Objectives: The precise location, origin and nature of nerve fibers innervating the urethral sphincter have not been clearly established. Classical anatomical studies based on cadaver dissections have provided conflicting results concerning the location of pudendal and autonomic nerve fibers. This study was designed to identify nerve fibers innervating the urethral sphincter and to provide a three-dimensional representation of their tissue relations in the female human fetus.
Materials and methods: Histology and immunohistochemistry (Masson's Trichromic, Luxol Fast Blue, Protein S 100 immunostaining and smooth fiber actin immunostaining) were performed on the external urethral sphincter of ten female fetuses with a crown-rump length of 112 to 340mm. Three-dimensional reconstructions of the urethral structure and innervation were obtained from serial sections using Surf Driver 3.5.3 software (David Moody and Scott Lozanoff).
Results: Three-dimensional reconstructions of the same sections with different stains demonstrated the precise structure of the muscle layers (smooth and striated muscle fibers) and nerve fibers (myelinated and unmyelinated) and their relations with the urethra and vaginal wall. The proximal third consisted of a circular smooth muscle sphincter, the middle third consisted of two circular layers of smooth and striated muscle fibers and the distal third consisted of a circular layer of smooth muscle fibers surrounded by an omega-shaped layer of striated muscle fibers. In the proximal third of the urethral sphincter, myelinated fibers were identified running with unmyelinated fibers from the pelvic plexus. These fibers were closely related to the lateral and anterior aspects of the vagina. Unmyelinated fibers entered the smooth muscle part of the sphincter at 4 o'clock and at 8 o'clock. Most myelinated fibers entered the sphincter at 3 o'clock and at 9 o'clock.
Conclusion: Histological and immunohistochemical three-dimensional reconstruction of the anatomical structures of the urethral sphincter provides a better understanding of the origin and nature of the Innervation participating in urinary continence. It provides a very informative view of the three-dimensional arrangement of sphincter muscle layers.