Surgical landmarks of the conduction system were histologically evaluated in 29 cardiac specimens. The distribution of the system was grossly inherent to the type of ventricular septal defect as classified by Soto and coworkers, although it varied individually. The relationship of the right bundle branch (RBB) to the papillary muscles was of surgical interest. In defects unrelated to maldevelopment of the septum of the conus, the RBB passed beneath or slightly anterior to upper accessory papillary muscles (AcPMs) and posterior to the medial papillary muscle (MPM), regardless of the subtype of defect. In defects caused by maldevelopment, such as tetralogy of Fallot, it passed anterior to the MPM. Such data support the hypothesis that the RBB descends beneath or anterior to embryological upper AcPMs, whatever the morphological role may be, because of the supposedly independent developmental origin of the MPM and AcPMs. The relationship between the RBB and upper AcPMs appeared further modified by the attitude of the trabecula septomarginalis. Our improved clinical results have demonstrated that such information offers a gross but practical guide for prevention of conduction disturbances.