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The vertebrate heart muscle (myocardium) develops from the first heart field (FHF) and expands by adding second heart field (SHF) cells. While both lineages exist already in teleosts, the primordial contributions of FHF and SHF to heart structure and function remain incompletely understood. Here we delineate the functional contribution of the FHF and SHF to the zebrafish heart using the  cis -regulatory elements of the  draculin  ( drl ) gene. The  drl  reporters initially delineate the lateral plate mesoderm, including heart progenitors. Subsequent myocardial  drl  reporter expression restricts to FHF descendants. We harnessed this unique feature to uncover that loss of  tbx5a  and  pitx2  affect relative FHF versus SHF contributions to the heart. High-resolution physiology reveals distinctive electrical properties of each heart field territory that define a functional boundary within the single zebrafish ventricle. Our data establish that the transcriptional program driving cardiac septation regulates physiologic ventricle partitioning, which successively provides mechanical advantages of sequential contraction.

Chamber identity programs drive early functional partitioning of the heart