Gluconeogenesis, the de novo formation of glucose from non-carbohydrate precursors, is confined to the proximal convoluted and proximal straight tubules of the mammalian kidney. Compared to liver, renal gluconeogenesis has different substrate requirements and responds to different regulatory stimuli. Stimuli in kidney include starvation, metabolic acidosis, glucocorticoid treatment, and, possibly, PTH and catecholamines. Regulation of gluconeogenic flux occurs at three or four key enzyme sites, particularly phosphoenolpyruvate carboxykinase (PEPCK) and fructose 1,6-bisphosphatase. Interest has focused on the relation among H+, Ca2+, and cyclic AMP in the hormonal regulation of gluconeogenesis. The importance of other putative regulators including fructose 2,6-bisphosphate remains to be determined.