Institute of Cytology, Russian Academy of Sciences, 194064 St. Petersburg, Russia.
Cell Biol Int. 2010 Dec;34(12):1163-70. doi: 10.1042/CBI20090434.
IGCs (interchromatin granule clusters), or nuclear speckles, are one of the most universal subnuclear organelles of eukaryotic cells. We have used insect oocytes to study the possible association of poly(A)+ RNA and some factors involved in mRNA export with IGCs. Oogenesis of the mecopteran, Panorpa communis, used as a model object, is characterized by a strict cessation of oocyte genome transcription activity towards the end of oocyte growth. Our previous studies on P. communis oocyte nuclei have shown that oocyte IGC counterparts in this species are very unusual, both in morphology and molecular composition, compared with the typical IGCs of mammalian somatic cells traditionally used as a model system. We have now used microinjections of 2'-O-Me(U)₂₂ probes conjugated with the fluorochrome TAMRA to localize poly(A)+ RNA in IGCs. RNA export proteins were also detected by immunofluorescent/confocal and immunogold labelling electron microscopy. We found that poly(A)+ RNA, hnRNPs A/B and NXF1 mRNA export factors are located in IGCs regardless of the transcriptional status of the nucleus. Our data support the idea of IGCs as universal and conserved nuclear domains that serve not only as splicing factor reservoirs, but also take part in mRNA retention and export.
染色质间颗粒簇(IGCs),又称核斑,是真核细胞最普遍的亚核细胞器之一。我们曾利用昆虫卵母细胞来研究多聚腺苷酸(poly(A)+)RNA 以及某些参与 mRNA 输出的因子与 IGCs 的可能关联。作为模式对象的脉翅目昆虫,麻皮䗛,其卵子发生过程的特点是在卵母细胞生长末期,卵母细胞基因组转录活性严格停止。我们之前对麻皮䗛卵母细胞核的研究表明,与传统上用作模型系统的哺乳动物体细胞的典型 IGCs 相比,该物种的卵母细胞 IGC 对应物在形态和分子组成方面都非常不寻常。现在,我们使用 2'-O-Me(U)₂₂ 探针与荧光染料 TAMRA 结合的显微注射来定位 IGCs 中的 poly(A)+ RNA。通过免疫荧光/共聚焦和免疫金标记电子显微镜,我们还检测到了 RNA 输出蛋白。我们发现,poly(A)+ RNA、hnRNPs A/B 和 NXF1 mRNA 输出因子无论核转录状态如何,都定位于 IGCs 中。我们的数据支持 IGCs 作为普遍存在和保守的核域的观点,这些核域不仅作为剪接因子库,还参与 mRNA 的保留和输出。