In most amoeboid cells, the main protein involved in motility is actin. Nematode sperm are an exception, and their amoeboid motility is based on major sperm protein (MSP). We have studied the localization of actin and MSP in spermatids and spermatozoa of Graphidium strigosum (Dujardin, 1845), a species which has elongate male germ cells in which organelles are easily identified. Electrophoreses of G. slrigosum sperm proteins indicate that the main protein band, about 15 kDa in molecular weight, is specifically recognized by an anti-MSP polyclonal antibody developed against MSP of Caenorhabditis elegans (Burke and Ward 1983). Actin is present in small quantities. Immunocytochemical observations reveal that actin and MSP have an identical localization in precise areas of the male germ cells. Spermatids are labelled as dots around a central unlabelled zone, and spermatozoa are labelled only at the level of the anterior cap. Observations in G. strigosum are similar to that previously obtained in Heligmosomoides polygyrus (Mansir and Justine 1996). Co-localization of actin and MSP in the anterior cap of the spermatozoon, the region associated with pseudopod production, does not demonstrate directly that actin is involved in amoeboid movements, but shows that the role of actin in the cytoskeleton of nematode sperm should be re-investigated.
Spermiogenesis and the spermatozoon were studied in the digenean Mesocoelium monas Rudolphi, 1819 (from the toad Bufo sp. in Gabon). An ultrastructural study revealed that spermiogenesis follows the usual pattern found in digeneans, i.e. proximo-distal fusion of axonemes with a median cytoplasmic process followed by elongation. The spermatozoon has two fully incorporated axonemes with the 9 +“1” trepaxonematan pattem. Indirect immunofluorescence localization of tubulin and fluorescent labelling of the nucleus were used to obtain additional information on the structure of the spermatozoon. It was thus shown that one of the axonemes is slightly shorter than the other (190 versus 220 pm) and that the filiform nucleus (65 pm in length) is located at the distal extremity of the spermatozoon (220 pm in length). Various monoclonal and polyclonal antibodies, specific to alpha, beta, acetylated-alpha, or general tubulin, were used and produced similar labelling.
Spermatological characters of the liver fluke Mediogonimus jourdanei Mas-Coma et Rocamora, 1978 were studied by means of transmission and scanning electron microscopy. Spermiogenesis begins with the formation of the differentiation zone containing two centrioles associated with striated rootlets and an intercentriolar body. These two centrioles originate two free flagella that undergo a 90° rotation before fusing with the median cytoplasmic process. Both nuclear and mitochondrial migrations toward the median cytoplasmic process occur before the proximodistal fusion of flagella. Finally, the constriction of the ring of arched membranes gives rise to the young spermatozoon. The mature sperm of M. jourdanei measures about 260 µm and presents two axonemes of different lengths with the typical pattern of the Trepaxonemata, two bundles of parallel cortical microtubules, one mitochondrion, a nucleus and granules of glycogen. An analysis of all the microphalloidean species studied to date emphasised some differences in certain characters found in Maritrema linguilla Jägerskiöld, 1908 and Ganeo tigrinum Mehra et Negi, 1928 in comparison to those in the remaining microphalloideans. The presence and variability of such ultrastructural characters according to family, superfamily or order have led several authors to propose their use in the analysis of trematode relationships and phylogeny. Therefore, apart from producing new data on the family Prosthogonimidae, the present study also compares the spermatological organization of M. jourdanei with other available ultrastructural studies focusing on the Microphalloidea.
Spermiogenesis and the ultrastructural organisation of the spermatozoon of the trypanorhynch cestode Aporhynchus menezesi Noever, Caira, Kuchta et Desjardins, 2010 are described by means of transmission electron microscopy. Type I spermiogenesis of A. menezesi starts with the formation of a differentiation zone containing two centrioles separated by an intercentriolar body constituted by five electron-dense plates. Each centriole gives rise to a free flagellum, which grows at an angle of 90° in relation to a median cytoplasmic process. The nucleus and cortical microtubules elongate along the spermatid body. Later, both flagella rotate and fuse with the median cytoplasmic process. At the final stage of spermiogenesis, the young spermatozoon is detached from the residual cytoplasm by a narrowing of the ring of arched membranes. The mature spermatozoon is a long and filiform cell, tapered at both ends, lacking mitochondria. It is characterized by the presence of two axonemes of the 9+'1' trepaxonematan pattern, the absence of crested bodies, the presence of parallel cortical microtubules and nucleus. This pattern corresponds to the type I spermatozoon of the eucestodes. The anterior extremity of the spermatozoon is characterized by the presence of an arc-like row of up to seven parallel cortical microtubules that partially surrounds the first axoneme. These anterior cortical microtubules are thicker than the posterior microtubules and, consequently, the sperm cell of A. menezesi exhibits two types of cortical microtubules. Another interesting aspect is the presence of α-glycogen rosettes. This spermatological pattern is similar to that observed in the spathebothriidean and diphyllobothriidean cestodes.
Ultrastructural characters of spermiogenesis and mature spermatozoon of Triaenorhina rectangula (Fuhrmann, 1908) are examined by transmission electron microscopy. Spermiogenesis follows the Bâ and Marchand's Type III spermiogenesis of cestodes. The process begins with the formation of a differentiation zone containing two centrioles and a cytoplasmic protrusion. The centrioles are associated with vestigial striated roots. One of the centrioles develops a free flagellum externally to the cytoplasmic protrusion. After a slight rotation, the free flagellum fuses with the cytoplasmic protrusion. In the final stage of spermiogenesis, a single crested body appears in the anterior part of the differentiating spermatozoon. The anterior extremity of the mature spermatozoon is characterised by an apical cone and a single crested body. The axoneme is of the 9+''1'' trepaxonematan type. A periaxonemal sheath and electron-dense rods are described in some parts of the mature spermatozoon. The nucleus is electron-dense and spirally coiled around the axoneme. The cortical microtubules are spirally arranged at an angle of about 40° to the spermatozoon axis. The present results show that the ultrastructural characters of spermiogenesis and mature spermatozoon of T. rectangula resemble most closely those in taeniids and metadilepidids. The comparison of these results with the only previous spermiological description of a paruterinid species reveals differences relative to the occurrence of filamentous rods of electron-dense material versus intracytoplasmic walls in the mature spermatozoon that may reflect the polyphyletic character of the Paruterinidae.
Ultrastructural analysis revealed that the spermatozoon of Discocotyle sagittata (Leuckart, 1842) is composed of two parallel axonemes, mitochondrion, nucleus and cortical microtubules. The nucleus, which occupies a central/distal position and has an unusual crescent-shaped profile, is slightly shorter than the mitochondrial rod. The two axonemes, which are of unequal length, and the cortical microtubules (up to 68 forming a continuous ring in the principal region) extend almost the entire length of the spermatozoon. A fold of the plasma membrane creates a unilateral flange or undulating membrane. Epifluorescence microscopy indicated that spermatogenesis gives rise to clusters of 64 spermatids connected to a common cytophore. Spermiogenesis and the structure of the filiform sperm of D. sagittata conform to the typical polyopisthocotylean pattern.
The present study describes the ultrastructure of the mature spermatozoon of Lecithocladium excisum (Rudolphi, 1819) (Digenea: Hemiuroidea: Hemiuridae) from the stomach of the marine teleost Scomber japonicus Houttuyn (Scombridae) captured in the Atlantic Ocean, off Dakar (Senegal). The ultrastructural organization of the spermatozoon of L. excisum follows the general model described in most digeneans. It presents two axonemes of the 9+'1' pattern of the Trepaxonemata, nucleus, mitochondrion and parallel cortical microtubules, among other characters. However, some particularities of the spermatozoon of L. excisum are (i) the presence of a membranous ornamentation not associated with cortical microtubules in its anterior extremity, (ii) the presence of a very reduced number of cortical microtubules located only in the ventral side of the spermatozoon and (iii) the absence of several structures described in most digeneans such as spine-like bodies and cytoplasmic expansions.