Several studies suggested that headache attacks and its frequency were mainly responsible for increased cardiovascular (CV) disease and mortality in patients with migraine with aura (MWA). Elevated serotonin level has been found to play a role in migraine attacks. Serotonin was best studied within the CV system for its role in the development of pulmonary hypertension, which had negative impact on right ventricular (RV) functions. Therefore, in this study we aimed to evaluate RV functions during headache attacks in MWA patients and its relation to attack frequency with the utility of 2-dimensional speckle tracking echocardiography (2D STE). Fifty-three patients with the diagnosis of MWA were enrolled in the study. All patients were evaluated by conventional and 2D-STE echocardiography and venous blood sampling for serotonin was obtained during headache-free period (HFP) and headache-attack period (HAP). Also, patients were divided into two groups according to attack frequency. Patients exhibited higher serotonin levels during HAP than HFP (p < 0.001). Regarding 2D-STE derived RV-free-strain parameters, patients had lower RV-free wall longitudinal speckle-tracking strain (RV-free-ST), RV-free systolic strain rate (RV-free-STR-S), RV-free early diastolic strain rate (RV-free-STR-E) and RV-free-STR-E/A ratio levels during HAP when compared with HFP (p 0.002, p 0.006, p < 0.001 and p 0.001, respectively). Thirty-one patients (58.4 %) had low-frequency attack. Patients with high-frequency attacks had increased serotonin levels (p 0.040) and decreased RV-free-ST, RV-free-STR-S, RV-free-STR-E and RV-free-STR-E/A ratio values during HAP when compared to low-frequency group (p 0.026, p 0.029, p 0.037 and p 0.019 respectively). This study demonstrated that migraine attacks, especially at higher frequencies, could have negative impact on RV systolic functions in MWA patients.