Conjunctival primary acquired melanosis (PAM) is a frequent precursor of conjunctival melanoma. Since there is indirect evidence that the conjunctiva is an estrogen-responsive tissue, and since it was suspected that estrogen has a role in the etiology of melanoma, we decided to evaluate whether PAM may be responsive to estrogen. Formalin-fixed, paraffin-embedded sections from 13 cases of PAM and 2 cases of conjunctival melanoma were immunostained with an estrogen-receptor (ER)-specific antibody. All lesions and the normal conjunctival tissue adjacent to the lesions were found to be ER negative. It is concluded that PAM and normal conjunctiva are not sensitive directly to estrogen. When considering previously reported data, it is conceivable that the normal conjunctiva, but not PAM, is indirectly affected by estrogen.