Generating Transgenic Reporter Lines for Studying Nervous System Development in the Cnidarian Nematostella vectensis

Methods Mol Biol. 2020:2047:45-57. doi: 10.1007/978-1-4939-9732-9_3.

Abstract

Neurons often display complex morphologies with long and fine processes that can be difficult to visualize, in particular in living animals. Transgenic reporter lines in which fluorescent proteins are expressed in defined populations of neurons are important tools that can overcome these difficulties. By using membrane-attached fluorescent proteins, such reporter transgenes can identify the complete outline of subsets of neurons or they can highlight the subcellular localization of fusion proteins, for example at pre- or postsynaptic sites. The relative stability of fluorescent proteins furthermore allows the tracing of the progeny of cells over time and can therefore provide information about potential roles of the gene whose regulatory elements are controlling the expression of the fluorescent protein. Here we describe the generation of transgenic reporter lines in the sea anemone Nematostella vectensis, a cnidarian model organism for studying the evolution of developmental processes. We also provide an overview of existing transgenic Nematostella lines that have been used to study conserved and derived aspects of nervous system development.

Keywords: Cnidaria; Fluorescent protein; Microinjection; Nematostella; Neurogenesis; Transgenic reporter.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified / growth & development
  • Genes, Reporter
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Nervous System / growth & development
  • Neurogenesis
  • Sea Anemones / genetics*
  • Sea Anemones / growth & development

Substances

  • Luminescent Proteins