title: Desensitization and re-sensitization of CGRP receptor function in human neuroblastoma SK-N-MC cells.
abstract: Calcitonin gene-related peptide (CGRP) is a highly potent vasodilator known to be involved in many physiological functions within the cardiovascular, gastrointestinal, immune, and nervous systems. This study assessed the desensitization of CGRP receptors by measuring agonist-mediated activation of adenylate cyclase in a model system employing human neuroblastoma-derived SK-N-MC cells. In these cells, we demonstrated that pre-incubation with CGRP (20 nM) induces a rapid desensitization of CGRP signaling (t(1/2)<or=3 min) by causing a decrease in potency and efficacy. CGRP's desensitization potency (DC(50)=0.29 nM) is similar to its activation potency on non-desensitized cells (EC(50)=0.20 nM). The desensitized receptors exhibited slow and incomplete re-sensitization upon removal of the pre-incubated ligand, resulting in 52-65% functional recovery after 3-5 h while CGRP binding sites were completely restored. Additional agonists within the calcitonin/CGRP family of peptides (calcitonin, amylin, adrenomedullin, and adrenomedullin 2) were compared to CGRP with regard to their ability to activate and desensitize CGRP receptors. Calcitonin and amylin did not cause receptor activation nor did they produce desensitization. Adrenomedullin and adrenomedullin 2 activated the receptors and produced desensitization, but at a slower rate and with a weaker desensitization potency than CGRP-induced desensitization. Adrenomedullin exhibited similar potency for receptor activation and desensitization, whereas adrenomedullin 2 has a 4-fold higher preference for receptor desensitization than for receptor activation. Activation and desensitization induced by CGRP, adrenomedullin and adrenomedullin 2 were blocked by the CGRP receptor antagonist CGRP8-37. These data indicate that CGRP receptors are desensitized by select peptides in the calcitonin/CGRP family. Slow recovery from the desensitized state may provide a strategy for timed modulation of the CGRP signaling pathway.
authors.affiliation: Bristol-Myers Squibb Co., 5 Research Parkway, Wallingford, CT 06492, United States. sokhom.pin@bms.com
authors.forename: Sokhom S
authors.initials: SS
authors.lastname: Pin
authors.forename: Cen
authors.initials: C
authors.lastname: Xu
authors.forename: Ben A
authors.initials: BA
authors.lastname: Bahr
mesh_terms: D053607:Adrenomedullin / Q000494:pharmacology; D000682:Amyloid / Q000494:pharmacology; D001932:Brain Neoplasms / Q000378:metabolism*; D002116:Calcitonin / Q000378:metabolism / Q000494:pharmacology; D015740:Calcitonin Gene-Related Peptide / Q000494:pharmacology; D000077221:Calcitonin Gene-Related Peptide Receptor Antagonists; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects; D003627:Data Interpretation, Statistical; D004298:Dopamine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D007457:Iodine Radioisotopes; D058228:Islet Amyloid Polypeptide; D007700:Kinetics; D009447:Neuroblastoma / Q000378:metabolism*; D018015:Receptors, Calcitonin Gene-Related Peptide / Q000819:agonists / Q000378:metabolism*