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Transcriptional Control of Development, Protein Synthesis, and Heat-Induced Heat Shock Protein 70 Synthesis in 2-Cell Bovine Embryos¹

^a Department of Dairy & Poultry Sciences, University of Florida, Gainesville, Florida 32611 ^b Department of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, People's Republic of China

Experiments were performed to evaluate the role of transcription in early development of bovine embryos. Two transcription inhibitors—5,6-dichloro-1-ß-D-ribofuranosylbenzimidazole (DRB) and actinomycin D—were used to test whether 1) the inhibitors alter the rate of early embryonic development and protein synthesis, 2) heat shock increases the steady-state amounts of mRNA for the inducible form of heat shock protein 70 (HSP70) in embryos, and 3) this latter effect is blocked by transcription inhibitors. Addition of either DRB or actinomycin D to culture medium beginning 8 h postinsemination (hpi) reduced the proportion of oocytes that had undergone cleavage by 32–34 hpi. Both transcription inhibitors also reduced the proportion of cleaved embryos that reached the 4-cell stage by 32–34 hpi. Incorporation of ³⁵S-labeled amino acids into de novo synthesized protein by bovine 2-cell embryos was lower for embryos cultured with DRB. Using reverse transcription-polymerase chain reaction, HSP70 mRNA in 2- and 4-cell embryos was increased by exposure to 42°C. Both inhibitors reduced amounts of HSP70 mRNA at 42°C. Results indicate that bovine embryos can undergo transcription in response to heat shock as early as the 2-cell stage. Moreover, the observations that transcription inhibitors reduce rates of cleavage and early development point out the importance of transcription for development from the earliest period of embryonic life.

¹ Research was supported in part by USDA NRICGP #96-35205-3728. This is Journal Series No. R-06800 of the Florida Agricultural Experiment Station.

² Correspondence: P.J. Hansen, Department of Dairy & Poultry Sciences, PO Box 110920, University of Florida, Gainesville, FL 32611-0920. FAX: 352 392 5595; hansen{at}dps.ufl.edu

³ Current address: Dept. of Veterinary Gynaecology & Obstetrics, Haryana Agricultural University, Hisar, Haryana, India.

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