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The amount of lactate and pyruvate production by kidney cortex homogenates (cell sap and microsomal fraction) from all intermediates of the glycolytic pathway has been investigated in normal and 3,3′,5-triiodo-L-thyronine-treated rats. In normal animals the lactate and pyruvate production increases steadily from glucose (154 nmoles × mg protein-1 × h-1) through phosphoenolpyruvate (31.850 nmoles × mg protein-1 h-1). The addition in vitro of triiodo-L-thyronine to kidney cortex homogenates has no significant effect on lactate production. In the animals treated for 5 days with 1 mg/kg body weight and per diem of triiodo-L-thyronine, the lactate and pyruvate formation from various intermediates of the Embden-Meyerhof pathway is significantly (P < 0.01) increased: from glucose 225% of the control value, from glucose 6-phosphate and fructose 6-phosphate, 192%, from fructose 1,6-bisphosphate and glyceraldehyde 3-phosphate, 132%, and from 3-phosphoglycerate and 2-phosphoglycerate, 120%. No significant increase in lactate and pyruvate production has been observed using either phosphoenolpyruvate or pyruvate as substrates. [ABSTRACT FROM AUTHOR]