Characterization of ANF receptors in cultured renal cortical vascular smooth muscle cells.
Because atrial natriuretic factor (ANF) has been demonstrated to decrease resistances in cortical renal vessels in vivo, we studied 125I-ANF binding and ANF-dependent guanosine 3',5'-cyclic monophosphate (cGMP) production in subcultured vascular smooth muscle cells (VSMC) prepared from the rabbit renal cortex. 125I-ANF specific binding at 4 degrees C represented 70% of total binding and reached a plateau at 30-60 min. Equilibrium saturation binding curves suggested one group of high-affinity receptor sites (KD = 78 +/- 16 pM, Bmax = 45 +/- 11 fmol/mg) but were compatible with several groups exhibiting close binding parameters. ANF, [Ala7,Ala23]ANF (a linear analogue), and C-ANF-(4-23) (a ligand of C receptors) inhibited 125I-ANF binding at 37 degrees C with nearly similar potencies. In contrast, at 4 degrees C, complete or nearly complete inhibition of binding was obtained with ANF and linear ANF, the latter exhibiting the weakest potency, whereas C-ANF-(4-23) displaced only 35% of the tracer. ANF markedly stimulated cGMP accumulation, with a threshold concentration of 10 pM and a stimulation of 115 times basal value at 0.1 microM. Linear ANF was also stimulatory with a much weaker potency. Around 25% of 125I-ANF bound to cell surface was internalized at 37 degrees C. Phenylarsine oxide partially inhibited internalization as well as the inhibitory potency of C-ANF-(4-23) on 125I-ANF binding. As shown by high-performance liquid chromatography extracellular 125I-ANF was rapidly degraded at 37 degrees C into its 125I-COOH-terminal tripeptide and 125I-Tyr.(ABSTRACT TRUNCATED AT 250 WORDS)
Béa
M L
ML
Institut National de la Santé et de la Recherche Médicale 64, Hôpital Tenon, Paris, France.
Dussaule
J C
JC
Bens
M
M
Ardaillou
R
R
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D001667:Binding, Competitive; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D007457:Iodine Radioisotopes; D007672:Kidney Cortex / Q000098:blood supply*; D007700:Kinetics; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D011817:Rabbits; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000378:metabolism*
D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor; D006152:Cyclic GMP; D015056:1-Methyl-3-isobutylxanthine