MERRIAM R W
J Biophys Biochem Cytol. 1961 Dec;11(3):559-70. doi: 10.1083/jcb.11.3.559.
Nuclei from nearly ripe eggs of Rana pipiens were isolated and cleaned in 0.1 M KCl. The whole nucleus was then digested to various degrees with ribonuclease or trypsin, followed by washing and fixation in either osmium tetroxide or potassium permanganate. The nuclear envelope was dissected off, placed on a grid, air dried, and compared with undigested controls in the electron microscope. Some envelopes were dehydrated, embedded in methacrylate, and sectioned. Annuli around "pores" are composed of a substance or substances, at least partially fibrillar, which is preserved by osmium but lost during permanganate fixation. Material within the "pores" is also preserved by osmium but partially lost after permanganate. No evidence of granules or tubules in the annuli was found in air dried mounts although a granular appearance could be seen in tangentially oriented thin sections. Thin sections of isolated envelopes give evidence of diffuse material within the "pores" as well as a more condensed diaphragm across their waists. In whole mounts of the envelope the total density within "pores" is relatively constant from "pore" to "pore." All material within "pores," including the condensed diaphragm, is removable by trypsin digestion. Wispy material from the "pore" structure projects into the nucleus and annular material extends into the cytoplasm. Both annular and diaphragm materials remain with the envelope when it is isolated and are thus considered a part of its structure, not merely evidences of material passing through. There is no evidence of ribonuclease-removable material in any part of the "pore" complex.
从豹蛙接近成熟的卵中分离出细胞核,并在0.1M氯化钾中进行清理。然后用核糖核酸酶或胰蛋白酶对整个细胞核进行不同程度的消化,接着用四氧化锇或高锰酸钾洗涤并固定。将核膜剥离,置于网格上,空气干燥,然后在电子显微镜下与未消化的对照进行比较。一些核膜进行脱水处理,包埋在甲基丙烯酸酯中并切片。“孔”周围的环由一种或多种物质组成,至少部分呈纤维状,这种物质能被锇保留,但在高锰酸钾固定过程中会丢失。“孔”内的物质也能被锇保留,但在高锰酸钾处理后会部分丢失。在空气干燥的标本中未发现环中有颗粒或小管的迹象,尽管在切向取向的薄片中可以看到颗粒状外观。分离出的核膜薄片显示“孔”内有弥散物质,以及横跨其腰部的更致密的隔膜。在核膜的整体标本中,“孔”内的总密度从一个“孔”到另一个“孔”相对恒定。“孔”内的所有物质,包括致密的隔膜,都可通过胰蛋白酶消化去除。来自“孔”结构的细丝状物质伸入细胞核,环状物质延伸到细胞质中。当核膜被分离时,环状和隔膜物质都与核膜相连,因此被认为是其结构的一部分,而不仅仅是物质通过的证据。在“孔”复合体的任何部分都没有核糖核酸酶可去除物质的证据。