Immunocontraceptive strategies have proved to be efficient in controlling fertility of various mammalian species. In the present study we have made the first steps towards the identification of Arvicola terrestris sperm antigens that could be used as targets in the development of a contraceptive vaccine to limit the proliferations of this pest rodent. Rabbit-raised polyclonal antisera directed against complete A. terrestris spermatozoa were used to identify and characterize on 2D-gels coupled with a MALDI-TOF mass spectrometry analysis A. terrestris sperm proteins. Amongst the proteins pinpointed by this approach some were further investigated based on their tissue- and/or sperm-specific expression, and their relevance to fertility or sperm/egg interaction. In parallel, three proteins that have been already reported in the literature to be appropriate targets for the development of contraceptive vaccines in other mammalian species have also been looked for in A. terrestris. With the selected protein targets, a reverse-PCR approach using degenerate primers was employed to amplify corresponding A. terrestris cDNAs. After conceptual translation and sequence alignment, different proteins were studied to determine zones with sufficient sequence divergence and of antigenic/immunogenic nature that could be used in future assays to immunize animals.