Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets
Summary: First-phase glucose-stimulated insulin secretion is mechanistically linked to type 2 diabetes, yet the underlying metabolism is difficult to discern due to significant islet-to-islet variability. Here, we miniaturize a fluorescence anisotropy immunoassay onto a microfluidic device to measur...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-10-01
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| Series: | Cell Reports: Methods |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2667237523002540 |
| _version_ | 1797649661036593152 |
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| author | Yufeng Wang Romario Regeenes Mahnoor Memon Jonathan V. Rocheleau |
| author_facet | Yufeng Wang Romario Regeenes Mahnoor Memon Jonathan V. Rocheleau |
| author_sort | Yufeng Wang |
| collection | DOAJ |
| description | Summary: First-phase glucose-stimulated insulin secretion is mechanistically linked to type 2 diabetes, yet the underlying metabolism is difficult to discern due to significant islet-to-islet variability. Here, we miniaturize a fluorescence anisotropy immunoassay onto a microfluidic device to measure C-peptide secretion from individual islets as a surrogate for insulin (InsC-chip). This method measures secretion from up to four islets at a time with ∼7 s resolution while providing an optical window for real-time live-cell imaging. Using the InsC-chip, we reveal two glucose-dependent peaks of insulin secretion (i.e., a double peak) within the classically defined 1st phase (<10 min). By combining real-time secretion and live-cell imaging, we show islets transition from glycolytic to oxidative phosphorylation (OxPhos)-driven metabolism at the nadir of the peaks. Overall, these data validate the InsC-chip to measure glucose-stimulated insulin secretion while revealing new dynamics in secretion defined by a shift in glucose metabolism. Motivation: Insulin secretion dynamics reflect islet β cell function and dysfunction in disease. Measurement of individual islet responses relative to metabolism is critical since functional heterogeneity has been reported in both rodents and humans. Therefore, we designed the InsC-chip to measure insulin secretion from multiple individual islets with high temporal resolution while leaving the tissue optically accessible for live-cell metabolic imaging. |
| first_indexed | 2024-03-11T15:50:01Z |
| format | Article |
| id | doaj.art-868d14e13e09472ba379a8c33823be9b |
| institution | Directory Open Access Journal |
| issn | 2667-2375 |
| language | English |
| last_indexed | 2024-03-11T15:50:01Z |
| publishDate | 2023-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports: Methods |
| spelling | doaj.art-868d14e13e09472ba379a8c33823be9b2023-10-26T04:18:33ZengElsevierCell Reports: Methods2667-23752023-10-01310100602Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual isletsYufeng Wang0Romario Regeenes1Mahnoor Memon2Jonathan V. Rocheleau3Advanced Diagnostics, Toronto General Hospital Research Institute, Toronto, ON M5G 1L7, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, CanadaAdvanced Diagnostics, Toronto General Hospital Research Institute, Toronto, ON M5G 1L7, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, CanadaAdvanced Diagnostics, Toronto General Hospital Research Institute, Toronto, ON M5G 1L7, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, CanadaAdvanced Diagnostics, Toronto General Hospital Research Institute, Toronto, ON M5G 1L7, Canada; Institute of Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, Canada; Departments of Medicine and Physiology, University of Toronto, Toronto, ON M5S 1A8, Canada; Corresponding authorSummary: First-phase glucose-stimulated insulin secretion is mechanistically linked to type 2 diabetes, yet the underlying metabolism is difficult to discern due to significant islet-to-islet variability. Here, we miniaturize a fluorescence anisotropy immunoassay onto a microfluidic device to measure C-peptide secretion from individual islets as a surrogate for insulin (InsC-chip). This method measures secretion from up to four islets at a time with ∼7 s resolution while providing an optical window for real-time live-cell imaging. Using the InsC-chip, we reveal two glucose-dependent peaks of insulin secretion (i.e., a double peak) within the classically defined 1st phase (<10 min). By combining real-time secretion and live-cell imaging, we show islets transition from glycolytic to oxidative phosphorylation (OxPhos)-driven metabolism at the nadir of the peaks. Overall, these data validate the InsC-chip to measure glucose-stimulated insulin secretion while revealing new dynamics in secretion defined by a shift in glucose metabolism. Motivation: Insulin secretion dynamics reflect islet β cell function and dysfunction in disease. Measurement of individual islet responses relative to metabolism is critical since functional heterogeneity has been reported in both rodents and humans. Therefore, we designed the InsC-chip to measure insulin secretion from multiple individual islets with high temporal resolution while leaving the tissue optically accessible for live-cell metabolic imaging.http://www.sciencedirect.com/science/article/pii/S2667237523002540CP: BiotechnologyCP: Metabolism |
| spellingShingle | Yufeng Wang Romario Regeenes Mahnoor Memon Jonathan V. Rocheleau Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets Cell Reports: Methods CP: Biotechnology CP: Metabolism |
| title | Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets |
| title_full | Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets |
| title_fullStr | Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets |
| title_full_unstemmed | Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets |
| title_short | Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets |
| title_sort | insulin c peptide secretion on a chip to measure the dynamics of secretion and metabolism from individual islets |
| topic | CP: Biotechnology CP: Metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2667237523002540 |
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