Mulligan P K, Rasch E M
Histochemistry. 1985;82(3):233-47. doi: 10.1007/BF00501400.
DNA replication patterns in the nurse and follicle cells of wild type and a female sterile mutant, fs(1)1304, of Drosophila melanogaster have been studied by DNA-Feulgen cytophotometry, using a cell dispersal technique that allowed the measurement of DNA amounts in individual nuclei from egg chambers of known developmental stages. DNA-Feulgen values associated with various ovarian nuclei from egg chambers at different stages of development were used to assess a base line DNA content for ovarian tissues and to estimate the extent of DNA replication in the nurse cells and follicle cells of growing and mature egg chambers. Our data show that both the nurse and follicle cells undergo multiple cycles of endonuclear DNA replication and that there may be selective amplification as well as underreplication by portions of the genome in these highly polyploid, ovarian cells. Alternative models are proposed to account for the DNA replication patterns observed. Comparisons of DNA-Feulgen levels in wild type ovarian nuclei with those found for the fs(1)1304 mutant and its heterozygote in the balanced stock fs/FM3, show that equivalent DNA levels are present in follicle cell nuclei from all three types of females. Nurse cell nuclei in the homozygous fs stock, however, fail to achieve the same high DNA levels observed in both fs/FM3 and wild type nurse cell nuclei. Although the nuclei of follicle cells in ovaries from fs/fs females appear morphologically like those surrounding egg chambers in wild type ovaries, nurse cell nuclei from mutant females show a more compacted organization of their chromatin than found for nurse cell nuclei from wild type ovaries at similar developmental stages. Our findings suggest that a major effect of the fs(1)1304 mutation may be on the coiling behavior of chromatin and the conformation of DNA-protein moieties in both nurse cell and follicle cell nuclei. These changes in chromatin structure apparently are manifest by perturbations in DNA replication patterns and normal gene function in these biosynthetically active cells.
利用细胞分散技术,通过DNA - 福尔根细胞光度法研究了野生型和果蝇雌性不育突变体fs(1)1304的滋养细胞和卵泡细胞中的DNA复制模式。该技术能够测量已知发育阶段卵室中单个细胞核的DNA含量。与不同发育阶段卵室的各种卵巢细胞核相关的DNA - 福尔根值,用于评估卵巢组织的基线DNA含量,并估计生长中和成熟卵室的滋养细胞和卵泡细胞中DNA复制的程度。我们的数据表明,滋养细胞和卵泡细胞都经历了核内DNA复制的多个周期,并且在这些高度多倍体的卵巢细胞中,基因组的部分区域可能存在选择性扩增以及复制不足。提出了替代模型来解释观察到的DNA复制模式。将野生型卵巢细胞核中的DNA - 福尔根水平与在平衡品系fs/FM3中的fs(1)1304突变体及其杂合子中发现的水平进行比较,结果表明,所有三种类型雌性的卵泡细胞核中都存在相当的DNA水平。然而,纯合fs品系中的滋养细胞核未能达到在fs/FM3和野生型滋养细胞核中观察到的相同高DNA水平。尽管fs/fs雌性卵巢中的卵泡细胞核在形态上看起来与野生型卵巢中围绕卵室的细胞核相似,但突变雌性的滋养细胞核显示出比相似发育阶段野生型卵巢的滋养细胞核更紧密的染色质组织。我们的研究结果表明,fs(1)1304突变的主要影响可能在于滋养细胞和卵泡细胞核中染色质的卷曲行为以及DNA - 蛋白质部分的构象。这些染色质结构的变化显然通过这些生物合成活跃细胞中DNA复制模式和正常基因功能的扰动而表现出来。