Whole-Brain Calcium Imaging in Larval Xenopus

Cold Spring Harb Protoc. 2020 Dec 1;2020(12). doi: 10.1101/pdb.prot106815.

Abstract

Sensory systems detect environmental stimuli and transform them into electrical activity patterns interpretable by the central nervous system. En route to higher brain centers, the initial sensory input is successively transformed by interposed secondary processing centers. Mapping the neuronal activity patterns at all of those stages is essential to understand sensory information processing. Larval Xenopus laevis is very well-suited for whole-brain imaging of neuronal activity. This is mainly due to its small size, transparency, and the accessibility of both peripheral and central parts of sensory systems. Here we describe a protocol for calcium imaging at several levels of the olfactory system using focal injection of chemical calcium indicator dyes or a Xenopus transgenic line with neuronal GCaMP6s expression. In combination with fast volumetric multiphoton microscopy, the calcium imaging methods described can provide detailed insight into spatiotemporal activity of entire brain regions at different stages of sensory information processing. Although the methods are broadly applicable to the central nervous system, in this work we focus on protocols for calcium imaging of glomeruli in the olfactory bulb and odor-responsive neurons in the olfactory amygdala.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Brain / metabolism*
  • Calcium / metabolism*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Larva / genetics
  • Larva / metabolism
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Odorants
  • Olfactory Bulb / cytology
  • Olfactory Bulb / metabolism
  • Olfactory Cortex / cytology
  • Olfactory Cortex / metabolism
  • Olfactory Receptor Neurons / metabolism
  • Olfactory Receptor Neurons / physiology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Smell / physiology
  • Xenopus laevis / genetics
  • Xenopus laevis / metabolism*
  • Xenopus laevis / physiology

Substances

  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Calcium