Primordial germ cells (PGCs) of Xenopus laevis are highly migratory. The last section of their migratory pathway is through the dorsal mesentery of the tadpole gut. This in vivo pathway is rich in fibronectin, a glycoprotein that promotes cell adhesion and migration in vitro. Isolated PGCs are associated with cells from the mesentery and with fibronectin. Treatment with trypsin removes both the mesentery cells and the fibronectin. The PGCs do not appear to resynthesize detectable fibronectin in vitro. In contrast, cultured adult mesentery epithelial cells synthesize large amounts of fibronectin and lay it down in subcellular fibrils that align with intracellular microfilament bundles. PGCs plated on cultured mesentery cell layers adhere to them, elongate and align with the microfilament bundles of the mesentery cells. PGCs adherent to mesentery cell layers are closely associated with fibronectin; moreover, F(ab)2 fragments of anti-Xenopus fibronectin IgG inhibit the adhesion and spreading of PGCs on the mesentery. These results indicate that PGCs can adhere to mesentery cells via fibronectin produced by the latter cells and suggest that fibronectin may be involved in the migration of PGCs in vivo.