Ammonium induces aberrant blastocyst differentiation, metabolism, pH regulation, gene expression and subsequently alters fetal development in the mouse.
The presence of ammonium in the culture medium has significant detrimental effects on the regulation of embryo physiology and genetics. Ammonium levels build up linearly over time in the culture medium when media containing amino acids are incubated at 37 degrees C. Ammonium in the culture media significantly reduces blastocyst cell number, decreases inner cell mass development, increases apoptosis, perturbs metabolism, impairs the ability of embryos to regulate intracellular pH, and alters the expression of the imprinted gene H19. In contrast, the rate of blastocyst development and blastocyst morphology appear to be normal. The transfer of blastocysts exposed to ammonium results in a significant reduction in the ability to establish a pregnancy. Furthermore, of those embryos that manage to implant, fetal growth is significantly impaired. Embryos exposed to 300 microM ammonium are retarded by 1.5 days developmentally at Day 15 of pregnancy. It is therefore essential that culture conditions for mammalian embryos are designed to minimize the buildup of ammonium to prevent abnormalities in embryo physiology, genetic regulation, pregnancy, and fetal development.
Lane
Michelle
M
Research Department, Colorado Center for Reproductive Medicine, Englewood, Colorado 80110, USA. michelle.lane@adelaide.edu.au
Gardner
David K
DK
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects* / Q000235:genetics; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D005314:Embryonic and Fetal Development / Q000187:drug effects; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008815:Mice, Inbred Strains; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000187:drug effects / Q000235:genetics
C094335:H19 long non-coding RNA; D000644:Quaternary Ammonium Compounds; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated