RRC ID 73960
Author Suzuki T, Kondo C, Kanamori T, Inouye S.
Title Video rate bioluminescence imaging of secretory proteins in living cells: localization, secretory frequency, and quantification.
Journal Anal Biochem
Abstract We have developed a method of video rate bioluminescence imaging to investigate protein secretion from a single mammalian cell and analyzed the localization, secretory frequency, and quantification of secreted protein. By detecting the luminescence signals from the Gaussia luciferase (GLase) reaction using a high-speed electron-multiplying charge-coupled device (EM-CCD) camera, video rate imaging was performed with a time resolution within 500 ms/image over 30 min in living cells. As a model study, we applied the method to visualize the glucose-stimulated insulin secretion from clustered pancreatic MIN6 β cells using the fused protein of GLase with preproinsulin. High-quality video images clearly showed that the glucose-stimulated insulin secretion from the clustered MIN6 β cells oscillated within a period of a few minutes over 10 min. In addition, the glibenclamide-induced insulin secretion from the clustered MIN6 β cells was visualized, suggesting that bioluminescence video rate imaging is a useful method for validating drug action in living cells.
Volume 415(2)
Pages 182-9
Published 2011-8-15
DOI 10.1016/j.ab.2011.03.039
PII S0003-2697(11)00226-0
PMID 21477579
MeSH Animals Copepoda / enzymology Glucose / pharmacology Glyburide / pharmacology Imidazoles / chemistry Imidazoles / pharmacology Insulin / analysis* Insulin / genetics Insulin / metabolism Insulin Secretion Insulin-Secreting Cells / metabolism* Luciferases / genetics Luciferases / metabolism Luminescent Measurements / methods* Mice Protein Precursors / genetics Protein Precursors / metabolism Pyrazines / chemistry Pyrazines / pharmacology Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Video Recording / methods*
IF 2.877
DNA material pJNC-hINS-GLuc (RDB19845)