Crack initiation plays a major role in very high cycle fatigue (VHCF) life, and the initiation of cracks is related to slip behavior. There is a need for improvement in the understanding of the influence of Ti-6Al-4V microstructures on VHCF performance and crack initiation modes. In this study, through an investigation of Ti-6Al-4V VHCF in equiaxed and bimodal microstructures, two different crack initiation modes were identified. The change in crack initiation mode is related to the variation in microtexture, for which a corresponding model is proposed. The VHCF performance of the bimodal microstructure is significantly improved compared to that of the equiaxed microstructure.