Abstract
Electroporation is an increasingly common technique used for exogenous gene expression in live animals, but protocols are largely limited to traditional laboratory organisms. The goal of this protocol is to test in vivo electroporation techniques in a diverse array of tadpole species. We explore electroporation efficiency in tissue-specific cells of five species from across three families of tropical frogs: poison frogs (Dendrobatidae), cryptic forest/poison frogs (Aromobatidae), and glassfrogs (Centrolenidae). These species are well known for their diverse social behaviors and intriguing physiologies that coordinate chemical defenses, aposematism, and/or tissue transparency. Specifically, we examine the effects of electrical pulse and injection parameters on species- and tissue-specific transfection of plasmid DNA in tadpoles. After electroporation of a plasmid encoding green fluorescent protein (GFP), we found strong GFP fluorescence within brain and muscle cells that increased with the amount of DNA injected and electrical pulse number. We discuss species-related challenges, troubleshooting, and outline ideas for improvement. Extending in vivo electroporation to non-model amphibian species could provide new opportunities for exploring topics in genetics, behavior, and organismal biology.
Funder
Division of Integrative Organismal Systems
New York Stem Cell Foundation
Gerstner Family Foundation, American Museum of Natural History
Division of Graduate Education
Publisher
Public Library of Science (PLoS)
Reference59 articles.
1. In vivo electroporation: a powerful and convenient means of nonviral gene transfer to tissues of living animals;T Muramatsu;Int J Mol Med,1998
2. Fundamentals of electroporative delivery of drugs and genes;E Neumann;Bioelectrochem Bioenerg,1999
3. Targeted electroporation in Xenopus tadpoles in vivo—from single cells to the entire brain;K Haas;Differentiation,2002
4. Electroporation-mediated gene delivery;JL Young;Adv Genet,2015