title: Quantitative impedimetric NPY-receptor activation monitoring and signal pathway profiling in living cells.
abstract: Label-free and non-invasive monitoring of receptor activation and identification of the involved signal pathways in living cells is an ongoing analytic challenge and a great opportunity for biosensoric systems. In this context, we developed an impedance spectroscopy-based system for the activation monitoring of NPY-receptors in living cells. Using an optimized interdigital electrode array for sensitive detection of cellular alterations, we were able for the first time to quantitatively detect the NPY-receptor activation directly without a secondary or enhancer reaction like cAMP-stimulation by forskolin. More strikingly, we could show that the impedimetric based NPY-receptor activation monitoring is not restricted to the Y1-receptor but also possible for the Y2- and Y5-receptor. Furthermore, we could monitor the NPY-receptor activation in different cell lines that natively express NPY-receptors and proof the specificity of the observed impedimetric effect by agonist/antagonist studies in recombinant NPY-receptor expressing cell lines. To clarify the nature of the observed impedimetric effect we performed an equivalent circuit analysis as well as analyzed the role of cell morphology and receptor internalization. Finally, an antagonist based extensive molecular signal pathway analysis revealed small alterations of the actin cytoskeleton as well as the inhibition of at least L-type calcium channels as major reasons for the observed NPY-induced impedance increase. Taken together, our novel impedance spectroscopy based NPY-receptor activation monitoring system offers the opportunity to identify signal pathways as well as for novel versatile agonist/antagonist screening systems for identification of novel therapeutics in the field of obesity and cancer.
authors.affiliation: Center for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: te Kamp
authors.affiliation: Center for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Ricco
authors.initials: R
authors.lastname: Lindner
authors.affiliation: Center for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Heinz-Georg
authors.initials: HG
authors.lastname: Jahnke
authors.affiliation: Center for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany.
authors.forename: Dana
authors.initials: D
authors.lastname: Krinke
authors.affiliation: Institute of Biochemistry, Universität Leipzig, Brüderstraße 34, D-04103 Leipzig, Germany.
authors.forename: Katja B
authors.initials: KB
authors.lastname: Kostelnik
authors.affiliation: Institute of Biochemistry, Universität Leipzig, Brüderstraße 34, D-04103 Leipzig, Germany.
authors.forename: Annette G
authors.initials: AG
authors.lastname: Beck-Sickinger
authors.affiliation: Center for Biotechnology and Biomedicine (BBZ), Universität Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany. Electronic address: andrea.robitzki@bbz.uni-leipzig.de.
authors.forename: Andrea A
authors.initials: AA
authors.lastname: Robitzki
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D015374:Biosensing Techniques; D020746:Calcium Channels, L-Type / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000737:chemistry / Q000378:metabolism; D058266:Dielectric Spectroscopy; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D017476:Receptors, Neuropeptide Y / Q000819:agonists / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D015398:Signal Transduction / Q000235:genetics*
chemical_list: D020746:Calcium Channels, L-Type; D017476:Receptors, Neuropeptide Y; D005576:Colforsin; D000242:Cyclic AMP
keywords: Impedance spectroscopy; Interdigital electrode array; Label-free activation monitoring; Neuropeptide Y receptor; Signal pathway identification