Quantification of Secretory Granule Exocytosis by TIRF Imaging and Capacitance Measurements

Abstract

Hormones and neurotransmitters are released from (neuro)endocrine cells by regulated exocytosis of secretory granules. During exocytosis, the granule membrane fuses with the plasma membrane, which allows release of the stored content into the bloodstream or the surrounding tissue. Here, we give a detailed description of two complementary methods to observe and quantify exocytosis in single cells: high-resolution TIRF microscopy and patch-clamp capacitance recordings. Precise stimulation of exocytosis is achieved by local pressure application or voltage-clamp depolarizations. While the chapter is focused on insulin-secreting cells as an accessible and disease-relevant model system, the methodology is applicable to a wide variety of secretory cells including chromaffin and PC12 cells.

Keywords

Chromaffin cells; Insulin release; Patch-clamp; PC12; Secretory vesicles

Published in

Methods in Molecular Biology
2023, volume: 2565, number: 2565, pages: 179-186
Title: Methods in Molecular Biology : Methods and Protocols
Publisher: Springer

SLU Authors

• Omar Hmeadi, Muhmmad

  • Uppsala University

UKÄ Subject classification

Cell and Molecular Biology

Publication identifier

• DOI: https://doi.org/10.1007/978-1-0716-2671-9_12
• ISBN: 978-1-0716-2670-2
• eISBN: 978-1-0716-2671-9

Permanent link to this page (URI)

https://res.slu.se/id/publ/131978