To clarify the role of the ovary in the sexual differentiation of prolactin and growth hormone cells, the anterior pituitary glands of mice ovariectomized before or after puberty were studied by a stereological morphometric technique with the electron microscope. A marked sex difference was observed in the relative proportions of these two types of cells in normal adult control animals. In male mice about 52% of anterior pituitary cells were growth hormone cells and about 10% were prolactin cells, while in female mice prolactin cells represented about 39% and growth hormone cells about 37% of the anterior pituitary cell population. Ovariectomy before puberty reduced the proportion of prolactin cells to about 10% and increased growth hormone cells to about 50% of the cell population. The size of prolactin cells and the development of their cell organelles was also reduced. Ovariectomy after puberty had less marked effects. These results suggest that ovariectomy before puberety induced the male phenotype by inhibition of the differentiation of prolactin cells and stimulation of the differentiation of growth hormone cells. The significance of these results in relation to the sexual differentiation of the pituitary gland is discussed.