We sought to determine how the mode of muscle metaboreflex activation influences the central hemodynamic response and cardiac inotropic and lusotropic function in healthy humans. Ten healthy males performed (i) isometric handgrip (IHG) with and without post-exercise ischemia (PEI) to examine the influence of isolated muscle metaboreflex activation and (ii) rhythmic handgrip (RHG) with and without ischemia to examine the influence of enhanced muscle metaboreflex activation. Heart rate (HR) and blood pressure (BP) were continuously monitored. Stroke volume (SV, Doppler echocardiography) was measured, cardiac output (CO = HR × SV) and total peripheral resistance (TPR = mean BP/CO) calculated, and indices of left ventricular systolic and diastolic function were obtained (tissue Doppler imaging). During isolated muscle metaboreflex activation with PEI following IHG, mean BP (+23 ± 3 mm Hg) and TPR were elevated from baseline (p < 0.05), whereas HR, SV, and CO were unchanged. Enhanced muscle metaboreceptor activation during ischemic RHG augmented the increase in mean BP, CO, and HR (p < 0.05 ischemic vs. free-flow RHG), whereas SV and TPR were unchanged from baseline. Neither isolated (PEI) nor enhanced muscle metaboreflex activation altered left ventricular systolic function (systolic myocardial velocity), but left atrial systolic function (late diastolic myocardial velocity) was enhanced. These findings indicate that the mode of muscle metaboreceptor activation (during vs. post handgrip) determines whether the resultant pressor response is flow (CO) or vasoconstriction (TPR) mediated, and that although left ventricular systolic function is unchanged, enhanced left atrial systolic function likely aids the preservation of SV during muscle metaboreflex engagement.