Fine structure of spermiogenesis with special reference to the spermatid morulae of the freshwater mussel Prisodon alatus (Bivalvia,Unionoidea)

Within the testicular cysts of the mussel Prisodon alatus are numerous somatic host cells described as Sertoli cells (SC), each containing a variable number of young spermatid morulae. Among them, several free spermatid morulae, spermatids, and spermatozoa were observed. Each free spermatid morula is surrounded by an external membrane. The early spermatids enclosed within the morulae have dense and homogeneous chromatin, and the cytoplasm occupies little space around the nucleus. Later, during spermiogenesis, the SC show lysis and disrupt to liberate the spermatid morulae. The membrane of the free morula is then disrupted, releasing the young spermatids. The SC disappear just after the appearance in the testis of a large number of free young spermatids. The nucleus of each free spermatid becomes gradually smaller and denser by the appearance of a granular pattern of condensed chromatin. During the maturation phase of the spermatids, the cytoplasm becomes more voluminous, and mitochondria and centrioles are more evident. Then, flagellogenesis occurs, and the nucleus gradually condenses into thicker strands. In the mature sperm, the apical zone has a disc-shaped acrosomal vesicle and the midpiece contains five mitochondria and two centrioles located at the same level. The flagellum has the common 9+2 microtubular pattern. The results are discussed with particular reference to Sertoli cells and clusters of spermatid morulae with those of species of closely related taxa in the bivalves. J. Morphol. 238:63–70, 1998. © 1998 Wiley-Liss, Inc.     

A scanning electron microscopical study of sperm development and activation in Arenicola marina L. (Annelida: Polychaeta)

Abstract

The lugworm, Arenicola marina L. has an annual cycle of reproduction with epidemic spawning and external fertilisation. The spermatozoa of Arenicola are unusual in that they are held immotile (as plates of several hundred cells known as morulae) in the coelomic fluid until activated just prior to spawning. Activation of Arenicola sperm is brought about by a sperm maturation factor (SMF) from the prostomium and can be carried out in vitro using an assay technique developed by Bentley (1985). Scanning electron microscopy is used here to examine the changes which occur during in vitro activation. This revealed that the bundles of flagella of inactive sperm become disorganised as flagella beating commences but the flagella at this stage are still bound together at their tips. The sperm heads then become separated from the cytophore and finally the distal binding of the flagella is broken to give free-swimming spermatozoa. Coelomocytes present in the coelomic fluid resorb unspawned gametes prior to the initiation of the next gametogenic phase.     

Ultrastructure of spermiogenesis in <Emphasis Type="Italic">Cristatella mucedo</Emphasis> Cuvier (Bryozoa: Phylactolaemata: Cristatellidae)

In Cristatella mucedo spermiogenesis occurs in a morula consisting of a large number of spermatids connected with a central cytophore. The mature sperm cell is filiform and consists of a head, a midpiece and a tail region, the latter two separated by a deep circular constriction. The comparatively short head contains a drop-shaped, bilaterally symmetrical and pointed nucleus capped by a minute acrosome. The single centriole is placed in a deep posterior invagination of the nucleus followed by the axoneme with the typical 9 + 2 pattern. The elongated midpiece is 0.9–1.1 μm thick and contains several helices of mitochondria surrounding the axoneme. The tail is thicker (1.3 μm) and richer in cytoplasm with many compact accumulations of an electron-dense substance lying peripherally and another less dense material wrapped around the axoneme. The course of the spermiogenesis and the fine structure of the sperm are very similar to that of Plumatella fungosa. Comparison with other species shows that the same sperm type is recognizable in four of the five families of Phylactolaemata and, provided it occurs also in the fifth family, the Stephanellidae, is a synapomorphy of the entire class.     

Electron microscopic study of spermatogenesis in the lung fluke (Paragonimus miyazakii)

Summary The characteristics of spermatogenesis in a type of pulmonary parasite, Paragonimus miyazakii have been observed using the electron microscope. Groups of several spermatocytes revealed mutual cytoplasmic connection. That degree of this fusion increased as spermatogenesis progressed, and finally developed into a so-called cytophore. Then, this cytophore remained joined with a spermatid by a short stalk until the spermatid changed into a sperm. The nucleus of the spermatid became elongated with a string-like arrangement of the chromatin, which, in turn, showed increased electron density. At the pole of the spermatid, linearly arranged microtubules developed just below the plasma membrane. Close to an elongated portion at the pole, two separate flagella start growing and later fuse with the sperm itself. In the sperm tail a couple of tail filament complexes, longitudinally oriented slender mitochondria, and a tubular structure were present.     

Spermatogenesis in Fasciola hepatica: an ultrastructural comparison of the effects of the anthelmintic,thiabendazole (“Fasinex”) and the microtubule inhibitor,tubulozole

Summary

The normal sequence of spermatogenesis in Fasciola hepatica has been established. The single-celled primary spermatogonium—after three mitotic divisons and one meiotic division, accompanied by incomplete cytokinesis—gives rise to a rosette of 32 spermatid cells from which the spermatozoa differentiate. Rosette formation begins to develop at the tertiary spermatogonial stage and leads to individual cells being joined together via a central “cytophore”. As revealed by transmission electron microscopy (TEM), spermatogenesis is readily disrupted by the active sulphoxide metabolite of triclabendazole (TCBZSX), a relatively new benzimidazole anthelmintic. Following short incubation times in this drug (3 h onwards), the spermatogonial stages become mitotically inactive and detached from their normal position close to the wall of the tubule. The rosette stages become increasingly disrupted, with fragmentation of the cytoplasmic cytophore. After longer time periods (12–24 h) the testis tubules were almost completely empty with few spermatogenic stages or mature spermatozoa present. The ultrastructural effects of TCBZ-SX bear some resemblance to those observed after treatment with tubulozole, a potent microtubule inhibitor. The results with the two drugs are discussed in relation to the mode of action of triclabendazole, which at present is unknown.     

COMPARATIVE SPERMATOLOGY OF THREE SPECIES OF DONAX (BIVALVIA) FROM SOUTH AFRICA

The fine structure of the sperm and spermatogenesis in threespecies of Donax (D. madagascariensis, D. sordidus and D. serra)are described. Although the morphology of the sperm of all speciesis very similar, each has unique features. Donax madagascariensisand D. sordidus reportedly hybridize in regions of sympatryand their spermatozoa are morphologically closer to one anotherthan to D. serra. All sperm are of the primitive type with ahead(about 2 µmu; long), mid-piece of four mitochondria andtail. The head comprises a barrel-shaped nucleus which is cappedby a small, complex acrosome. The structure of the acrosomeis typical of heterodont bivalves. During spermatogenesis thepattern of nuclear chromatin condensation is granular. Glycogenfirst appears in the cytoplasm of spermatids, and in the maturesperm is sited in the mid-piece and base of the acrosome. (Received 15 May 1989; accepted 25 June 1989)     

Identification and distribution of actin in spermatogenic cells and spermatozoa of the rabbit

Using a monoclonal antibody as a highly specific probe and a seminal particle-free fraction of rabbit ejaculated spermatozoa, actin has been localized in the postacrosomal region of mature rabbit spermatozoa. The sperm actin has been extracted and identified on two-dimensional PAGE immunoblots as a single spot of pI = 5.45 and Mr = 43,000. Rabbit sperm actin is present in a nonfilamentous form and is not removed by removing the plasma membrane. Unlike mature spermatozoa, however, filamentous actin is present in spermatogenic cells, as determined by rhodamine phalloidin staining. Starting as diffusely distributed in spermatocytes, actin accumulates in the subacrosomal space and appears as a band in conjunction with the developing acrosome. This band lengthens throughout the spermatid stage and becomes continuous with the postacrosomal region staining in testicular spermatozoa. Actin may therefore function during spermatogenesis to both shape the acrosome to the nucleus and to anchor inner acrosomal membrane proteins.     

Spermatogenesis in Boccardiella hamata (Polychaeta: Spionidae) from the Sea of Japan: sperm formation mechanisms as characteristics for future taxonomic revision

Reunov, A.A., Yurchenko, O.V., Alexandrova, Y.N. and Radashevsky, V.I. 2009. Spermatogenesis in Boccardiella hamata (Polychaeta: Spionidae) from the Sea of Japan: sperm formation mechanisms as characteristics for future taxonomic revision. —Acta Zoologica (Stockholm) 91 : 477–456. To characterize novel features that will be useful in the discussion and validation of the spionid polychaete Boccardiella hamata from the Sea of Japan, the successive stages of spermatogenesis were described and illustrated. Spermatogonia, spermatocytes and early spermatids are aflagellar cells that develop synchronously in clusters united by a cytophore. At the middle spermatid stage, the clusters undergo disintegration and spermatids produce flagella and float separately in coelomic fluid as they transform into sperm. Spermatozoa are filiform. The ring‐shaped storage platelets are located along the anterior nuclear area. The nucleus is cupped by a conical acrosome. A nuclear plate is present between the acrosome and nucleus. The nucleus is a cylinder with the implantation fossa throughout its length and with the anterior part of the flagellum inside the fossa. There is only one centriole, serving as a basal body of the flagellum, situated in close vicinity of the acrosomal area. A collar of four mitochondria is located under the nuclear base. The ultrastructure of B. hamata spermatozoa from the Sea of Japan appears to be close to that of B. hamata from Florida described by Rice (Microscopic Anatomy of Invertebrates, Wiley‐Liss, Inc., New York, 1992), suggesting species identity of the samples from the two regions. However, more detailed study of Florida’s B. hamata sperm is required for a reliable conclusion concerning the similarity of these two polychaetes. In addition to sperm structure, features such as the cytophore‐assigned pattern of spermatogenic cell development, the synchronous pattern of cell divisions, the non‐flagellate early spermatogenic stages, and the vesicle amalgamation that drives meiotic cell cytokinesis and spermatid diorthosis will likely be useful in future testing of the validity of B. hamata and sibling species throughout the world.     

Development and submicroscopic anatomy of male gametes in Halammovortex nigrifrons (Plathelminthes, Rhabdocoela, ”Dalyellioida”): phylogenetic implications and functional aspects

The steps of spermiogenesis and the submicroscopic anatomy of male gametes in Halammovortex nigrifrons are described. During spermiogenesis the cytophore develops pseudopod-like extensions, and bung-like deposits of dark material become attached to the basal bodies of the cilia. During the phase of cell elongation, cilia stay near the edge of the cytophore. Spermatozoa bear two free cilia or flagella. The axonemata are equipped with glycogen islets appearing at regular spaces. The sperm body is characterized by dot-like dense granules linearly arranged, intense glycogen aggregations in a channel-shaped deposition and giant dense bodies. Events of spermiogenesis and the features of mature male gametes in H. nigrifrons corroborate the hypothesis of the existence of a monophylum within the Rhabdocoela encompassing several, but not all taxa of the ”Typhloplanoida” and ”Dalyellioida”. The Dalyelliidae (including the species of the Temnocephalida) belong to this monophylum.     

Poor centrosomal function of cat testicular spermatozoa impairs embryo development in vitro after intracytoplasmic sperm injection

In the domestic cat, morula-blastocyst formation in vitro is compromised after intracytoplasmic sperm injection (ICSI) with testicular compared to ejaculated spermatozoa. The aim of this study was to determine the cellular basis of the lower developmental potential of testicular spermatozoa. Specifically, we examined the influence of sperm DNA fragmentation (evaluated by TUNEL assay) and centrosomal function (assessed by sperm aster formation after ICSI) on first-cleavage timing, developmental rate, and morula-blastocyst formation. Because the incidences of DNA fragmentation were not different between testicular and ejaculated sperm suspensions, DNA integrity was not the origin of the reduced developmental potential of testicular spermatozoa. After ICSI, proportions of fertilized and cleaved oocytes were similar and not influenced by sperm source. However, observations made at 5 h postactivation clearly demonstrated that 1) zygotes generally contained a large sperm aster after ICSI with ejaculated spermatozoa, a phenomenon never observed with testicular spermatozoa, and 2) proportions of zygotes with short or absent sperm asters were higher after ICSI with testicular spermatozoa than using ejaculated spermatozoa. The poor pattern of aster formation arose from the testicular sperm centrosome, which contributed to a delayed first cleavage, a slower developmental rate, and a reduced formation of morulae and blastocysts compared to ejaculated spermatozoa. When a testicular sperm centrosome was replaced by a centrosome from an ejaculated spermatozoon, kinetics of first cell cycle as well as embryo development quality significantly improved and were comparable to data from ejaculated spermatozoa. Results demonstrate for the first time in mammals that maturity of the cat sperm centrosome (likely via epididymal transit) contributes to an enhanced ability of the spermatozoon to produce embryos that develop normally to the morula and blastocyst stages.     

Ultrastructure of sperm development in the free-living marine nematode Metachromadora itoi (Chromadoria, Desmodorida)

The spermatogenesis of the free‐living marine nematode Metachromadora itoi was studied with electron microscopy. Spermatocytes and early spermatids have no cytoplasmic components specific for nematodes, i.e. membranous organelles (MO) and fibrous bodies (FB). The late spermatids are subdivided into the residual body and the main cell body with a centrally located nucleus devoid of a nuclear envelope. A pair of 9 × 2 centrioles is associated with the nuclei of spermatids and spermatozoa. The nucleus of the mature spermatid is surrounded by a thick mass of radially arranged FB delimited externally by a discontinuous layer of mitochondria, which underlie a thin ectoplasm. Sperm development is accompanied by transfer of FB matter through the mitochondrion layer into the ectoplasm. The immature spermatozoa from the testis have the centrally located nucleus surrounded by a transparent halo with remnants of FB. The halo is delimited by a sphere of mitochondria that underlie the thick fibrous ectoplasm, a derivative of the FB. In the mature spermatozoa the ectoplasm is transformed into the prominent unpolarized pseudopod. The central nucleus is surrounded by a transparent halo and a sphere of mitochondria, which underlie the pseudopod. MO were not found throughout spermatogenesis. In general, spermatogenesis in M. itoi differs from that observed in many nematodes but resembles in some details the sperm development in some chromadorid and tylenchomorph nematodes. The phylogenetic importance of this sperm development is discussed.     

Ultrastructure of sperm development and mature sperm morphology in three species of commensal bivalves (Mollusca: Galeommatoidea)

Ultrastructural features of the ovotestes, spermatogenesis, and the mature sperm are described for three galeommatid bivalves, Divariscintilla yoyo, Divariscintilla troglodytes, and Scintilla sp., from stomatopod burrows in eastern Florida. All three species yielded similar results except with respect to mature sperm dimensions. The ovotestis contains three types of somatic cells within the testicular portion: flattened myoepithelial cells defining the outer acinal wall; underlying pleomorphic follicle cells containing abundant glycogen deposits; and scattered, amoeboid cells containing lysosomal-like inclusions which are closely associated with developing sperm. Early spermatogenesis is typical of that reported from other bivalves. In contrast, the late stages of spermiogenesis involve the migration and gradual rotation of the acrosomal vesicle, resulting in a mature acrosome tilted about 70° from the long axis of the cell. The mature sperm possesses an elongated, slightly curved nucleus; a subterminal, concave acrosome with a nipple-like central projection; five spherical mitochondria and two centnoles in the middlepiece; and a long flagellum. The rotational asymmetry and the presence of perimitochondrial glycogen deposits in these sperm are unusual in the Bivalvia and may be associated with fertilization specializations and larval brooding common among galeommatoideans.     

Spermiogenesis and modified sperm morphology in the "seepworm" Methanoaricia dendrobranchiata (Polychaeta: Orbiniidae) from a methane seep environment in the Gulf of Mexico: implications for fertilization biology

Spermatogenesis and mature sperm morphology have been described along with limited observations of the ovary in Methanoaricia dendrobranchiata, an orbiniid polychaete associated with dense populations of the mussel Bathymodiolus childressi at brine pools on the Louisiana slope, Gulf of Mexico. The species is gonochoric with gonads serially repeated in numerous segments and each associated with a nexus of blood vessels at the base of the parapodia. In the female, synchronous, intraovarian egg development occurs with the release from the ovary of large, yolky eggs into the coelom at first meiotic metaphase. Sperm develop in the coelom as free-floating, plasmodial clones interconnected via an anuclear cytophore. At the end of spermiogenesis, mature spermatozoa float freely in the coelom. The mature spermatozoon differs significantly from that of shallow-water orbiniid species by possessing an elongated nucleus and a greatly elongated and curved acrosome reaching 19.5 microm in length. The spermatozoon resembles an ent-aquasperm and may not fertilize the eggs directly in seawater in the classical manner. We hypothesize that the unusual spermatozoon morphology in this species has evolved due to the hypoxic environment in which the adults live and that fertilization biology is likely modified in some way to minimize sperm exposure to high levels of hydrogen sulfide. An analysis of life-history features in shallow-water orbiniids is used to infer reproductive features in M. dendrobranchiata that could not be directly documented.     

Differentiation of binuclear spermatozoa in mice

In an electron microscopy study of abnormal spermatogenesis in mice, we have found that two discrete haploid nuclei may be located in a single spermatid cytoplasm after the second meiotic division. The spermatid continues to differentiate and forms a binucleate spermatozoon with both nuclei separately packaged within the sperm head. The Golgi apparatus of the double spermatid forms a single proacrosome that attaches to both nuclei. Apparently, one acrosomal structure differentiates to cover and compartmentalize the two haploid nuclei within the sperm head. Chromatin condensation appears normal. The head morphology and number of flagella vary in mature spermatozoa produced by this process. This work demonstrates one pathway by which polyploid spermatids continue to differentiate to spermatozoa after failure of cytoplasmic division or possibly cellular fusion.     

Ultrastructure of sperm and spermatogenesis of Lobatostoma manteri (Trematoda, Aspidogastrea)

Mature sperm has two axonemes of the 9 + '1' pattern incorporated in the sperm body, a row of peripheral microtubules interrupted along part of the sperm by the axonemes, some microtubules in the interior of the sperm and a long lateral extension (lobe) of the sperm body, an elongate nucleus and mitochondrion, and many dense rod-like structures. A supporting rod extends underneath a specialized region consisting of alternating thin and thick transverse rows of irregular dense patches, and with surface ridges around (all or) most of the surface of the sperm. Primary spermatocytes in the prophase of the first meiotic division have synaptonemal complex(es), and are rich in mitochondria. In early spermiogenesis, mitochondria are arranged around the surface of the nucleus, a dense layer appears at one pole of the nucleus, close to an apposed dense layer at the cell membrane in which a row of microtubules develops. The intercentriolar (= central) body develops close to the nucleus. The fully developed intercentriolar body has a regular striation and is located perpendicular and close to the surface of the nucleus. Two flagella extend into the space surrounding the outgoing median process, their basal bodies are located perpendicular to the intercentriolar body and their cross-striated rootlets extend along the surface of the rounded nucleus. At a later stage, rootlets and flagella become more parallel with the intercentriolar body, the nucleus and the fused mitochondria migrate into the median process, and the flagella become incorporated into the median process (= sperm body). The outgrowing spermatozoa are connected to the cytoplasm of the cytophore by dense arching membranes. Finally, rootlets of flagella are resorbed and the spermatozoa are pinched off close to the basal bodies. Two species (Lobatostoma and Multicotyle) of the same family differ strongly in the type of spermiogenesis, although their mature sperm is of the same basic type, i.e. spermiogenesis is not necessarily more useful for phylogenetic considerations than sperm structure.     

Spermatogenesis and spermiogenesis in Microcotyle sp. (Microcotylidae, Monogenea)

Ultrastructural observations of spermatogenesis and spermiogenesis in Microcotyle sp. a microcotylid monogenean parasite from the gills of Hypostomus sp., are described. The spermatogonia were irregularly shaped, forming a peripheral layer of cells; spermatocytes were larger than spermatogonia and a nuclear synaptonemal complex was observed; young spermatids were joined by a central cytophore forming rosettes. Spermiogenesis was characterized by the outgrowth of a cytoplasmic protuberance, the zone of differentiation, containing the basal bodies, separated by an intercentriolar body, from which flagella grow out and fuse posteriorly with the median process. Cross sections of the anterior and the middle regions of spermatozoa revealed nuclei, mitochondria, peripheral microtubules, and paired axonemes each with a 9+1 pattern.     

Phagocytosis of sperm by follicle cells of the carnivorous sponge Asbestopluma occidentalis (Porifera, Demospongiae)

During spermatogenesis of the carnivorous sponge Asbestopluma occidentalis, follicle cells that lined the spermatocysts phagocytosed unreleased mature sperm. Such follicle cells are part of the complex envelope that limits spermatocysts of A. occidentalis, which is also comprised of a collagen layer, a thick layer of intertwined cells, and spicules. Follicle cells showed vesicles containing single phagocytosed spermatozoa within their cytoplasm. Additionally, lipids and other inclusions were observed within the cytoplasm of follicle cells. It is likely that follicle cells recapture nutrients by phagocytosing spermatozoa and use them to form lipids and other inclusions. Such sperm phagocytosis is usually performed in higher invertebrates and vertebrates by Sertoli cells that are located in the testis wall. While Sertoli cells develop a wide range of functions such as creating a blood-testis barrier, providing crucial factors to ensure correct progression of spermatogenesis, and phagocytosis of aberrant, degenerating, and unreleased sperm cells, sponge follicle cells may only display phagocytotic activity on spermatogenic cells.     

Sperm of Doradidae (Teleostei: Siluriformes)

Spermatic characteristics were studied in 10 species representing several distinct groups within the catfish family Doradidae. Interestingly, different types of spermatogenesis, spermiogenesis and spermatozoa are correlated with intrafamilial groups previously proposed for Doradidae. Semi-cystic spermatogenesis, modified Type III spermiogenesis, and biflagellate sperm appear to be unique within Doradidae to the subfamily Astrodoradinae. Other doradid species have sperm with a single flagellum, cystic spermatogenesis, and spermiogenesis of Type I (Pterodoras granulosus, Rhinodoras dorbignyi), Type I modified (Oxydoras kneri), or Type III (Trachydoras paraguayensis). Doradids have an external mode of fertilization, and share a few spermatic characteristics, such as cystic spermatogenesis, Type I spermiogenesis and uniflagellate sperm, with its sister group Auchenipteridae, a family exhibiting sperm modifications associated with insemination and internal fertilization. Semi-cystic spermatogenesis and biflagellate spermatozoa are also found in Aspredinidae, and corroborate recent proposals that Aspredinidae and Doradoidea (Doradidae + Auchenipteridae) are sister groups and that Astrodoradinae occupies a basal position within Doradidae. The co-occurrence in various catfish families of semi-cystic spermatogenesis and either biflagellate spermatozoa (Aspredinidae, Cetopsidae, Doradidae, Malapturidae, Nematogenyidae) or uniflagellate sperm with two axonemes (Ariidae) reinforces the suggestion that such characteristics are correlated. Semi-cystic spermatogenesis and biflagellate sperm may represent ancestral conditions for Loricarioidei and Siluroidei of Siluriformes as they occur in putatively basal members of each suborder, Nematogenyidae and Cetopsidae, respectively. However, if semi-cystic spermatogenesis and biflagellate sperm are ancestral for Siluriformes, cystic spermatogenesis and uniflagellate sperm have arisen independently in multiple lineages including Diplomystidae, sister group to Siluroidei.     

Structure of the testis and spermatogenesis of the viviparous teleost Poecilia mexicana (Poeciliidae) from an active sulfur spring cave in Southern Mexico

We describe the histological characteristics of the testis and spermatogenesis of the cave molly Poecilia mexicana, a viviparous teleost inhabiting a sulfur spring cave, Cueva del Azufre, in Tabasco, Southern Mexico. P. mexicana has elongate spermatogonial restricted testes with spermatogonia arranged in the testicular periphery. Germ cell development occurs within spermatocysts. As spermatogenesis proceeds, the spermatocysts move longitudinally from the periphery of the testis to the efferent duct system, where mature spermatozoa are released. The efferent duct system consists of short efferent duct branches connected to a main efferent duct, opened into the genital pore. Spermatogenesis consisted of the following stages: spermatogonia (A and B), spermatocytes (primary and secondary), spermatids, and spermatozoa. The spermatozoa are situated within spermatocysts, with their heads oriented toward the periphery and flagella toward the center. Once in the efferent duct system, mature spermatozoa are packaged as unencapsulated sperm bundles, that is, spermatozeugmata. We suggest that the histological characteristics of the testis and spermatogenesis of P. mexicana from the Cueva del Azufre, and the viviparous condition where the spermatozoa enter in the female without been in the water, have allowed them to invade sulfurous and/or subterranean environments in Southern Mexico, without requiring complex morphofunctional changes in the testis or the spermatogenetic process.