Mott M R, Callen H G
J Cell Sci. 1975 Mar;17(3):241-61. doi: 10.1242/jcs.17.3.241.
The ultrastructure of lampbrush chromosomes has been examined in sections of end-embedded spread preparations, where the nuclear sap was dispersed prior to fixation, and in oocyte nuclei fixed entire, in 60-kV and i-MV electron microscopes. In spread preparations the axial chromomeres are seen to be organized as regularly spaced, unravelled skeins of DNP, each with a skein width of some 30 nm, though in some chromomeres there are regions where the DNP is much more densely packed. In both unravelled and dense regions, the 'ultimate' DNP fibres, wherever they can be identified, appear to be some 5 nm wide and thrown into loose coils. The unravelled state, although it clearly reflects an orderly packaging of the non-transcribing DNP, is an artifact of preparation; in sections of entire nuclei all chromomeres are seen to consist of DNP fibrils in the more densely packed state. The interchromeric fibril is single, and some 10 nm or less wide; it shows no sign of transcriptional activity. In sections of end-embedded preparations the RNPmatrix of most lateral loops, where transcription occurs, is seen to be made up of particles, each uniformly some 30 nm in diameter and strung together in linear array. These RNP particles are equally evident in sections of whole nuclei. In many loops the strings of particles are wound back on themselves to form regularly spaced, dense aggregates, each some 200-300 nm wide or wider; the larger aggregates can be resolved in the light micrpscope. The RNP particles are of the same dimensions throughout the lengths of individual lateral loops, and of substantially the same dimensions in loops of different gross morphologies. It is suggested that as each successive short length of RNA is transcribed from loop axis DNA, a protein associates with this RNA and winds it up to form a 'manageable' package, allowing transcription to proceed.
在末端包埋铺展制片的切片中,以及在60千伏和1兆伏电子显微镜下对完整固定的卵母细胞核进行观察,研究了灯刷染色体的超微结构。在铺展制片中,可以看到轴染色粒是由规则间隔、解旋的脱氧核糖核蛋白(DNP)细丝组成,每条细丝宽度约为30纳米,不过在一些染色粒中存在DNP更为密集堆积的区域。在解旋区域和密集区域,无论何处能够识别,“最终”的DNP纤维似乎宽约5纳米,并呈松散螺旋状。解旋状态虽然清楚地反映了非转录DNP的有序包装,但却是制片的人为产物;在完整细胞核的切片中,所有染色粒都由处于更密集堆积状态的DNP纤维组成。染色粒间纤维是单一的,宽度约为10纳米或更小;未显示转录活性的迹象。在末端包埋制片的切片中,大多数发生转录的侧环的核糖核蛋白(RNP)基质由颗粒组成,每个颗粒直径均匀约为30纳米,并以线性排列串在一起。这些RNP颗粒在完整细胞核的切片中同样明显。在许多环中,颗粒串自身回绕形成规则间隔的密集聚集体,每个聚集体宽约200 - 300纳米或更宽;较大的聚集体在光学显微镜下可以分辨。RNP颗粒在单个侧环的整个长度上尺寸相同,并且在不同总体形态的环中尺寸基本相同。有人提出,随着RNA的每一段连续短片段从环轴DNA转录而来,一种蛋白质与该RNA结合并将其缠绕起来形成一个“便于处理”的包裹,从而使转录得以进行。