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RESEARCH ARTICLE
Contents Vol 18(5)

Metabolic regulation of in-vitro-produced bovine embryos. II. Effects of phenazine ethosulfate, sodium azide and 2,4-dinitrophenol during post-compaction development on glucose metabolism and lipid accumulation

Jose Fernando De La Torre-Sanchez A C , David K. Gardner B , Kimberly Preis A B , John Gibbons B D and George E. Seidel Jr A E
+ Author Affiliations
- Author Affiliations

A Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523-1683, USA.

B Colorado Center for Reproductive Medicine, 799 E. Hampden Av., Englewood, CO 80113, USA.

C Present address: Centro de Investigación Regional ‘Pacifico Centro’, INIFAP. Interior Parque Los Colomos S/N Col. Providencia. GuadalaJara, Jalisco, Mexico. CP 44660.

D Present address: Department of Animal and Veterinary Science, Clemson University, Clemson, SC 29634-0311, USA.

E Corresponding author. Email: gseidel@colostate.edu

Reproduction, Fertility and Development 18(5) 597-607 https://doi.org/10.1071/RD05064
Submitted: 8 June 2005  Accepted: 7 March 2006   Published: 24 May 2006

Abstract

The objective was to compare effects of three metabolic regulators on development of post-compaction bovine embryos. In-vitro-produced 8- to 16-cell embryos were allocated to treatments for 72 h in G2.2 medium as follows: 0.3 µm phenazine ethosulfate (PES); 27 µm sodium azide (NaN3); 30 µm 2,4-dinitrophenol (DNP); and control, no regulator. Treatments responded similarly for blastocyst rates and embryo quality responses (P > 0.1). The PES treatment resulted in higher glucose metabolism than the NaN3 treatment (18.5 v. 14.5 pmol per embryo per h, P < 0.05), and both did not differ from DNP or the control. The PES treatment tended to result in more flux of glucose through the pentose phosphate pathway (PPP) than the control (50.5 v. 21.5%, P < 0.11). The NaN3 treatment caused more glucose uptake than the PES treatment (38.9 v. 13.1 pmol per embryo per h, P < 0.01), but neither differed from the control or DNP treatment (P > 0.1). Glycolysis for the PES treatment was 187%, which was higher than any of the other groups (88–94%; P < 0.01). There were fewer medium + large lipid granules in the cytoplasm of PES-treated embryos than any other group, including the in vitro control (P < 0.01). However, in vivo control embryos had still fewer large and medium-sized lipid granules (P < 0.01) than the PES treatment. Developmental competence to Day 14 after embryo transfer was similar among treatments. The PES treatment increased glucose metabolism, tended to increase the PPP flux of glucose and clearly reduced accumulation of lipids in embryos produced in the chemically defined media used. Use of PES in culture media may be a promising approach to improving in vitro production of embryos.


Acknowledgments

Numerous students and personnel in the Embryo Transfer Laboratory at Colorado State University assisted with these experiments. This research was funded in part by the Colorado State University Experiment Station via United States Department of Agriculture Regional Project W-1171 and the National Research Initiative grant no. 2003-35203-13705 of the United States Department of Agriculture Cooperative State Research, Education and Extension Service. J. F. De La Torre-Sanchez was supported by the Consejo Nacional de Ciencia y Tecnología (CONACYT) and the Instituto Nacional de Investigaciones Forestales, Agricolas y Pecuarias (INIFAP) Mexico.


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