Spatial distribution of motor endplates affects the shape of the electrical activity recorded from muscle. In order to provide information for realistic models of action potential propagation within muscles, we assembled three-dimensional maps of the motor endplates of the rat medial gastrocnemius (MGM) and lateral gastrocnemius (MGL) muscles. The maps were assembled from histological cross sections stained for acetylcholinesterase activity. Within MGL, the motor endplates formed three columns along its longitudinal axis. Within MGM, the motor endplates were arranged in a leaf-like body that shifted obliquely from proximal to distal. As inferred from the proximo-distal distribution of the cross-sectional projection area, the majority of the motor endplates were concentrated in the middle of MGL and in the distal third of MGM. Regions of maximal motor endplate concentration are considered most suitable for injections of neuroactive substances, such as neuronal tracers. The assembled maps of the gastrocnemius muscles can be used as guides for such injections within the motor endplate zones.