Abstract

The stimulation of alpha-2 adrenergic receptors in the kidney produces diuresis. In this study, alpha-2 adrenergic receptors were examined in inner medullary collecting duct (IMCD) cells isolated from the rabbit kidney. The equilibrium binding of [3H] rauwolscine to IMCD cell homogenate was measured after incubation for 45 min at 25 degrees C in the absence (total binding) and presence (nonspecific binding) of 100 microM phentolamine. The specific binding of [3H]rauwolscine was saturable with a Bmax of 170 fmol/mg of protein and Kd of 4.18 nM. The displacement of [3H]rauwolscine binding to IMCD cells by adrenergic antagonists and agonists displayed the following order of potency: phentolamine > yohimbine > clonidine > oxymetazoline > azepexole > propranolol > prazosin > alpha-methyl norepinephrine > epinephrine. Activation of alpha-2 adrenergic receptors by azepexole decreased vasopressin- and forskolin-stimulated cAMP formation in the IMCD cells. Because IMCD cells in the kidney have a hypertonic environment made up of urea and NaCl, we also determined the effects of a mixture of NaCl and urea (1200 mOsm/kg water) on the binding of [3H]rauwolscine and cAMP formation in the IMCD cells. The hypertonicity increased the Kd and Bmax of [3H]rauwolscine binding to 15.2 nM and 240 fmol/mg of protein, respectively. In addition, hypertonicity decreased forskolin-stimulated cAMP formation in IMCD cells. It is concluded that: 1) high-affinity specific alpha-2 adrenergic receptors are present in the rabbit IMCD; 2) selective alpha-2 agonists inhibit vasopressin- and forskolin-stimulated cAMP formation in rabbit IMCD and 3) the effects of adrenergic drugs on the IMCD in vivo will depend on the renal medullary osmolality that itself depends on the state of the hydration of the animal.