{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T08:53:06Z","timestamp":1700211186952},"reference-count":37,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2010,1,26]],"date-time":"2010-01-26T00:00:00Z","timestamp":1264464000000},"content-version":"vor","delay-in-days":5535,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microcirculation"],"published-print":{"date-parts":[[1994,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p><jats:bold>Objective:<\/jats:bold> This study involved the pharmacological detection and characterization of binding sites for the neuromodulator neuropeptide Y (NPY) in an <jats:italic>in vitro<\/jats:italic> preparation of capillary endothelial cells derived from bovine adrenal medulla.<\/jats:p><jats:p><jats:bold>Methods:<\/jats:bold> Equilibrium binding assays were conducted on intact cells with <jats:sup>125<\/jats:sup>I Bolton\u2010Hunter labeled NPY (<jats:sup>125<\/jats:sup>I\u2010BH\u2010NPY). The specificity of the high\u2010affinity binding site was evaluated in competition experiments with cold NPY, (Leu<jats:sup>31<\/jats:sup>, Pro<jats:sup>34<\/jats:sup>)NPY (a Y<jats:sub>1<\/jats:sub> receptor ligand, Y<jats:sub>1<\/jats:sub>RL), NPY<jats:sub>13\u201336<\/jats:sub> (a Y<jats:sub>2<\/jats:sub> receptor ligand, Y<jats:sub>2<\/jats:sub>RL), and two other members of the pancreatic polypeptide\u2010fold (PP\u2010fold) family: peptide YY (PYY) and avian pancreatic polypeptide (APP). Forskolin\u2010stimulated adenylate cyclase activity was assessed to detect the participation of this second messenger pathway in the neuromodulator action at the studied cell preparation.<\/jats:p><jats:p><jats:bold>Results:<\/jats:bold> Nonlinear regression analysis of the binding data indicated the existence of high\u2010affinity binding sites with an equilibrium dissociation constant (<jats:italic>K<jats:sub>d<\/jats:sub><\/jats:italic>) value of 39.00 \u00b1 12.84 nM and a maximal binding (<jats:italic>B<\/jats:italic><jats:sub>max<\/jats:sub>) of 489.89 \u00b1 155.49 fmol\/10<jats:sup>6<\/jats:sup> cells (mean \u00b1 SE, <jats:italic>n<\/jats:italic> = 6). NPY, Y<jats:sub>1<\/jats:sub>RL, and PYY displayed a concentration that inhibits the specific binding by 50% IC<jats:sub>50<\/jats:sub> (nM) values of 4.06 \u00b1 1.66 (<jats:italic>n<\/jats:italic> = 4), 2.94 \u00b1 0.75 (<jats:italic>n<\/jats:italic> = 5), and 18.36 \u00b1 10.36 (<jats:italic>n<\/jats:italic> = 3), respectively. APP and Y<jats:sub>2<\/jats:sub>RL were unable to compete with <jats:sup>125<\/jats:sup>I\u2010NPY in the concentration range 0.001\u20131 \u03bcM. Further evaluation of second messenger pathways suggested that NPY binding sites in this model are coupled to the inhibition of adenylate cyclase. NPY significantly inhibited the forskolin\u2010stimulated adenosine cyclic 3\u2032,5\u2032\u2010(hydrogen phosphate) (cAMP) accumulation with a maximal effect of 37.03 \u00b1 6.28%, <jats:italic>n<\/jats:italic> = 5 and an IC<jats:sub>50<\/jats:sub> of 5.96 \u00b1 1.87 nM. The Y<jats:sub>1<\/jats:sub>RL produced a comparable response (IC<jats:sub>50<\/jats:sub> = 5.35 \u00b1 1.39 nM, <jats:italic>n<\/jats:italic> = 4; maximal inhibition of 61.05 \u00b1 13.03%) and Y<jats:sub>2<\/jats:sub>LR had no detectable effect at a similar concentration range.<\/jats:p><jats:p><jats:bold>Conclusions:<\/jats:bold> The results demonstrate the existence of a Y<jats:sub>1<\/jats:sub> receptor in the adrenal medulla capillary endothelial cells, which may be relevant to the postjunctional effect of NPY on this gland.<\/jats:p>","DOI":"10.3109\/10739689409146753","type":"journal-article","created":{"date-parts":[[2008,4,24]],"date-time":"2008-04-24T10:34:27Z","timestamp":1209033267000},"page":"267-273","source":"Crossref","is-referenced-by-count":8,"title":["Specific <sup>125<\/sup>I Neuropeptide Y Binding to Intact Cultured Bovine Adrenal Medulla Capillary Endothelial Cells"],"prefix":"10.1111","volume":"1","author":[{"given":"P.","family":"SANABRIA","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W. 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