title: Uptake of betaine into mouse cumulus-oocyte complexes via the SLC7A6 isoform of y+L transporter.
abstract: Betaine (N,N,N-trimethylglycine) has previously been shown to function in cell volume homeostasis in early mouse embryos and also to be a key donor to the methyl pool in the blastocyst. A betaine transporter (SLC6A20A or SIT1) has been shown to be activated after fertilization, but there is no saturable betaine uptake in mouse oocytes or eggs. Unexpectedly, the same high level of betaine is present in mature metaphase II (MII) eggs as is found in one-cell embryos despite the lack of transport in oocytes or eggs. Significant saturable betaine transport is, however, present in intact cumulus-oocyte complexes (COCs). This transport system has an affinity for betaine of ∼227 μM. The inhibition profile indicates that betaine transport by COCs could be completely blocked by methionine, proline, leucine, lysine, and arginine, and transport is dependent on Na(+) but not Cl(-). This is consistent with transport by a y+L-type amino acid transport system. Both transcripts and protein of one y+L isoform, SLC7A6 (y+LAT2), are present in COCs, with little or no expression in isolated germinal vesicle (GV)-stage oocytes, MII eggs, or one-cell embryos. Betaine accumulated by COCs is transferred into the enclosed GV oocyte, which requires functional gap junctions. Thus, at least a portion of the endogenous betaine in MII eggs could be derived from transport into cumulus cells and subsequent transfer into the enclosed oocyte before gap junction closure during meiotic maturation. The oocyte-derived betaine then could be regulated and supplemented by the SIT1 transporter that arises in the embryo after fertilization.
authors.affiliation: Ottawa Hospital Research Institute, Ottawa, Ontario, Canada.
authors.forename: Hannah E
authors.initials: HE
authors.lastname: Corbett
authors.forename: Chantal D
authors.initials: CD
authors.lastname: Dubé
authors.forename: Sandy
authors.initials: S
authors.lastname: Slow
authors.forename: Michael
authors.initials: M
authors.lastname: Lever
authors.forename: Jacquetta M
authors.initials: JM
authors.lastname: Trasler
authors.forename: Jay M
authors.initials: JM
authors.lastname: Baltz
mesh_terms: D026923:Amino Acid Transport Systems, Basic / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001622:Betaine / Q000378:metabolism* / Q000493:pharmacokinetics; D001692:Biological Transport / Q000502:physiology; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D054885:Cumulus Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D005306:Fertilization / Q000502:physiology; D050485:GABA Plasma Membrane Transport Proteins; D017629:Gap Junctions / Q000378:metabolism; D007477:Ions / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D011247:Pregnancy; D014316:Tritium
chemical_list: D026923:Amino Acid Transport Systems, Basic; D000596:Amino Acids; D002352:Carrier Proteins; D050485:GABA Plasma Membrane Transport Proteins; D007477:Ions; C542311:LAT3 protein, mouse; D014316:Tritium; C072207:betaine plasma membrane transport proteins; D001622:Betaine
keywords: betaine; cell volume; cumulus cells; oocyte; transporters