Two-cell stage mouse embryos appear to lack mechanisms for alleviating intracellular acid loads.
Mouse embryos at the two-cell stage, like other cells, can recover from an intracellular acid-load. Our previous work has shown, surprisingly, that there is no contribution to this recovery by Na+/H+ antiport activity. Here we show that the recovery similarly is not affected by inhibition of other known intracellular pH (pHi) regulatory mechanisms. Specifically, the recovery is unaffected by lack of external Na+, inhibition of anion exchange, or lack of bicarbonate, which eliminates the Na(+)-dependent HCO3-/Cl- exchanger as a possible mechanisms. These conditions also eliminate any possible Na+,HCO3- cotransporter operating to relieve acid-loading. Recovery is unaffected similarly by nonspecific inhibitors of H(+)-ATPase activity. These observations lead to the conclusion that recovery from acid-load is a passive process in the two-cell mouse embryo. Similarly, the mean base-line pHi (6.84) is not dependent on known pHi regulatory mechanisms. The embryos exhibit a marked intracellular alkalinization when exposed to Cl(-)-free medium in the presence of bicarbonate. This response is eliminated by an inhibitor of anion exchange and by lack of bicarbonate, but is independent of Na+. These results indicate that there is probably a Na(+)-independent HCO3-/Cl- exchanger active in these cells, presumably functioning to alleviate alkaline loads.
Baltz
J M
JM
Laboratory of Human Reproduction and Reproductive Biology, Harvard Medical School, Boston, Massachusetts 02115.
Biggers
J D
JD
Lechene
C
C
D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D027963:Chloride-Bicarbonate Antiporters; D004622:Embryo, Mammalian / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors; D017923:Sodium-Hydrogen Exchangers
D002352:Carrier Proteins; D027963:Chloride-Bicarbonate Antiporters; D008565:Membrane Proteins; D017923:Sodium-Hydrogen Exchangers; D006180:Proton-Translocating ATPases