title: Commonly used fertility drugs, a diet supplement, and stress force AMPK-dependent block of stemness and development in cultured mammalian embryos.
abstract: OBJECTIVE
  The purpose of the present study is to test whether metformin, aspirin, or diet supplement (DS) BioResponse-3,3'-Diindolylmethane (BR-DIM) can induce AMP-activated protein kinase (AMPK)-dependent potency loss in cultured embryos and whether metformin (Met) + Aspirin (Asa) or BR-DIM causes an AMPK-dependent decrease in embryonic development.
  
  
  METHODS
  The methods used were as follows: culture post-thaw mouse zygotes to the two-cell embryo stage and test effects after 1-h AMPK agonists' (e.g., Met, Asa, BR-DIM, control hyperosmotic stress) exposure on AMPK-dependent loss of Oct4 and/or Rex1 nuclear potency factors, confirm AMPK dependence by reversing potency loss in two-cell-stage embryos with AMPK inhibitor compound C (CC), test whether Met + Asa (i.e., co-added) or DS BR-DIM decreases development of two-cell to blastocyst stage in an AMPK-dependent (CC-sensitive) manner, and evaluate the level of Rex1 and Oct4 nuclear fluorescence in two-cell-stage embryos and rate of two-cell-stage embryo development to blastocysts.
  
  
  RESULTS
  Met, Asa, BR-DIM, or hyperosmotic sorbitol stress induces rapid ~50-85 % Rex1 and/or Oct4 protein loss in two-cell embryos. This loss is ~60-90 % reversible by co-culture with AMPK inhibitor CC. Embryo development from two-cell to blastocyst stage is decreased in culture with either Met + Asa or BR-DIM, and this is either >90 or ~60 % reversible with CC, respectively.
  
  
  CONCLUSIONS
  These experimental designs here showed that Met-, Asa-, BR-DIM-, or sorbitol stress-induced rapid potency loss in two-cell embryos is AMPK dependent as suggested by inhibition of Rex1 and/or Oct4 protein loss with an AMPK inhibitor. The DS BR-DIM or fertility drugs (e.g., Met + Asa) that are used to enhance maternal metabolism to support fertility can also chronically slow embryo growth and block development in an AMPK-dependent manner.
authors.affiliation: CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI, 48201, USA. abolnick@med.wayne.edu.
authors.forename: Alan
authors.initials: A
authors.lastname: Bolnick
authors.affiliation: CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI, 48201, USA.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Abdulhasan
authors.affiliation: CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI, 48201, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Kilburn
authors.affiliation: Fertility and Surgical Associates of California, Thousand Oaks, CA, 91361, USA.
authors.forename: Yufen
authors.initials: Y
authors.lastname: Xie
authors.affiliation: EmbryoTech Laboratories, 140 Hale Street, Haverhill, MA, 01830, USA.
authors.forename: Mindie
authors.initials: M
authors.lastname: Howard
authors.affiliation: Ob/Gyn, IVF Clinic, University Physician Group, Wayne State University School of Medicine, 26400 W 12 Mile Road, Suite 140, Southfield, MI, 48034, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Andresen
authors.affiliation: University of Utah, 201 Presidents Circle, Salt Lake City, UT, 84112, USA.
authors.forename: Alexandra M
authors.initials: AM
authors.lastname: Shamir
authors.affiliation: CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI, 48201, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Dai
authors.affiliation: CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI, 48201, USA.
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Puscheck
authors.affiliation: CS Mott Center for Human Growth and Development, Department of Ob/Gyn, Reproductive Endocrinology and Infertility, Wayne State University School of Medicine, 275 East Hancock, Detroit, MI, 48201, USA.|Program for Reproductive Sciences and Department of Physiology, Wayne State University School of Medicine, Detroit, MI, 48201, USA.|Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, MI, 48201, USA.|Institutes for Environmental Health Science, Wayne State University School of Medicine, Detroit, MI, 48201, USA.|Department of Biology, University of Windsor, Windsor, ON, N9B 3P4, Canada.
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Rappolee
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D000818:Animals; D001241:Aspirin / Q000494:pharmacology*; D019587:Dietary Supplements; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000166:cytology*; D047108:Embryonic Development / Q000187:drug effects*; D005299:Fertility Agents / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology*; D008687:Metformin / Q000494:pharmacology*; D051379:Mice; D013012:Sorbitol / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology; D013312:Stress, Physiological
chemical_list: D005299:Fertility Agents; D007211:Indoles; D013012:Sorbitol; D008687:Metformin; D055372:AMP-Activated Protein Kinases; D001241:Aspirin; C016392:3,3'-diindolylmethane
keywords: Diet supplement; Embryo culture; Embryo development; Embryo quality; Fertility drugs