title: Real-time monitoring of intracellular signal transduction in PC12 cells by two-dimensional surface plasmon resonance imager.
abstract: Real-time observation of intracellular process of signal transduction is very useful for biomedical and pharmaceutical applications as well as for basic research work of cell biology. The conventional methods used to observe intracellular reactions have not been convenient with several steps such as labeling and washing steps prior to the readout. Consequently, there is a critical need for label-free observation techniques for monitoring intracellular reactions. For feasible and reagentless observation of intracellular alterations in real time, we examined the use of a high-resolution two-dimensional surface plasmon resonance (2D-SPR) imager for monitoring of intracellular signal transduction that was mainly translocation of protein kinase C via local refractive index change in PC12 cells adhered on a gold sensor slide without any indicator reagent. PC12 cells were stimulated with KCl and phorbol-12-myristate-13-acetate (PMA, a protein kinase C [PKC] activator) at different concentrations in order to induce intracellular PKC translocation. 2D-SPR signal (reflection intensity change) is very consistent with the cellular response normally detected for these stimulants. Our results suggest that complex intracellular reactions could be real-time monitored and characterized by the 2D-SPR imager. It is further expected that signal transmission that was followed by the translocation of signaling proteins could be observed at the single cell level with the high-resolution 2D-SPR imager.
authors.affiliation: Field of Life, Information and System Sciences, Graduate School of Science and Engineering for Research, University of Toyama, Toyama 930-8555, Japan; Major of Biological Information Systems Science, Graduate School of Innovative Life Science for Education, University of Toyama, Toyama 930-8555, Japan. Electronic address: hshinoha@eng.u-toyama.ac.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shinohara
authors.forename: Yhutarou
authors.initials: Y
authors.lastname: Sakai
authors.forename: Tanveer Ahamd
authors.initials: TA
authors.lastname: Mir
mesh_terms: D000818:Animals; D016716:PC12 Cells / Q000378:metabolism*; D010703:Phorbol Esters / Q000378:metabolism; D011189:Potassium Chloride / Q000378:metabolism; D011493:Protein Kinase C / Q000032:analysis / Q000378:metabolism; D021381:Protein Transport; D051381:Rats; D015398:Signal Transduction; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D010703:Phorbol Esters; C119222:phorbol-12-myristate; D011189:Potassium Chloride; D011493:Protein Kinase C
keywords: 2D–SPR imaging; PC12 cell; Signal transduction