Involvement of free cholesterol and high-density lipoprotein in development and resistance of the preimplantation bovine embryo to heat shock.

Development of the mammalian preimplantation embryo is susceptible to disruption by elevated temperature. The molecular and biochemical bases for developmental, genetic, and other differences in embryonic resistance to heat shock are largely not known. Here we tested the hypothesis that increasing free cholesterol content could improve embryonic resistance to heat shock. Culture of bovine embryos at 41.0°C for 15 h beginning at 30 h after insemination (1- to 2-cell stage) reduced development to the blastocyst stage. Reduction in embryonic cholesterol content by culture with methyl-β-cyclodextrin (MBCD) reduced development. This effect of MBCD could be abrogated in 1 of 2 experiments if the molecule was loaded with cholesterol before addition to culture medium. Even though culture with cholesterol-loaded MBCD increased free cholesterol content, it did not increase resistance of embryos to heat shock. Treatment of embryos with cholesterol-loaded high density lipoprotein (HDL) increased embryonic resistance to heat shock even though it slightly reduced embryo cholesterol content. It is likely that other actions of HDL (e.g., protection from free radicals) were responsible for the thermoprotective properties of this molecule. A final experiment was performed to determine whether the increased resistance of embryos at d 5 of development to heat shock as compared with the 2-cell embryo was due to changes in free cholesterol content. However, there was no significant difference in cholesterol content between 2-cell embryos and d 5 embryos that were > 16 cells in development. In conclusion, raising cholesterol content does not improve embryonic survival in response to heat shock. Depletion of cholesterol, in contrast, reduces competence of embryos to develop to the blastocyst stage. High density lipoprotein is thermoprotective to embryos and probably acts through a mechanism independent of its actions on embryonic content of free cholesterol.