Targeted electroporation in Xenopus tadpoles in vivo--from single cells to the entire brain.
Electroporation is becoming more popular as a technique for transfecting neurons within intact tissues. One of the advantages of electroporation over other transfection techniques is the ability to precisely target an area for transfection. Here we highlight this advantage by describing methods to restrict transfection to either a single cell, clusters of cells, or to include large portions of the brain of the intact Xenopus tadpole. Electroporation is also an effective means of gene delivery in the retina. We have developed these techniques to examine the effects of regulated gene expression on various neuronal properties, including structural plasticity and synaptic transmission. Restriction of transfection to individual cells aids in imaging of neuronal morphology, while bulk cell transfection allows examination of the affects of gene expression on populations of cells by biochemical assays, imaging, and electrophysiological recording.
Haas
Kurt
K
haas@cshl.org
Jensen
Kendall
K
Sin
Wun Chey
WC
Foa
Lisa
L
Cline
Hollis T
HT
D000818:Animals; D001921:Brain / Q000502:physiology*; D003911:Dextrans / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives*; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D007814:Larva; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D009474:Neurons / Q000502:physiology; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis / Q000235:genetics
D003911:Dextrans; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D009841:Oligonucleotides; D011994:Recombinant Proteins; C015219:fluorescein isothiocyanate dextran; D049452:Green Fluorescent Proteins; C024098:6-carboxyfluorescein; D016650:Fluorescein-5-isothiocyanate