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.

从我们的2年研究中获得以下结果：1。肌肉和卵子蛋白的二维电泳分析成功地用于鉴定Oryzias属的物种。 2。发现Medaka的孵化酶由两个不同的蛋白质HCE和LCE组成，并且两个成分纯化并表征了。 3。Medaka的肌肉LDH -A_4和-B_4同工酶被纯化并部分表征。 4。关于Oryzias属的杂交不育指出了几种可能性。 5。使用胰蛋白酶 - 基因和BRDU方法建立了在染色体中获得连续带的方法。 6。成功地应用了C频带和非带方法，以用于O. curvinotus和O. minutillusb的染色体。 7.日本Medaka被确认由4个主要人群组成。 NEI的遗传距离估计为0.3至0.5，这对于体内变化来说是相当大的值。 8.尝试在D-RR，DA和韩国Medaka中开始建立近交菌株。 9。到目前为止，通过使用多态性在8种标记酶方面建立了对近交菌株的遗传监测方法。 10。从Medaka特异性基准测试系统获得的B等位基因的新6个突变是由我们自己使用伽马射线和乙基硝基库（一种潜在的化学诱变剂）所开发的。在这些新的突变体中，眼睛表面的黑色素蛋白的程度小于B和B^p Poci的突变体中的范围，这表明所有新分离的6个突变甚至对B和B <@1p>@D1都具有隐性，也可能是B基因座的底部等位基因。 11。MLR分析程序12。从比较肿瘤学的角度来看，Medaka中赞助商以及化学诱导的肿瘤。