Characterization of glucose transport in preimplantation mouse embryos

Abstract

Membrane transport of glucose divorced from metabolism, was analysed in 2-cell embryos, morulae and blastocysts in the preimplantation mouse. A non-metabolizable radiolabelled analogue, 3,0 methyl D-glucose (3OMG) was used, and glucose was used as well in morulae and blastocysts; incubation times were < or = 5 min. Uptake occurred by combination of a non-saturable process, resistant to cytochalasin-B, and a facilitated process exhibiting classic Michaelis-Menten kinetics. The rate constant for the non-saturable component increased from 1.22 +/- 0.12 pL embryo-1 min-1 in 2-cell embryos to 2.08 +/- 0.44 pL embryo-1 min-1 in blastocysts, determined using 3OMG. The Km values of the saturable component for 3OMG at 22 degrees C were relatively constant at about 6.5 mM in 2-cell embryos, morulae and blastocysts. At 37 degrees C, the Km increased from 6 mM in 2-cell embryos to 17 mM in blastocysts. Vmax increased about five-fold during development from the 2-cell stage to the morula stage and about three-fold during development to the blastocyst. The Km values for glucose in morulae and blastocysts were constant at about 1.3 mM at 37 degrees C. Uptake of 3OMG in blastocysts was inhibited by glucose and stimulated by incubation in glucose-free medium. There was no kinetic evidence for the participation of multiple saturable components in uptake by blastocysts or morulae. This supports the observation that the glucose transporter GLUT2, which is first expressed at the 8-cell stage to supplement GLUT1 expressed in the oocyte, does not contribute to the uptake of environmental glucose and is, therefore, probably restricted in expression to abcoelic membrane areas of the trophectoderm. Together with the known values of glucose in uterine fluid, the kinetic data indicate that most glucose enters the trophectoderm by this GLUT1 at a rate which directly reflects the external glucose concentrations. The activity increased on a cellular basis as development proceeded, suggesting increased activity to meet the increasing metabolic requirements of the blastocyst for glucose.