title: Video rate bioluminescence imaging of secretory proteins in living cells: localization, secretory frequency, and quantification.
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.
authors.affiliation: Department of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya 464-8650, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Suzuki
authors.forename: Chihiro
authors.initials: C
authors.lastname: Kondo
authors.forename: Takao
authors.initials: T
authors.lastname: Kanamori
authors.forename: Satoshi
authors.initials: S
authors.lastname: Inouye
mesh_terms: D000818:Animals; D033342:Copepoda / Q000201:enzymology; D005947:Glucose / Q000494:pharmacology; D005905:Glyburide / Q000494:pharmacology; D007093:Imidazoles / Q000737:chemistry / Q000494:pharmacology; D007328:Insulin / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D011719:Pyrazines / Q000737:chemistry / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014741:Video Recording / Q000379:methods*
chemical_list: D007093:Imidazoles; D007328:Insulin; D011498:Protein Precursors; D011719:Pyrazines; D011993:Recombinant Fusion Proteins; C017144:coelenterazine; C013737:preproinsulin; D008156:Luciferases; D005947:Glucose; D005905:Glyburide