The location and dynamics of small nuclear ribonucleoproteins (snRNPs) were studied in salivary gland polytene chromosomes of Chironomus tentans by immunofluorescence with specific snRNP antibodies. Monoclonal antibody against the snRNP Sm antigens reacted at all sites of transcription (puffs and Balbiani rings). The amount of snRNP immunofluorescence was strictly dependent on transcription, increasing in parallel with gene activation and decreasing upon repression. Identical patterns of localization and transcriptional dependence were observed with antibodies specific for U1 or U2 snRNPs. These latter results show that the involvement of U1 and U2 snRNPs in transcription-related processes involves a high proportion, rather than small subsets, of active gene loci. In addition, the colocalization of U1 and U2 snRNPs at loci known to contain only one messenger RNA transcription unit (e.g. Balbiani ring 2) raises the possibility that both of these snRNPs interact with the same transcript. Finally, the lack of immunofluorescence at repressed loci indicates that snRNPs are not structural components of the chromatin (DNP) fiber, and also shows that unused snRNPs are not stored in chromatin. These latter points, and the growing evidence for the involvement of U1 snRNP in splicing, suggest that nascent pre-mRNA is the major chromosomal binding site for snRNPs.