Selection Mechanisms for Mutants As Studied by the Medaka Specific-Locus Test System.

通过青鳉特异性基因座测试系统研究的突变体选择机制。

基本信息

批准号：
01440093
负责人：
SHIMA Akihiro
金额：
$ 11.39万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1991
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01440093/
关键词：
Tester Medaka Specific-locus mutation Radiation Developmental process Selection Dominant lethals Malformation

项目摘要

1. The total cumulative number of the Medaka embryos examined at the end of this project reached 488, 890. This number of embryos corresponds to a total of 1, 198, 2-79 loci.2. The wild type male Medaka were exposed to gamma-rays, fission neutron or ethylnitrosourea. These treated males were allowed to mate with non-treated female tester Medaka which are homozygous recessive at the 3 loci, i. e. b/b ; gu/gu ; lf/lf.3. The gamma-irradiations were done using a 111-TBq ^<137>Cs gamma source located in the Research Center for Nuclear Science and Engineering, The University of Tokyo. The dose-rate used was 0.95 Gy/min.4. The irradiations with fission neutron were performed in the UTR-Kinki reactor (Atomic Energy Research Institute, Kinki University, in collaboration with Prof. S. Kondo and Dr. T. Ito) operated at 1 Watt with the following conditions : dDn/dt=16.9 cGy/hr, dDg/dt=24.8 cGy/hr.5. The treatments of the Medaka with ethylnitrosourea (ENU) were done by allowing the male Medaka to s … More wim in 1 mM aqueous solution of ENU (pH=5.8) at 27ﾟC for 2 hours, following by extensive rinse with tap water.6. The spontaneous total mutation 5 rate was 4.2x10^<-5>/locus, while the spontaneous viable mutation rate was 5.5x10^<-6>/locus, a value vdry close to that of the mouse (8.1x10^<-6>/locus, Ressell & Kelly (1982)).7. The induced rates of viable mutation per unit dose (rad) of gamma-rays for the Medaka sperm, spegmatids and spermatogonia were found to be respectively 176, 51 and 7.5 (x10^<-8>/locus).8. Based on the above-obtained values, the doubling gamma-ray dose for the induction of viable mutation was estimated to be 3.2, 12 and 98 rad (cGy) for sperm, spermatids and spermatogonia, respectively.9. Wg have established a method to obtain cell lines from individual Medaka embryos with malformation of various severities which is expected to result in dominant lethal mutation. Surprisingly, cell lines from moribund malformed : embryos tended to grow over a year more actively than those from the normal' controls without malformation.10. We also established an AP-PCR method to examine induced DNA damages in male getm cells, which is expected to give more direct clue to study the mechanisms of embryonic selection during development. Less

1。在该项目结束时检查的Medaka胚胎的总数达到488，890。此胚胎数量对应于1，198，2-79 Loci.2。野生型男性Medaka暴露于伽马射线，裂变中性或乙基硝基乙酸酯。这些经过治疗的雄性被允许与未处理的女性测试仪Medaka交配，这些男性测试师Medaka在3个基因座中是纯合隐性的。 e。 b/b; gu/gu; LF/LF.3。使用位于东京大学核科学与工程研究中心的111-TBQ ^<137> CS伽马来源进行伽马 - iRrAdiations。使用的剂量率为0.95 Gy/min.4。裂变中性的辐照在UTR-KINKI反应堆（Kinki University，Kinki University，Kinki University，与S. Kondo教授和T. Ito博士合作）中以1瓦的操作与以下条件：DDN/DT = 16.9 CGY/HR，DDG/DDG/DT = 24.8 CGY/HR.5。通过允许雄性Medaka在27°C的ENU（pH = 5.8）中以1 mm水溶液在27°C中进行2小时，通过大量的用自来水冲洗，用雄性Medaka进行了Medaka的治疗方法。6。赞助的总突变5率为4.2x10^<-5>/locus，而赞助的可行突变率为5.5x10^<-6>/lotus，一个值接近小鼠的值VDRY（8.1x10^<-6>/locus，Ressell＆Kelly（Ressell＆Kelly（1982）））。7。发现Medaka精子，Spegmatids和精子的伽马射线每单位剂量（RAD）的诱导速率分别为176、51和7.5（X10^<-8>/locus）.8。基于上述值，对于精子，精子，精子和精子分别为3.2、12和98 RAD（CGY），升高伽马射线剂量为3.2、12和98 RAD（CGY）9。 WG已经建立了一种从单个Medaka胚胎中获取细胞系的方法，其畸形的各种严重程度有望导致占主导地位致死突变。出人意料的是，莫里博德（Moribund）的细胞系畸形：胚胎比没有畸形的正常对照组的胚胎更积极地生长一年多。10。我们还建立了一种AP-PCR方法，用于检查雄性GETM细胞中的DNA损伤，该方法有望提供更多直接的线索来研究发育过程中胚胎选择的机制。较少的