title: Electroporation-based methods for in vivo, whole mount and primary culture analysis of zebrafish brain development.
abstract: BACKGROUND
  Electroporation is a technique for the introduction of nucleic acids and other macromolecules into cells. In chick embryos it has been a particularly powerful technique for the spatial and temporal control of gene expression in developmental studies. Electroporation methods have also been reported for Xenopus, zebrafish, and mouse.
  
  
  RESULTS
  We present a new protocol for zebrafish brain electroporation. Using a simple set-up with fixed spaced electrodes and microinjection equipment, it is possible to electroporate 50 to 100 embryos in 1 hour with no lethality and consistently high levels of transgene expression in numerous cells. Transfected cells in the zebrafish brain are amenable to in vivo time lapse imaging. Explants containing transfected neurons can be cultured for in vitro analysis. We also present a simple enzymatic method to isolate whole brains from fixed zebrafish for immunocytochemistry.
  
  
  CONCLUSIONS
  Building on previously described methods, we have optimized several parameters to allow for highly efficient unilateral or bilateral transgenesis of a large number of cells in the zebrafish brain. This method is simple and provides consistently high levels of transgenesis for large numbers of embryos.
authors.affiliation: Developmental Neurobiology Group, Temasek Life Sciences Laboratory, Research Link, National University of Singapore, 117604, Singapore. michaelh@tll.org.sg
authors.forename: Michael
authors.initials: M
authors.lastname: Hendricks
authors.forename: Suresh
authors.initials: S
authors.lastname: Jesuthasan
mesh_terms: D000818:Animals; D001921:Brain / Q000196:embryology* / Q000254:growth & development / Q000601:surgery*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000196:embryology / Q000601:surgery; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D018715:Microscopy, Video / Q000379:methods; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods; D009696:Nucleic Acids / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection / Q000295:instrumentation / Q000379:methods; D015027:Zebrafish / Q000196:embryology* / Q000254:growth & development / Q000601:surgery*
chemical_list: D009696:Nucleic Acids