BLOOM W, LEIDER R J
J Cell Biol. 1962 May;13(2):269-301. doi: 10.1083/jcb.13.2.269.
Chromosome segments of urodele cells lose some substance after irradiation with about 10(-1) ergs/micro(2) of heterochromatic ultraviolet light. These segments stain faintly or negatively with the Feulgen and pyronine-methyl-green methods and weakly with the Alfert-Gesch-wind stain for basic protein. In the living cells, Perry found in these chromosome segments a decrease of 50 to 60 per cent in absorption at 2400, 2600, and 2800 A, i.e., in the region of intense chromosomal absorption that is maximal at 2600 A. Apparently the material lost contains DNA (?DNP) and we call the process DNA-steresis. In such cells, fixed in neutral formalin in Tyrode's solution and stained with phosphotungstic acid, electron microscopy shows that the unirradiated parts of the chromosomes consist of (a) a homogeneous or finely fibrillar material (component-A) filling the meshes of (b) an irregular network with bars 40 to 300 A in diameter, some of which continue into a similar interchromosomal network. DNA-steretic portions of the chromosomes consist mainly of this network and only small amounts of component-A, which presumably contains the DNA. We have not been able to demonstrate DNA-steresis with the electron microscope after primary fixation with OsO(4) or KMnO(4). Structural changes due to DNA-steresis are compared with certain nuclear changes in the mitotic cycle.
蝾螈细胞的染色体片段在用约10⁻¹尔格/微米²的异染色质紫外线照射后会失去一些物质。这些片段用福尔根染色法和派洛宁-甲基绿染色法染色很浅或呈阴性,用阿尔费特-格施温德碱性蛋白染色法染色较弱。在活细胞中,佩里发现这些染色体片段在2400、2600和2800埃处的吸收减少了50%至60%,即在2600埃处达到最大值的强烈染色体吸收区域。显然,丢失的物质含有DNA(?脱氧核糖核蛋白),我们将这个过程称为DNA缺失。在这样的细胞中,用台氏液中的中性福尔马林固定并用磷钨酸染色,电子显微镜显示染色体未照射的部分由(a)一种均匀或细纤维状物质(成分A)组成,填充在(b)一个不规则网络的网孔中,该网络的条带直径为40至300埃,其中一些延伸到类似的染色体间网络。染色体的DNA缺失部分主要由这个网络组成,只有少量的成分A,推测成分A含有DNA。在用四氧化锇或高锰酸钾进行初次固定后,我们无法用电子显微镜证明DNA缺失。将DNA缺失引起的结构变化与有丝分裂周期中的某些核变化进行了比较。