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
在本研究中,我们利用结构生物学技术,即三维和功能体内核磁共振观察技术,发展了一种新的环境内分泌干扰物(EDs)对家畜生殖和遗传毒性及致畸性的评估方法。(1)活体评估:应用新的活体内核磁共振波谱技术和^<;31>;P和^1H-核磁共振成像技术,准确和非侵入性地评估环境内分泌干扰物的致畸作用。以极低剂量(0.001~1,000 ng/kg体重)给妊娠小鼠注射雌二醇(天然雌激素、己烯雌酚、合成雌激素、双酚A、植物雌激素等),在体内无创性、定量地获取胚胎的P-核磁共振谱,以评价能量代谢。胚胎ATP水平显著下降,但常规组织学和生化方法未检测到明显的毒理学变化。因此,体内核磁共振技术具有高度的敏感性(至少是常规方法的1000倍),对于EDs和许多环境污染物的毒理学评估是有用的。(2)体外评估:使用培养的卵母细胞、精子、受精卵和胚胎来评估EDs和环境污染物的生殖和遗传毒性。用激光共聚焦显微镜非侵入性实时成像技术显示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