Ca2+ imaging of synaptic compartments using subcellularly targeted GCaMP8f in Drosophila

STAR Protoc. 2024 Mar 15;5(1):102832. doi: 10.1016/j.xpro.2023.102832. Epub 2024 Jan 9.

Abstract

GCaMP8f is a sensitive genetically encoded Ca2+ indicator that enables imaging of neuronal activity. Here, we present a protocol to perform Ca2+ imaging of the Drosophila neuromuscular junction using GCaMP8f targeted to pre- or postsynaptic compartments. We describe ratiometric Ca2+ imaging using GCaMP8f fused to mScarlet and synaptotagmin that reveals Ca2+ dynamics at presynaptic terminals. We then detail "quantal" imaging of miniature transmission events using GCaMP8f targeted to postsynaptic compartments by fusion to a PDZ-binding motif. For complete details on the use and execution of this protocol, please refer to Li et al.,1 Han et al.,2 Perry et al.,3 and Han et al.4.

Keywords: Cell Biology; Genetics; Microscopy; Model Organisms; Neuroscience.

MeSH terms

  • Animals
  • Drosophila Proteins* / genetics
  • Drosophila* / physiology
  • Neuromuscular Junction / physiology
  • Neurons
  • Presynaptic Terminals / physiology

Substances

  • Drosophila Proteins