Selective staining of X chromosome segments in Schistocerca gregaria after denaturation and reassociation procedures

Abstract

The DNA of fixed mitotic and meiotic chromosomes and of spermatides of Schistocerca gregaria males was heat denaturated and then differentially reassociated in a Giemsa buffer or in acridine orange buffer solution. After this procedure, two to three large, selectively stained regions are seen in the X chromosome of spermatocytes and spermatides. Denaturation and reassociation experiments have shown that after differential reassociation such a selective stainability of chromosome regions is characteristic for the presence of fast-reassociating, i.e., repetitive DNA (Stockert and Lisanti, 1972). The possible presence of repetitive DNA in the X chromosome regions concerned can not be the only reason for the occurrence of the heavily stained segments after reassociation because (1) these segments are obtained in positively heteropycnotic X chromosomes, but not in negatively heteropycnotic Xs and (2) they do not occur in positively heteropycnotic X chromosomes when the histones have been extracted before the denaturation and reassociation processes. Contrary to the latter statement, the heavily stained X chromosomal regions are preserved when the histones are removed after the denaturation and reassociation steps. — It is assumed that the heavily stained X chromosome segments represent DNA reassociation complexes which are only formed if histones are present. It is discussed whether the formation of the X chromosome complexes depends on a specific chromatin configuration within positively heteropycnotic X chromosomes.