Puvion E, Viron A, Assens C, Leduc E H, Jeanteur P
J Ultrastruct Res. 1984 May;87(2):180-9. doi: 10.1016/s0022-5320(84)80077-5.
Small nuclear ribonucleoproteins (snRNPs) containing U1, U2, U4, U5, and U6 small nuclear RNAs were detected by ultrastructural immunocytochemistry in the nuclei of isolated rat hepatocytes using Fab fragments of anti-Sm and anti-RNP autoantibodies. Their localization was carried out in normal cells and in cells treated with two drugs, the adenosine analog DRB and CdCl2, which alter the number and distribution of nuclear RNP components. It was found that more precise determination of the distribution of these small RNAs could be obtained by using two complementary procedures in parallel rather than either one alone. They consisted of an indirect immunoperoxidase labeling carried out before embedment and an indirect immunogold labeling applied to thin sections of cells embedded in Lowicryl K4M. The results indicate that snRNPs are associated with all extranucleolar perichromatin fibrils and granules and interchromatin fibrils, which confirms that they occur in structures involved in the synthesis and processing of hnRNA. The snRNPs are not associated with nucleolar perichromatin granules induced by DRB, which confirms that there may be two kinds of perichromatin granules. The snRNPs are also associated with the still enigmatic interchromatin granules which apparently do not contain hnRNA but at least in DRB-treated cells, also contain ribosomal RNA.
利用抗Sm和抗RNP自身抗体的Fab片段，通过超微结构免疫细胞化学方法，在分离的大鼠肝细胞的细胞核中检测到了含有U1、U2、U4、U5和U6小核RNA的小核核糖核蛋白（snRNP）。在正常细胞以及用两种药物（腺苷类似物DRB和CdCl2）处理的细胞中进行了它们的定位，这两种药物会改变核RNP成分的数量和分布。结果发现，通过并行使用两种互补方法而非单独使用其中一种方法，可以更精确地确定这些小RNA的分布。这两种方法包括在包埋前进行的间接免疫过氧化物酶标记，以及应用于包埋在Lowicryl K4M中的细胞薄切片的间接免疫金标记。结果表明，snRNP与所有核仁外周染色质纤维和颗粒以及染色质间纤维相关，这证实了它们存在于参与hnRNA合成和加工的结构中。snRNP与DRB诱导的核仁外周染色质颗粒不相关，这证实了可能存在两种外周染色质颗粒。snRNP还与仍然神秘的染色质间颗粒相关，这些颗粒显然不含有hnRNA，但至少在DRB处理的细胞中，也含有核糖体RNA。
