ED和许多环境污染物的毒理学评估。(2)体外评估:采用体外培养的卵母细胞、精子、受精卵和胚胎评价了EDs和环境污染物的生殖和遗传毒性。采用激光共聚焦显微镜观察DNA烷基化、类固醇激素受体mRNA表达异常。
英文摘要
In the present investigation, we developed a novel assessment method for reproductive and genetic toxicity and teratogenicity of environmental endocrine disrupters (EDs) in farm animals using structural biological techniques, i.e. three dimensional and functional in vivo nuclear magnetic resonance (NMR) observation techniques.(1) In vivo assessment : the novel in vivo ^<31>P-and ^1H-NMR spectoroscopy and ^<31>P-and ^1H-NMR imaging techniques for accurate and noninvasive assessment of teratogenicity induced by EDs were applied. Mice with pregnancy were administered EDs (estradiol 17beta as natural estrogen, diethylstilbestrol, synthetic estrogen, used as positive control reagent, bisphenol A, phyto estrogens, and so on) at extremely low dose (0.001-1,000 ng/kg of body weight), and then in vivo ^<31>P-NMR spectra of embryos were acquired noninvasively and quantitatively to evaluate the energy metabolism (^<31>P-ATP). A significant decrease in embryo ATP level was seen, but no significant toxicological changes were detected by conventional histological and biochemical methods. Thus, in vivo NMR techniques are highly sensitive (at least 1,000-fold more sensitive than conventional methods) and are useful for toxicological assessment of EDs and many environmental pollutants.(2) In vitro assessment : Cultured oocytes, sperm, fertilized eggs and embryos were used to assess the reproductive and genetic toxicity of EDs and environmental pollutants. DNA arkylation, abnormalities of steroid hormone receptor mRNA expression were visualized by noninvasive-real time imaging techniques using a confocal laser microscopy.
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Noboru Manabe et al.: "Reproductive Biology Update; Regulatory mechanisms of granulosa cell apoptosis in porcine ovarian follicle atresia." Nakanishi Pub.Co., Kyoto, Japan, 498 (1998)
Noboru Manabe 等人:“生殖生物学更新;猪卵巢卵泡闭锁中颗粒细胞凋亡的调节机制。”
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通讯作者:
Mizuho Nakayama,Noboru Manabe,Hajime Miyamoto et al.: "Species specific differences in apoptotic cell localization in granulosa and theca interna cells during follicular atresia in porcine and bovine ovaries."Journal of Reproduction and Development. 46. 1
Mizuho Nakayama、Noboru Manabe、Hajime Miyamoto 等人:“猪和牛卵巢卵泡闭锁期间颗粒细胞和卵泡膜内细胞凋亡细胞定位的物种特异性差异。”生殖与发育杂志。
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Noboru Manabe,Akira Myoumoto,Chiemi Tajima,Hajime Miyamoto et al.: "Immunochemical characteristics of a novel cell death receptor and a decoy receptor on granulosa cells of porcine ovarian follicles."Cytotechnology. 33・2. 189-201 (2000)
Noboru Manabe、Akira Myoumoto、Chiemi Tajima、Hajime Miyamoto 等:“猪卵巢卵泡颗粒细胞上的新型细胞死亡受体和诱饵受体的免疫化学特征”。《细胞技术》33・2。
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Miki Sugimoto et al.: "Ultrastructural changes in granulosa cells in porcine anttal follicles under going atresia indicate apoptotic cell death." Journal of Reproduction and Development. 44. 7-14 (1998)
Miki Sugimoto 等人:“闭锁的猪蚁卵泡中颗粒细胞的超微结构变化表明细胞凋亡。”
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眞鍋 昇ら: "Histochemical examination of cellular carbohydrate chains which change during ovarian follicular atresia in the porcine ovary." Journal of Japan Embryo Transfer Society. 19・1. 101-110 (1997)
Noboru Manabe 等:“猪卵巢卵泡闭锁期间细胞碳水化合物链的组织化学检查”,日本胚胎移植学会杂志 19・1(1997)。
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共 63 条
Real-time assessment technique for reproductive toxicity of environmental disruptors by nuclear magnetic resonance microscopy
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批准号:14360167
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项目类别:Grant-in-Aid for Scientific Research (B)
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资助金额:$9.22万
-
财政年份:2002
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负责人:MIYAMOTO Hajime
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依托单位:
Regulation mechanisms of germinal cell selection in mammals and its artificial control
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批准号:11460127
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项目类别:Grant-in-Aid for Scientific Research (B)
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资助金额:$9.66万
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财政年份:1999
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负责人:MIYAMOTO Hajime
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依托单位:
Regulatory mechanisms of granulosa cell apoptosis in porcine ovarian follicle atresia
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批准号:08456141
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项目类别:Grant-in-Aid for Scientific Research (B)
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资助金额:$4.8万
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财政年份:1996
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负责人:MIYAMOTO Hajime
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依托单位:
A novel assessment technique of reproductive and genetic toxicity in domestic animals using a nuclear magnetic resonance instrument
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批准号:06556048
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项目类别:Grant-in-Aid for Scientific Research (A)
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资助金额:$6.02万
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财政年份:1994
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负责人:MIYAMOTO Hajime
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依托单位:
Artificial control of oocyte maturation by in vitro perfused organ culture of the ovaries of farm animals
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批准号:05454112
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项目类别:Grant-in-Aid for General Scientific Research (B)
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资助金额:$4.29万
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财政年份:1993
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负责人:MIYAMOTO Hajime
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依托单位:
Development of cryopreservation method of the farm animal ovary
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批准号:02660274
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项目类别:Grant-in-Aid for General Scientific Research (C)
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资助金额:$1.54万
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财政年份:1990
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负责人:MIYAMOTO Hajime
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依托单位:
Efficient use of oocytes by the perfusion of farm animal ovary
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批准号:62560267
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项目类别:Grant-in-Aid for General Scientific Research (C)
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资助金额:$1.34万
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财政年份:1987
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负责人:MIYAMOTO Hajime
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依托单位:
海外基金