Human retinal pigment epithelial cells possess muscarinic receptors coupled to calcium mobilization.
Human retinal pigment epithelial (RPE) cells in culture demonstrated saturable specific binding of [3H]quinuclidinyl benzilate (QNB). Specific binding represents about 75% of total binding. Scatchard analysis yields a Kd of 0.178 nM and Bmax of 42 fmol/mg protein. Atropine and carbachol show typical displacement curves, and a Hill plot has a slope of 0.96, suggesting a homogeneous population of receptors. Muscarinic agonists have no effect on intracellular cyclic adenosine monophosphate levels in RPE cells measured by radioimmunoassay, nor do they alter the isoproterenol-induced stimulation of adenylate cyclase. However, both acetylcholine and carbachol cause a rapid increase in intracellular calcium concentration measured by the fluorescent indicator quin 2. Atropine reverses the calcium rise when added after agonist and prevents the rise when added prior to agonist. These data suggest that human RPE cells possess muscarinic receptors coupled to calcium mobilization.
Friedman
Z
Z
Department of Ophthalmology, University of Virginia School of Medicine, Charlottesville 22908.
Hackett
S F
SF
Campochiaro
P A
PA
D001285:Atropine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D004848:Epithelium / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D011813:Quinuclidinyl Benzilate / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000378:metabolism*; D012160:Retina / Q000378:metabolism*
D011976:Receptors, Muscarinic; D011813:Quinuclidinyl Benzilate; D001285:Atropine; D002217:Carbachol; D002118:Calcium