Astrocytes are coupled via gap junction channels, predominantly formed by connexin-43 (Cx43), and contribute to neuronal function in the normal and diseased brain. In this study, we demonstrate that epidermal growth factor (EGF), applied to cortical astrocytes, results in a decrease in the expression of Cx43 mRNA and protein. We have further shown that the decrease is associated with the receptor tyrosine kinase pathway and the MEK inhibitor prevents EGF-stimulated down-regulation of Cx43 expression. These findings demonstrate a previously unknown function of EGF on cultured astrocytes, which may be relevant to the regulation of astrocytic growth and differentiation.