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The cytochemical nature of the ultrastructural nucleolar transformation previously observed in the vaginal epithelium of the ovariectomized mouse after local application of oestradiol-17β was investigated using a recently developed ultrastructural staining technique. Oestradiol treatment induced ribonucleic acid (RNA) synthesis, especially in the nucleolus but also in the nuclear chromatic region and ribosomes. Actinomycin D administered simultaneously with oestradiol reduced the oestrogen-induced nucleolar response to the control level. These findings are discussed with special reference to the mode of action of oestrogens.