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A Ratiometric Sensor for Imaging Insulin Secretion in Single β Cells
Martina Schifferer1, Dmytro A Yushchenko2, Frank Stein1
1Interdisciplinary Chemistry Group, Cell Biology & Biophysics Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Cell Chemical Biology
|April 4, 2017
Summary
Researchers developed RINS1, a novel reporter for visualizing insulin secretion from single cells. This tool enables precise monitoring of glucose-stimulated insulin secretion and drug responses in beta cells.
Area of Science:
- Biochemistry
- Cell Biology
- Endocrinology
Background:
- Accurate measurement of insulin secretion from single cells is crucial for understanding diabetes.
- Existing methods for monitoring insulin release lack reliability and single-cell resolution.
Purpose of the Study:
- To develop a genetically encoded reporter system for real-time visualization and quantification of insulin secretion at the single-cell level.
- To assess the utility of the reporter for studying glucose-stimulated insulin secretion and drug responses.
Main Methods:
- Development of RINS1, a reporter fusing proinsulin with superfolder GFP (sfGFP) and mCherry.
- Expression and validation of RINS1 in MIN6 and primary mouse beta cells.
- Utilizing total internal reflection fluorescence microscopy to monitor insulin release.
Main Results:
- RINS1 demonstrated proper processing and yielded single-labeled insulin species in beta cells.
- Preferential release of the insulin-sfGFP construct over the mCherry-C-peptide was observed upon stimulation.
- RINS1 enabled ratiometric monitoring of glucose-stimulated insulin secretion and pulsatile insulin release in response to tolbutamide.
Conclusions:
- RINS1 is a reliable tool for single-cell analysis of insulin secretion dynamics.
- The reporter facilitates investigations into the mechanisms of insulin release and the efficacy of antidiabetic drugs.