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

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

Research output: Contribution to Journal › Article › Academic › peer-review

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).

Original language	English
Article number	100325
Pages (from-to)	1-18
Number of pages	18
Journal	STAR Protocols
Volume	2
Issue number	1
Early online date	3 Feb 2021
DOIs	https://doi.org/10.1016/j.xpro.2021.100325
Publication status	Published - 19 Mar 2021

Bibliographical note

Funding Information:
The authors thank Robbert Zalm for cloning and producing lentiviral particles, Desiree Schut and Lisa Laan for astrocyte and neuronal cultures, Joke Wortel for animal breeding, Joost Hoetjes for genotyping, and members of the CNCR DCV project team for fruitful discussions. This work is supported by an ERC Advanced Grant ( 322966 ) of the European Union (to M.V.)

Funding Information:
The authors thank Robbert Zalm for cloning and producing lentiviral particles, Desiree Schut and Lisa Laan for astrocyte and neuronal cultures, Joke Wortel for animal breeding, Joost Hoetjes for genotyping, and members of the CNCR DCV project team for fruitful discussions. This work is supported by an ERC Advanced Grant (322966) of the European Union (to M.V.), R.I.H. and A.M. acquired data. R.I.H. A.M. C.M.P. M.V. and R.F.T. designed figures and wrote the protocol. The authors declare no competing interests.

Publisher Copyright:
© 2021 The Author(s)

Funding

The authors thank Robbert Zalm for cloning and producing lentiviral particles, Desiree Schut and Lisa Laan for astrocyte and neuronal cultures, Joke Wortel for animal breeding, Joost Hoetjes for genotyping, and members of the CNCR DCV project team for fruitful discussions. This work is supported by an ERC Advanced Grant ( 322966 ) of the European Union (to M.V.) The authors thank Robbert Zalm for cloning and producing lentiviral particles, Desiree Schut and Lisa Laan for astrocyte and neuronal cultures, Joke Wortel for animal breeding, Joost Hoetjes for genotyping, and members of the CNCR DCV project team for fruitful discussions. This work is supported by an ERC Advanced Grant (322966) of the European Union (to M.V.), R.I.H. and A.M. acquired data. R.I.H. A.M. C.M.P. M.V. and R.F.T. designed figures and wrote the protocol. The authors declare no competing interests.

Funders	Funder number
CNCR
European Commission
European Research Council	322966

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Cell culture
Cell membrane
Microscopy
Molecular/chemical probes
Neuroscience

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10.1016/j.xpro.2021.100325

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Cite this

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title = "Quantitative analysis of dense-core vesicle fusion in rodent CNS neurons",

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).",

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

Abstract

Bibliographical note

Funding

UN SDGs

Keywords

Access to Document

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