論文 - 詳細
| RRC ID | 57785 |
|---|---|
| 著者 | Shigeto H, Ono T, Ikeda T, Hirota R, Ishida T, Kuroda A, Funabashi H. |
| タイトル | Insulin sensor cells for the analysis of insulin secretion responses in single living pancreatic β cells. |
| ジャーナル | Analyst |
| Abstract |
Investigation of the functions of insulin-secreting cells in response to glucose in single-living cells is essential for improving our knowledge on the pathogenesis of diabetes. Therefore, it is desired to develop a new convenient method that enables the direct detection of insulin secreted from single-living cells. Here, insulin-sensor-cells expressing a protein-based insulin-detecting probe immobilized on the extracellular membrane were developed to evaluate the insulin-secretion response in single-living pancreatic β cells. The protein-based insulin-detecting probe (NαLY) was composed of a bioluminescent protein (nano-luc), the αCT segment of the insulin receptor, L1 and CR domains of the insulin receptor, and a fluorescent protein (YPet). NαLY exhibited a bioluminescence resonance energy transfer (BRET) signal in response to insulin; thus, cells of Hepa1-6 line were genetically engineered to express NαLY on the extracellular membrane. The cells were found to act as insulin-sensor-cells, exhibiting a BRET signal in response to insulin. When the insulin-sensor-cells and pancreatic β cells (MIN6 cell line) were cocultured and stimulated with glucose, insulin-sensor-cells nearby pancreatic β cells showed the spike-shaped BRET signal response, whereas the insulin-sensor-cells close to one pancreatic β cell did not exhibit such signal response. However, all the insulin-sensor-cells showed a gradual increase in BRET signals, which were presumably attributed to the increase in insulin concentrations in the culture dish, confirming the function of these insulin-sensor-cells. Therefore, we demonstrated that heterogenetic insulin secretion in single-living pancreatic β cells could be measured directly using the insulin sensor cells. |
| 巻・号 | 144(12) |
| ページ | 3765-3772 |
| 公開日 | 2019-6-21 |
| DOI | 10.1039/c9an00405j |
| PMID | 31089611 |
| MeSH | Animals Bioluminescence Resonance Energy Transfer Techniques / methods* Biosensing Techniques / methods* Cell Line, Tumor Coculture Techniques / methods Fluorescence Glucose / metabolism Insulin / analysis* Insulin-Secreting Cells / metabolism* Luminescent Proteins / genetics Luminescent Proteins / metabolism Mice Protein Engineering / methods Receptor, Insulin / genetics Receptor, Insulin / metabolism Single-Cell Analysis / methods* |
| IF | 3.978 |
| 引用数 | 2 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 3 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | Hepa 1-6(RCB1638) |