Lafarga M, Berciano M T, Andres M A, Testillano P S
Departamento de Anatomía y Biología Celular, Facultad de Medicina, Santander, Spain.
J Neurocytol. 1994 Aug;23(8):500-13. doi: 10.1007/BF01184073.
This study was designed to determine the effects of cycloheximide, a protein synthesis inhibitor that interferes with rRNA synthesis and processing, on the nucleoli and coiled bodies of supraoptic nucleus neurons from normally-hydrated and osmotically-stimulated rats. The number of nucleoli and the nucleolar size were estimated on smear preparations of previously silver-impregnated supraoptic nucleus. No significant differences were registered in the mean number of nucleoli per cell in cycloheximide-treated rats. The number of nucleoli per neuron remained constant, at about 1.3, in all animal groups, suggesting that the nucleoli number is strictly regulated in differentiated neurons. By contrast, a significant reduction in the average nucleolar volume of supraoptic nucleus neurons was detected in cycloheximide-treated groups of rats in comparison with their equivalent non-treated groups. By electron microscopy, most nucleoli and coiled bodies of supraoptic nucleus neurons exhibited cycloheximide-induced alterations in their fine structure and configuration. Nucleolar changes included the occurrence of a few large fibrillar centres, the formation of microspherules and small intranucleolar vacuoles or dilated interstices, and the partial segregation of nucleolar components coupled with the transformation of reticulated nucleoli--a nucleolar configuration characteristic of supraoptic nucleus neurons of non-cycloheximide-treated rats--into compact ones. The redistribution of nucleolar components might reflect the interference with rDNA transcription, and also supports the hypothesis that the normal assembly of these components into the nucleolus depends upon ongoing nucleolar transcription. Concerning coiled bodies, most of them revealed ultrastructural alterations, particularly segregation of the amorphous matrix, compactation of coiled threads and formation of coiled body-derived dense bodies of fibrillar nature. Moreover, cycloheximide also induced the formation of smaller dense bodies--here referred to as dense microbodies--which presumably represent a distinct nuclear entity different from coiled bodies. Ultrastructural silver staining of nuclear bodies showed a selective silver reaction on the dense fibrillar component of normal and altered coiled bodies, as well as on the dense microbodies. The possible relationship between the nucleolus and both coiled bodies and dense microbodies is discussed.
本研究旨在确定放线菌酮(一种干扰rRNA合成与加工的蛋白质合成抑制剂)对正常水合及渗透压刺激大鼠视上核神经元的核仁及卷曲小体的影响。通过对先前经银浸染的视上核涂片制备物进行观察,估算核仁数量及核仁大小。在经放线菌酮处理的大鼠中,每个细胞的核仁平均数无显著差异。在所有动物组中,每个神经元的核仁数量保持恒定,约为1.3个,这表明在分化的神经元中核仁数量受到严格调控。相比之下,与未处理的等效组相比,经放线菌酮处理的大鼠组中视上核神经元的平均核仁体积显著减小。通过电子显微镜观察,视上核神经元的大多数核仁及卷曲小体在其精细结构和形态上呈现出放线菌酮诱导的改变。核仁变化包括出现一些大的纤维中心、微球粒和小的核仁内空泡或扩张的间隙形成,以及核仁成分的部分分离,同时伴有网状核仁(未用放线菌酮处理的大鼠视上核神经元的核仁形态特征)转变为致密核仁。核仁成分的重新分布可能反映了对rDNA转录的干扰,也支持了这样一种假说,即这些成分正常组装到核仁中依赖于正在进行的核仁转录。关于卷曲小体,它们中的大多数显示出超微结构改变,特别是无定形基质的分离、卷曲丝的致密化以及形成纤维性质的卷曲小体衍生致密体。此外,放线菌酮还诱导形成较小的致密体——这里称为致密微体——其可能代表一种与卷曲小体不同的独特核实体。核体的超微结构银染色显示在正常和改变的卷曲小体以及致密微体的致密纤维成分上有选择性的银反应。文中讨论了核仁与卷曲小体及致密微体之间可能的关系。