It has been reported that α2-adrenoceptors importantly contribute to the urinary output, renin release and water and sodium excretion. We recently revealed that yohimbine, α2-adrenoceptor antagonist, has protective effect on renal ischemia/reperfusion and cisplatin-induced acute kidney injury in rat. However, the localization of α2-adrenoceptor subtypes in kidney is unclear. The present study aimed to investigate the localization of α2-adrenoceptor subtypes in rat kidney. We used 8 weeks Sprague Dawley rats. Using immunofluorescence, both α2A and α2B-adrenoceptors expressed basolateral, but not apical, membrane of the epithelial cells of the proximal tubules. We also showed that α2A and α2B-adrenoceptors were neither expressed in the glomeruli nor in the collecting duct, thin ascending limb and vasa recta. On the other hands, we revealed that α2C-adrenoceptors localized in the glomeruli, lumen of the cortical and medullary collecting duct. Thus, we demonstrated that the α2-adrenoceptor subtypes were variously distributed in rat kidney.