Regional differences in composition of the perforatorium and outer periacrosomal layer of the rat spermatozoon as revealed by immunocytochemistry

The rat perforatorium is the part of the perinuclear theca that underlies the acrosomic system. It appears to be composed of several polypeptides. The main objective of this study was to determine the distribution of seven of these perforatorial polypeptides in the head of the rat spermatozoon. For this purpose, polyclonal antibodies were affinity purified from these polypeptides and tested 1) for their distribution on electron-microscope sections of late spermatids and spermatozoa by immunogold labeling and 2) for their specificity on Western blots of denatured perforatorial polypeptides by immunoblotting. Immunoblotting showed that all seven of the prominent perforatorial polypeptides had epitopes in common. Immunogold labeling of spermatozoa showed that antibodies against the 13, 13.4, and 16 kDa polypeptides were restricted in their localization to the thicker apical portion of the perforatorium and to the inner zone of the ventral spur. However, antibodies against the 34, 43, 57, and 63 kDa polypeptides reacted with the entire perforatorium but, in addition, reacted with the inner part of the ventral spur and with a portion of the "outer periacrosomal layer" lying between the plasma membrane and the outer acrosomal membrane. These results suggest 1) that there are regional differences in protein composition of the perforatorium, of the outer periacrosomal layer, and of the postacrosomal dense lamina; and 2) that perforatorial polypeptides may not necessarily be restricted to the subacrosomal region, but may also compose portions of the outer periacrosomal layer and postacrosomal dense lamina. Based on both immunoblotting and immunocytochemical results, using an antiactin monoclonal antibody that recognizes all known isoforms of actin, actin was not detected in the perforatorium of step 19 spermatids or spermatozoa. Actin, however, together with the seven perforatorial polypeptides tested, was present in the subacrosomal space of elongating spermatids before the process of condensation of the perforatorium takes place.     

Spermiogenesis and sperm structure in the crabUca tangeri (Crustacea,Brachyura), with special reference to the acrosome differentiation

Summary Early spermatids of the crabUca tangeri consists of the nucleus of granular chromatin and the cytoplasm, which contains a proacrosomal vesicle in close association with membrane lamellae. In the mid spermatids an invagination of the acrosomal vesicle membrane gives rise to the formation of the perforatorium, a spindle-shaped tubule which encloses tubular membranous structures. The pair of centrioles located at the base of the acrosome is not directly involved in perforatorial differentiation. The acrosomal vesicle shows a heterogeneous content composed of the operculum, the thickened ring, and three layers of different materials concentrically arranged around the perforatorium. During the late spermatid stage the nuclear profile differentiates numerous slender arms and the chromatin arranges into fibers. Membranous tubules from the cytoplasm become incorporated into the tubular structures of the perforatorium. The mature spermatozoon has the typical structure of the branchyuran sperm, with a complex acrosome, cupped by the nucleus, and a thin cytoplasmic band intervening between the former main elements. The centrioles are degenerate. The nuclear arms are unusually numerous (more than 20) and lack microtubules or microtubular derivatives.     

Differentiation of the acrosomal complex in ostrich (Struthio camelus) spermatids

The acrosomal complex of ostrich sperm consists of a small, cone-shaped acrosome and a slender, cylindrical perforatorium housed within a deep endonuclear canal. The perforatorium is almost exclusively endonuclear in location and is only covered by the acrosome at its point of origin in the apical subacrosomal space. The development of the acrosome is generally similar to that described in other non-passerine birds. Small proacrosomal granules (vesicles) emanating from the Golgi apparatus coalesce to form a large, membrane-bound acrosomal vesicle filled with homogeneous, electron-dense material. The acrosomal vesicle attaches to the nucleus via a shallow depression and subsequently collapses to form the typical cap-like acrosome of non-passerine birds. In ostrich spermatids the endonuclear canal becomes obvious when the collapsed acrosomal vesicle has assumed a dumbbell-shaped appearance. The perforatorium, which originates from moderately electron-dense material contained within the apical subacrosomal space, expands within the deepening endonuclear canal. The material of the perforatorium does not originate in the form of an obvious granule as in chicken and budgerigar spermatids. Indications are that in ostrich spermatids the developing acrosome plays a role in the shaping of the tip of the nucleus. The perforatorium, however, appears to represent a residual structure that has no specifically identified function. © 1996 Wiley-Liss, Inc.     

Structural changes in sperm from the fiddler crab, Uca tangeri (Crustacea, Brachyura), during the acrosome reaction.

The acrosome reaction (AR) was induced in sperm from the brachyuran crustacean Uca tangeri either by mixing male and female gametes in filtered seawater or by treating the spermatozoa with the divalent cation ionophore A23187. This latter method provided a sufficient number of reacted spermatozoa to allow a detailed ultrastructural study of the AR. The process consists of two separate phases: a) initial release of the acrosomal vesicle contents, and b) further elongation of the acrosomal filament, which causes reversal of the rigid capsule limiting the acrosomal vesicle contents. The elongate acrosomal filament consists of an apical perforatorium and a basal columnar structure called here the proximal piece. The former derives from the perforatorium of the uninduced sperm stage with only small ultrastructural changes. The proximal piece forms from myelin-like membrane layers which are initially distributed all around the subacrosomal region and then accumulate in a column at the perforatorial base, thus promoting a sudden forward projection of the perforatorium. The AR in brachyurans is thought to be a passive mechanism that utilizes the negative pressure exerted on the nucleus--caused by emptying of the acrosomal vesicle--for an organized accumulation of membrane-rich material immediately behind the perforatorium, with the final result of the raising of a 3 microns long acrosomal filament.     

Sperm ultrastructure of the spider crab Maja brachydactyla (Decapoda: Brachyura)

This study describes the morphology of the sperm cell of Maja brachydactyla, with emphasis on localizing actin and tubulin. The spermatozoon of M. brachydactyla is similar in appearance and organization to other brachyuran spermatozoa. The spermatozoon is a globular cell composed of a central acrosome, which is surrounded by a thin layer of cytoplasm and a cup‐shaped nucleus with four radiating lateral arms. The acrosome is a subspheroidal vesicle composed of three concentric zones surrounded by a capsule. The acrosome is apically covered by an operculum. The perforatorium penetrates the center of the acrosome and has granular material partially composed of actin. The cytoplasm contains one centriole in the subacrosomal region. A cytoplasmic ring encircles the acrosome in the subapical region of the cell and contains the structures‐organelles complex (SO‐complex), which is composed of a membrane system, mitochondria with few cristae, and microtubules. In the nucleus, slightly condensed chromatin extends along the lateral arms, in which no microtubules have been observed. Chromatin fibers aggregate in certain areas and are often associated with the SO‐complex. During the acrosomal reaction, the acrosome could provide support for the penetration of the sperm nucleus, the SO‐complex could serve as an anchor point for chromatin, and the lateral arms could play an important role triggering the acrosomal reaction, while slightly decondensed chromatin may be necessary for the deformation of the nucleus. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Spermatozoal ultrastructure in four genera of Homolidae (crustacea,decapoda): Exemplified byHomologenus sp.,Latreillopsis sp.,Homolomannia sibogae andParomolosis boasi

The spermatozoa ofHomologenus sp.,Latreillopsis sp.,Homolomannia sibogae andParomolopsis boasi confirm characteristics of a distinctive homolid spermatozoon previously established forHomola sp.,Paromola sp. andParomola petterdi. Homolid features are (1) moderate anteroposterior depression of the acrosome (ratio of length: width 0.4–0.6) as in lyreidine raninids (0.5), depression being greater in dromiids and dynomenids (both 0.3); (2) the capitate form of the perforatorium, shared with dromiids, dynomenids and lyreidine raninids; (3)the autapomorphic spiked-wheel form of the anterior expansion of the perforatorium; (4) horizontal zonation of the acrosome is possibly a unique synapomorphy of homolids with dromiids and dynomenids, and therefore an autapomorphy of the dromioid-homolid assemblage. In dromiids the posterior zone is proportionately the larger, while in homolids the anterior zone is the larger. The anterior zone is complexly subdivided in dynomenids; (5) the autapomorphic presence of numerous radial arranged extension of the acrosomal operculum into the perforatorium; (6) presence of nuclear arms, a symplesiomorphy of all investigated crabs, but small or questionably sometimes absent in Dromiidae; (7) absence of microtubules from the nuclear arms, as in dromiids, raninids, higher heterotremes and thoracotremes; (8) transient presence of a posterior median process of the nucleus. The process is not seen in dromiids but occurs in anomurans and lower heterotremes; (9) apical perforation of the operculum, also seen, apparently symplesiomorphically, in dromiids, raninids, and lower heterotreme families; (10) absence of an acrosome ray zone, probably homoplasic with absence in raninids; (11) location of most of the cytoplasm, including tortuous membranes and degenerating mitochondria, below the acrosome, also seen inLyreidus; (12) presence, in at least some species, of centrioles, unknown in dromiids and raninids and variable in occurrence in heterotremes.     

Ultrastructure of spermatogenesis in Cerastoderma glaucum (Cardiacea) and Spisula subtruncata (Mactracea)

Summary

In Cerastoderma glaucum, Sertoli cells are rich in lipids, glycogen and lysosomes, and premeiotic cells exhibited nuage, a prominent Golgi complex and endoplasmic reticulum cisternae encircling the nucleus. The Golgi complex gives rise to proacrosomal vesicles during mid-spermiogenesis, and the round acrosomal vesicle, with a dense fibrillar core, migrates laterally while linked to the plasma membrane as it develops the subacrosomal material. In its final position, the vesicle becomes cap-shaped (0.6 μm) and differentiates into apical light and basal dense regions. The elongated and helicoidal nucleus (8–9.9 μm) has a thin tip (0.3 μm) that invades the subacrosomal space, and in the midpiece (0.8 μm) two of the four mitochondria extend laterally to the nucleus (1.5–2.1 μm). In Spisula subtruncata, Sertoli cells are rich in lipids, glycogen and phagocytosed sperm. Premeiotic cells exhibit nuage, a prominent Golgi complex that gives rise to proacrosomal vesicles from the leptotene stage and a flagellimi that is extruded at zygotene. The acrosomal vesicle forms during the round spermatid stage and differentiates into a large and dense basal region and an apical light region. It then migrates while linked to the plasma membrane by its apical pole. Development of the subacrosomal perforatorium is associated with nuage materials and endoplasmic reticulum vesicles. The mature cap-shaped (0.6 μm) acrosomal vesicle exhibits a large apical and irregular region with floccular contents and a basal dense region. The round nucleus becomes barrel-shaped (1.5 μm) and the midpiece (0.8 μm), with four mitochondria, contains a few glycogen particles.     

An Ultrastructural Study of Spermatozoa of the Majidae with Particular Reference to the Aberrant Spermatozoon of Macropodia longirostris (Crustacea,Decapoda, Brachyura)

A total of 17 species, in 14 genera of majids have been examined for sperm ultrastructure. The present account describes the sperm of six of these species, in two subfamilies: Pisinae—Sphenocarcinus orbiculatus and Sphenocarcinus stuckiae and Inachinae—Cyrtomaia furici, Grypacheus hyalinus, Platymaia rebierei and Macropodia longirostris. M. longirostris has the only eubrachyuran sperm in which the acrosome is known to depart radically from a subspheroidal form. The acrosome is semilunar in shape and is bordered by a very thin layer of cytoplasm and an unusually uniform, narrow band of chromatin. The apical surface of the acrosome is almost flat, though slightly concave, whereas the posterior surface forms a hemisphere, and is almost completely occupied by the thin, centrally perforate, electron dense operculum. The bulk of the acrosome consists of a homogeneous, moderately electron dense outer acrosome zone. This surrounds a small inner acrosome zone internal to which is an ellipsoidal, pale perforatorium capped by a central acrosome zone. Majid sperm are distinguished by a flattened and/or centrally depressed operculum; a further characteristic is that the pointed perforatorium is relatively short and frequently does not reach the operculum. They vary inter alia with regard to presence or absence of a posterior median process and, apparently, of centrioles and of microtubules in the nuclear arms, and in the number of these arms. Perforation of the operculum, seen in the Pisinae, is not constant in the Inachinae. Spermatozoal ultrastructure offers no certain support for a close relationship of majids with parthenopids or hymenosomatids.     

The subacrosomal granule and its evolution during spermiogenesis in a lizard

Summary Some aspects of spermiogenesis have been studied in the testis of the teiid lizard Cnemidophorus lemniscatus lemniscatus by electron microscopy. Shortly after the acrosomal vesicle is lodged in a nuclear concavity of the spermatid, a dense granule differentiates in the center of the subacrosomal space. It is cone-shaped and shows a longitudinal striation. Its base applies to the acrosomal membrane and, through this, to the acrosomal granule. Its rounded vertex causes a depression of the nuclear membranes which, initially juxtaposed, separates at this point to form a vesicle. The granule develops and becomes a rod when spermiogenesis is advanced and the subacrosomal space has taken the form of a secondary cap. The rod is cylindrical, retains its original striation and has a convex acrosomal end. It encloses the vesicle formed by the nuclear envelope in its base and follows the apex of the nucleus. Meanwhile, the acrosomal granule loses its identity and the acrosomal cap is filled with a dense substance, in which a fringe of translucent material differentiates. This fringe lies in the dorsal and apical margins of the acrosome and is incompletely divided by longitudinal crests of the dense acrosomal substance. A projection of the Sertoli cell forms an accessory cap which envelops the acrosome and is in turn covered by the cytoplasm of the spermatid, constituting an intricate association. Two reflex membranes underlie the plasmalemma in the outer surface of the projection of the Sertoli cell. They are continuous with one another at their ends and with the cell membrane in the edge of pores. In the peripheral cytoplasm of the spermatid facing the accessory cap, numerous microtubules run longitudinally. By means of thin membranes some are interconnected or connected with the plasmalemma, from which they seem to originate.This research forms part of project N. 31.26.S1-0244 supported by the Consejo Nacional de Investigaciones Científicas y Tecnológicas     

Ultrastructure of spermiogenesis of Philippia (Psilaxis) oxytropis,with special reference to the taxonomic position of the architectonicidae (Gastropoda)

Summary Spermiogenesis of the architectonicid Philippia (Psilaxis) oxytropis was studied using transmission electron microscopy. Both spermatids and mature sperm of Philippia show features comparable to sperm/spermatids of euthyneuran gastropods (opisthobranchs, pulmonates) and not mesogastropods (with which the Architectonicidae are commonly grouped). These features include: (1) Accumulation of dense material on the outer membrane of anterior of the early spermatid nucleus — this material probably incorporated into the acrosome; (2) Structure of the unattached and attached spermatid acrosome (apical vesicle, acrosomal pedestal) accompanied by curved (transient) support structures; (3) Formation of the midpiece by individual mitochondrial wrapping around the axonemal complex, and the subsequent fusion and metamorphosis of the mitochondria to form the midpiece; (4) Presence of periodically banded coarse fibres surrounding the axonemal doublets and intra-axonemal rows of granules. A glycogen piece occurs posterior to the midpiece but is a feature observed in both euspermatozoa of mesogastropods (and neogastropods) and in sperm of some euthyneurans.Despite the lack of paracrystalline material or glycogen helices within the midpiece (both usually associated with sperm of euthyneurans), the features of spermiogenesis and sperm listed indicate that the Architectonicidae may be more appropriately referable to the Euthyneura than the Prosobranchia.Abbreviations a acrosome - ap anterior region of acrosomal pedestal - as support structures of spermatid acrosome - av apical vesicle of acrosome (acrosomal vesicle of un-attached acrosome) - ax axoneme - b basal region of acrosomal pedestal - c centriole - cf coarse fibres - cr cristal derivative of midpiece - db intra-axonemal dense granules - drs dense ring structure - gg glycogen granules - gp glycogen piece - G Golgi complex - m mitochondrion - mt microtubules - n nucleus - pm plasma membrane - sGv small Golgi vesicles     

北草蜥精子的超微结构--兼评不同类群蜥蜴精子形态的差异

应用透射电镜对北草蜥精子的超微结构研究结果表明,北草蜥精子头部顶体囊始终呈圆形,由皮质和髓质组成;顶体囊单侧脊的皮质与髓质问具电子透亮区;穿孔器1个,无穿孔器基板;具顶体下腔;细胞核长形,核内小管缺,核前电子透亮区缺,核肩圆。尾部颈段具片层结构。中段短,多层膜结构缺;纵切面上具2层线粒体;横切面上每圈线粒体6个;2组致密体,具连续的环状结构;线粒体与环状结构的排列模式：rs1／mi1、rs2／mi2;纤维鞘伸人中段,具终环。主段前面部分具薄的细胞质颗粒区;纤维3和8至主段前端消失;轴丝呈“9＋2”型。蜥蜴科内不同种类的线粒体数目不同,但都具有2组致密体。不同类群蜥蜴的顶体囊、顶体下腔、核前电子透亮区、穿孔器基板、核肩,以及线粒体与致密体的数目和排列方式等精子超微结构特征都为研究蜥蜴的系统发生提供了辅助信息。     

The Ultrastructure of the Spermatozoa of Squamata—I. Scincidae,Gekkonidae and Pygopodidae (Reptilia)

Abstract Squamate autapomorphies seen in sperm of the Scincidae (e.g. Ctenotus robustus, Carlia pectoralis, Cryptoblepharus virgatus, and Lampropholis delicata) are penetration of the fibrous sheath of the axoneme into the midpiece, and the paracrystalline subacrosomal cone. Sphenomorphus group spermatozoa (e.g. Ctenotus) and the Egernia group (Tiliqua) differ from the more derived Eugongylus group (C. virgatus, L. delicata and C. pectoralis) in that the acrosome is elongate and apically depressed; the perforatorium is strongly oblique; the midpiece is relatively short, with four dense ring structures in longitudinal succession; mitochondria are columnar; and enlarged peripheral fibres 3 and 8 do not show the gross anterior enlargement seen in Carlia and Lampropholis. Heteronotia binoei (Gekkonidae) sperm have no epinuclear electron-lucent region; nuclear shoulders are smooth, as in sphenomorph but not Eugongylus group skinks; mitochondria are columnar; unlike skinks, the median surfaces of the mitochondria are indented by triangular, sometimes longitudinally, interconnected dense bodies. In Lialis burtonis (Pygopodidae) sperm, the perforatorium extends virtually to the tip of the fore-shortened apically domed acrosome; nuclear shoulders are absent; the mitochondria alternate singly or in groups with one or more dense bodies which also form an interrupted collar around the distal centriole. Spermatozoal ultrastructure suggests that a common ancestry of snakes and pygopods deserves consideration.     

Development of the acrosomic system of the spermatozoon in the Norwegian lemming (Lemmus lemmus)

Summary An electronmicroscopic study on the development of the acrosome system of the spermatozoon in the Norwegian lemming (Lemmus lemmus) has been performed. The proacrosomal granules were found to be few and to consist of a dense center and a less dense periphery, and the acrosomal vesicle to contain stainable material of the same structure but of lower density than that of the acrosomal granule. Like in the guinea pig, two zones of different density exist throughout acrosome development and are visible also in mature spermatozoa. A large osmiophilic formation consisting of saccular, tubular, and lamellar structures, was found between the apex of the condensed nucleus and the acrosome and was identified as perforatorium.We are greatly thankful to Dr. Antti Telkkä, and Mr. Mauri Nyholm, M. Sc. for expert advise in electronmicroscopy and Mr. P. Lehtimäki for photographic help. For the supply of the lemmings we are indebted to Prof. K. Lagerspetz, and Mr. O. Hissa, M. Sc. This work is related to a series of studies on the biology of the Norwegian lemming (for previous works see Asp et al.).     

Ultrastructural study of spermiogenesis in a rare desert amphisbaenian Diplometopon zarudnyi

Spermiogenesis, in particular the head differentiation of Diplometopon zarudnyi, was studied at the ultrastructural level by Transmission Electron Microscope (TEM). The process includes acrosomal vesicle development, nuclear elongation, chromatin condensation and exclusion of excess cytoplasm. In stage I, the proacrosomal vesicle occurs next to a shallow fossa of the nucleus, and a dense acrosomal granule forms beneath it. This step commences with an acrosome vesicle forming from Golgi transport vesicles; simultaneously, the nucleus begins to move eccentrically. In stage II, the round proacrosomal vesicle is flattened by projection of the nuclear fossa, and the dense acrosomal granule diffuses into the vesicle as the fibrous layer forms the subacrosomal cone. Circular manchettes surrounded by mitochondria develop around the nucleus, and the chromatin coagulates into small granules. The movement of the nucleus causes rearrangement of the cytoplasm. The nucleus has uniform diffuse chromatin with small indices of heterochromatin. The subacrosome space develops early, enlarges during elongation, and accumulates a thick layer of dark staining granules. In stage III, the front of the elongating nucleus protrudes out of the spermatid and is covered by the flat acrosome; coarse granules replace the small ones within the nucleus. One endonuclear canal is present where the perforatorium resides. In stage IV, the chromatin concentrates to dense homogeneous phase. The circular manchette is reorganized longitudinally. The Sertoli process covers the acrosome and the residues of the cytoplasmic lobes are removed. In stage V, the sperm head matures.     

The ultrastructure of the spermatozoon ofParadynomene tuberculata Sakai, 1963 (Crustacea,Brachyura, Dynomenidae): Synapomorphies with dromiid sperm

The dynomenid spermatozoon, exemplified here byParadynomene tuberculata, resembles the spermatozoa of the Dromiidae, Homolidae and lyreidine raninoids and differs markedly from those of other crabs (the heterotreme, thoracotremes, raninines and raninoidines) in the depressed, discoidal form of the acrosome and the capitate form of the perforatorium. Four or five apparent dynomenid—dromiid sperm synapomorphies are recognizable. (1) Dynomenids (P. tuberculata) and dromiids differ from homolids and lyreidines in the greater depression of the acrosome (ratio of length to width=0.3); (2) the capitate head of the perforatorium is bilaterally prolonged inP. tuberculata as in dromiids though symmetrical in homolids; (3) dynomenid and dromiid sperm lack the—albeit variably developed—posterior median process of the nucleus seen in homolids, anomurans, raninoids and lower heterotremes; (4)P. tuberculata, like dromiids and less distinctly homolids, has an apical protuberance of subopercular material through the opercular perforation, unknown in other crabs, being distinct from the apical button of thoracotreme sperm; (5) a less certain synapomorphy is the anterolateral electron-pale peripheral zone of the acrosome. These synapomorphies endorse a sister-group relationship of dynomenids and dromiids,P. tuberculata sperm differs notably from the sperm of dromiids in the more complex zonation of the acrosome. The perforatorium lacks the radial rays (“spiked wheel”) of homolid sperm and does not show the “amoeboid” form seen in lyreidines. Absence of internal corrugations of the perforatorial chamber is a major difference from all examined raninids. Centrioles are only very tentatively identifiable. Nuclear arms are absent in glutaraldehyde fixed spermatozoa ofP. tuberculata and have not been observed in the dromiidPetalomera lateralis but are present as three small radial vertices in the dromiidDromidiopsis edwardsi and in homolids.P. tuberculata resemblesPetalomera lateralis in the large size of the sperm nucleus relative to the acrosome compared withD. edwardsi and homolids.     

Ultrastructure of spermiogenesis in the American alligator,Alligator mississippiensis (Reptilia,Crocodylia, Alligatoridae)

Testicular samples were collected to describe the ultrastructure of spermiogenisis in Alligator mississipiensis (American Alligator). Spermiogenesis commences with an acrosome vesicle forming from Golgi transport vesicles. An acrosome granule forms during vesicle contact with the nucleus, and remains posterior until mid to late elongation when it diffuses uniformly throughout the acrosomal lumen. The nucleus has uniform diffuse chromatin with small indices of heterochromatin, and the condensation of DNA is granular. The subacrosome space develops early, enlarges during elongation, and accumulates a thick layer of dark staining granules. Once the acrosome has completed its development, the nucleus of the early elongating spermatid becomes associated with the cell membrane flattening the acrosome vesicle on the apical surface of the nucleus, which aids in the migration of the acrosomal shoulders laterally. One endonuclear canal is present where the perforatorium resides. A prominent longitudinal manchette is associated with the nuclei of late elongating spermatids, and less numerous circular microtubules are observed close to the acrosome complex. The microtubule doublets of the midpiece axoneme are surrounded by a layer of dense staining granular material. The mitochondria of the midpiece abut the proximal centriole resulting in a very short neck region, and possess tubular cristae internally and concentric layers of cristae superficially. A fibrous sheath surrounds only the axoneme of the principal piece. Characters not previously described during spermiogenesis in any other amniote are observed and include (1) an endoplasmic reticulum cap during early acrosome development, (2) a concentric ring of endoplasmic reticulum around the nucleus of early to middle elongating spermatids, (3) a band of endoplasmic reticulum around the acrosome complex of late developing elongate spermatids, and (4) midpiece mitochondria that have both tubular and concentric layers of cristae. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Fine structure of acrosome biogenesis and of mature sperm in the bivalve molluscs<Emphasis Type="Italic"> Glycymeris</Emphasis> sp. (Pteriomorphia) and<Emphasis Type="Italic"> Eurhomalea rufa</Emphasis> (Heterodonta)

Proacrosomal vesicles form during the pachytene stage, being synthetized by the Golgi complex in Glycymeris sp., and by both the Golgi and the rough endoplasmic reticulum in Eurhomalea rufa. During early spermiogenesis, a single acrosomal vesicle forms and its apex becomes linked to the plasma membrane while it migrates. In Glycymeris sp., the acrosomal vesicle then turns cap-shaped (1.8 μm) and acquires a complex substructure. In E. rufa, proacrosomal vesicles differentiate their contents while still at the premeiotic stage; as the acrosomal vesicle matures and its contents further differentiate, it elongates and becomes longer than the nucleus (3.2 μm), while the subacrosomal space develops a perforatorium. Before condensation, chromatin turns fibrillar in Glycymeris sp., whereas it acquires a cordonal pattern in E. rufa. Accordingly, the sperm nucleus of Glycymeris sp. is conical and elongated (8.3 μm), and that of E. rufa is short and ovoid (1.1 μm). In the midpiece (Glycymeris sp.: 1.1 μm; E. rufa: 0.8 μm), both species have four mitochondria encircling two linked orthogonal (Glycymeris sp.) or orthogonal and tilted (30–40°; E. rufa) centrioles. In comparison with other Arcoida species, sperm of Glycymeris sp. appear distinct due to the presence of an elongated nucleus, a highly differentiated acrosome, and four instead of five mitochondria. The same occurs with E. rufa regarding other Veneracea species, with the acrosome of the mature sperm strongly resembling that of the recent Mytilinae. Electronic Publication     

Spermiogenesis in the Oligochaetoid Polychaete Questa (Annelida, Questidae)

The spermatids are connected to a central cytophore by cytoplasmic bridges and are polarized in the sequence: "empty cytoplasm"; uncondensed nucleus; mitochondria which surround the distal region of the nucleus and the centrioles; axoneme; posterolateral to the base of the axoneme, the Golgi apparatus and (when secreted) the acrosomal rudiment. The dome-shaped acrosome vesicle elongates progressively as it migrates to the tip of the elongating and condensing nucleus; subacrosomal material gives rise to an almost equally long, tubular, thick-walled perforatorium. After the acrosome has greatly elongated, the mitochondria are reduced to two, which lose their rounded form and invest the growing axoneme to give a very elongate midpiece. Transfer of materials from nucleus to mitochondria is discussed. Microtubules surrounding the acrosome and nucleus disappear by maturity, but those internal to the mitochrondria apparently persist as the accessory microtubules, unique in the Annelida, which surround the 9 + 2 axoneme. Microvilli of the egg envelope, which have tetrads of terminal branches (epivitelline projections) resembling epicuticular projections, are less than 1 μm long, whereas the mature acrosome exceeds 5 μm. This suggests that the correlation seen in oligochaetes is absent.     

The ultrastructure of the spermatozoa of Epipedobates flavopictus (Amphibia,Anura, Dendrobatidae), with comments on its evolutionary significance

We describe, for the first time, the spermatozoon ultrastructure of a dendrobatid frog, Epipedobates flavopictus. Mature spermatozoa of E. flavopictus are filiform, with a moderately curved head and a proportionally short tail. The acrosomal vesicle is a conical structure that covers the nucleus for a considerable distance. A homogeneous subacrosomal cone lies between the acrosome vesicle and the nucleus. The nucleus contains a nuclear space at its anterior end, and electron-lucent spaces and inclusions. No perforatorium is present. In the midpiece, the proximal centriole is housed inside a deep nuclear fossa. Mitochondria are scattered around the posterior end of the nucleus and inside the undulating membrane in the anterior portion of the tail. In transverse section the tail is formed by an U-shaped axial fiber connected to the axoneme through an axial sheath, which supports the undulating membrane. The juxta-axonemal fiber is absent. The spermatozoon of E. flavopictus has several characteristics not observed before in any anurans, such as a curved axial fiber, absence of a juxta-axonemal fiber, and presence of mitochondria in the typical undulating membrane. Our results endorse the view that, in anurans, the conical perforatorium and subacrosomal cone are homologous and that Dendrobatidae should be grouped within Bufonoidea rather than Ranoidea.