α2C-Adrenoceptors modulatel-DOPA uptake in opossum kidney cells and in the mouse kidney

Abstract

Targeted deletion or selective pharmacological inhibition of α2C-adrenoceptors in mice results in increased brain tissue levels of dopamine and its precursor l-3,4-dihydroxyphenylalanine (l-DOPA), without significant changes in l-DOPA synthesis. l-DOPA uptake is considered the rate-limiting step in dopamine synthesis in the kidney. Since α2C-adrenoceptors may influence the transport of l-DOPA, we investigated the effect of α2C-adrenoceptor activation on l-DOPA uptake in a kidney cell line (opossum kidney cells). l-DOPA and dopamine kidney tissue levels in α2C-adrenoceptor knockout (α2CKO) mice and in mice treated with the selective α2C-adrenoceptor antagonist JP-1302 were also evaluated. The α2-adrenoceptor agonist medetomidine (0.1–1,000 nM) produced a concentration-dependent decrease in l-DOPA uptake in opossum kidney cells (IC50: 2.5 ± 0.5 nM and maximal effect: 28 ± 5% of inhibition). This effect was abolished by a preincubation with JP-1302 (300 nM). Furthermore, the effect of medetomidine (100 nM) was abolished by a preincubation with U-0126 (10 μM), a MEK1/2 inhibitor. Kidney tissue levels of l-DOPA were significantly higher in α2CKO mice compared with wild-type mice (wild-type mice: 58 ± 2 pmol/g tissue and α2CKO mice: 81 ± 15 pmol/g tissue, P < 0.05) and in mice treated with JP-1302 (3 μmol/kg body wt) compared with control mice (control mice: 62 ± 2 pmol/g tissue and JP-1302-treated mice: 75 ± 1 pmol/g tissue, P < 0.05), both without significant changes in dopamine kidney tissue levels. However, mice treated with JP-1302 on a high-salt diet presented significantly higher dopamine levels in the kidney and urine compared with control animals on a high-salt diet. In conclusion, in a kidney cell line, α2C-adrenoceptor activation inhibits l-DOPA uptake, and in mice, deletion or blockade of α2C-adrenoceptors increases l-DOPA kidney tissue levels.