The membrane surface of polarized epithelial cells can be separated in apical and basolateral domains that differ in molecular composition and function. Components of the cytoskeleton obviously play an important role in both generation and maintenance of epithelial polarity. The microtubular system is uniformly aligned in enterocytes and pancreatic acinar cells with the minus ends of the microtubules located apically and the plus ends basally. Drug-induced disassembly of microtubules results in mistargeting of pancreatic zymogen granules and of apical but not basolateral membrane proteins of enterocytes. Association of zymogen granules with the minus end motor protein, cytoplasmic dynein, and components of its receptor, dynactin, indicates that microtubules are important for vectorial delivery of apical but not basolateral post-Golgi vesicles. The lateral membrane domain of the gut epithelium is scaffolded by a spectrin-based membrane cytoskeleton consisting basically of actin, spectrin, and ankyrin. Ankyrin binds to different integral membrane proteins, such as the sodium pump in glandular and kidney epithelial cells or to the anion exchanger (AE2) of gastric parietal cells, thereby probably playing a critical role in maintenance and/or generation of the polarized distribution of these basolateral membrane proteins. Scattered epithelial cells sharing apical cytoskeletal features of gustatory receptor cells were identified as brush cells (tufted cells). These cells are rich in nitric oxide synthase and contain in their apical brush border the gustatory trimeric G-protein, alpha-gustducin, indicating that brush cells are involved in chemoreceptive signaling.