Plasmodium spp. development within mosquito vectors includes invasive ookinete and sporozoites stages, which display gliding movements during invasion of the midgut and salivary glands, respectively. Sporozoite gliding has been well documented, and several proteins released during sporozoite's locomotion have been described. However, proteins on the ookinete gliding trails are poorly described. In the present study, we documented that 2 proteins, Pbs25 and circumsporozoite thrombospondin-related protein (CTRP), are released during Plasmodium berghei ookinete gliding on a solid surface substrate.