A novel technique for determining the collagen fibre orientation pattern of cross-sections of cortical bone was used to study mid-diaphyseal sections from the equine radius. Several in vivo strain gauge studies have demonstrated that this bone is loaded in bending during locomotion in such a way that the cranial cortex is consistently subjected to longitudinal tensile strains and the caudal cortex to longitudinal compressive strains. Twenty-three radii from 17 horses were studied. All the bones obtained from adult horses exhibited a consistent pattern of collagen fibre orientation across the cortex. The cranial cortex, subjected to intermittent tension, and the lateral and medial cortices, through which the neutral axis passes, contained predominantly longitudinally oriented collagen fibres. The caudal cortex, subjected to longitudinal compression during life, contained predominantly oblique/transverse collagen. This pattern was less evident in bones from foals. Microscopic analysis of the bones studied showed that primary lamellar bone was composed of predominantly longitudinal collagen fibres, irrespective of cortex. However, there was a strong relationship between cortical location and fibre orientation within remodelled bone. Secondary osteons which formed in the caudal (compressive) cortex contained predominantly oblique/transverse collagen, while those which formed elsewhere contained longitudinal collagen. This observation explained the developmental appearance of the characteristic macroscopic pattern of collagen fibre orientation across the whole cortex in the adult. These findings provide evidence for the existence of a relationship between the mechanical function of a bone with its architecture, and now demonstrate that it extends to the molecular level.