Human nonfunctioning pituitary adenomas (NFPA) may produce CG in addition to the classical glycoprotein hormones (LH, FSH, and TSH). The aim of the present study was to localize LH beta, FSH beta, TSH beta, alpha-subunit (alpha SU), CG, and its beta-subunit (beta SU) in NFPA using a highly specific immunohistochemical technique. Nine NFPA, obtained at surgery, were processed for both electron microscopy and immunohistochemistry. Three tumors resulted oncocytomas, and six were null cell. Using an immunofluorescence technique, all tumors were positive for at least one glycoprotein; in particular, seven adenomas were markedly positive for CG beta, whereas only two were positive for the intact CG. No association among LH beta, FSH beta, and CG beta positivity could be demonstrated in the different adenomas. In the seven tumors positive for both CG beta and alpha SU, double fluorescence labeling demonstrated that six cases localized CG beta and alpha SU in different cells, but only one tumor showed the two subunits colocalized in the same cells. These data confirm that pituitary tumors synthesize both alpha SU and beta SU of glycoprotein hormones; in particular, the present study indicates that the majority of NFPA is able to synthesize CG, particularly its beta SU. Moreover, the localization of CG beta and alpha SU in different tumoral cells might account for the preferential expression of beta SU and alpha SU instead of the intact hormonal molecules in NFPA.