title: Bovine oocytes and early embryos express mRNA encoding glycerol kinase but addition of glycerol to the culture media interferes with oocyte maturation.
abstract: Glycerol plays multi-functional roles in cellular physiology. Other than forming the backbone molecule for glycerophospholipid and triglyceride (TG), glycerol acts as an energy substrate for glycolysis. Spermatozoa are known to utilize glycerol for energy production, but there are no reports of this in oocytes. In this study, the value of glycerol as an energy substrate for bovine oocyte maturation (Exp. 1) and the gene expression of glycerol kinase (GK), an enzyme crucial for cellular glycerol utilization, in bovine oocytes and early embryos (Exp. 2) were examined. In Exp. 1, in vitro maturation (IVM) was conducted using synthetic oviduct fluid supplemented with/without glucose (1.5 mM) and/or glycerol (1.0 mM), and maturation rate, degree of cumulus expansion, glucose consumption and lactate production by cumulus-oocyte complexes (COC) were examined. In Exp. 2, to examine the developmental expression of GK mRNA, cumulus cells, oocytes and embryos at the 2-, 8- and 16-cell, morula, expanded blastocyst and hatched blastocyst stages were obtained in separate experiments, and the expression of GK mRNA was quantified using a real-time PCR. Glycerol did not support oocyte maturation or cumulus expansion. Addition of glycerol to glucose-supplemented media significantly decreased the maturation rate. Expression of GK mRNA was very low in cumulus cells, whereas an appreciable level of the transcript was observed in the oocytes. GK mRNA was detected in embryos at all the stages examined, and its expression significantly increased at the morula stage. These results indicate that glycerol, at least at the present concentration, is not beneficial as a constituent of the medium for bovine oocyte maturation. However, the appreciable levels of GK mRNA found in the oocyte and embryo imply a physiological role for glycerol in bovine oocyte maturation and embryo development.
authors.affiliation: Department of Agriculture and Life Science, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan.
authors.forename: Sumika
authors.initials: S
authors.lastname: Okawara
authors.forename: Seizo
authors.initials: S
authors.lastname: Hamano
authors.forename: Masafumi
authors.initials: M
authors.lastname: Tetsuka
mesh_terms: D000818:Animals; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D003470:Culture Media; D005260:Female; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000494:pharmacology*; D005991:Glycerol Kinase / Q000096:biosynthesis / Q000235:genetics*; D006651:Histocytochemistry / Q000662:veterinary; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D009865:Oocytes / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
chemical_list: D003470:Culture Media; D012333:RNA, Messenger; D019344:Lactic Acid; D005991:Glycerol Kinase; D005947:Glucose; D005990:Glycerol