Kirschner R H, Rusli M, Martin T E
J Cell Biol. 1977 Jan;72(1):118-32. doi: 10.1083/jcb.72.1.118.
We have used high resolution scanning electron microscopy (SEM) to study the nuclear envelope components of isolated mouse liver nuclei. The surfaces of intact nuclei are covered by closely packed ribosomes which are distinguishable by SEM from nuclear pore complexes. After removal of nuclear membranes with the nonionic detergent Triton X-100, the pore complexes remain attached to an underlying, peripheral nuclear lamina, as described by others. The surface of this dense lamina is composed of particulate granules, 75-150 A in diameter, which are contiguous over the entire periphery. We did not observe the pore-to-pore fibril network suggested by other investigators, but such a structure might be the framework upon which the dense lamina is formed. Morphometric analysis of pores and pore complexes shows their size, structure, and density to be similar to that of other mammalian cells. In addition, several types of pore complex-associated structures, not previously reported by other electron microscope (EM) techniques, are observed by SEM. Our studies suggest that the major role of the dense lamina is associated with the distribution, stability, and perhaps, biogenesis of nuclear pore complexes. Treatment of isolated nuclei with a combination of Triton X-100 and sodium deoxycholate removes membranes, dense lamina, and nuclear pore complexes. The resulting "chromatin nuclei" retain their integrity despite the absence of any limiting peripheral structures.
我们利用高分辨率扫描电子显微镜(SEM)研究了分离的小鼠肝细胞核的核膜成分。完整细胞核的表面覆盖着紧密排列的核糖体,通过扫描电子显微镜可将其与核孔复合体区分开来。如其他人所描述的那样,用非离子去污剂Triton X-100去除核膜后,核孔复合体仍附着在下方的外周核纤层上。这种致密核纤层的表面由直径为75 - 150埃的颗粒组成,它们在整个周边连续排列。我们没有观察到其他研究者所提出的孔间纤维网络,但这样的结构可能是形成致密核纤层的框架。对核孔和核孔复合体的形态计量分析表明,它们的大小、结构和密度与其他哺乳动物细胞相似。此外,通过扫描电子显微镜观察到了几种其他电子显微镜技术以前未报道过的与核孔复合体相关的结构。我们的研究表明,致密核纤层的主要作用与核孔复合体的分布、稳定性以及可能的生物发生有关。用Triton X-100和脱氧胆酸钠的组合处理分离的细胞核可去除膜、致密核纤层和核孔复合体。尽管没有任何限制外周结构,但由此产生的“染色质核”仍保持其完整性。