Effects of oxygen tension and IGF-I on HIF-1α protein expression in mouse blastocysts

Abstract

Purpose: Hypoxia inducible factors (HIFs) are key regulators of oxygen homeostasis in response to reduced oxygenation in somatic cells. In addition, HIF-1α protein can be also induced by insulin-like growth factor I (IGF-I) treatment in various cell lines under normoxic condition. However, the expression and function of HIF-1α in embryogenesis are still unclear. Therefore, the objectives of this study were to examine the expression of HIF-1α in mouse blastocysts cultured under hypoxic and normoxic conditions, and to determine whether oxygen tension and IGF-I influence embryonic development through stimulation of HIF-1α expression. Methods: Mouse embryos were cultured from the 1-cell to blastocyst stage under 5 % or 20 % O2 in both the absence and presence of IGF-I. Results: The embryonic development rates to the blastocyst stage were not affected by oxygen tension or IGF-I treatment. HIF-1α protein was localized to the cytoplasm of blastocysts, and its levels were independent of oxygen concentration or IGF-I treatment. Blastocysts cultured under 5 % O2 exhibited significantly higher total cell numbers (83.4 ± 18.1) and lower apoptotic index (3.7 ± 1.5) than those cultured under 20 % O2 (67.4 ± 15.6) (6.9 ± 3.5) (P < 0.05). IGF-I reduced the apoptotic index in both oxygen conditions, but a significant decrease was detected in the 20 % O 2 group. Conclusions: HIF-1α may not be a major mediator that responds to change in oxygen tension within blastocysts, inconsistent with that of somatic cells. Supplementation of culture media with IGF-I has been shown to promote embryo development by an anti-Apoptotic effect, instead of increasing HIF-1α protein expression. © 2012 Springer Science+Business Media New York.