Cellular localization of Babesia bovis rhoptry-associated protein 1 in the merozoite stage with immuno-electron microscopy

We examined the detailed cellular localization of the rhoptry-associated protein-1 (RAP-1) of Babesia bovis merozoites by the post-embedding method of immuno-electron microscopy (IEM) using the anti-RAP-1 monoclonal antibody (mAb) 1C1 in order to substantiate the result in our previous report by an indirect immunofluorescent antibody test and the pre-embedding method of IEM. RAP-1 slightly distributed only in cytoplasm of merozoites, especially around the nucleus, but not in the rhoptry organelle at an early stage after invasion into a red blood cell. Then, the RAP-1 was accumulated in the rhoptry organelle and was also found in the iRBC cytoplasm, which suggests that synthesis and release of RAP-1 may occur in the iRBC. Finally, the RAP-1 was found within and around merozoites after the breakdown of the iRBC. The results of the present study suggest that the RAP-1 of B. bovis merozoites functions not only in the invasion into RBC but also in the escape from iRBC.