Popken Jens, Graf Alexander, Krebs Stefan, Blum Helmut, Schmid Volker J, Strauss Axel, Guengoer Tuna, Zakhartchenko Valeri, Wolf Eckhard, Cremer Thomas
Division of Anthropology and Human Genetics, Biocenter, LMU Munich, Planegg-Martinsried, Germany; Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.
Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, Munich, Germany.
PLoS One. 2015 May 1;10(5):e0124619. doi: 10.1371/journal.pone.0124619. eCollection 2015.
The present study demonstrates a major remodeling of the nuclear envelope and its underlying lamina during bovine preimplantation development. Up to the onset of major embryonic genome activation (MGA) at the 8-cell stage nuclei showed a non-uniform distribution of nuclear pore complexes (NPCs). NPCs were exclusively present at sites where DNA contacted the nuclear lamina. Extended regions of the lamina, which were not contacted by DNA, lacked NPCs. In post-MGA nuclei the whole lamina was contacted rather uniformly by DNA. Accordingly, NPCs became uniformly distributed throughout the entire nuclear envelope. These findings shed new light on the conditions which control the integration of NPCs into the nuclear envelope. The switch from maternal to embryonic production of mRNAs was accompanied by multiple invaginations covered with NPCs, which may serve the increased demands of mRNA export and protein import. Other invaginations, as well as interior nuclear segments and vesicles without contact to the nuclear envelope, were exclusively positive for lamin B. Since the abundance of these invaginations and vesicles increased in concert with a massive nuclear volume reduction, we suggest that they reflect a mechanism for fitting the nuclear envelope and its lamina to a shrinking nuclear size during bovine preimplantation development. In addition, a deposit of extranuclear clusters of NUP153 (a marker for NPCs) without associated lamin B was frequently observed from the zygote stage up to MGA. Corresponding RNA-Seq data revealed deposits of spliced, maternally provided NUP153 mRNA and little unspliced, newly synthesized RNA prior to MGA, which increased strongly at the initiation of embryonic expression of NUP153 at MGA.
本研究表明,在牛胚胎植入前发育过程中,核膜及其下方的核纤层发生了重大重塑。在8细胞期主要胚胎基因组激活(MGA)开始之前,细胞核显示核孔复合体(NPC)分布不均匀。NPC仅存在于DNA与核纤层接触的部位。核纤层未与DNA接触的延伸区域没有NPC。在MGA后的细胞核中,整个核纤层与DNA的接触较为均匀。因此,NPC在整个核膜中均匀分布。这些发现为控制NPC整合到核膜中的条件提供了新的线索。从母体mRNA产生向胚胎mRNA产生的转变伴随着多个被NPC覆盖的内陷,这可能满足了mRNA输出和蛋白质输入增加的需求。其他内陷以及与核膜无接触的核内部分和囊泡仅对核纤层蛋白B呈阳性。由于这些内陷和囊泡的丰度随着核体积的大幅减小而增加,我们认为它们反映了一种机制,即在牛胚胎植入前发育过程中使核膜及其核纤层适应不断缩小的核大小。此外,从合子期到MGA阶段,经常观察到无相关核纤层蛋白B的核外NUP153(NPC的标志物)簇沉积。相应的RNA测序数据显示,在MGA之前有剪接的、由母体提供的NUP153 mRNA沉积,新合成的未剪接RNA很少,而在MGA时NUP153胚胎表达开始时,新合成的RNA大量增加。