Whole mount electron microscopy of polytene chromosomes from Drosophila melanogaster.

Abstract

A simple method of whole mount electron microscopy has been developed to study the fine structural organization of Drosophila melanogaster polytene chromosomes. This method preserves the structural features of these chromosomes and may be of use for rapid gene localization at the ultrastructural level. Chromosome bands were electron dense regions composed of closely packed groups of chromomeres; thin bands consisted of a single row of transverse chromomeres, while thicker bands were composed of two to several rows of chromomeres. Interband regions contained relatively straight chromatin fibres which traversed the interband zone either singly or in bundles of several fibres. The interband chromatin fibres were generally 130 A in diameter or thicker, and appeared to be composed of two or more thinner (80-90 A) fibres. Presumptive puff regions were characterized by extended chromatin fibres having a longitudinal orientation, however some transverse rows of chromomeres were also seen in these regions, suggesting that not all of the chromomeres in a band may be involved in puff formation. The chromatin fibres inthe puffs were frequently thinner than those found in the interband regions. In stretched chromosomes, the chromatin fibres were drawn out into a mass of parallel fibres without any distinction between band and interband regions, supporting the hypothesis that individual chromatids are continuous through both band and interband regions and probably extend throughout the whole chromosome. No core fibres were observed in any of the polytene chromosomes studied.