Use of a fluorescent stain for visualization of nuclear material in living oocytes and early embryos.
Hoechst dyes 33342 and 33258 were used to visualize pronuclei and nuclei of early preimplantation embryos. Murine one-cell zygotes exposed to dye stained rapidly over a range of concentrations (0, 0.02, 0.04, 0.1 or 0.2 micrograms/50 microliter of media). Development to morula and blastocyst in vitro was reduced (39/70, 56%; p less than 0.05) compared to controls (44/57, 77%) but not completely blocked. Porcine and bovine zygotes and embryos could also be stained but required incubation times up to 4 hr. Porcine embryos exposed to Hoechst 33342 had limited (p less than 0.01) in vitro development (29/74, 39%) compared to unstained controls (49/64, 76%). Hoechst dyes stain embryos from different species but suitably adjusted incubation times are required. Limited preimplantation development in vitro may be expected following staining and exposure to ultraviolet light.
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D000818:Animals; D001562:Benzimidazoles; D006690:Bisbenzimidazole; D002455:Cell Division; D002467:Cell Nucleus / Q000648:ultrastructure*; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005456:Fluorescent Dyes; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000166:cytology*
D001562:Benzimidazoles; D005456:Fluorescent Dyes; D006690:Bisbenzimidazole; C017807:bisbenzimide ethoxide trihydrochloride