Does fertilization in the filarial nematode Dirofilaria immitis occur through endocytosis of spermatozoa?

Information on the ultrastructural details of fertilization in filarial nematodes are still unavailable. Here we report new data on this process in Dirofilaria immits, the heartworm of dogs and cats. Electron microscopy allowed us to observe oocytes engulfing spermatozoa through an endocytosis-like process. We also observed spermatozoa inside the oocytes which still possessed their plasma membrane and which were crearly enveloped by a further membrane, likely derived from the endocytosis process. At this stage, at the interface between the sperm membrane and the endocytotic membrane (vacuolar space), we observed fiocculent material in the proximity of the membranous organelles (MOs) of the sperm. In the proximity of the MOs, we also observed the enlargement of the vacuolar space. Other images showed the dissolution of the sperm membrane, and the release of nuclear masses and organelles in the egg cytoplasm. We did not observe the fusion of lysosomes to the endocytotic vacuoles. In addition, the lysis of the sperm organelles has never been observed inside the vacuoles containing the whole sperm. Thus we suggest that the degradation of the endocytotic and sperm plasma membranes is determined by material released by the MOs. Since we did not observe the entry of sperm into the oocytes by other mechanisms, we also suggest that endocytosis is the normal process used by the spermatozoon to get into the egg cytoplasm in D. immitis. Finally, during our observations of the seminal receptacle we did not observe any structure in the spermatozoa which could be interpreted as an intracellular bacterium. This is consistent with previous results indicating that the bacterium Wolbachia in filarial nematodes is not transmitted through the sperm.