The distal nephron plays a capital role in the fine regulation of sodium reabsorption. Compared to the cortical collecting duct, much less information is available on the hormonal regulation of sodium transporter genes in the distal convoluted tubule (DCT), where the thiazide-sensitive Na⁺-Cl^- cotransporter (NCC) is the major entry pathway for Na⁺. The aim of the study was to characterize the in vitro effects of aldosterone (ALDO) (1µM) and cyclic AMP (8Br-cAMP) (0.5 mM) on mouse DCT (mDCT), by using an immortalized mDCT cell line. Western blot analysis and semiquantitative RT-PCR were performed to analyze the expression of genes involved in sodium transport. The mDCTcell line expressed the 11-hydroxysteroid dehydrogenase type 2 gene and both the mineralocorticoid and glucocorticoid receptor genes, suggesting ALDO responsiveness. In this sense, we found that mDCT cells expressed the amiloride-sensitive sodium channel (ENaC) and responded to aldosterone by up-regulating the alpha subunit protein. Similarly, ₁ Na⁺-K⁺ ATPase protein was up-regulated by ALDO and 8Br-cAMP. In addition, the aldosterone intermediate gene sgk1 mRNA was increased in response to both ALDO and 8Br-cAMP and the transcription factor HNF3 mRNA was induced by 8Br-cAMP. With respect to NCC regulation, although ALDO induced NCC protein levels in mice in vivo, neither ALDO nor 8Br-cAMP significantly induced the NCC mRNA or protein levels in mDCT cells. These results suggest that in mDCT, ALDO and cAMP modulate some downstream mediators and effectors in vitro, but do not influence the expression of NCC in this cell model.