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Development of the Japanese Medaka as a Laboratory Animal with Genetic and Environmental Control.

日本青鳉作为遗传和环境控制实验动物的开发。

基本信息

批准号：
01304054
负责人：
SHIMA Akihiro
金额：
$ 7.68万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Co-operative Research (A)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

The following results were obtained from our 2 years' study :1. The two dimensional electrophoretic analysis of muscle and egg proteins was successfully used for identification of species of the genus oryzias. 2. The hatching enzyme of the Medaka was found to consist of two different proteases, HCE and LCE, and the two components were purified and characterized. 3. The muscle LDH-A_4 and ーB_4 isozymes of the Medaka were purified and partially characterized. 4. Several possibilities were pointed out with regard to the hybrid sterility in the genus Oryzias. 5. The method to obtain continuous bands in chromosomes was established using trypsinーGiemsa as well as BrdU method. 6. The C banding and NOR banding methods were successfully applied for the chromosomes of O. curvinotus and O. minutillusB. 7. The Japanese Medaka was definitively confirmed to consist of 4 major populations. The Nei's genetic distance was estimated to be 0.3 to 0.5, which was a fairly large value for intraspecies variation. 8. The attempt was started and continued to establish inbred strains in dーrr, Da and Korean Medaka. 9. The method for genetic monitoring of the inbred strains thus far obtained was established by the use of polymorphisms with regard to 8 marker enzymes. 10. The new 6 mutations at the b allele were isolated from F_1 progeny obtained by the Medaka specific-locus test system that has been developed by ourselves using gamma-rays and ethylnitrosoures, a potent chemical mutagen. In these new mutants, the extent of melanizatin of eyes body surface was less than that in the mutants at the b and b^p Poci, indicating that all the 6 mutations newly isolated were recessive even to b and b<@1p>@D1, possibly the bottom allele at the b locus. 11. The MLR assay procedure was developed using leukocytes of the goldfish, and was started being applied for the Medaka. 12. The spontaneous as well as chemically induced tumors in the Medaka were characterized from a view point of comparative oncology.

经过两年的研究，我们得到了以下结果： 1．肌肉和卵蛋白的二维电泳分析已成功用于稻属物种的鉴定。 2.发现青鳉的孵化酶由两种不同的蛋白酶HCE和LCE组成，并对这两种组分进行了纯化和表征。 3. 青鳉肌肉LDH-A_4和ーB_4同工酶的纯化和部分表征。 4. 指出了稻属杂种不育的几种可能性。 5.建立了利用胰蛋白酶ーGiemsa和BrdU法获得染色体连续条带的方法。 6. C显带和NOR显带方法成功应用于O. curvinotus和O. minutillusB染色体。 7. 日本青鳉被明确证实由 4 个主要种群组成。 Nei的遗传距离估计为0.3至0.5，这对于种内变异来说是一个相当大的值。 8. 尝试开始并继续建立dーrr、Da和韩国青鳉近交系。 9.利用8个标记酶的多态性建立了对迄今为止获得的近交系进行遗传监测的方法。 10. b等位基因的新6个突变是从F_1后代中分离出来的，该系统是通过我们自己开发的青鳉特异性基因座测试系统使用伽马射线和乙基亚硝基（一种有效的化学诱变剂）获得的。在这些新突变体中，眼体表面黑色素的程度小于b和b^p Poci突变体，表明新分离的6个突变体甚至对b和b<@1p>@D1（可能是b基因座的底部等位基因）都是隐性的。 11. MLR 测定程序是使用金鱼的白细胞开发的，并开始应用于青鳉。 12. 从比较肿瘤学的角度对青鳉中的自发肿瘤和化学诱发肿瘤进行了表征。