OBJECTIVE The intrinsic urethral sphincter is composed of adjacent striated and smooth muscle. We studied the sequential expression of smooth and striated muscle proteins to gain insight into the ontogeny of intrinsic sphincter development. METHODS The intrinsic urethral sphincters of timed Fischer 344 rat embryos at 14, 16 and 18 days of gestation, neonates on postnatal day 1 and adult animals were examined. Serial sections of the urethra and adjacent levator ani muscles were studied histologically with hematoxylin and eosin, anti-alpha-smooth muscle actin, anti-alpha-sarcomeric actin and antistriated muscle myosin heavy chain antibodies. RESULTS The intrinsic urethral sphincter was identified within the periurethral mesenchyma as early as day 14 of gestation. Although striated myotubules were identified within the urethra by hematoxylin and eosin staining starting on postnatal day 1, striated muscle myosin heavy chain protein was absent in the embryonic and neonatal development of the sphincter, and it was expressed only in the mature myotubule of adults. alpha-Smooth muscle actin was expressed throughout the urethral sphincter of embryonic and neonatal animals. In adults alpha-smooth muscle actin was confined to the smooth muscle component of the urethra. Co-expression of alpha-smooth and alpha-sarcomeric muscle actin by the striated sphincter myotubule was noted only in neonates. CONCLUSIONS Development of the intrinsic urethral sphincter is characterized by sequential expression of well characterized muscle marker proteins. The co-expression of smooth and striated muscle markers by developing sphincter myotubule suggests the possibility that trans-differentiation of smooth to striated muscle occurs in the developing genitourinary tract.