We report the expression pattern of hemidesmosome-associated proteins laminin-5, composed of alpha 3 beta 3 gamma 2 chains, and HD1 in the developing mouse and human intestine, an organ in which variations in structure and function parallel morphogenesis and differentiation. Immunocytochemistry analysis revealed the coexpression of laminin-5 and HD1 at the basal pole of differentiating epithelial cells. Distinct noticeable variations occurring in the location of laminin alpha 3 chain in development of mouse gut were stressed by the reverse transcriptase-polymerase chain reaction data. A peculiar finding was also the location of laminin gamma 2 chain in the intestinal muscle coat. The cellular origin of laminin gamma 2 chain was examined by immunocytochemistry on interspecies hybrid intestines with specific antibodies recognizing mouse antigens. Complementary and sequential production of laminin gamma 2 chain was observed, by epithelial cells as establishment of the basement membrane occurs and by mesenchymal cells in the more differentiated organ. These results support the concept of mesenchymal involvement in deposition of basement membrane molecules, a crucial process for intestinal differentiation. Taken together these data provide the first evidence for the coexpression of hemidesmosome-associated proteins in the gut, a non-stratified tissue.