High-Throughput Calcium Imaging Screen of Toxins' Function in Dissociated Sensory Neurons

Methods Mol Biol. 2020:2068:275-282. doi: 10.1007/978-1-4939-9845-6_15.

Abstract

Many toxins from a variety of venomous animals and plants have evolved to target neuronal ion channels and receptors. However, a significant obstacle in the study of these toxins is the finding and characterization of their specific molecular target. Here, we describe a method for fast and efficient screening of venom and toxin activity using live-cell calcium imaging. We describe the use of Fura-2, a calcium indictor that changes its fluorescence properties in response to intracellular calcium elevations, to measure the activity of neurons from the dorsal root and trigeminal ganglia. Calcium imaging is an efficient technique for testing many of the venom's components on large numbers of neurons simultaneously. This technique offers a novel tool for low-cost and rapid characterization of functionally active toxins and their target receptors.

Keywords: Calcium imaging; Fluorescent microscopy; Fura-2AM; High-throughput screening; Sensory neurons.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Calcium / chemistry
  • Calcium / metabolism*
  • Fura-2 / chemistry
  • Ganglia, Spinal / diagnostic imaging
  • High-Throughput Screening Assays / methods*
  • Microscopy, Fluorescence
  • Sensory Receptor Cells / metabolism*
  • Trigeminal Ganglion / diagnostic imaging

Substances

  • Calcium
  • Fura-2