Simpson Pat, Schneiderman Howard A
Center of Pathobiology, University of California, Irvine, 92664, Irvine, California.
Wilehm Roux Arch Dev Biol. 1975 Sep;178(3):247-275. doi: 10.1007/BF00848432.
A method of isolating temperature-sensitive (ts) mutations blocking clone development, based on the analysis of twin spots produced by X-ray induced somatic recombination is reported. From this screen 10 ts mutations were recovered which caused an absence of the lethal-bearing clone at the restrictive temperature. Eight of these mutations were analyzed. Seven proved to be autonomous ts cell lethals and one was an autonomous ts mutation which reversibly affected cell division and growth of imaginal disc cells and growth of larval cells. The effects on development of one of the cell lethal mutations,l(1)ts-504, are described. Heat pulses (29°C) 24-72 hrs long caused a high frequency (up to 90%) of morphologically abnormal animals. The abnormalities observed were of two major kinds: deficiencies and duplications of imaginal disc derivatives. In addition, alterations of tarsal segmentations occurred. Heat pulses to larvae also delayed pupariation and eclosion by as much as four days. In general, longer pulses led to a greater delay in pupariation and eclosion and a higher frequency of deficiencies and duplications than shorter pulses. Exposure to restrictive temperature early in larval development delayed pupariation and resulted in mostly normal animals; exposure during the second and early third larval instar also delayed pupariation and led to a high frequency of duplications; exposure later in larval life, i.e. mid and late third larval instar, caused no delay in pupariation but led to a high frequency of deficiencies. These results can be explained by the occurrence of areas of cell death, which can be seen in the imaginal discs of larvae exposed to restrictive temperature by staining with trypan blue. This conclusion is further supported by the observation in gynandromorphs of duplications of female nonmutant tissue. These results are discussed in relation to current theories of pattern formation.
报道了一种基于对X射线诱导的体细胞重组产生的双斑分析来分离阻碍克隆发育的温度敏感(ts)突变的方法。通过该筛选,获得了10个ts突变,这些突变在限制温度下导致携带致死基因的克隆缺失。对其中8个突变进行了分析。7个被证明是自主ts细胞致死突变,1个是自主ts突变,它可逆地影响成虫盘细胞的细胞分裂和生长以及幼虫细胞的生长。描述了其中一个细胞致死突变l(1)ts - 504对发育的影响。长达24 - 72小时的热脉冲(29°C)导致形态异常动物的高频出现(高达90%)。观察到的异常主要有两种:成虫盘衍生物的缺失和重复。此外,跗节节段发生改变。对幼虫的热脉冲还使化蛹和羽化延迟多达四天。一般来说,较长的脉冲比较短的脉冲导致化蛹和羽化的延迟更大,缺失和重复的频率更高。在幼虫发育早期暴露于限制温度会延迟化蛹,产生的大多是正常动物;在幼虫第二龄期和第三龄期早期暴露也会延迟化蛹并导致高频率的重复;在幼虫后期,即第三龄期中期和后期暴露,不会延迟化蛹,但会导致高频率的缺失。这些结果可以通过细胞死亡区域的出现来解释,通过用台盼蓝染色可以在暴露于限制温度的幼虫成虫盘中看到这些区域。在雌雄嵌合体中观察到的雌性非突变组织重复进一步支持了这一结论。结合当前的模式形成理论对这些结果进行了讨论。