Myoepithelial cells (MECs) are contractile elements showing a combined epithelial and smooth muscle phenotype. Among the numerous immunohistochemical markers employed to detect MECs, smooth muscle actin (SMA) is the most widely used. Recently, other markers of smooth muscle differentiation have been demonstrated in MECs, such as calponin, heavy caldesmon (h-caldesmon), and smooth muscle myosin heavy chain (SMM-HC). In the present study normal salivary, mammary, and sweat glands have been studied with four markers of smooth muscle differentiation (SMA, calponin, h-caldesmon, and SMM-HC). The four markers were differentially expressed in the various types of glands. In parotid glands MECs mainly expressed calponin and caldesmon; in submandibular and in cutaneous apocrine and eccrine glands, MECs strongly expressed SMA, calponin, and caldesmon; in minor salivary glands all four markers were equally strongly expressed; and in mammary glands SMA, calponin, and SMM-HC were present both in periductal and periacinar MECs while caldesmon was present in periductal MECs only. In addition to MECs, SMA stained stromal myofibroblasts, sometimes hampering the identification of MECs. Among the other markers, calponin stained only rare stromal myofibroblasts, while caldesmon and SMM-HC were confined to MECs. In conclusion, these latter markers are very useful for identifying MECs. It is suggested that the differential expression of smooth muscle contractile proteins might reflect different functions of MECs in the various sites. Int J Surg Pathol 8(1):29-37, 2000