We studied the surgical anatomy of 14 hearts with isolated ventricular septal defects and the precise relationship of the atrioventricular conduction axis to their rims. Ten of these hearts were investigated by serial sectioning of their atrioventricular conduction systems. All defects were divided into perimembranous or muscular types as previously suggested by Soto et al. The distinguishing feature of the perimembranous defect was that the central fibrous body formed part of its rim, this fact indicated that the conduction tissue was always to the right of the surgeon's hand when the defect was approached through the right atrium. The precise relationship of the ventricular conduction tissues varied depending upon whether the perimembranous defect extended into the inlet, trabecular or outlet components of the muscular septum. The nonbranching bundle was closest to the septal rim in the inlet and trabecular defects, frequently being buried in a remnant of the interventricular membranous septum. However, in these defects the nonbranching and branching bundles were also found in other specimens remote from the septal crest, yet were carried on the left ventricular aspect of the septum. In perimembranous inlet defects, the penetrating bundle detoured deep into the central fibrous body. In perimembranous outlet defects, the conduction tissues were remote from the septal crest. In contrast to these findings, the conduction tissues were away from the edges of the defects in trabecular and outlet muscular defects. In conclusion, in the perimembranous inlet defect, the placement of the sutures on the annulus of tricuspid valve should be avoided in view of its abnormal long penetrating bundle.