Diez J L, Santa-Cruz M C, Aller P
Chromosoma. 1977 Jun 23;61(4):369-79. doi: 10.1007/BF00288620.
Modifications in the synthesis of salivary gland RNA were induced by treatments with 10 microgram/ml cycloheximide (CHM) on 4th instar larvae of Chironomus pallidivitattus. After 3, 6 and 24 h CHM treatment, RNA was labeled "in vitro", by incubating the salivary glands in a medium containing H3-uridine. The electrophoretical analyses corresponding to the 3 and 6 h treatment showed a stimulation of the non-ribosomal components of the newly synthesized RNA, while preribosomal RNA synthesis appeared depressed. This fact was also confirmed at cytological level, since autoradiograms made after 3 h of CHM treatment showed a reduced H3-uridine label over the nucleolus and an increase of diffuse labeling over the chromosomes. Longer treatments (24 h) causes a considerable inhibition of the synthesis of all RNA species. The role played by protein synthesis inhibition in the aforementioned effects is discussed.--Some of the morphological implications of CHM treatment, such as modifications of the nucleolar structure (nucleolar segregation) are also reported. The use of a squash technique based on glutaraldehyde fixation of the salivary glands, considerably facilitates such studies.
用10微克/毫升环己酰亚胺（CHM）处理苍白摇蚊四龄幼虫，可诱导唾液腺RNA合成发生改变。在CHM处理3、6和24小时后，通过将唾液腺在含有H3 - 尿苷的培养基中孵育来“体外”标记RNA。对应于3小时和6小时处理的电泳分析表明，新合成RNA的非核糖体成分受到刺激，而前核糖体RNA合成似乎受到抑制。这一事实在细胞学水平上也得到了证实，因为CHM处理3小时后制作的放射自显影片显示核仁上的H3 - 尿苷标记减少，而染色体上的弥散标记增加。更长时间的处理（24小时）会导致所有RNA种类的合成受到相当大的抑制。讨论了蛋白质合成抑制在上述效应中所起的作用。还报告了CHM处理的一些形态学影响，如核仁结构的改变（核仁分离）。基于唾液腺戊二醛固定的压片技术的使用，极大地促进了此类研究。
