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Electroporation as a tool to study in vivo spinal cord regeneration
Author(s) -
Echeverri K.,
Tanaka E.M.
Publication year - 2003
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.10238
Subject(s) - spinal cord , biology , electroporation , ependymal cell , differential interference contrast microscopy , regeneration (biology) , anatomy , microbiology and biotechnology , neuroscience , green fluorescent protein , fluorescence microscope , blastema , transfection , microscopy , pathology , gene , fluorescence , genetics , medicine , physics , quantum mechanics
Tailed amphibians such as axolotls and newts have the unique ability to fully regenerate a functional spinal cord throughout life. Where the cells come from and how they form the new structure is still poorly understood. Here, we describe the development of a technique that allows the visualization of cells in the living animal during spinal cord regeneration. A microelectrode needle is inserted into the lumen of the spinal cord and short rapid pulses are applied to transfer the plasmids encoding the green or red fluorescent proteins into ependymal cells close to the plane of amputation. The use of small, transparent axolotls permits imaging with epifluorescence and differential interference contrast microscopy to track the transfected cells as they contribute to the spinal cord. This technique promises to be useful in understanding how neural progenitors are recruited to the regenerating spinal cord and opens up the possibility of testing gene function during this process. Developmental Dynamics 226:418–425, 2003. © 2003 Wiley‐Liss, Inc.