Detailed information on how an epithelial differentiation occurred in the developing eustachian tube and middle ear would be helpful in understanding both normal physiology and pathology of the tubotympanum. This study was undertaken to establish patterns of laminin and E-cadherin in the embryonic mouse eustachian tube and middle ear by use of immunohistochemistry at a stage when epithelial differentiation is taking place. This study was also designed to clarify the role of the middle ear mesenchyme. During the development of the eustachian tube, relatively high immunoreactivity to laminin was observed in the epithelium at gestational days 16 and 17, when the developments of ciliated and secretory cells were first observed. At the time of birth, in contrast to epithelium of the eustachian tube, epithelium of the middle ear cavity showed predominant expression of laminin and E-cadherin. These findings suggest that the expressions of laminin and E-cadherin may be correlated with maturation of the epithelium in the eustachian tube and middle ear and that the epithelial differentiation of the developing murine eustachian tube and middle ear may be controlled by epithelial-mesenchymal interaction and cell-to-cell interaction.