Sperm ultrastructure and a new type of spermiogenesis in two species of Pimelodidae, with a comparative review of sperm ultrastructure in Siluriformes (Teleostei: Ostariophysi)

During spermiogenesis, the spermatids of the pimelodid species Pimelodus maculatus and Pseudoplatystoma fasciatum show a central flagellum development, no rotation of the nucleus, and no nuclear fossa formation, in contrast to all previously described spermatids of Teleostei. These characteristics are interpreted as belonging to a new type of spermiogenesis, named here type III, which is peculiar to the family Pimelodidae. In P. maculatus and P. fasciatum, spermatozoa possess a spherical head and no acrosome; their nucleus contains highly condensed, homogeneous chromatin with small electron-lucent areas; and a nuclear fossa is not present. The centriolar complex lies close to the nucleus. The midpiece is small, has no true cytoplasmic channel, and contains many elongate and interconnected vesicles. Several spherical to oblong mitochondria are located around the centriolar complex. The flagellum displays the classical axoneme (9+2) and no lateral fins. Only minor differences were observed among the pimelodid species and genera. Otherwise, spermiogenesis and spermatozoa in the two species of Pimelodidae studied exhibit many characteristics that are not found in other siluriform families, mainly the type III spermiogenesis.     

Fine structure of the germinating cells during the spermiogenesis of Arion rufus (Mollusca,Pulmonate Gastropoda)

Changes in spermatozoan ultrastructure have been studied during spermiogenesis of the slug Arion rufus (Gastropoda, Pulmonata, Stylommatophora). The ovotestis was investigated during the male stage, definite by the presence of spermatozoa. Some peculiar characteristics are shown by early spermatids: Around the nucleus, the nuclear envelope presents two thick layers located on opposite sides, the apical and basal plates, that will determine the antero-posterior axis of the spermatid. The chromatin, first dispersed throughout the nucleoplasm gives later on thick filaments which become attached over the inner surface of these plates. The chromatin filaments are then arranged parallel to the antero-posterior axis as the nucleus elongates. The position of the plates determines the antero-posterior axis of the spermatid. In the mature spermatozoa, the chromatin is more condensed and the nucleus presents an helical organization. The acrosome and flagellum are respectively attached externally to the center of the apical and basal plates. The acrosome consists of a membrane-bound vesicle and forms a column of homogeneous material. In the middle piece, the mitochondria have been transformed into a mitochondrial derivate by the way of a complicated metamorphosis. The axoneme is surrounded by three mitochondrial helices but only one of them contains glycogene granules. © 1996 Wiley-Liss, Inc.     

Ultrastructure of the spermatogenesis of the cockle Anadara granosa L. (Bivalvia: Arcidae)

In this paper spermatogenesis and sperm ultrastructure of the cockle Anadara granosa are studied using transmission electron microscopy. The spermatocyte presents electron-dense vesicles and the arising axoneme that begins to form the flagellum. During spermatid differentiation, proacrosomal vesicles appear to migrate towards the presumptive anterior pole of the nucleus; eventually these vesicles become acrosome. The spermatozoon of Anadara granosa is of the primitive type. The acrosome, situated at the apex of the nucleus, is cap-shaped and deeply invaginated at the inner side. The spherical nucleus of the spermatozoon contains dense granular chromatin and shows invagination at the posterior poles. The centriole shows the classic nine triplets of microtubules. The middle piece consists of the centriolar complex surrounded by five giant mitochondria. It is shown that the ultrastructure of spermatozoa and spermiogenesis of Anadara granosa reveals a number of features that are common among bivalves. Received: 29 September 1998 / Received in revised form: 20 May 1999 / Accepted: 14 June 1999     

Ultrastructural study of spermatogenesis in the free-living marine nematode <Emphasis Type="Italic">Paracyatholaimus pugettensis</Emphasis> Weiser et Hopper, 1967 (Chromadorida: Cyatholaimidae)

Spermatogenesis and the morphology of mature sperm in the free-living chromadorid Paracyatholaimus pugettensis from the Sea of Japan were studied using transmission electron microscopy. In spermatocytes fibrous bodies (FBs) appear; in spermatids, the synthetic apparatus is located in the residual body, whereas the main cell body (MCB) houses the nucleus, mitochondria, and FBs. The nucleus of the spermatid consists of a loose fibrous chromatin that is not surrounded by a nuclear envelope; centrioles lie in the perinuclear cytoplasm. The plasma membrane of the spermatid MCB forms numerous filopodia. Immature spermatozoa from the proximal part of the testis are polygonal cells with a central nucleus. The latter is surrounded by mitochondria and FBs with poorly defined boundaries. The immature spermatozoa bear lamellipodia all along their surface. Mature spermatozoa are polarized cells with an anterior pseudopodium, which is filled with filaments that make up the cytoskeleton; the MCB houses a nucleus that is surrounded by mitochondria and osmiphilic bodies. In many ultrastructural characteristics, the spermatozoa of P. Pugettensis are similar to those of most nematode species studied so far (i.e., they are ameboid, have no acrosome, axoneme, or nuclear envelope). On the other hand, as in other chromadorids, no aberrant membrane organelles were observed during spermatogenesis of P. Pugettensis.Original Russian Text Copyright © 2004 by Biologiya Morya, Zograf, Yushin.     

Spermatogenesis in a Free-Living Marine Nematode Neochromadora poecilosoma (Chromadorida,Chromadoridae) Studied Using TEM

Spermatogenesis and the structure of mature spermatozoa were studied using TEM in a free-living marine chromadorid nematode Neochromadora poecilosoma from the Sea of Japan. In spermatocytes, fibrous bodies (FB) develop; in spermatids, the synthetic apparatus lies in the residual body, while the nucleus, mitochondria, and FB are located in the main cell body (MCB). The nucleus consists of a diffuse chromatin of fibrous structure, which is not enclosed in a nuclear envelope. In the spermatid stage, the development of FB is completed, and immature spermatozoa from the proximal region of the testis do not show any structural differences from the MCB of spermatids. The mature spermatozoa are polarized cells. They attach to the uterus wall by a pseudopod filled with filaments of the cytoskeleton; in the MCB of spermatozoon, there is a nucleus surrounded by mitochondria and osmiophilic bodies. The spermatozoa of N. poecilosoma show typical ultrastructure features of sperm cells found in most studied nematodes (amoeboid nature and the absence of axoneme, acrosome, and nuclear envelope). However, no aberrant organelles characteristic of nematode spermatozoa were found throughout sperm development in N. poecilosoma and other chromadorids.     

The spermiogenesis and the early spermatozoa of <Emphasis Type="Italic">Armorloricus elegans</Emphasis> (Loricifera,Nanaloricidae)

The spermiogenesis consisting of five spermatid stages and the early spermatozoon has been investigated in Armorloricus elegans (Loricifera) with the use of transmission electron microscopy. The male reproductive system consists of three parts; testes, vasa deferentia and seminal vesicles. Caudally, the two seminal vesicles merge together in a ciliated duct and the excretory/gonadal—and digestive systems continue through the recto-urogenital canal, which opens via the lateral gonopores and the temporarily closed anal system. Spermiogenesis mainly occurs in the testes, whereas further maturation of the late spermatids and early spermatozoa occurs in the vasa deferentia and seminal vesicles. A maturation gradient (from spermatocytes to spermatozoa) is found from the posterior peripheral part of the testes to the anterior periphery and then centrally. During spermiogenesis the round nucleus becomes more osmiophilic and condensation of chromatin occurs. Later the nucleus elongates until it becomes rod-shaped in the early spermatozoa. In the second spermatid stage, a large vesicle is formed by saccules developed from the Golgi complex. This vesicle develops further and consists of three different osmiophilic parts with some crystal-like structures inside and is on the outside almost entirely surrounded by thick striated filaments. In the mid-piece the flagellum has a typical 9 × 2 + 2 axoneme and the two mitochondria are fused into a single sheet surrounding the flagellum. In the early spermatozoon stage an acrosomal-like cap structure with an acrosome filament appears proximal to the protruded rod-shaped nucleus. This cap is not formed by the Golgi complex and therefore might not be a true acrosome. Comparing the early spermatozoa of A. elegans with other cycloneuralians has shown some similarities with especially Kinorhyncha and Priapulida. These similarities are thought to be plesiomorphic.     

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.     

Spermiogenesis in the spider,Pisaurina Sp.: A fine structure study

A fine structure study of spermatids and spermatozoa of the spider, Pisaurina sp. demonstrates that early spermiogenesis is similar to other flagellate spermatozoa. An acrosome forms from a Golgi-derived, acrosomal vesicle, a perforatorium indents acromosome and nucleus, a flagellum with a three-plus-nine tubule substructure is formed and nuclear chromatin condenses during spermiogenesis. Divergence from typical spermatozoa includes the presence of a three-tubule substructure of the central flagellar shaft, progressive rounding-up of late spermatids with concomitant incorporation of previously formed flagellum. This evidence is presented in terms of its possible functional significance in fertilization and gamete fusion in spiders.     

Spermatogenesis and sperm ultrastructure in three species of polydora and in streblospio benedicti (Polychaeta: Spionidae)

Summary Spermatogenesis was studied at the ultrastructural level in Polydora ligni, P. websteri, P. socialis and Streblospio benedicti. Spermatogonia, spermatocytes, spermatids and mature sperm are described. In all four species, meiosis occurs in the coelom following release of spermatogonia from the gonad. In Polydora spp., chromatin condensation is lamellar with no microtubules present during nuclear elongation. In S. benedicti, chromatin condensation is fibrous with a manchette of microtubules present around the nucleus. In all four species, the acrosome forms from a Golgi-derived vesicle situated at the base of spermatids. The acrosome in Polydora spp. is conical with a distinctive substructure whereas the S. benedicti acrosome is long and spiral. The implantation fossa is short in all species except P. ligni. All four species have elongated sperm heads. The middlepiece as well as the nucleus is elongated in Polydora spp. whereas S. benedicti has a long nucleus but a short middlepiece. Platelet-shaped electron-dense bodies are present throughout the nuclear region and middlepiece of Polydora spp. and the nuclear region of S. benedicti. These membrane-bounded bodies may be energy storage organelles. The use of ultrastructural data in analysis of sibling species complexes is discussed.Contribution Number 203 from Harbor Branch Foundation, Inc.     

Seasonal ultrastructural modifications of the seminiferous epithelium in twoEulemur species:E. fulvus andE. macaco

A comparative ultrastructural study of the seminiferous epithelium was conducted during the mating and non-mating seasons of twoEulemur species:E. fulvus andE. macaco. The ultrastructure of the junctional complexes of the Sertoli cells, and the modifications in the spermatids during spermiogenesis are reported. Acridine orange staining of the sperm cells of these animals showed that the chromatin compaction was complete in all spermatozoa.     

Ultrastructure of spermiogenesis in Plagioscion squamosissimus (Teleostei, Perciformes, Sciaenidae)

Spermiogenesis in Plagioscion squamosissimus occurs in cysts. It involves a gradual differentiation process of spermatids that is characterized mainly by chromatin compaction in the nucleus and formation of the flagellum, resulting in the spermatozoa, the smallest germ cells. At the end of spermiogenesis, the cysts open and release the newly formed spermatozoa into the lumen of the seminiferous tubules. The spermatozoa do not have an acrosome and are divided into head, midpiece, and tail or flagellum. The spermatozoa of P. squamosissimus are of perciform type with the flagellum parallel to the nucleus and the centrioles located outside the nuclear notch.     

Spermiogenesis in the rainbow trout (Salmo gairdneri)

In an ultrastructural study on the spermiogenesis of the rainbow trout (Salmo gairdneri R.) four spermatogenetic stages were identified. In young round spermatids, the nuclear chromatin was first heterogeneous (euchromatin and heterochromatin). Subsequently, it became more homogeneous and started to condense in the form of coarse granules and fibers and then into fibrils associated in ribbon-like elements which eventually partly fused together. During early spermiogenesis, a juxtanuclear vacuole appeared in the area where the nuclear envelope was specialized due to condensation of material between the two envelopes and a slight accumulation of nuclear material. This area was finally located in the anterior part of spermatids and spermatozoa; it probably plays a role during fertilization. A flagellar rootlet appeared early in spermiogenesis; it may play a role in the attachment of the flagellum to the nucleus since it persisted until the centriolar complex was definitively fixed in the implantation fossa. The flagellum did not display a plasma membrane and was first located in the cytoplasm, but when it was later extruded from the cell, it acquired a membrane. The cytoplasm was rich in ribosomes (free or in small groups) but poor in membranous organelles. The few mitochondria polarized around the centriolar complex were finally organized into an annular mid-piece. The spermatids remained connected by intercellular bridges until the end of spermiogenesis. The complexity of trout spermiogenesis is intermediate between that in poecilids and that in carp and pike, which have very simple spermatozoa. The role of the material from the nucleus and the cytoplasm reaching the Sertoli cell in the control of spermatogenesis has been discussed.     

Nuclear morphogenesis during spermiogenesis in the nudibranch molluscHypselodoris tricolor (Gastropoda,opisthobranchia)

An electron microscope study was carried out on Hypselodoris tricolor spermatids to describe the development of the nuclear morphogenesis and investigate the possible cause(s) of the change in the shape of the spermatid nucleus during spermiogenesis. Three different stages may be distinguished in the course of the nuclear morphogenesis on the basis of the morphology and inner organization of the nucleus. Stage 1 spermatid nuclei are spherical or ovoid in shape and the nucleoplasm finely granular in appearance. Stage 2 nuclei exhibit a disc- or cup-shaped morphology, and the chromatin forms short, thin filaments. During stage 3, a progressive nuclear elongation takes place, accompanied by chromatin rearrangement, first into fibers and then into lamellae, both formations helically oriented. A row of microtubules attached to the nuclear envelope completely surrounds the nucleus. Interestingly, the microtubules always lie parallel to the chromatin fibers adjacent to them. Late stage 3 spermatids show the highest degree of chromatin condensation and lack the manchette at the end of spermiogenesis. Our findings indicate the existence of a clear influence exerted on the chromatin by the manchette microtubules, which appear to be involved in determining the specific pattern of chromatin condensation in Hypselodoris tricolor.     

Ultrastructure of testes and spermatogenesis in the trombiculid mite,Hirsutiella zachvatkini (Schluger)

Summary

Spermatogenesis and sperm ultrastructure of the trombiculid mite Hirsutiella zachvatkini (Schluger 1948) have been investigated using transmission electron microscopy and compared with other arachnids studied. Sperm differentiation takes place in groups of synchronously developed germ cells of the two large sac-like paired testes. Each testis is composed of a secretory epithelium, which occupies their medio-ventral regions, and of a germinative epithelium situated in the latero-dorsal parts of testes together with large somatic cells. The germ cells are represented on sections by spermatogonia, spermatocytes, early, middle and late spermatids, and mature spermatozoa. Spermatocytes and spermatids contain two centrioles, which disappear afterwards, and a small Golgi-like structure forming an acrosomal cistema. Mature spermatozoa, which lie both within the meshes of somatic cells and also free in the lumen of testes, are compact oval aflagellate cells provided with peripheral channels. They also contain an acrosome, flattened between the cell membrane and the round electron-dense chromatin body, an oval body of lesser density lying in close proximity to the chromatin body, and a group of 5–7 mitochondria with spherically arranged cristae situated immediately behind the nuclear bodies. An acrosomal filament may be sometimes seen beneath the acrosome in the middle spermatids and disappears in the mature spermatozoa. These findings show that the mode of differentiation and pattern of organization of the male sex cells in trombiculid mites are of rather primitive type compared with other acarine spermatozoa.     

Spermiogenesis and spermatozoa ultrastructure in Salminus and Brycon, two primitive genera in Characidae (Teleostei: Ostariophysi: Characiformes)

In Salminus, spermiogenesis is cystic and gives origin to a type I aquasperm. Spermatid differentiation is characterized by chromatin condensed into thick fibres, nuclear rotation, nuclear fossa formation, cytoplasmic channel formation, mitochondrial fusion producing long and ramified mitochondria, and the presence of several membranous concentric rings around the plasma membrane that encircles the cytoplasmic channel. In Salminus and Brycon, spermatozoa are very similar. They exhibit a spherical nucleus and chromatin condensed into fibre clusters, and a deep nuclear fossa. They show a long midpiece with few elongate mitochondria at the initial region and a cytoplasmic channel completely encircled by one or two membranous concentric rings. The flagellar axis is perpendicular to the nucleus and exhibits the classic axoneme (9 + 2). The very strong similarity observed between Salminus and Brycon spermatozoa supports the hypothesis that these subfamilies are likely to have a monophyletic origin.     

Spermatogenesis and sperm structure in Carpoglyphus lactis (L.) (Acari: Astigmata)

Testes, spermatogenesis and spermatozoa are described in the mite Carpoglyphus lactis (L.), the first representative of the Hemisarcoptoidea superfamily studied ultrastructurally. Paired testes are located posteriorly in the idiosoma, with germaria situated dorsolaterally. The germarium consists of a compact group of spermatogonia; no testicular central cell was found. The remainder of the gonad is occupied by germ cells in different stages of spermatogenesis, distributed separately rather than in cysts, and embedded in a few large somatic cells filling the remaining space. Spermatocytes are covered by a spongy layer, a product of the Golgi apparatus. Spermatids are anucleate. Their chromatin condenses into granular and then tubular threads. As spermiogenesis progresses, the spongy layer assembles at a single site and forms a structure termed the spongy body; mitochondria become electron dense, elongate and gather forming a bundle; a narrow ER cistern, promptly transforming into a dense lamella, appears between the mitochondria and chromatin. Mature spermatozoa are small, highly electron-dense cells interdigitating with others via superficial protrusions. They possess chromatin threads, electron-dense lamella and mitochondria, but do not have an acrosome.Our results support the monophyly of Astigmata, but do not explain the phylogenetic affinities of Hemisarcoptoidea to other superfamilies of astigmatic mites.     

Cloning and Functional Analysis of Histones H3 and H4 in Nuclear Shaping during Spermatogenesis of the Chinese Mitten Crab,Eriocheir sinensis

During spermatogenesis in most animals, the basic proteins associated with DNA are continuously changing and somatic-typed histones are partly replaced by sperm-specific histones, which are then successively replaced by transition proteins and protamines. With the replacement of sperm nuclear basic proteins, nuclei progressively undergo chromatin condensation. The Chinese Mitten Crab (Eriocheir sinensis) is also known as the hairy crab or river crab (phylum Arthropoda, subphylum Crustacea, order Decapoda, and family Grapsidae). The spermatozoa of this species are aflagellate, and each has a spherical acrosome surrounded by a cup-shaped nucleus, peculiar to brachyurans. An interesting characteristic of the E. sinensis sperm nucleus is its lack of electron-dense chromatin. However, its formation is not clear. In this study, sequences encoding histones H3 and H4 were cloned by polymerase chain reaction amplification. Western blotting indicated that H3 and H4 existed in the sperm nuclei. Immunofluorescence and ultrastructural immunocytochemistry demonstrated that histones H3 and H4 were both present in the nuclei of spermatogonia, spermatocytes, spermatids and mature spermatozoa. The nuclear labeling density of histone H4 decreased in sperm nuclei, while histone H3 labeling was not changed significantly. Quantitative real-time PCR showed that the mRNA expression levels of histones H3 and H4 were higher at mitotic and meiotic stages than in later spermiogenesis. Our study demonstrates that the mature sperm nuclei of E. sinensis contain histones H3 and H4. This is the first report that the mature sperm nucleus of E. sinensis contains histones H3 and H4. This finding extends the study of sperm histones of E. sinensis and provides some basic data for exploring how decapod crustaceans form uncondensed sperm chromatin.     

On sperm ultrastructure,spermiogenesis and the spermatophore of Heterolaophonte minuta (Copepoda,Harpacticoida)

Summary The spermatophore, mature spermatozoon and spermiogenesis of Heterolaophonte minuta have been investigated by light and electron microscopy. The spermatophore contains three different secretions which are responsible for the discharge of the contents of the spermatophore, the formation of the fertilization tube and the storage of the spermatozoa. The spermatozoon represents a type new for the Copepoda. It is a filiform cell about 25 m in length, ellipsoid in transverse section and tapered at the posterior end. The elongated nucleus contains chromatin fibrils and does not possess a nuclear envelope. Posterior to the nucleus, six mitochondria are placed one after the other. The posterior part of the spermatozoon contains parallel pseudomembranes. The gamete is not helically twisted and is without a flagellum and centrioles. The most remarkable feature of the spermatozoon is an osmiophilic cap in front of the nucleus. This cap corresponds to the acrosome of the spermatozoon. Early stages of spermiogenesis take place in the testis, where the spermatids are incorporated into accessory cells. The origin of the chromatin fibrils and the glycocalyx, as well as the breakdown of the nuclear envelope and centrioles, represent the final steps of spermiogenesis which occur in the vas deferens.     

Semicystic spermatogenesis and biflagellate spermatozoon ultrastructure in the Nile electric catfish Malapterurus electricus (Teleostei: Siluriformes: Malapteruridae)

Spermatogenesis and spermatozoon ultrastructure in the Nile electric catfish Malapterurus electricus are described using scanning and transmission electron microscopy. Although the testis organization conforms to the ‘unrestricted’ spermatogonial type, the species has a rare type of spermatogenesis not previously described among catfishes, ‘semicystic’, in which the cyst ruptures before the spermatozoon stage. Spermiogenesis also involves some peculiar features such as condensation of the chromatin in the posterior part of the nucleus to form a compact electron‐dense mass with some irregular electron‐lucent lacunae, while the uppermost part of the nucleus is a loose electron‐lucent area, absence of the nuclear rotation and, as a consequence, the centriolar complex and the initial segment of each flagellum arise directly in a position perpendicular to the basal pole of the nucleus, and occurrence of numerous vesicles in the midpiece. In addition, spermiogenesis includes migration of the diplosome and mitochondria to the basal pole of the nucleus, formation of two moderate nuclear fossae, each of which contains the centriolar complex, development of two independent flagella and elimination of the excess cytoplasm. The mature spermatozoon has a more or less round head with no acrosome or acrosomal vesicle, a long midpiece with numerous mitochondria and vesicles and two long tails or flagella having the classical axoneme structure of 9 + 2 microtubular doublet pattern and with no lateral fins and membranous compartment. These findings suggest that the ultrastructural features of spermiogenesis and spermatozoa of M. electricus are synapomorphies of types I and II spermiogenesis and spermiogenesis is closely similar to the type described in the Nile catfish Chrysichthys auratus.