EM autoradiographic studies on polytene nuclei of Drosophila melanogaster

Abstract

Replication in the chromocentre heterochromatin of salivary gland polytene nuclei of Drosophila melanogaster has been examined by ³H-thymidine EM autoradiography. In vitro pulse labelling of salivary glands from late third instar larvae showed that the chromocentre heterochromatin replicates in synchrony with the euchromatin in the nucleus. Within the chromocentre region, the central compact mass, identified earlier as the alpha heterochromatin, did not incorporate ³H-thymidine at any stage of the S, while the surrounding beta heterochromatin was always labelled in nuclei with labelled euchromatin. In a second set of experiments, growing larvae from just after hatching till late third instar stages, were fed on food containing ³H-thymidine, and at the end of larval life, the incorporation in salivary gland nuclei was examined by EM autoradiography. A grain density analysis of the EM autoradiographs revealed that the alpha heterochromatin does not replicate at all from after hatching till late third instar while the beta heterochromatin replicates as much as the euchromatin. Non-replication of the alpha heterochromatin provides the explanation for the lowered amount of heterochromatin in the polytene nuclei compared to their diploid counterparts. Implications of these observations on the organization of chromocentre heterochromatin in polytene nuclei and its homology to the heterochromatic regions in mitotic chromosomes are discussed.