Protocol

Mounting of Embryos, Larvae, and Pupae for Live Drosophila Dendritic Arborization Neuron Imaging

  1. Adrian W. Moore1,3
  1. 1Laboratory for Neurodiversity, RIKEN Center for Brain Science, Wako-shi, 350-0106, Japan
  2. 2Department of Cellular and Molecular Biology, Institute for Translational Medicine, University of Liverpool, Liverpool L69 3BX, United Kingdom
  1. 3Correspondence: adrian.moore{at}riken.jp

Abstract

Live imaging approaches are essential for monitoring how neurons go through a coordinated series of differentiation steps in their native mechanical and chemical environment. These imaging approaches also allow the study of dynamic subcellular processes such as cytoskeleton remodeling and the movement of organelles. Drosophila dendritic arborization (da) neurons are a powerful experimental system for studying the dendrite arbor in live animals. da neurons are located on the internal surface of the body wall and, therefore, are easily accessible for imaging. Moreover, many genetic tools target da neurons to disrupt genes or proteins of interest and allow the investigator to visualize fluorescent markers and endogenously tagged proteins in the neurons. This protocol introduces methods for preparing and mounting intact Drosophila embryos, larvae, and pupae, allowing live imaging of dynamic cellular processes in da neurons.

Footnotes

  • From the Drosophila Neurobiology collection, edited by Bing Zhang, Ellie Heckscher, Alex C. Keene, and Scott Waddell.