Generation of extramitochondrial reducing power in gluconeogenesis.

1. Kidney-cortex slices incubated with pyruvate formed glucose and lactate in relatively large and approximately equimolar quantities. The formation of these products involves two exclusively cytoplasmic NADH(2)-requiring reductions, catalysed by lactate dehydrogenase and triose phosphate dehydrogenase. From the rates of glucose and lactate formation it can be calculated that over 1000mu-moles of NADH(2) must have been produced in the cytoplasm/g. dry wt. of tissue/hr. 2. When lactate is a gluconeogenic precursor the required NADH(2) is generated in the cytoplasm, but, when a substrate more highly oxidized than glucose, such as pyruvate, is the precursor, there is no direct cytoplasmic source of NADH(2). Quantitative data on the fate of pyruvate are in accord with the conclusion that the NADH(2) was primarily formed intramitochondrially by the dehydrogenases of cell respiration, with pyruvate as the major substrate. 3. Similar observations and conclusions apply to experiments with mouse-liver slices incubated with pyruvate, serine or aspartate. 4. Addition of ethanol, which increases the formation of NADH(2) in the cytoplasm, increased the formation from pyruvate of lactate but not of glucose. 5. In view of the low permeability of mitochondria for NAD and NADH(2) it must be postulated that special carrier mechanisms transfer the reducing equivalents of intramitochondrially generated NADH(2) to the cytoplasm. Reasons are given in support of the assumption that the malate-oxaloacetate system acts as the carrier. 6. Various aspects of the generation of reducing power and its transfer from mitochondria to cytoplasm are discussed.