BIOMAT Database Logo BIOMAT Database Logo

Bulk electroporation of retinal ganglion cells in live Xenopus tadpoles. 2013

Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline

Individual neurons in the developing nervous system of Xenopus laevis can be visualized by the targeted delivery of a fluorophore. The fluorophore can be delivered as a fluorescent dye or DNA that encodes a fluorescent protein. Local iontophoresis is a method that works well for transfer of fluorescent dye to retinal ganglion cells (RGCs) in the eye, but it does not give a high yield for delivery of DNA. This is largely because the degree of pigmentation of the eyes, even in albino strains, makes it difficult to visualize RGC somata during pipette positioning. Bulk retinal electroporation is a better approach for delivery of plasmid DNA to RGC. The method described here works best in tadpoles older than stage 42.

UI MeSH Term Description Entries
D007814 Larva Wormlike or grublike stage, following the egg in the life cycle of insects, worms, and other metamorphosing animals. Maggots,Tadpoles,Larvae,Maggot,Tadpole
D008164 Luminescent Proteins Proteins which are involved in the phenomenon of light emission in living systems. Included are the "enzymatic" and "non-enzymatic" types of system with or without the presence of oxygen or co-factors. Bioluminescent Protein,Bioluminescent Proteins,Luminescent Protein,Photoprotein,Photoproteins,Protein, Bioluminescent,Protein, Luminescent,Proteins, Bioluminescent,Proteins, Luminescent
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012165 Retinal Ganglion Cells Neurons of the innermost layer of the retina, the internal plexiform layer. They are of variable sizes and shapes, and their axons project via the OPTIC NERVE to the brain. A small subset of these cells act as photoreceptors with projections to the SUPRACHIASMATIC NUCLEUS, the center for regulating CIRCADIAN RHYTHM. Cell, Retinal Ganglion,Cells, Retinal Ganglion,Ganglion Cell, Retinal,Ganglion Cells, Retinal,Retinal Ganglion Cell
D013194 Staining and Labeling The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts. Histological Labeling,Staining,Histological Labelings,Labeling and Staining,Labeling, Histological,Labelings, Histological,Stainings
D014982 Xenopus laevis The commonest and widest ranging species of the clawed "frog" (Xenopus) in Africa. This species is used extensively in research. There is now a significant population in California derived from escaped laboratory animals. Platanna,X. laevis,Platannas,X. laevi
D018014 Gene Transfer Techniques The introduction of functional (usually cloned) GENES into cells. A variety of techniques and naturally occurring processes are used for the gene transfer such as cell hybridization, LIPOSOMES or microcell-mediated gene transfer, ELECTROPORATION, chromosome-mediated gene transfer, TRANSFECTION, and GENETIC TRANSDUCTION. Gene transfer may result in genetically transformed cells and individual organisms. Gene Delivery Systems,Gene Transfer Technique,Transgenesis,Delivery System, Gene,Delivery Systems, Gene,Gene Delivery System,Technique, Gene Transfer,Techniques, Gene Transfer,Transfer Technique, Gene,Transfer Techniques, Gene
D018274 Electroporation A technique in which electric pulses, in kilovolts per centimeter and of microsecond-to-millisecond duration, cause a loss of the semipermeability of CELL MEMBRANES, thus leading to ion leakage, escape of metabolites, and increased uptake by cells of drugs, molecular probes, and DNA. Depending on the dosage, the formation of openings in the cell membranes caused by the electric pulses may or may not be reversible. Electric Field-Mediated Cell Permeabilization,Irreversible Electroporation,Reversible Electroporation,Electropermeabilisation,Electric Field Mediated Cell Permeabilization,Electroporation, Irreversible,Electroporation, Reversible

Related Publications

Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Transfection of mouse retinal ganglion cells by in vivo electroporation.
April 2011, Journal of visualized experiments : JoVE,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Targeted electroporation in Xenopus tadpoles in vivo--from single cells to the entire brain.
June 2002, Differentiation; research in biological diversity,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Generating Retinal Injury Models in Xenopus Tadpoles.
October 2023, Journal of visualized experiments : JoVE,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Development of retinal central projection in Xenopus tadpoles.
January 1986, Progress in clinical and biological research,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Development of neuronal specificity in retinal ganglion cells of Xenopus.
February 1968, Developmental biology,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Gene delivery into mouse retinal ganglion cells by in utero electroporation.
September 2007, BMC developmental biology,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Morphological classification of retinal ganglion cells in adult Xenopus laevis.
January 1988, Anatomy and embryology,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Bulk Electroporation-Mediated Gene Transfer into Xenopus Tadpole Brain.
October 2018, Cold Spring Harbor protocols,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Action potentials of embryonic dorsal root ganglion neurones in Xenopus tadpoles.
October 1978, The Journal of physiology,
Edward S Ruthazer, and Anne Schohl, and Neil Schwartz, and Aydin Tavakoli, and Marc Tremblay, and Hollis T Cline
Development and stability of postional information in Xenopus retinal ganglion cells.
April 1972, Proceedings of the National Academy of Sciences of the United States of America,
Copied contents to your clipboard!