Effects of DMSO on the structure and function of polytene chromosomes of Chironomus

Dimethylsulfoxide (DMSO) controlled puff induction and repression (or non-induction) in larval polytene chromosomes of Chironomus tentans were studied for the case of the Balbiani rings (BR). A characteristic reaction pattern, involving BR 1, BR 2, and BR 3, all in salivary gland chromosome IV was found. In vivo exposure of 4th instar larvae (not prepupae) to 10%n DMSO at 18 degrees C first evokes an over-stimulation of BR 3 while DMSO-stimulation of puffing at BR 1 and BR 2 always follows that of BR 3. After removal of the drug, a rapid uniform collapse of all puffs occurs, thus more or less restoring the banding pattern of all previously decondensed chromosome segments. Recovery proceeds as BR's and other puffs reappear. By observing the restoration, one can locate the site from which a BR (puff) originates. BR 2, which is normally the most active non-ribosomal gene locus in untreated larvae, here serves as an example. As the sizes of BR 3, BR 1 and BR 2 change, so do the quantities of the transcriptional products in these gene loci (and vice versa), as estimated electron-microscopically in ultrathin sections and autoradiographically in squash preparations. In autoradiograms, the DMSO-stimulated BRs exhibit the most dense concentration of silver grains and therefore the highest rate of transcriptional activity. In DMSO-repressed BRs (and other puffs) the transcription of the locus specific genes is not completely shut off. In chromosomes from nuclei with high labelling intensities the repressed BRs (and other puffs) always exhibit a low level of 3H-uridine incorporation in vivo. The absence of cytologically visible BR (puff) formation therefore does not necessarily indicate complete transcriptional inactivity. Typically, before the stage of puff formation the 3H-uridine labelling first appears in the interband-like regions.