Diabetic nephropathies are characterized by glycogen accumulation in distal tubular cells that eventually leads to their apoptosis. The present study aims to determine if adiponectin and AMPK are involved in the regulation of glycogen synthase in these structures. Western blots of isolated distal tubules revealed the presence of ADIPOR1, catalytic AMPK sub-units 1 and 2, their phosphorylated active forms and the glycogen-binding AMPK sub-unit 2. ADIPOR2 was not detected. Expression levels of ADIPOR1, AMPK 1, AMPK 2 and AMPK 2 were increased in streptozotocin-treated diabetic rats while phosphorylated active AMPK levels were strongly decreased. Immunohistochemistry revealed the presence of adiponectin receptor ADIPOR1 on the luminal portion of distal tubules and TAL cells. Catalytic sub-units 1 and 2, their phosphorylated active forms and the glycogen-binding sub-unit 2 were also found in the same cells, confirming immunoblot results. In vitro, AICAR (2 mM) and globular adiponectin (10 µg/ml) activated catalytic AMPK in distal tubules isolated from kidneys of normal rats but much weakly in those from diabetic rats. Glycogen synthase (GS) inhibition paralleled AMPK activation, in both groups of animals: active GS levels were low in control animals and elevated in diabetic ones. Finally, glucose-6-phosphate, an allosteric activator of GS was also increased in diabetes. These results demonstrate that in distal tubular cells, adiponectin through luminal ADIPOR1 activates AMPK, leading to the inhibition of GS. During hyperglycaemia, this regulation is altered which may explain, at least in part, the accumulation of large glycogen deposits.