Aims <p>In elite athletes, differentiation between physiological and pathological cardiac remodelling can be challenging. Left ventricular (LV) strain has become part of the standard in echocardiography. Data on right ventricular (RV), left atrial (LA) and right atrial (RA) strain are sparse in elite athletes.</p> Methods <p>Transthoracic echocardiography including longitudinal strain analysis of all cardiac chambers was performed using a GE Vivid iq in 112 male elite professional cyclists aged 24–32&#xa0;years. All athletes were actively participating in top level international races and included race champions and Olympic medallists. The results were compared to published reference values of healthy males of similar age.</p> Results <p>LV average global longitudinal strain (LV aGLS) was –21.7% (–24.9 to –18.5), significantly higher (<i>p</i> &lt; 0.0001) than the reference. RV free wall strain (RV FWS) was –26.2% (–32.3 to –20.1), and not different (<i>p</i> = 0.36) from the reference. Compared to the reference, LA reservoir strain (LASr), LA conduit strain (LAScd), RA reservoir strain (RASr) and RA conduit strain (RAScd) were higher (all <i>p</i> &lt; 0.0001), whereas LA and RA contractile strain (LASct and RASct) were not (both <i>p</i> = 0.2). Strain values were independent of respective chamber sizes. There was an inverse association between the left ventricular mass index and LASr (<i>p</i> = 0.010) and LAScd (<i>p</i> = 0.042).</p> Conclusion <p>Strain parameters are independent of the extent of hypertrophy in elite cyclists. An underlying myocardial disease may be suspected in case of lower-than-expected longitudinal strain of any chamber.</p> Graphical abstract <p></p>