Newly hatched domestic chicks serve as ideal models for studies of the neural basis of behavioral plasticity, particularly for understanding the mechanisms of learning such as filial imprinting. To elucidate the molecular basis and gene functions involved in learning, we developed an in-vivo gene-transfer system in the brain of a living chick using electroporation. When green fluorescent protein-encoding plasmids were transfected to a chick brain, green fluorescence was clearly observed, and expression at the protein level was confirmed by immunoblotting. Most of the transfected brain cells were neuronal cells with dendrites. This neuron-selective electroporation system will facilitate the analysis of gene functions in the living chick brain and provide further clues as to the molecular mechanisms of avian learning.