Hatamie Amir, He Xiulan, Ewing Andrew, Rorsman Patrik
Department of Physiology, Sahlgrenska Academy, University of Gothenburg, Medicinaregatan 11-13, 41390 Gothenburg, Sweden.
Department of Chemistry and Molecular Biology, University of Gothenburg, Kemivägen 10, 412 96, Gothenburg, Sweden.
ACS Meas Sci Au. 2024 Oct 10;4(6):629-637. doi: 10.1021/acsmeasuresciau.4c00058. eCollection 2024 Dec 18.
Single cell Amperometry (SCA) is a powerful, sensitive, high temporal resolution electrochemical technique used to quantify secreted molecular messengers from individual cells and vesicles. This technique has been extensively applied to study the process of exocytosis, and it has also been applied, albeit less frequently, to investigate insulin exocytosis from single pancreatic beta cells. Insufficient insulin release can lead to diabetes, a chronic lifestyle disorder that affects millions of people worldwide. This review aims to summarize and highlight electrochemical measurements of insulin via monitoring its secretion from beta cells by SCA with micro- and nanoelectrodes since the 1990s and to explain how and why serotonin is used as a proxy for monitoring insulin during exocytosis from single beta cells. Finally, we describe how the combination of SCA measurements with the intracellular vesicle impact electrochemical cytometry (IVIEC) technique has led to important findings regarding fractional release types in beta cells. These findings, reported recently, have opened a new window in the study of pore formation, exocytosis from single vesicles, and the mechanisms of insulin secretion. This sensitive cellular electroanalysis approach should help in the development of novel therapeutic strategies targeting diabetes in the future.
单细胞安培法(SCA)是一种强大、灵敏且具有高时间分辨率的电化学技术,用于定量单个细胞和囊泡分泌的分子信使。该技术已被广泛应用于研究胞吐作用过程,也较少被用于研究单个胰腺β细胞的胰岛素胞吐作用。胰岛素释放不足会导致糖尿病,这是一种影响全球数百万人的慢性生活方式疾病。本综述旨在总结和强调自20世纪90年代以来,通过使用微电极和纳米电极的SCA监测β细胞分泌胰岛素的电化学测量方法,并解释在单个β细胞胞吐过程中,为何以及如何使用5-羟色胺作为监测胰岛素的替代物。最后,我们描述了SCA测量与细胞内囊泡冲击电化学细胞术(IVIEC)技术的结合如何带来了关于β细胞中分数释放类型的重要发现。这些最近报道的发现为孔形成、单个囊泡的胞吐作用以及胰岛素分泌机制的研究打开了一扇新窗口。这种灵敏的细胞电分析方法应有助于未来开发针对糖尿病的新型治疗策略。