Binucleate and biflagellate spermatozoa in Tricholepidion gertschi Wygodzynsky (Insecta, Zygentoma)

The phenomenon of sperm pairing is known from some species of the apterygotan insect order Zygentoma, and has been described as the close apposition of two sperm cells. When released from the testes, they are single cells; pairing taking place in the deferent ducts. In a study of the relic species Tricholepidion gertschi, Zygentoma, sperm pairing was found to be due to a true fusion of two partners along their entire sperm head regions. The spermatozoon thus formed has two acrosomes, two nuclei and two separate sperm tails. The biflagellate spermatozoon swims with coordinated movements of its two flagella only when the two flagella lie close together but is totally uncoordinated when separate. The spermatozoon is about 50 microm long, thus much shorter than those of related apterygotan species. The mechanism of sperm cell fusion is unclear, although it appears that a 55-nm wide layer of electron dense substance, here termed the peripheral lamina, may play a role in delimiting the extent of sperm fusion.     

The spermatozoon of the Old Endemic Australo‐Papuan and Philippine rodents – its morphological diversity and evolution

Breed, W.G. and Leigh, C.M. 2010. The spermatozoon of the Old Endemic Australo‐Papuan and Philippine rodents – its morphological diversity and evolution.—Acta Zoologica (Stockholm) 91 : 279–294 The spermatozoon of most murine rodents contains a head in which there is a characteristic apical hook, whereas most old endemic Australian murines, which are part of a broader group of species that also occur in New Guinea and the Philippines, have a far more complex sperm form with two additional ventral processes. Here we ask the question: what is the sperm morphology of the New Guinea and Philippines species and what are the trends in evolutionary changes of sperm form within this group? The results show that, within New Guinea, most species have a highly complex sperm morphology like the Australian rodents, but within the Pogonomys Division some species have a simpler sperm morphology with no ventral processes. Amongst the Philippines species, many have a sperm head with a single apical hook, but in three Apomys species the sperm head contains two additional small ventral processes, with two others having cockle‐shaped sperm heads. When these findings are plotted on a molecular phylogeny, the results suggest that independent and convergent evolution of highly complex sperm heads containing two ventral processes has evolved in several separate lineages. These accessory structures may support the sperm head apical hook during egg coat penetration.     

The spermatozoon of Eurasian murine rodents: Its morphological diversity and evolution

The murine rodents are the most speciose subfamily of mammals. Here the morphology of the spermatozoon, as determined by scanning and transmission electron microscopy of representative species from four Eurasian clades, is described. Much interspecific variability in all components of the spermatozoon was found to occur, although most species have a bilaterally flattened sperm head with a single apical hook of variable length and orientation. Ultrastructural observations indicate that this apical hook invariably contains a nuclear projection as well as a large extension of the subacrosomal cytoskeleton, as a perforatorium rostrally, and a complex asymmetrical acrosomal extension. These spermatozoa also have relatively long tails that are attached to the lower concave surface of the sperm head. Uniquely, in species in the Apodemus clade, the apical hook is orientated caudally. In a few species a highly derived sperm head morphotype that does not contain an apical hook is present. These sperm heads vary in morphology from being globular in two species of Bandicota, to bilaterally flattened and paddle-shaped in Tokudaia and Micromys. In spermatozoa of the latter two genera the subacrosomal cytoskeleton, which is less extensive than in species with a hooked sperm head, forms an apical extension, but that is not the case in Bandicota. In all species where the sperm head lacks an apical hook the acrosome is more symmetrical. The sperm tail is much shorter in these species, with attachment to the head occurring on the ventral surface in Tokudaia and basal in Micromys and the two species of Bandicota. As the sperm head morphotype with a complex apical hook is present in all the major clades of murine rodents, it is likely to be a plesiomorphic character within each of these clades, with the nonhooked sperm heads, which vary greatly in structure between species of the different lineages, probably being independently derived. The ultrastructural organization of the sperm head of Bandicota, but not those of Micromys or Tokudaia, suggest divergence in some of the morphological events associated with sperm-egg interaction at the time of fertilization.     

暗褐蝈螽与优雅蝈螽精子超微结构的比较研究(直翅目,螽斯科)

研究了暗褐蝈螽Gampsocleis sedakovii(Fischer von Waldheim)和优雅蝈螽G.gratiosa Brunner von Wattenwyl精子的超微结构。这两种蝈螽精子头部的顶体复合体由顶体外层、顶体本体和顶体组成,顶体复合体位于细胞核前端,并包裹部分细胞核;颈部具5纵层细胞器;尾部鞭毛轴丝为典型的9+9+2型,线粒体衍生体部分晶状化。暗褐蝈螽精子较短,顶体复合体夹角较大,精子鞭毛横切面直径稍大;优雅蝈螽精子稍长,顶体复合体夹角较小,精子鞭毛横切面直径较小,两种精子超微结构差异不显著,其生殖隔离机制有待进一步研究。     

Behavior of the spermatozoon during sperm-blastomere fusion and its significance for fertilization (Saccoglossus kowalevskii: hemichordata)

Summary Association of spermatozoa with blastomeres removed from the egg envelope was studied in order to investigate sperm membrane behavior under circumstances unlike those attending fertilization of normal envelope-enclosed eggs. Under the altered circumstances all parts of the surface of the sperm cell were afforded the opportunity of meeting blastomere plasma membrane. Nevertheless, in each case the spermatozoon first became activated, an acrosomal tubule formed, and it was this organelle alone which fused and established continuity with the blastomere plasma membrane. This behavior of the spermatozoon with respect to the denuded blastomere parallels the behavior of the sperm cell with respect to the envelope-enclosed egg at fertilization. Since the sperm cell behaves so similarly in two situations which are so different with respect to what the spermatozoon encounters, this behavior is considered to reflect the inherent nature of the spermatozoon, and analysis of this behavior in relation to a blastomere is considered to be valid also in relation to an egg during fertilization. It is concluded that membrane fusion, involving the acrosomal tubule of an activated spermatozoon, is the only means by which gametic union is established and that these are obligatory rather than fortuitous features of sperm-egg association in Saccoglossus. It is suggested that these features are probably obligatory also in other species which exhibit this pattern of sperm-egg association.This investigation was supported by Research Grant HD-00007 from the National Institutes of Health, United States Public Health Service.     

Ultrastructure of spermiogenesis and synthesis of enigmatic cytoplasmic inclusions in the spirally shaped spermatozoon of the polychaete Spio setosa (Annelida: Spionidae)

The sperm of Spio setosa (Polychaeta, Spionidae) are known to be very unusual in form; here, spermiogenesis and the structure of the spermatozoon in this species are described by transmission electron microscopy. While spermiogenesis is similar to that described for many other polychaetes, two notable exceptions to this process include the synthesis of abundant ring‐shaped and tubular, membrane‐bounded cytoplasmic inclusions in the midpiece, and the differentiation of a spirally shaped sperm head. Spermatids develop as free‐floating tetrads in the male's coelom. A microtubular manchette does not develop during chromatin condensation and nuclear elongation, and the spiral acrosome forms as a single Golgi‐derived vesicle that migrates anteriorly to become housed in a deep anterior nuclear fossa. Early in spermiogenesis, numerous Golgi‐derived, membrane‐bounded cytoplasmic inclusions appear in the cytoplasm; these ultimately occupy the sperm midpiece only. The mature spermatozoon in the male has a 15‐μm‐long head consisting of a nucleus coiled like a spring and a spiral acrosome with differentiated substructure, the posterior two thirds of which sits in an anterior nuclear fossa. The midpiece is wider than the rest of the spermatozoon and contains 9–10 spherical mitochondria surrounding the two centrioles, as well as numerous membrane‐bounded conoid and tubular cytoplasmic inclusions. The axoneme has a 9 + 2 arrangement of microtubules. By contrast, stored sperm in the female's seminal receptacles have lost the midpiece inclusions but contain an abundance of glycogen. The function of the midpiece inclusions remains unresolved, and the significance of their absence in stored sperm within the seminal receptacle and the appearance of midpiece glycogen stores remains unclear and requires additional investigation.     

Intracytoplasmic injection of porcine, bovine, mouse, or human spermatozoon into porcine oocytes

We determined the incidence of activation, male pronuclear formation, and apposition of pronuclei in porcine oocytes following intracytoplasmic injection of various porcine sperm components and foreign species spermatozoa, such as that of cattle, mouse or human. The porcine oocytes were activated by injection of a spermatozoon or an isolated sperm head. In contrast, injection of either sperm tail or a trypsin- or NaOH-treated sperm head failed to induce oocyte activation. Because injection of mouse, bovine, or human spermatozoon activated porcine oocytes, the sperm-borne activation factor(s) is not strictly species-specific. Male pronuclear formation and pronuclear apposition were observed in porcine oocytes following injection of porcine, bovine, mouse or human spermatozoa. Electrical stimulation following sperm cell injection did not enhance the incidence of male pronuclear formation or pronuclear apposition compared with sperm cell injection alone (P > 0.1). Following porcine sperm injection, the microtubular aster was organized from the neck of the spermatozoon, and filled the whole cytoplasm. In contrast, following injection of bovine, mouse, or human spermatozoon, the maternal-derived microtubules were organized from the cortex to the center of the oocytes, which seems to move both pronuclei to the center of oocytes. Cleavage to the two-cell stage was observed at 19-21 hr after injection of porcine spermatozoon. However, none of the oocytes following injection of mouse, bovine, or human spermatozoa developed to the mitotic metaphase or the two-cell stage. These results suggested that the oocyte activating factor(s) is present in the perinuclear material and that it is not species-specific for the porcine oocyte. Self-organized microtubules seemed to move the pronuclei into center of oocytes when foreign species spermatozoa were injected into porcine oocytes.     

THE SPERMATOZOON OF ARTHROPODA: ZOOTERMOPSIS NEVADENSIS AND ISOPTERAN SPERM PHYLOGENY

In this paper, the ultrastructure of the spermatozoon of Zootermopsis nevadensis (Isoptera, Hodotermitidae) and of some Rhinotermitidae and Termitidae is described. Zootermopsis sperm is rod like, aflagellate, immotile, and without an acrosome; it is composed of a filiform nucleus encircled by a monolayered microtubular manchette, and a few mitochondria. This spermatozoon was previously thought to be flagellate, and therefore the most primitive in Isoptera: our present study suggests a new phylogenetical position for Hodotermitidae. All the species of Rhinotermitidae and Termitidae studied by us show a similar spheroidal sperm model, devoid of acrosome, flagellum and manchette at spermatid stage, and are made up of only a round nucleus, two mitochondria and a centriole. This widely distributed model seems to be the more evolved in the order. The nature of sperm evolution in the Isoptera is considered.     

Ultrastructure of the spermatozoon of Centroderma spinosissima (Stossich, 1886) (Digenea: Mesometridae) and its phylogenetic potential

The Mesometridae includes only five genera and eight species. The available data on the ultrastructure of sperm cells of mesometrid species referred to two species only, Elstia stossichianum and Wardula capitellata. The present study revealed the ultrastructure of the spermatozoon of a third genus and third species of Mesometridae, Centroderma spinosissima. The mature spermatozoon of C. spinosissima presents two axonemes with different lengths of the Ehlers’ 9 + ‘1’ trepaxonematan pattern, a nucleus, two mitochondria, two bundles of parallel cortical microtubules, external ornamentation of the plasma membrane, a lateral expansion, spine-like bodies, cytoplasmic ornamented buttons and granules of glycogen. The spermatozoon of C. spinosissima is similar to those of the previously studied mesometrids. However, some peculiarities such as the presence of two mitochondria, the disposition of the external ornamentation of the plasma membrane and the morphology of the posterior spermatozoon extremity, characterize the male gamete of C. spinosissima. Moreover, the presence of cytoplasmic ornamented buttons is a characteristic found only in the mature spermatozoon of mesometrids and it probably represents an autapomorphy for this family.     

Trypsin-like enzymes during fertilization in the shrimp Rhynchocinetes typus

The spermatozoon of Rhynchocinetes typus is atypical because it is nonmotile and lacks head and tail. The body has a rigid spike. Neither an acrosome-like structure nor changes during gamete interaction which could be interpreted as an acrosome reaction have been observed in this species. Nevertheless, the spermatozoon exerts a lytic effect on the extracellular envelope of the egg, and in this way it penetrates through egg-coats, forming a channel. In this research we found that crude spermatozoa extracts analyzed by gelatin SDS-PAGE developed one band of protease activity that was completely inhibited by SBTI (soybean trypsin inhibitor) and pAB (p-aminobenzamidine). In sperm extracts an enzymatic activity was determined, using BAEE (N-benzoil-L-arginine ethyl ester), but not ATEE, as substrate. This activity was inhibited by SBTI and pAB. We observed that in vitro fertilization was inhibited by spermatozoon incubation with the trypsin inhibitors SBTI, PMSF (phenyl-methanesulphonyl fluoride), and pAB. Additionally, we observed that when whole isolated egg-coats were incubated with sperm extract and then analyzed by SDS-PAGE, one band of the egg-coats disappeared. These results have been interpreted as sperm trypsin-like enzyme participation in R. typus sperm passage through the egg-coats. Mol. Reprod. Dev. 46:581–586, 1997. © 1997 Wiley-Liss, Inc.     

Morphological observations on sperm–egg interactions during in vivo fertilization in the dasyurid marsupial Sminthopsis crassicaudata

The aim of the present study was to determine the morphological changes that take place in the male and female gametes during in vivo fertilization in the Australian marsupial, the fat-tailed dunnart, Sminthopsis crassicaudata. Plastic sections were cut of sperm and eggs recovered from the oviducts of recently mated individuals, and light microscopy of thick, and transmission EM of thin, sections was carried out. It was found that, before penetration of the zona, the spermatozoon came to lie along the outer surface with its rostral tip forming a depression in the zona substance. During penetration, zona material was packed tightly around the spermatozoon, and no large hole was formed. A spermatozoon within the perivitelline space had made contact with the oolemma by way of its apical tip. In a spermatozoon partly incorporated into the ooplasm, fusion appeared to have taken place between its plasma membrane and that of the oolemma. Mucoid coat material became deposited outside the zona at this time; its existence and/or the release of cortical granule content probably prevented polyspermy. Once inside the egg cytoplasm, the sperm head sometimes travelled a considerable distance before chromatin decondensation occurred. In addition, it appeared to rotate somewhat on its axis at this time. Finally, some membranous structures were found around two condensed sperm heads in the ooplasm, which may have been part of the pronuclear envelope. Thus this study on in vivo fertilization in the dunnart documents, for the first time, some aspects of fertilization in an Australian marsupial as seen with the transmission electron microscope; it indicates a few differences from those previously found for the American opossum.     

Ultrastructure comparée de la spermiogenèse et du spermatozoïde de trois espèces de Paravortex (Rhabdocoela, "Dalyellioida", Graffillidae), Turbellariés parasites intestinaux de Mollusques

Three species of the genus Paravortex were studied. Spermiogenesis involves progressive lengthening of the spermatid. No centriole nor axoneme were found. Thc mature spermatozoon is threadlike, 30μm long and aflagellate. It contains a nucleus, two longitudinal parallel mitochondria, numerous dense bodies limited by a membrane, a row of cortical microtubules and granules. The spatial arrangement of the microtubules wah deduced from a statistical analysis of their number in transverse section. The microtubules are continuous along the length of the spermatozoon and are arranged along a long cone. The granules, not limited by a membrane, are about 25 nm in diameter. They are arranged in a semicircle and represent a new structure for sperm of Platyhclminthes. A statistical comparison demonstrated that two species may be distinguished by the number of their cortical sperm microtubules. This is the tirst ultrastructural study of an aflagellate spermatozoon in the Rhabdocoela. A rickettsia was found in P. cardii .     

Sperm structure and phylogeny of Astigmata

The Astigmata, a large and variable group, is still a subject of taxonomic dispute. Particularly, their origin from ancestors of the lower oribatid mites (e.g., Malaconothroidea) seems well documented by many lines of evidence. The structure of spermatozoa has been successfully applied to phylogenetic investigations in many animal groups. The aim of our study was to provide new data on spermatozoon structure in Astigmata and to consider its appropriateness in phylogenetic studies. The study reveals information on spermatozoa in 17 species of Astigmata (11 species studied for the first time) extending our knowledge to 18 species (one species known only from the literature) representing 12 families and 7 superfamilies. Spermatozoa have the same basic structure in all species: cells are multiform and the chromatin forms thin threads embedded directly in the cytoplasm; the acrosome is absent. The cytoplasm in most species contains electron-dense lamellae, varying in both number and arrangement within the cell. In Sarcoptoidea, electron-dense tubules in contact with lamellae margins were also observed in Psoroptidae (Psoroptes equi), whereas in two representatives of Sarcoptidae (Notoedres cati and Sarcoptes scabiei), only electron-dense tubules were found. In two species, Canestrinia sellnicki (Canestrinioidea: Canestriniidae) and Scutulanyssus obscurus (Analgoidea: Pteronyssidae), neither lamellae nor tubules were present. The mitochondria in a spermatozoon are usually gathered at the cell periphery and their structure is usually modified to form so-called mitochondrial derivatives. The chromatin threads are an autapomorphy strongly supporting the monophyly of Astigmata. As spermatozoa vary considerably between species in Astigmata, we deduce that sperm structure may be useful for phylogenetic analyses within the group. Several conclusions concerning the affinities within Astigmata are presented. Spermatology seems to be unhelpful, however, in questions on the origin of Astigmata (particularly for Astigmata-Oribatida relationships), since their sperm do not possess synapomorphies with sperm of the remaining groups of Acariformes, i.e., Endeostigmata, Prostigmata, and Oribatida.     

Ultrastructural studies of spermatozoa in three bivalve species with notes on evolution of elongated sperm nucleus in primitive spermatozoa

Sperm ultrastructure and spermiogenesis of the three bivalve species Musculus discors, Nucula sulcata, and Dreissena polymorpha have been studied. During spermatid differentiation in Musculus discors and Nucula sulcata the nucleus attains an elongated rod-like shape. The spermatozoon from Nucula sulcata was found to have a cup-shaped acrosome and five mitochondria surrounding two centrioles in the middle piece. The spermatozoa from Musculus discors has a long complex acrosome. From the distal centriole striated processes extend and attach to the plasma membrane. The spermatozoon of the fresh water species Dreissena polymorpha agrees in all main features with those of other invertebrate groups with external fertilization. It is thus of the primitive type with barrel-shaped nucleus and four to five mitochondria1 spheres in the middle piece. The acrosome is a prominant, complex structure at the apex of the mature spermatozoon. A comparison of sperm ultrastructure among bivalves indicates that there is a certain correlation between the evolution of the elongated sperm nucleus and large, yolk-rich eggs. In species with an elongated sperm nucleus the increased egg size has often led to a lecithotrophic or direct development. The elongated nucleus is a slight modification of the primitive type. There is a great variation in acrosome structure among bivalve spermatozoa, reflecting diverging functional demands at fertilization of the eggs.     

Interspecific variation of sperm morphology in the Australian rodent genus Zyzomys

The sperm head morphology and tail length of two species of Australian rock rats, Zyzomys argurus and Zyzomys pedunculatus, are presented. In Z. argurus the sperm head has an apical hook together with two ventral processes extending from the upper concave surface that are largely composed of cytoskeletal material, and the sperm tail is about 135 µm in length. By contrast, in Z. pedunculatus the sperm head is paddle‐shaped with the nucleus capped by an acrosome that has a large apical segment and is surrounded by a thin layer of cytoskeletal material, and the sperm tail is only around 85 µm in length. Since the structure of the spermatozoon of Z. argurus is similar to that of most of the old endemic Australian rodents it is presumed to be the ancestral condition within the Zyzomys genus with that of Z. pedunculatus being highly derived and showing convergence with the sperm structure in some other orders of mammals.     

Immotile,aflagellate spermatozoa in Ptiliidae coleopterans

The first aflagellate and immotile Coleopteran spermatozoon is described in a group of species belonging to the family Ptiliidae. The mature spermatozoon is devoid of flagellum, centrioles and mitochondria, includes a three-layered acrosomal complex (extraacrosomal layer, acrosome and perforatorium) and a long nucleus made up of two regions of different densities. A compact submembranary capsule and a thick glycocalyx are also present. Motility organelles are absent during the whole spermiogenesis, which is not regressive. The new structures peculiar for this type of sperma are designed for protection. No other sperm models with these characteristics have been described so far.     

The Spermatozoon in Tardigrades: Evolution and Relationships with the Environment

The spermatozoon ultrastructure of seven species of tardigrades, one heterotardigrade echiniscid and six eutardigrades, was analysed and described. Tardigrade species were collected from freshwater sediments, moss and leaf litter. Aphylogenetic evaluation was made and relationships of spermatozoon shape and habitat were discussed. The spermatozoon morphology of the moss-dwelling echiniscid(heterotardigrade) was very similar to that found in marine species of the same order. This indicates that in Echiniscoididae change of habitat had little influence on sperm structural organisation. In contrast, eutardigrade spermatozoa have developed several different structural organisations that may be related to family and/or habitat.     

Spermiophagic activity in the female genital system of some species of terrestrial isopods (Crustacea,Halophilosciidae)

The females of some species of the family Halophilosciidae receive in the course of mating a quantity of sperm considerably redundant with respect to the number of eggs that can be fertilized; this is possible thanks to the peculiar morpho-functional organization that characterizes their genital system and that allows them to store the sperm not only in the great seminal receptacle but also within the ovary. While most of the sperm stay free in the lumen of the seminal receptacle, a part of those present in the ovary undergoes a process of capture by the follicular cells with consequent internalization within endocellular cavities. This process concerns exclusively the immotile tail, that characterizes the peculiar spermatozoon of the isopods and which is essentially of proteic nature. After their capture the sperm tails undergo a gradual process of digestion, which seems to be apparently realized without the intervention of lysosomes. The possible role of this spermiophagic activity might be to represent a significant trophic paternal investment aimed at improving the fitness of the female and of the offsprings.     

Oocyte activation: lessons from human infertility

During fertilization, the spermatozoon penetrates through the cumulus cells and the zona pellucida that surrounds the oocyte, before it binds and fuses with the oocyte plasma membrane to induce activation. In vitro fertilization (IVF) studies performed in non-human mammals have contributed extensive knowledge regarding the mechanisms by which the spermatozoon activates the meiotic-arrested oocyte to resume meiosis, cleave and develop into an embryo. Although IVF has been used extensively for treating subfertile couples, not all of them were able to benefit from this procedure. In intracytoplasmic sperm injection (ICSI), one viable spermatozoon only is sufficient for successful fertilization of a single oocyte. Moreover, the injected fertilizing spermatozoon bypasses several physiological barriers, compared with IVF, which together could explain the high success rate for this procedure. ICSI has also allowed the identification of sperm components that are required for successful fertilization.     

Ultrastructural studies on the hypothalamic neurosecretory neurons of the rat

The spermatozoa of two closely related species of ophiuroids, Ophiocoma echinata and Ophiocoma wendti, were examined ultrastructurally. Morphologically, these spermatozoa resemble those of other non-echinoid echinoderms. The acrosomal complex, completely contained within an anterior fossa in the spherical nucleus, consists of a membrane-limited acrosomal vesicle and periacrosomal material. Events of the acrosomal reaction in O. echinata and O. wendti are presented. In both species, the reaction results in the establishment of an extracellular coat of acrosomal vesicle origin on the anterior surface of the spermatozoon. The possible role of this extracellular coat in the species-specific binding of sperm and ova is discusses. The origin of acrosomal tubule membrane is elucidated.