Focal electroporation in zebrafish embryos and larvae

Methods Mol Biol. 2009:546:145-51. doi: 10.1007/978-1-60327-977-2_10.

Abstract

A method is described that allows the introduction by electroporation of either small dyes or larger RNA, DNA, or morpholino constructs into single cells or small groups of cells in zebrafish embryos or larvae. The dye or construct is delivered to cells via a patch-like microelectrode that also delivers the electroporation stimulus train. This technique allows the experimenter to target cells of their choice at a particular time of development and at a particular location in the embryo, and is useful for fate mapping, analysing neuronal organisation, ectopic expression and gene knockdown experiments.

MeSH terms

  • Animals
  • Animals, Genetically Modified / embryology
  • Animals, Genetically Modified / genetics
  • Animals, Genetically Modified / metabolism
  • DNA / genetics
  • DNA / metabolism
  • Developmental Biology / methods*
  • Electroporation / methods*
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / drug effects
  • Fluorescent Dyes / pharmacology
  • Larva / cytology
  • Larva / drug effects
  • Microscopy
  • RNA / genetics
  • RNA / metabolism
  • Rhodamines / pharmacology
  • Time Factors
  • Zebrafish / embryology*
  • Zebrafish / genetics*
  • Zebrafish / metabolism

Substances

  • Fluorescent Dyes
  • Rhodamines
  • tetramethylrhodamine
  • RNA
  • DNA