Cardiogenic differentiation mandates robust metabolic signaling and information exchange between mitochondria and cytosolic/ nuclear compartments to ensure developmental programming and an energetic continuum that sustains the function of Trichostatin A HDAC inhibitor nascent cardiomyocytes. Underlying the transition from low-BMS-354825 energy requiring pluripotency into a cardiac phenotype is a switch in energy metabolism, from anaerobic glycolysis to more efficient mitochondrial oxidative phosphorylation. Glycolytic and creatine kinase network formation provides energetic connectivity between expanding mitochondrial clusters and ATP-utilization cellular sites. Despite advances in decoding the dynamics of major ATP production and distribution processes during lineage specification, metabolic signaling circuits responsible for integration of energetic events with cardiogenic programming remain largely unknown. Adenylate kinase phosphorelays are recognized facilitators of metabolic signaling, optimizing intracellular energetic communication and local ATP supply. The unique property of adenylate kinase catalysis to transfer both b- and c-phosphoryls doubles the energetic potential of the ATP molecule, and provides a thermodynamically efficient mechanism for high-energy phosphoryl transport from mitochondria to myofibrils and the cell nucleus. Recent studies indicate that mitochondrial adenylate kinase is required for unfolded protein response and that AK2 deficiency compromises embryonic development and hematopoiesis by interfering with mitochondrial ATP/ADP exchange. In this regard, the stressresponsive adenylate kinase isoform network, coupled with AMP signaling through AMP-activated kinase, provides highfidelity surveillance of energy metabolism to sustain the balance of energy supply and demand. The metabolic sensor AMPK appears essential for embryonic development, maintaining cell polarity and cell cycle progression, and the upstream kinase LKB1 is critical for cardiac development, and in hematopoietic stem cell maintenance and cell division. However, the contribution of the adenylate kinase/AMPK tandem in stem cell cardiac differentiation has not been determined. Here, we uncovered a developmental deployment and upregulation of the integrated adenylate kinase and AMP-AMPK signaling system underlying the execution of cardiogenic programming during embryonic stem cell differentiation. Nuclear translocation of adenylate kinase and p-AMPK supported energydependent cell division, and facilitated asymmetric differentiation leading to cardiac specification. Targeted knockdown of the adenylate kinase-dependent energetic and AMP signaling cascade disrupted maturation of mitochondrial networks and myofibrillogenesis, precluding formation and function of organized cardiac beating structures.