Phosphocreatine protects transgenic mouse liver expressing creatine kinase from hypoxia and ischemia.
Creatine kinase (CK) is normally found at high levels in muscle and brain and catalyzes the reaction phosphocreatine (PCr) + MgADP + H+<==>creatine (Cr) + MgATP. CK is not normally found at high levels in liver. A line of transgenic mice that express high levels of the BB-dimer of CK (CKB) in liver has allowed us to assess the role of CKB during periods of low oxygen stress. During 40 min of ischemia of normal perfused livers at 25 degrees C, ATP levels are depleted, and pH decreases to 6.6. pH recovers to a preischemic level after 30 min of reperfusion of normal livers; however, P(i) levels are significantly higher and ATP levels significantly lower than preischemic values. In transgenic liver with an initial PCr-to-ATP ratio of 4.5, ATP levels are maintained until PCr is markedly depleted. pH remains at preischemic levels for 16 min of ischemia of transgenic livers. During this length of ischemia in normal livers, pH has dropped to 6.9. pH, P(i), and ATP levels return to preischemic values within 30 min of reperfusion in transgenic livers containing PCr and CK. During 90 min of hypoxia of normal perfused livers at 37 degrees C, ATP is depleted. After 15 min of hypoxia of normal livers, there is a significant increase in the release of lactate dehydrogenase (LDH). In transgenic livers, ATP is maintained, and no increase in LDH release is observed for up to 90 min, depending on the level of PCr before hypoxia. These results demonstrate the role of CKB in buffering ATP levels and regulating intracellular pH during periods of low oxygen stress.(ABSTRACT TRUNCATED AT 250 WORDS)
Miller
K
K
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.
Halow
J
J
Koretsky
A P
AP
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D003402:Creatine Kinase / Q000096:biosynthesis* / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D000860:Hypoxia / Q000378:metabolism / Q000517:prevention & control*; D007511:Ischemia / Q000378:metabolism / Q000517:prevention & control*; D007527:Isoenzymes; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000098:blood supply / Q000201:enzymology / Q000502:physiology*; D008112:Liver Glycogen / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D051379:Mice; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D010725:Phosphocreatine / Q000378:metabolism*; D015424:Reperfusion; D013997:Time Factors
D007527:Isoenzymes; D008112:Liver Glycogen; D010725:Phosphocreatine; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase