Cardiac glycoside ouabain, when bound to cardiac Na+K+-ATPase (sodium pump), inhibits the pump leading to positive inotropy. It also stimulates sodium pump interacting with other proteins and activates several signaling pathways including phosphatidylinositol 3-kinases (PI3K) /Akt and ERK pathways (1,2). These events originate in the plasma membrane caveolae (3,4). In cardiomyocytes, caveolins 1, 2 and 3 are present. This study aims to examine the role of caveolin-1 in Na+K+-ATPase mediated cardiac signaling and cardiac function using wild type (WT) and caveolin-1 knockout (cav-1KO) mice. Ouabain-induced PI3K/Akt and ERK activation and – caused cardiac hypertrophy were reduced markedly in cav-1 KO cardiomyocytes. Additionally, association of the Na+K+-ATPase α1 subunit and caveolin-3 was decreased in cav-1 KO cardiomyocytes. Moreover, ouabain-induced interaction of the α1 subunit of Na+K+-ATPase and PI3K was also reduced in cav-1 KO cardiomyocytes. Deletion of cav-1 reduced protein content of PI3K/Akt and the β1 subunit of Na+K+-ATPase in caveolar fractions in cardiomyocytes. Transient ouabain infusion increased cardiac contractility in WT and KO mice monitored through cardiac catheterization. Surprisingly, the effect of ouabain on cardiac contractilities was not reduced when cav-1 was absent. This was further confirmed with isolated working hearts from WT and cav-1 KO mice. Our data indicate that ouabain-induced cardiac signaling and hypertrophy is mediated by a mechanism through interaction of cav-1 and Na+K+-ATPase. However, ouabain exerted caveolin-1-independent cardiac contractility.