Tubule fragments were isolated by collagenase treatment of guinea pig kidney cortex. 2. 3':5'-Cyclic AMP increased gluconeogenesis from lactate, pyruvate, malate and fructose. 3. Noradrenaline decreased gluconeogenesis from lactate, pyruvate, 2-oxoglutarate and fructose. 4. Angiotensin II slightly, but significantly, increased gluconeogenesis from lactate. 5. Gluconeogenesis from glycerol as sole substrate was negligible. Gluconeogenesis from combinations of glycerol together with either lactate, pyruvate, 2-oxoglutarate or malate was appreciably greater than the sum of the glucose output observed when these substrates were added singly.