Differential replication of heterochromatin in polytene chromosomes of the Old World screw-worm fly,Chrysomya bezziana (Diptera: Calliphoridae), analysed by fluorescence staining

Abstract

The distribution and replication of heterochromatin in polytene trichogen chromosomes of the Old World screw-worm fly,Chrysomya bezziana, were studied using fluorescent staining techniques. Quinacrine and distamycin-DAPI, which selectively stain AT-rich DNA, and chromomycin, specific for GC-rich sequences, were used. Bright quinacrine and DA-DAPI fluorescence was found in the sex chromosome body and in all autosomal centromere regions. Chromomycin (CMA) staining results in very little bright fluorescence of the sex chromosome body and autosomal centromeric regions, but many bright bands of varying morphology are distributed in autosomal arms. The expected negative CMA staining of quinacrine and DA-DAPI bright regions was not found. The lack of reciprocal staining patterns may result from changes in the higher order chromatin structure of polytene chromosomes, or intercalation of divergent heterochromatic sequences. Comparison of the different staining techniques in mitotic and polytene cells shows that heterochromatin is differentially under-replicated, so that the proportions of the distinct fluorescent-specific chromatin changes during polytenization. CMA staining within autosomal arms suggests that repeated sequences intercalated in euchromatin are co-replicated during polytenization. The numerous fluorescent markers described also provide further morphological features for use in comparative cytological analysis ofC. bezziana.