This video demonstrates how to visualize axonal pathways of genetically defined groups of neurons in the embryonic chick spinal cord utilizing in ovo electroporation of reporter genes under the control of specific enhancer elements.
I. Electroporation
Egg handling
Preparations
Windowing and electroporation
II. Spinal cord open-book preparation
Detachment of the spinal cord from the embryo
Fixation
Immunohistochemistry
Mounting open book for imaging
Electroporation of plasmid DNA into the chick embryo evolves as a powerful technique for in vivo ectopic expression. The combination of specific enhancers, and chick electroporation provides a quick and efficient tool for deciphering axonal pathways of a genetically defined group of neurons1,3,5. Utilizing the Cre/LoxP and the Gal4/UAS amplification systems can augment the levels and duration of expression. The emerging picture is of a complex divergence of axonal cues that arises from interneuron subpopulations, defined by the direction of their axonal projections. In addition, simultaneous molecular and spatial restricted labeling of two neuronal populations can be attained. Thus, providing information about the architecture of neuronal circuits 3.
This work was supported by grants to AK from the Israel Science Foundation, The Israel ministry of health, and DFG (German Research Foundation).