Stable transgenesis in Astyanax mexicanus using the Tol2 transposase system

Dev Dyn. 2019 Aug;248(8):679-687. doi: 10.1002/dvdy.32. Epub 2019 Apr 15.

Abstract

Background: Astyanax mexicanus is a well-established fish model system for evolutionary and developmental biology research. These fish exist as surface forms that inhabit rivers and 30 different populations of cavefish. Despite important progress in the deployment of new technologies, deep mechanistic insights into the genetic basis of evolution, development, and behavior have been limited by a lack of transgenic lines commonly used in genetic model systems.

Results: Here, we expand the toolkit of transgenesis by characterizing two novel stable transgenic lines that were generated using the highly efficient Tol2 system, commonly used to generate transgenic zebrafish. A stable transgenic line consisting of the zebrafish ubiquitin promoter expresses enhanced green fluorescent protein ubiquitously throughout development in a surface population of Astyanax. To define specific cell-types, a Cntnap2-mCherry construct labels lateral line mechanosensory neurons in zebrafish. Strikingly, both constructs appear to label the predicted cell types, suggesting many genetic tools and defined promoter regions in zebrafish are directly transferrable to cavefish.

Conclusion: The lines provide proof-of-principle for the application of Tol2 transgenic technology in A. mexicanus. Expansion on these initial transgenic lines will provide a platform to address broadly important problems in the quest to bridge the genotype-phenotype gap.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified / genetics
  • Fishes
  • Gene Transfer Techniques*
  • Green Fluorescent Proteins / genetics
  • Lateral Line System
  • Methods
  • Models, Animal
  • Promoter Regions, Genetic
  • Proof of Concept Study
  • Transposases*
  • Ubiquitin / genetics
  • Zebrafish / genetics

Substances

  • Ubiquitin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Transposases