Detection of genome impairment in bovine early embryos by autoradiography: a methodological note

Abstract

The applicability of Pavlok's method characterising the nuclear status of early preimplantation bovine embryos by nuclear labelling pattern after a short pulse of [5-³H]uridine (revealing in situ detection of RNA transcription at the onset of the major embryonic transcription) was tested on experimentally irradiated 8- to 16-cell bovine embryos. After [5-³H]uridine labelling the semi-thin sections of these embryos were analysed by autoradiography for intranuclear distribution of newly synthesised RNA expected to be influenced by increasing doses of irradiation by gamma rays from a ⁶⁰Co source. In control embryos, the labelling was homogeneously distributed in nucleoplasm and in nucleoli. The expected effects were clearly detected already in embryos irradiated with a dose of 2 Gy, in which low-level RNA synthesis was localised mostly at the periphery of the nucleus, the nuclear centre being without labelling. A detailed analysis of consecutive sections of embryos from all groups of irradiated and control embryos, using an arbitrary scale considering these effects, confirmed the detectability of the threshold level of genome impairment.