Cohen Merav, Tzur Yonatan B, Neufeld Esther, Feinstein Naomi, Delannoy Michael R, Wilson Katherine L, Gruenbaum Yosef
Department of Genetics, The Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.
J Struct Biol. 2002 Oct-Dec;140(1-3):232-40. doi: 10.1016/s1047-8477(02)00516-6.
Nuclear membranes and nuclear pore complexes (NPCs) are conserved in both animals and plants. However, the lamina composition and the dimensions of NPCs vary between plants, yeast, and vertebrates. In this study, we established a protocol that preserves the structure of Caenorhabditis elegans embryonic cells for high-resolution studies with thin-section transmission electron microscopy (TEM). We show that the NPCs are bigger in C. elegans embryos than in yeast, with dimensions similar to those in higher eukaryotes. We also localized the C. elegans nuclear envelope proteins Ce-lamin and Ce-emerin by pre-embedding gold labeling immunoelectron microscopy. Both proteins are present at or near the inner nuclear membrane. A fraction of Ce-lamin, but not Ce-emerin, is present in the nuclear interior. Removing the nuclear membranes leaves both Ce-lamin and Ce-emerin associated with the chromatin. Eliminating the single lamin protein caused cell death as visualized by characteristic changes in nuclear architecture including condensation of chromatin, clustering of NPCs, membrane blebbing, and the presence of vesicles inside the nucleus. Taken together, these results show evolutionarily conserved protein localization, interactions, and functions of the C. elegans nuclear envelope.
核膜和核孔复合体(NPCs)在动物和植物中都是保守的。然而,核纤层的组成以及NPCs的尺寸在植物、酵母和脊椎动物之间存在差异。在本研究中,我们建立了一种方案,可保留秀丽隐杆线虫胚胎细胞的结构,用于通过超薄切片透射电子显微镜(TEM)进行高分辨率研究。我们发现,秀丽隐杆线虫胚胎中的NPCs比酵母中的更大,其尺寸与高等真核生物中的相似。我们还通过包埋前金标免疫电子显微镜对秀丽隐杆线虫的核膜蛋白Ce-核纤层蛋白和Ce-emerin进行了定位。这两种蛋白都存在于内核膜或其附近。一部分Ce-核纤层蛋白存在于核内部,但Ce-emerin不存在。去除核膜后,Ce-核纤层蛋白和Ce-emerin都与染色质相关。去除单一的核纤层蛋白会导致细胞死亡,这可通过核结构的特征性变化观察到,包括染色质凝聚、NPCs聚集、膜泡形成以及核内出现囊泡。综上所述,这些结果表明秀丽隐杆线虫核膜在蛋白质定位、相互作用和功能方面具有进化保守性。