In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development

This experiment demonstrates in ovo electroporation of chick midbrain to study the effect of exogenous gene expression on midbrain development and dopaminergic neuron differentiation. First, inject the midbrain of hamburger Hamilton, stage 11 chick embryos with desired plasmid, DNA, to specifically target exogenous DNA into the developing midbrain vesicle. Then using custom made small electrodes, electroporated the midbrain vesicle to deliver the exogenous DNA specifically into the midbrain neural progenitors.

Next, harvest stage 23 embryos with good M cherry expression in the midbrain and generate optimal midbrain transverse sections. Ultimately, immuno staining analysis for dopaminergic neuron markers such as tyrosine hydroxylase can show the differentiation of neural progenitors into mature dopaminergic neurons induced by the expression of genes of interest. The main advantage of this technique over other existing methods of in ovo electroporation is that this protocol can achieve membrane specific electroporation to study mirin dopaminergic neuron differentiation.