Novel cell line selectively expressing neuropeptide Y-Y2 receptors

Neuropeptide Y (NPY) recognition by the human neuroblastoma cell lines SiMa, Kelly, SH-SY5Y, CHP-234, and MHH-NB-11 was analyzed in radioactive binding assays using tritiated NPY. For the cell lines CHP-234 and MHH-NB-11 binding of 3H]propionyl-NPY was observed with Kd-values of 0.64 +/- 0.07 nM and 0.53 +/- 0.12 nM, respectively, determined by saturation analysis with non-linear regression. The receptor subtype was determined by competition analysis using the subtype selective NPY analogues Leu31, Pro34]-NPY (NPY-Y1, NPY-Y5), Ahx(5-24)]-NPY (NPY-Y2), Ala31, Aib32]-NPY (NPY-Y5), NPY 3-36] (NPY-Y2, NPY-Y5), and NPY 13-36] (NPY-Y2). Both cell lines, CHP-234 and MHH-NB-11, the latter one being characterized for NPY receptors for the first time, showed exclusive expression of NPY-Y2 receptors. In both cell lines binding of NPY induced signal transduction, which was monitored as reduction of forskolin-induced cAMP production in an ELISA.