Quantitative analysis of dense-core vesicle fusion in rodent CNS neurons

Alessandro Moro; Rein I Hoogstraaten; Claudia M Persoon; Matthijs Verhage; Ruud F Toonen

doi:10.1016/j.xpro.2021.100325

Quantitative analysis of dense-core vesicle fusion in rodent CNS neurons

STAR Protoc. 2021 Feb 3;2(1):100325. doi: 10.1016/j.xpro.2021.100325. eCollection 2021 Mar 19.

Authors

Alessandro Moro¹, Rein I Hoogstraaten^{2

1}, Claudia M Persoon^{2

1}, Matthijs Verhage^{2

1}, Ruud F Toonen²

Affiliations

¹ Department of Clinical Genetics, UMC Amsterdam, the Netherlands.
² Department of Functional Genomics, Center for Neurogenomics and Cognitive Research (CNCR), Vrije Universiteit (VU) Amsterdam, de Boelelaan 1085, 1081 HV Amsterdam, the Netherlands.

Abstract

Neuropeptides are essential signaling molecules secreted by dense-core vesicles (DCVs). They contribute to information processing in the brain, controlling a variety of physiological conditions. Defective neuropeptide signaling is implicated in several psychiatric disorders. Here, we provide a protocol for the quantitative analysis of DCV fusion events in rodent neurons using pH-sensitive DCV fusion probes and custom-written analysis algorithms. This method can be used to study DCV fusion mechanisms and is easily adapted to investigate fusion principles of other secretory organelles. For complete details on the use and execution of this protocol, please refer to Persoon et al. (2019).

Keywords: Cell culture; Cell membrane; Microscopy; Molecular/chemical probes; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Algorithms*
Animals
Dense Core Vesicles / genetics
Dense Core Vesicles / metabolism*
Genes, Reporter
Hippocampus / metabolism*
Mice
Neurons / metabolism*
Synapses / genetics
Synapses / metabolism*