Imaging synaptic vesicle exocytosis-endocytosis with pH-sensitive fluorescent proteins

Olusoji A T Afuwape, Ege T. Kavalali

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

The introduction of pHluorin, a pH-sensitive GFP, by Miesenbock and colleagues provided a versatile tool to studies of vesicle trafficking, in particular synaptic vesicle exocytosis and endocytosis. By tagging pHluorin to the luminal region of the synaptic vesicular protein synaptobrevin (also called VAMP, vesicleassociated membrane protein) or other synaptic vesicle-specific proteins such as the vesicular glutamate transporter-1, we are able to directly track synaptic vesicle endocytosis in response to stimuli in a molecularly specific manner. Here, we describe the process of imaging synaptic vesicle endocytosis in response to extracellular stimulation in dissociated neuronal cultures of hippocampal neurons obtained from rats—also applicable to mice—using pHluorin-tagged vesicular glutamate transporter-1 as a reporter.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages187-200
Number of pages14
Volume1474
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1474
ISSN (Print)10643745

Keywords

  • Endocytosis
  • Live-cell imaging
  • Microscopy
  • PHluorin
  • Synaptic vesicles

ASJC Scopus subject areas

  • Medicine(all)
  • Molecular Biology
  • Genetics

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  • Cite this

    Afuwape, O. A. T., & Kavalali, E. T. (2016). Imaging synaptic vesicle exocytosis-endocytosis with pH-sensitive fluorescent proteins. In Methods in Molecular Biology (Vol. 1474, pp. 187-200). (Methods in Molecular Biology; Vol. 1474). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6352-2_11