Members of the integrin family of adhesion receptors mediate both cell-cell and cell-matrix interactions and have been shown to play vital roles in embryonic development, wound healing, metastasis, and other biological processes. The integrin ␣91 is a receptor for the extracellular matrix proteins osteopontin and tenacsin C and the cell surface immunoglobulin vascular cell adhesion molecule-1. This receptor is widely expressed in smooth muscle, hepatocytes, and some epithelia. To examine the in vivo function of ␣91, we have generated mice lacking expression of the ␣9 subunit. Mice homozygous for a null mutation in the ␣9 subunit gene appear normal at birth but develop respiratory failure and die between 6 and 12 days of age. The respiratory failure is caused by an accumulation of large volumes of pleural fluid which is rich in triglyceride, cholesterol, and lymphocytes. ␣9 ؊/؊ mice also develop edema and lymphocytic infiltration in the chest wall that appears to originate around lymphatics. ␣9 protein is transiently expressed in the developing thoracic duct at embryonic day 14, but expression is rapidly lost during later stages of development. Our results suggest that the ␣9 integrin is required for the normal development of the lymphatic system, including the thoracic duct, and that ␣9 deficiency could be one cause of congenital chylothorax.Integrins are heterodimeric receptors for extracellular matrix and cell surface counterreceptors, which play important roles in embryonic development, inflammation, wound healing, and tumorigenesis (8, 9, 15). The integrin 1 subunit pairs with at least 12 ␣ subunits, forming the largest subfamily of integrins. Ablation of the 1 gene produces early embryonic lethality (3, 17), and most null mutations described for individual 1-associated ␣ subunits cause severe but individually distinct developmental phenotypes (10).␣91 is a member of the 1 family that recognizes tenascin C (23, 26), osteopontin (16,24), and vasular cell adhesion molecule-1 (VCAM-1) (20) as ligands. Immunostaining of mouse tissue has shown that this integrin is expressed in skeletal and cardiac muscle, visceral smooth muscle, hepatocytes, airway epithelium, squamous epithelium, and choroid plexus epithelium (13,21). Wound healing experiments performed on mouse corneas (18,19) suggest that ␣9 may play a role in corneal epithelial migration and differentiation. In mouse embryos, ␣9 expression was not detected prior to embryonic day 12.5 (E12.5) (21), suggesting that this integrin is unlikely to play a general role in the earliest stages of tissue morphogenesis.In vitro experiments demonstrate that ␣91 mediates cell adhesion as well as cell migration on all three known ligands (16,20,26). In addition, ␣9-transfected SW480 cells use ␣91 to proliferate on tenascin-C (25), and these effects are associated with phosphorylation of the integrin signaling intermediates focal adhesion kinase and paxillin and with activation of the mitogen-activated protein kinase isoform, erk2. However, the in vivo function(s) ...