Ultrastructure of spermatogenesis and spermatozoa of Cephalodasys maximus (Gastrotricha,Macrodasyida)

Summary Spermatogenesis and sperm ultrastructure of the macrodasyidan gastrotrich Cephalodasys maximus are described by means of transmission electron microscopy. The filiform sperm consists of an acrosomal accessory structure and an acrosomal vesicle, both being surrounded by spiralled material. The successive nuclear helix encloses the spiral-shaped mitochondrion and the axoneme of the flagellum is accompanied by dense strings, three helical elements and peripheral microtubules. During spermiogenesis the acrosomal accessory structure develops first and moves into a cell projection, where the spiral around this acrosomal rod forms. A nuclear section with condensed chromatin and one single fused large mitochondrion follow into the extension, becoming helical. A connecting clasp between nucleus and flagellum shortens to a cap-like structure. Parallel to the acrosomal and nuclear projection the flagellum develops where the spiralled elements and the basal plate form in succession, while the basal body shrinks.     

Ultrastructural analysis of spermiogenesis in Iguana iguana (Reptilia: Sauria: Iguanidae)

Spermiogenesis in the lizard, Iguana iguana, was studied by transmission and scanning electron microscopy. During this process, structures such as the acrosomal complex in the spermatid head and the axonemal complex in the mid and principal pieces of the flagellum are formed. The nuclear content is initially compacted into thick, longitudinal chromatin filaments. Nuclear shape is determined by further compaction and by the manchette, a layer of microtubules surrounding the head. The acrosomal complex originates from Golgi vesicles and the interaction between the proacrosomal vesicle and the nucleus. The midpiece consists of a pair of centrioles, surrounded by a fibrous sheath and rings of simple and modified mitochondria. The centrioles sustain the axoneme that appears at the end of the midpiece. The axoneme extends throughout the principal piece of the flagellum with the 9 + 2 pattern, still surrounded by the fibrous sheath. In the endpiece, the axoneme continues, surrounded only by the plasma membrane. In the lumen of seminiferous tubules, immature spermatozoa retain abundant residual cytoplasm.     

Sperm Bundles in the Seminal Vesicle of the Crematogaster victima (Smith) Adult Males (Hymenoptera: Formicidae)

This study establishes the presence of spermatodesm in the seminal vesicles of sexually mature males of Crematogaster victima (Smith). In this species, the spermatozoa are maintained together by an extracellular matrix in which the acrosomal regions are embedded. This characteristic has not yet been observed in any other Aculeata. However, the sperm morphology in this species is similar to that described for other ants. The spermatozoa measure on average 100 μm in length, and the number of sperm per bundle is up to 256. They are composed of a head formed by the acrosome and nucleus; this is followed by the flagellum, which is formed by the centriolar adjunct, an axoneme with a 9?+?9?+?2 microtubule pattern, two mitochondrial derivatives, and two accessory bodies. The acrosome is formed by the acrosomal vesicle and perforatorium. The nucleus is filled with compact chromatin with many areas of thick and non-compacted filaments. Both mitochondrial derivatives have the same shape and diameters. The presence of sperm bundles in sexually mature males differentiates C. victima from other ants; however, the similarities in the sperm ultrastructure support the monophyly of this insect group.     

Testicular,spermatogenesis and sperm morphology in Martarega bentoi (Heteroptera: Notonectidae)

The testicular, spermatogenesis and sperm morphology of the backswimmer Martarega bentoi was described using light and transmission electron microscopy. In this species, a pair of testes, two deferent ducts, two different pairs of accessory glands, and an ejaculatory duct form the male reproductive system. Each testis consists of two testicular follicles, which are arranged side by side in snail shape. The follicles are filled with cysts at different stages of spermatogenesis, but in the same cyst the germ cells (up to 64) are in the same stage. At the end of spermatogenesis, the sperm cells are very long, with the flagellum measuring approximately 2500 μm in length, the nucleus only 19 μm, and the acrosome, with two distinct regions, 300 μm. The flagellum is composed of an axoneme, with a 9 + 9 + 2 microtubular pattern, and 2 asymmetric mitochondrial derivatives (MDs). These have the anterior ends inserted into two cavities at the nucleus base, exhibit two paracrystalline inclusions, and have bridges linking them to the axoneme. Few spermatozoa per cyst, asymmetry in size and shape of the MDs, as well as their insertion at the nuclear base are characteristics considered derived, and that differentiate the sperm of M. bentoi from those of the Nepomorpha, Belostomatidae and Nepidae.     

Structure and ultrastructure of the spermatozoa of Bephratelloides pomorum (Fabricius) (Hymenoptera: Eurytomidae)

The spermatozoa of Bephratelloides pomorum are very long and fine. Each spermatozoon measures about 620 μm in length by 0.38 μm in diameter and, when seen under the light microscope, appears to be wavy along its entire length. The head, which is approximately 105 μm, comprises a small acrosome and a nucleus. The acrosome is made up of a cone-shaped acrosomal vesicle surrounding the perforatorium and the anterior end of the nucleus. Innumerable filaments radiate from it. The perforatorium has a diameter equal to that of the nucleus at their junction, where it fits with a concave base onto the rounded nuclear tip. The nucleus is helicoidal and completely filled with homogeneous compact chromatin. It is attached to the tail by a very long and quite electron-dense centriolar adjunct that extends anteriorly from the centriole in a spiral around the nucleus for approximately 8.5 μm. The tail consists of an axoneme with the 9+9+2 microtubule arrangement pitched in a long helix, as well as a pair of spiraling mitochondrial derivatives (with regularly arranged cristae) that coil around the axoneme, and two small accessory bodies. As well as the spiraling of the nucleus, mitochondrial derivatives and axonemal microtubules, the sperm of B. pomorum present other very different morphological features. These features include the acrosome and centriolar adjunct, both of which differentiate the spermatozoa from the majority of sperm found in other Hymenoptera. In addition these structural variations demonstrate that the sperm of chalcidoids provide characteristics that can certainly prove useful for future phylogenetic analysis at the subfamily level and, possibly, the genus too.     

Polymorphism of spermatozoa in Largus rufipennis Laporte 1832 (Heteroptera: Pyrrhocoroidea: Largidae)

The production of polymorphic spermatozoa has been registered in various insect orders such as Diptera, Lepidoptera, and Hemiptera. In this work, morphology of two types of spermatozoa produced by Largus rufipennis was reported for the first time in the Largidae family. For this, techniques including optical and transmission electron microscopy were used. Spermatozoa measured, on the average, 260 and 200 μm, and both types possessed a nucleus measuring on the average 65 μm. No ultrastructural differences were observed between the two spermatozoa types from L. rufipennis. The head region is composed of an acrosome, a nucleus, and part of the centriolar adjunct. The centriolar adjunct is in parallel with the nucleus and followed by mitochondrial derivates. The flagellum consists of an axoneme (9 + 9 + 2 microtubules) and two mitochondrial derivatives; no other accessory bodies were observed. The mitochondrial derivatives are symmetric in size and diameter. A similar quantity of the two spermatozoa types was observed in the seminal vesicle (57% of the large type and 43% of the small type), while in the spermatheca of the female, the larger spermatozoa were preferentially stored (87%). These results permit discussions concerning of the species biology reproduction, most specifically sperm competition strategies.     

Spermiogenesis and ultrastructure of spermatozoa in Saxipendium coronatum (Hemichordata,Enteropneusta), with consideration of their relation to reproduction and dispersal

Summary Sperm ultrastructure and spermiogenesis of the enteropneust hemichordate Saxipendium coronatum conforms to the general pattern of the prototype spermatozoon found in many phyla. The sperm is about 29 m long, including head, middle piece, and tail. The Saxipendium spermatozoon has some unique features. The head is pyramidal in shape and the nucleus has four frontal ridges radiating from the base of the acrosomal region. The acrosome is composed of a large acrosomal vesicle surrounded by periacrosomal material. The acrosomal region projects about 1 m in front of the nucleus and has a width at the base of 1.5 m. The middle piece is dish-shaped and contains a large mitochondrial mass surrounding the centriolar region. The centriolar region is partially located in a centriolar fossa at the basal part of the nucleus. In spermatids, an anchoring fiber apparatus is observed surrounding the centriolar region. The distal ends of the fibers are attached to the plasmalemma by electron-dense thickenings. The tail is a simple flagellum. The sperm of Saxipendium and the small eggs found in the female suggest non-specialized external fertilization and embryogeny leading to a planktotrophic larva. The main results of the fine structure of the spermatozoon in Saxipendium are summarized in Fig. 12.Abbreviations used in the figures an antrum - av acrosomal vesicle - ax axoneme - d distal centriole - ep epidermis - f flagellum - gp gonopore - m mitochondrion - mp middle piece - n nucleus - p proximal centriole - per periacrosomal material - sp sperm - te testis - vac vacuolated cells     

Sperm structure in the scissurellid gastropod Sinezona sp. (Prosobranchia, Pleurotomarioidea)

Spermatozoa of Sinezona sp. (Scissurcllidac) arc examined ultrastructurally and compared with spermatozoa of other vetigastropods. Sinezona sperm are characterized by the following features: (1) a squat-conical acrosomal vesicle; (2) subacrosomal deposits–one forming a perinuclear sheath and the second forming a rod which almost penetrates the nucleus; (3) prominent nuclear invaginations enclosing the axial rod and centriolar regions; (4) a midpiece consisting of axoneme surrounded by a single, sleeve-like mitochondrion; (5) an annulus occurring posterior to the midpiece; and (6) a flagellum. Spermatozoa of Sinezona sp. are clearly modified in structure in comparison with spermatozoa of other pleurotomarioids and, more generally, most other vetigastropods (these retain multiple, spherical mitochondria and, with rare exceptions, do not have the ccntrioles located in a nuclear invagination).     

Morphology and fine structure of Acipenser persicus (Acipenseridae, Chondrostei) spermatozoon: Inter-species comparison in Acipenseriformes

This study describes morphology and fine structure of the Persian sturgeon (Acipenser persicus) (Acipenseridae, Chondrostei) spermatozoon. The results show that the spermatozoon of A. persicus is differentiated into an elongated head (length: mean±SD: 7.1±0.5μm) with an acrosome (length: 1.2±0.2μm), a cylindrical midpiece (length: 1.8±0.5μm), a flagellum (length: 50.3±5.9μm) and a total length of 59.2±6.2μm. Ten posterolateral projections (PLPs) arise from the posterior edge of the acrosome and there were 3 endonuclear canals that traversed the nucleus from the acrosomal end to the basal nuclear fossa region. Three to six mitochondria were in peripheral midpiece and the proximal and distal centrioles were located near to "implantation fossa" and basement of the flagellum. The axoneme has a typical eukaryotic structure composed of 9 peripheral microtubules and a central pair of single microtubule surrounded by the plasma membrane. Lateral fins were observed along the flagellum. The fins started and ended at 0.5-1μm from midpiece and at 4-6μm from the end of flagellum. There were significant differences in the size of almost all measured morphological parameters between males and flagellar, midpiece and nucleus characters were more isolated parameters that can be considered for detecting inter-individual variations. This study showed that sperm morphology and fine structure are similar among sturgeon species, but the dimensions of the parameters may differ.     

The spermatozoon of Mengenilla moldrzyki (Strepsiptera,Mengenillidae): Ultrastructure and phylogenetic considerations

Even though the spermatozoa of several strepsipteran species were described earlier, no data were available for the basal family Mengenillidae. Well-fixed material of the recently described Tunisian species Mengenilla moldrzyki was used for a detailed examination of the sperm ultrastructure. The total length is c. 30 μm. The head region contains a conical acrosome vesicle (0.3-0.35 μm) and an elongated nucleus (7.3 μm) with dense chromatin. Some granular material along with a uniformely dense centriole adjunct and two mitochondrial derivatives are visible at the posterior end of the nucleus. The material of the centriole adjunct does not extend along the flagellum and accessory bodies are absent. The mitochondrial derivatives are elongated structures crossed by a longitudinal crista but lacking parallel transverse cristae and paracrystalline material in the dense matrix. The mitochondrial derivatives gradually reduce their size and end at the most posterior tail region. The flagellar axoneme has a 9 + 9 + 2 pattern and originates beneath the nucleus. In the terminal tail region the axoneme gradually disintegrates. Despite the extreme specialization of the endoparasitc group, strepsipteran spermatozoa are mostly characterized by plesiomorphies. The pattern within the order is largely uniform, but Mengenilla displays several apomorphic features compared to the presumptive strepsipteran groundplan (e.g., absence of crystallizations and cristae in the mitochondrial derivatives). The subdivision of the intertubular material into two compartments with a dense beak-like structure adhering to the tubular wall supports a clade Coleopterida (=Strepsiptera + Coleoptera) + Neuropterida.     

Sperm ultrastructure in honeycomb (foam) oysters (Mollusca,Bivalvia, Gryphaeidae,Pycnodontinae): comparison with other Ostreoidea and taxonomic implications

Sperm ultrastructural features of the honeycomb (foam) oysters Hyotissa hyotis, H. sinensis, and H. mcgintyi (Gryphaeidae) are described and compared with other Ostreoidea and more generally with other pteriomorphian Bivalvia. Spermatozoa of H. sinensis and H. mcgintyi (the type species of Parahyotissa Harry 1985) exhibit (1) a broad, low‐conical acrosomal vesicle; (2) subacrosomal material (very electron‐dense granular material and an almost electron‐lucent axial rod); (3) a spheroidal nucleus with a wide anterior invagination (filled with subacrosomal components); (4) a midpiece composed of four spherical mitochondria surrounding a pair of centrioles (rootlet associated with proximal centriole); and (5) a flagellum. Sperm of Hyotissa hyotis (type species of Hyotissa Stenzel 1971) differ markedly from those of H. sinensis and H. mcgintyi, in having (1) a conical acrosomal vesicle showing coarse granular texture anteriorly; (2) a very electron‐dense axial rod; (3) a barrel‐shaped nucleus with a long, narrow anterior invagination (filled with both subacrosomal components) and a basal invagination partly housing the proximal centriole; and (4) five midpiece mitochondria and no proximal centriolar rootlet. Results indicate that H. sinensis should be relocated to another genus, possibly a revised genus Parahyotissa, and also show that the sperm of H. sinensis and H. mcgintyi show many similarities to those of the Ostreidae, with the exception that the ‘axial rod’ component of the subacrosomal material is less electron‐dense than the surrounding substance (more dense in Ostreidae, as in H. hyotis). No family defining sperm features of the Gryphaeidae can be identified.     

Spermiogenesis in Odontophrynus cultripes (Amphibia,Anura, Leptodactylidae): Ultrastructural and cytochemical studies of proteins using E-PTA

Spermiogenesis in the South American leptodactylid frog Odontophrynus cultripes was analyzed ultrastructurally. The spermatids undergo morphological modification while still enclosed in microtubule-rich processes of Sertoli cells. Electron-dense plates resembling junctional structures appear in regions at which the spermatids lie in close contact with the surface of Sertoli cell processes. Spermatid differentiation can be divided into five distinct stages based mainly on chromatin condensation. In the late stages, the densely compacted chromatin loses reactivity to ethanolic phosphotungstic acid (E-PTA). Helical arrangements of microtubules appear in the cytoplasm that surrounds the spermatid nucleus after the second stage. The acrosomal vesicle differentiates into a cone-shaped acrosome that caps the anterior region of the nucleus. The connecting piece, located in the flagellum implantation zone, has transverse striations, and is continuous with the axial rod. The tail is formed by a 9 + 2 axoneme, an undulating membrane, and an axial rod that is rich in basic proteins as demonstrated by E-PTA staining.     

Ultrastructure of spermatozoa and spermiogenesis inSpirula spirula (L.): systematic importance and comparison with other cephalopods

Spermatozoa and spermiogenesis in the deep-water cephalopodSpirula sprirula (L.) are examined using transmission electron microscopy. Mature spermatozoa (taken from spermatophores) are elongate cells 115–120 μm long, composed of a conical acrosomal vesicle, cylindrical nucleus (6.8–7 μm long), flagellum and a loose mitochondrial sleeve — the latter concealing the proximal 6–8 μm of the flagellum. The acrosomal vesicle is 2.8 μm long with fibro-granular contents and an electron-lucent apical zone. Subacrosomal material, organized as closely packed granules, fills a basal invagination of the acrosomal vesicle. In early spermatids the flagellum is derived from a triplet substructure centriole positioned close to the developing nuclear invagination. As flagellum formation proceeds, the acrosomal vesicle (produced evidently through Golgi secretion) attaches to the condensing nucleus. Spermatids are connected by cytoplasmic bridges throughout their development, and exhibit a perinuclear sheath of microtubules from the onset of the fibrous stage of nuclear condensation (mid-, late spermatids). In mid-spermatids, mitochondria collect posterior to the nucleus and subsequently are packed into a cylindrical extension of the plasma membrane to form the periflagellar mitochondrial sleeve. These features of spermiogenesis and mature spermatozoa ofSpirula clearly associate the Spirulidae with the Sepiida, Teuthida and Sepiolida — particularly with the latter order. However, pending results of a thorough review of coleoid sperm morphology, the Spirulidae are here included in their own order — Spirulida (of Reitner & Engeser, 1982) — rather than in either the Sepiida or Sepiolida.     

Ultrastructure of euspermatozoa and paraspermatozoa in the volutid snail <Emphasis Type="Italic">Adelomelon ancilla</Emphasis> (Mollusca: Caenogastropoda)

The ultrastructure of the euspermatozoa and the paraspermatozoa is investigated in Adelomelon ancilla, through histological section observed by transmission electron microscopy. Euspermatozoa of A. ancilla consists of: (1) a conical acrosomal vesicle (with a short basal invagination, constricted anteriorly) which is flattened at the apex and associated with an axial rod, a centrally perforated basal plate and a short accessory membrane, (2) a rod-shaped, solid and highly electron-dense nucleus (with a short basal fossa containing a centriolar complex and a initial portion of a 9 + 2 axoneme), (3) an elongate midpiece consisting of the axoneme sheathed by 5–6 helical mitochondrial elements each exhibiting a dense U-shaped outer layer, (4) an elongate glycogen piece (where the axoneme is sheathed by nine tracts of glycogen granules), (5) a dense annulus at the junction of the midpiece and glycogen piece, and (6) a short free tail region (where the axoneme is surrounded only by plasma membrane). We observed a parasperm in A. ancilla. This is vermiform in shape and is composed of multiple axonemes and extensive cytoplasm with numerous vesicles, and mitochondria are scattered inside the axonemes. Sperm of A. ancilla is characterized by the euspermatozoa type 2 and the paraspermatozoa morphology belongs to type 5. The U shaped electrodense mitochondrial element in the midpiece of the eusperm and the constriction in the acrosomal vesicle present in A. ancilla are exclusive. We suggest that these characteristics could have taxonomic importance, because these was observed in other volutids and have not been observed in the rest of caenogastropods studies. We consider that the morphology of paraspermatozoa in A. ancilla corresponds to the “lancet” type.     

Ultrastructural analysis of spermiogenesis in Admetus pomilio (Arachnida,amblypygi)

The mature spermatozoon of Admetus pomilio is a spherical cell containing nucleus and tightly coiled flagellum. In early spermatids the Golgi apparatus forms the acrosomal vesicle and at the opposite side the distal centriole gives rise to the axonemal complex of the sperm tail. As the nucleus elongates, chromatin forms twisted filaments and the spermatid nucleus takes on a helical form. Microtubules are juxtaposed with the nucleus envelope, which is separated from a central chromatin mass by an electron lucid region. A long perforatorium, located on the border of the chromatin mass, runs helically in the nucleus from the centriolar region to subacrosomal space. During tail elongation, the anterior part of the axoneme is surrounded by a long, spiral mitochondrial sheath. In the late spermatid, chromatin filaments appear twisted and become aggregated. The nucleus and flagellum undergo further contortions in which the nucleus coils and the flagellum winds up into the body of the cell and coils in a regular fashion. The mitochondrial sheath surrounds about 2/3 of the 9 + 3 axoneme. These features of spermatid ultrastructure resemble those in the primitive Liphistiomorpha.     

Ultrastructure of Spermatozoa and Spermatids in Bonellia viridis and Hamingia arctica (Echiura) With Some Phylogenetic Considerations

Spermatozoa of the echiurans Bonellia viridis and Hamingia arctica show a similar ultrastructure. They are of a modified type. The head consists of a roughly cylindrical nucleus, which has a cover of electron-dense material. The acrosome is very large and consists of an acrosomal vesicle and a rod-shaped perforatorium or acrosomal rod. In close association with the nucleus, one or two mitochondria are found forming an irregular ring around the posterior tip of the nucleus and the centriolar apparatus. There are two centrioles, the proximal one with the conventional triplet microtubular structure. The tail flagellum is about 50 μm long and has the 9+2 axonemal structure. The oblique attachment of the acrosome to the anterior part of the nucleus gives the spermatozoon a bilateral symmetry. However, in the nuclear morphology, the arrangement of electron-dense material around the nucleus, in the mitochondria, and in the attachment of the tail flagellum, the spermatozoon shows asymmetric organization. The sperm structure in bonelliids is unique but its genesis and the morphology of the mitochondrial midpiece support the theory that the echiurans are related to the annelids. The main results of the study are summarized in Fig. 11.     

Sperm ultrastructure and spermiogenesis in two Exogone species (Polychaeta, Syllidae, Exogoninae)

Abstract. The spermatozoa of Exogone naidina and E. dispar are characterized by a prominent bell-shaped acrosome, a spheroidal nucleus, and a conventional flagellum. During spermiogenesis, the acrosomal vesicle undergoes conspicuous modifications leading to its final bell shape with a posterior opening. The subacrosomal material initially shows radiating filaments but in mature sperms it appears as a meshwork of electron-opaque material. The acrosomal axis is oblique with respect to the main longitudinal sperm axis. The chromatin is arranged in electron-opaque strands in the early spermatids, then becomes amorphous, and is finally organized in filaments in mature sperms. Centrioles are orthogonally arranged beneath the nucleus and fibers radiate from the distal centriole to contact the plasma membrane and the single mitochondrion. The latter is located eccentrically on the side of the nucleus opposite the acrosome. A disk-shaped structure is evident beneath the distal centriole. The flagellar axoneme has a 9+2 microtubule pattern. A conspicuous glycocalyx surrounds the flagellar plasma membrane, and an electron-lucent space is present between these two structures at the distal tip of the flagellum. We compare the sperm morphology of these two species of Exogone with that described in other members of the subfamily Exogoninae. The fine structure of these two species supports the occurrence of an ent-aquasperm type within Exogoninae, in accordance with the brood strategy present within this subfamily. The mode of reproduction is of taxonomic importance for defining subfamilies within Syllidae, and is likely also of phylogenetic significance. Because epitoky is probably plesiomorphic, the ent-aquasperm type found in Exogoninae can be considered a derived feature within Syllidae.