Aldehydes are ubiquitous pollutants generated during the combustion of organic materials, and are present in air, water, and food. Several aldehydes are also endogenous products of lipid peroxidation and by-products of drug metabolism. Despite well-documented high reactivity of unsaturated aldehydes, little is known regarding their cardiovascular effects and their role in cardiac pathology. Accordingly, we examined the myocardial effects of the model unsaturated aldehyde acrolein. In closed-chest mice, intravenous acrolein (0.5 mg/kg) induced rapid but reversible LV dilatation and dysfunction. In mouse myocytes, micromolar acrolein acutely depressed myofilament Ca²⁺ responsiveness without altering catecholamine sensitivity, similar to the phenotype of stunned myocardium. Immunoblotting revealed increased acrolein-protein adducts and protein-carbonyls in both acrolein-exposed myocardium (1.8-fold increase, p < 0.002) and myocytes (6.4-fold increase, p < 0.02). Both the contractile dysfunction and adduct formation were markedly attenuated by pretreatment with the thiol donor N-acetylcysteine (5 mM). Two-dimensional gel electrophoresis and MALDI-TOF/MS analysis revealed two groups of adducted proteins: sarcomeric/cytoskeletal proteins (cardiac &#945;-actin, desmin, myosin light polypeptide 3) and energy metabolism proteins (mitochondrial creatine kinase 2, ATP synthase), indicating site-specific protein modification that was confirmed by immunohistochemical co-localization. We conclude that direct exposure to acrolein induces selective myofilament impairment which may be in part related to the modification of proteins involved in myocardial contraction and energy metabolism. Myocardial dysfunction induced by acrolein and related aldehydes may be symptomatic of toxicological states associated with ambient/occupational exposures or drug toxicity. Moreover, aldehydes such as acrolein may mediate cardiac dysfunction in pathologies characterized by high oxidative stress.