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CHANGES IN ACCESSIBILITY OF DNA TO VARIOUS FLUOROCHROMES DURING SPERMATOGENESIS
Citation:
Evenson, D., Z. Darzynkiewicz, L. Jost, F. Janca, AND B. Ballachey. CHANGES IN ACCESSIBILITY OF DNA TO VARIOUS FLUOROCHROMES DURING SPERMATOGENESIS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-86/163 (NTIS PB87114609), 1986.
Description:
Accessibility of mouse testicular and vas deferens (vas) sperm cell DNA to acridine orange, propidium iodide, ellipticine, Hoechst 33342, mithramycin, chromomycin A3, 4'6-diamidino-2-phenylindole (DAPI), and 7-amino-actinomycin D (7-amino-AMD) was determined by flow cytometry. Permealized cells were either stained directly or after pretreatment with 0.06 N HCI. For histone-containing tetraploid, diploid, and round spermatid cells, HCI extraction of nuclear proteins caused an approximately sixfold increase of 7-amino-AMD stainability but had no significant effect on DAPI stainability. For these same cell types, the stainability with other intercalating (acridine orange, propidium iodide, ellipticine) and externally binding (Hoechst 33342, mithramycin, chromomycin A3) dyes was increased by 1.6- to 4.0-fold after HCI treatment. In sharp contrast, HCI treatment of vas sperm did not increase the staining level of 7-amino-AMD, DAPI, or propidium iodide but did increase the staining level for the other intercalating dyes (1.3- to 1.5-fold) and external dyes (1.3- to 1.9-fold).