Effects of cycloheximide on the structural organization of the nucleolus and the coiled body in normal and stimulated supraoptic neurons of the rat

Summary

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.