A novel assessment technique of reproductive and genetic toxicity in domestic animals using a nuclear magnetic resonance instrument

使用核磁共振仪评估家畜生殖和遗传毒性的新型技术

基本信息

批准号：
06556048
负责人：
MIYAMOTO Hajime
金额：
$ 6.02万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

项目摘要

In vivo and in vitro phosphorus-31 and proton nuclear magnetic resonance (NMR) spectra of the ovary, liver, skeletal muscle and embryo of experimental and farm animals were acquired using a house-made surface coil, conventional round coil, and/or imaging coils. Beta-ATP/inorganic phosphate ratio which represents the high energy state and intracellular pH etc.were observed in vivo. Many environmental toxicants were identified and quantified without any extraction processes from these organs and embryos using a NMR spectroscopy. Proton NMR experiments were done by a modification of conventional techniques, and 31P NMR experiments were performed as follows : During the NMR examination, the sample was maintained in a probe chamber at controlled air temperature of 25. C.In.vivo 31P NMR spectra of the steady-state organs within intact animals were obtained using a JNM-alpha-400 FT-NMR spectromenter (JEOL,Tokyo, Japan) equipped with 89-mm bore super-conducting magnet (9.20 Tesla ; Oxford Inst … More ruments, Bedford, MA,USA). The animal was mounted on a surface coil probe (two-turns, 2.0cm diameter) with was tuned to 31P at 161.7MHz and used in conjunction with a ferrite screen composed of strips of computer tape (Scotch 700 ; Three M Co., St.Louis, MO) to eliminate 31P NMR signals arising from superficial muscles (7). Field homogeneity was controlled by adjusting shim parameters using a water 1H signal derived from the organ sample. Acquisition conditions were set as follows to obtain fully relaxd signals : the pulse width was 20 micro-seconds, the pulse repetition time was 3.0s, and 200 scans were accumulated. Thus, it took 5-25 min for each measurement. After exponential multiplication, the accumulated free induction decays were subjected to Fourier transformation. Individual spectral peak areas were calculated by computer integration with JEOL software (JEOL), and the creatine phosphate (CP) peak was used as a positional reference (0 ppm). The relative amount of each compound was converted to absolute concentration by comparison with the data of an external reference (20mM methylene diphosphonate in water) in a spherical bulb. The present study showed that NMR spectroscopy is a novel assessment tools for assessment of reproductive and genetic toxicity in experimental and domestic animals. Less

使用自制表面线圈、传统圆形线圈和/或成像技术获取实验动物和农场动物的卵巢、肝脏、骨骼肌和胚胎的体内和体外磷 31 和质子核磁共振 (NMR) 谱体内观察到代表高能量状态的β-ATP/无机磷酸盐比率和细胞内pH值等，无需从这些器官和胚胎中进行任何提取过程即可鉴定和定量。质子NMR实验通过对常规技术的修改进行，并且31P NMR实验进行如下：在NMR检查期间，将样品保持在25°C.In.vivo 31P的受控空气温度下的探针室中。完整动物内稳态器官的 NMR 谱是使用配备有89 毫米孔径超导磁体（9.20 特斯拉；牛津研究所，美国马萨诸塞州贝德福德）将动物安装在表面线圈探针（两匝，直径 2.0 厘米）上，并在 161.7 处调谐至 31P。 MHz 并与由计算机胶带条组成的铁氧体屏蔽（Scotch 700；Three M Co.，St.Louis，MO）结合使用，以消除来自浅层肌肉的 31P NMR 信号通过使用源自器官样品的水 1H 信号调整匀场参数来控制，以获得完全松弛的信号：脉冲宽度为 20 微秒。，脉冲重复时间为3.0s，累计扫描200次，因此每次测量需要5-25分钟，经过指数倍增后，进行累计的自由感应衰变。通过与 JEOL 软件 (JEOL) 的计算机集成计算单个光谱峰面积，并使用磷酸肌酸 (CP) 峰作为位置参考 (0 ppm)。与球形灯泡中的外部参考（水中的 20mM 二磷酸亚甲基酯）的数据进行比较本研究表明，核磁共振波谱是一种用于评估实验动物和家养动物的生殖和遗传毒性的新型评估工具。较少的

项目成果

期刊论文数量（34）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

眞鍋昇ら: "Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in the pig ovaries" Experientia. 52. 647-651 (1996)

Noboru Manabe 等人：“细胞凋亡发生在猪卵巢闭锁窦卵泡的颗粒细胞中，而不是卵丘细胞”Experientia 52. 647-651 (1996)。

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眞鍋昇ら: "Effect of the mycelial extract of cultured cordyceps sinensis on in vivo hepatic energy metabolism in the mouse." Journal of Pharmacology. 70. 85-88 (1996)

Noboru Manabe 等人：“培养冬虫夏草菌丝体提取物对小鼠体内肝脏能量代谢的影响。药理学杂志 70. 85-88 (1996)”

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Noboru Manabe, Yuzuru Imai, Yoshihiro Kimura, Akira Myoumoto, Miki Sugimoto, Hajime Miyamoto, Yoshinori Okamura, and Manabu Fukumoto: "Ca2+/Mg2+-dependent endonuclease but not ca2+Dependent, mg2+-Dependent or cation-independent endonuclease is involved in

Noboru Manabe、Yuzuru Imai、Yoshihiro Kimura、Akira Myoumoto、Miki Sugimoto、Hajime Miyamoto、Yoshinori Okamura 和 Manabu Fukumoto：“Ca2 / Mg2 依赖性核酸内切酶而非 ca2 依赖性、mg2 依赖性或非阳离子依赖性核酸内切酶参与参与