New fluorescence reactions in DNA cytochemistry. 1. Microscopic and spectroscopic studies on nonrigid fluorochromes

Some nonrigid DNA-binding antibiotics and fluorochromes that recognize adenine-thymine (AT) sequences are widely applied in biomedical research, but the microscopic use, spectral characteristics and DNA binding modes of other similar compounds have been overlooked or scarcely explored. After treatment with thioflavine T, auramine O and G, curcumin, bis-aminophenyl-oxadiazol, berenil and distamycin A, a bright DNA-dependent fluorescence reaction was found in the chromatin of interphase nuclei, meiotic and polytene chromosomes, spermatozoa heads and kinetoplasts of Trypanosoma cruzi epimastigotes. Nucleoli and basophilic cytoplasm showed low or no fluorescence; the highest emission occurred in the AT-rich kinetoplast DNA. When bound to DNA or in the presence of alpha-cyclodextrin and viscous solvents or cosolutes, nonrigid compounds revealed a striking enhancement of fluorescence. The results indicate that these new or poorly known fluorochromes bind selectively to DNA-containing structures and that the minor groove from AT-rich DNA regions could represent the specific and highly fluorescent binding site.