In renal epithelia, vasopressin influences salt and water transport chiefly via V2 receptors (V2R). A combination of vasopressin-induced effects along distinct portions of the nephron and collecting duct system may help to balance the net effects of antidiuresis in cortex and medulla. Previous studies on the intrarenal distribution of V2R have been inconclusive with respect to segment- and cell type-related V2R expression. Our study therefore aimed to present a high-resolution analysis of V2R mRNA expression in rat, mouse, and human kidney epithelia, supplemented with immunohistochemical data. Cell types of the renal tubule were identified histochemically using specific markers. Pronounced V2R signal in thick ascending limb (TAL) was corroborated functionally; phosphorylation of the Na⁺,K⁺,2Cl^- -cotransporter 2 (NKCC2) was established in cultured rbTAL cells and in rats with diabetes insipidus (DI) that were treated with the V2R agonist, desmopressin. We have found solid V2R mRNA expression in medullary TAL (MTAL), macula densa, connecting tubule (CNT), cortical (CCD) and medullary collecting duct (MCD), and weaker expression in cortical TAL (CTAL) and distal convoluted tubule (DCT) in all three species. Additional V2R immunostaining of kidneys and rbTAL cells was confirmatory. In agreement with strong V2R expression in MTAL, DI kidneys and cultured TAL cells revealed sharp, selective increases in NKCC2 phosphorylation upon desmopressin treatment. Macula densa cells constitutively showed strong NKCC2 phosphorylation. Results suggest comparably significant effects of vasopressin-induced V2R-signalling in MTAL and in CNT/CD principal cells across the studied species. Strong V2R expression in macula densa may be related with tubulo-vascular signal transfer.