Video rate bioluminescence imaging of secretory proteins in living cells: localization, secretory frequency, and quantification |
| |
Authors: | Suzuki Takahiro Kondo Chihiro Kanamori Takao Inouye Satoshi |
| |
Affiliation: | aDepartment of Biochemistry, School of Dentistry, Aichi-Gakuin University, Nagoya 464-8650, Japan;bYokohama Research Center, JNC Corporation, Yokohama 236-8605, Japan |
| |
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. |
| |
Keywords: | Exocytosis Gaussia luciferase EM-CCD camera Insulin oscillations Diabetes |
本文献已被 ScienceDirect PubMed 等数据库收录! |
|