Detection and evaluation of chromosomal aberrations induced by high doses of gamma-irradiation using immunogold-silver painting of prematurely condensed chromosomes.
We developed a method for detecting chromosomal aberrations after irradiation with high doses of gamma-rays. At high dose levels, few of the irradiated cells are able to enter mitosis so that it is difficult to obtain enough mitotic chromosomes using a conventional colcemid block. Therefore, okadaic acid was used to condense prematurely the chromosomes of interphase cells. Even after irradiation with doses > 40 Gy, okadaic acid was able to force sufficient numbers of cells to condense their chromosomes. Segments of chromosomes were then detected by chromosome painting using a gold-conjugated antibody followed by silver enhancement. This simple method allows highly damaged chromosomes to be detected with great sensitivity.
Gotoh
E
E
Division of Genetic Resources, National Institute of Health Japan, Tokyo, Japan.
Asakawa
Y
Y
D002453:Cell Cycle; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D002875:Chromosomes / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects*; D003703:Demecolcine / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005720:Gamma Rays; D006801:Humans; D007150:Immunohistochemistry; D008214:Lymphocytes; D019319:Okadaic Acid / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D011829:Radiation Dosage
D004791:Enzyme Inhibitors; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D003703:Demecolcine