Sperm and spermatozeugma ultrastructure in the inseminating species Tyttocharax cochui, T. tambopatensis, and Scopaeocharax rhinodus (Pisces: Teleostei: Characidae: Glandulocaudinae: Xenurobryconini)

This article presents the scanning and transmission electron microscopy of the spermatozoa and sperm packets of three inseminating species of the glandulocaudine tribe Xenurobryconini. All three species, Scopaeocharax rhinodus, Tyttocharax cochui, and T. tambopatensis produce unencapsulated sperm packets (= spermatozeugmata) of similar morphology. The external anterior surface of each spermatozeugma is comprised of elongate sperm heads arranged in parallel, and the posterior part is made up of tightly packed flagella. The interior of the anterior portion consists of alternating layers of sperm heads and flagella. The remarkable integrity of each packet appears to be maintained through an electron-dense secretion seen among all parts of the cells. Spermatozeugma formation takes place within the spermatocysts at the end of spermiogenesis and at spermiation fully formed packets are released. Morphology of the mature spermatozoa was similar in all three species. Each nucleus is elongate, flattened along most of its length, and tapers at either end. The two centrioles are nearly parallel to one another and are located just anterior to the nucleus. Elongate mitochondria are located along the nucleus. The single flagellum, which lacks axonemal fins, is initially contained within a short cytoplasmic collar. Accessory microtubules run parallel to the long axis of the nucleus just beneath the plasma membrane. During spermiogenesis, no nuclear rotation occurs and the cytoplasmic canal containing the flagellum elongates along with the nucleus. However, prior to spermiation all but the anterior portion of the collar degenerates. The sperm modifications observed in these species are discussed as adaptations to the unique reproductive habit of insemination.     

STUDIES OF SPERMIOGENESIS IN A NEMATODE, NIPPOSTRONGYLUS BRASILIENSIS

The fine structure of the developing spermatids and the mature sperm of Nippostrongylus brasiliensis was investigated. Immature spermatids are found at one end of the tubelike testis, and the mature sperm at the other. The spermatid has a prominent nucleus, with the chromatin clumped at the margin. It also contains a pair of centrioles, located near the nucleus. The cytoplasm is filled with ribosomal clusters, but it lacks an organized Golgi area or endoplasmic reticulum. Besides the normal mitochondria, the spermatid has specialized mitochondrionlike inclusions with dense matrix, few broad cristae, and a crystalloid structure always facing the nucleus. As spermiogenesis proceeds, the nucleus elongates, comes to lie at one end, and later evaginates to form a separate head structure, leaving the mitochondria and other cytoplasmic organelles in a broad cytoplasmic region. The nuclear material becomes filamentous and spiral, and the centrioles come to lie at one end near the junction of the head and the cytoplasmic portion of the sperm. Microtubules are found in the cytoplasmic region extending from the tubelike nucleus. The specialized mitochondria are about eighteen in number, and are arranged in rows in staggered groups of three around the microtubules in the cytoplasmic region. The mature sperm is aflagellate and lacks an acrosome. No movement of the sperm was ever observed.     

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.     

Ultrastructure of spermiogenesis and the spermatozoon in Castrada cristatispina Papi, 1951 (Platyhelminthes, Rhabdocoela, Typhloplanida)

Spermiogenesis in Castrada cristatispina begins with the formation of a zone of differentiation containing two centrioles with associated striated rootlets and an intercentriolar body between them. The centrioles give rise to two parallel, free flagella of the Trepaxonemata 9 + '1' pattern, growing out in opposite directions. Spermatids undergo a latero-ventral rotation of the flagella and a subsequent disto-proximal rotation of centrioles, and a distal cytoplasmic projection appears. The former rotation involves the compression of a row of microtubules and allows the recognition of a ventral side and a dorsal side. At the end of the differentiation, the centrioles and cortical microtubules lie parallel to the sperm axis. The modifications of the intercentriolar body and the migration of the nucleus and the centrioles toward the distal projection are described. The mature spermatozoon of C. cristatispina is filiform, tapered at both ends and shares several features with the other Rhabdocoela gametes. Nevertheless, the posterior extremity is capped by an electron-dense material. A gradient between mitochondria and dense bodies exists along the sperm axis. This study has enable us a phylogenetic approach of the Rhabdocoela through a comparison of the ultrastructural features of C. cristatispina with the other Rhabdocoela taxa. We propose the disto-proximal rotation of centrioles as a synapomorphy of the Rhabdocoela.     

Flagellar gyration and midpiece rotation during extension of the acrosomal process of Thyone sperm: how and why this occurs

The midpiece of Thyone sperm contains a large mitochondrion and a centriolar pair. Associated with one of the pair, i.e., the basal body of the flagellum, are satellite structures which apparently anchor the flagellar axoneme to the mitochondrion and to the plasma membrane covering the midpiece. Immediately before and as the acrosomal process elongates, the flagellum and the midpiece begin to rotate at 1-2 rotations per second even though the head of the sperm, by being firmly attached on its lateral surfaces to the coverslip, does not rotate at all. This rotation is not observed in the absence of flagellar beating whose frequency is much greater than that of its gyration. To understand how the midpiece rotates relative to the sperm head, it is first necessary to realize that in Thyone the flagellar axoneme projects at an acute angle to the principal axis of the sperm and is bent towards one side of this axis. Thus movement of the flagellum induces the sperm to tumble or yaw in solution. If the head is stuck, the midpiece will rotate because all that connects the sperm head to the midpiece is the plasma membrane, a liquid-like layer. A finger-like projection extends from the proximal centriole into an indentation in the basal end of the nucleus. In contrast to the asymmetry of the flagellum, this indentation is situated exactly on the principal axis of the sperm and, along with the finger-like projection, acts as a biological bearing to maintain the orderly rotation of the midpiece. The biological purpose of flagellar gyration during fertilization is discussed.     

Giant,accordioned sperm acrosomes of the greater bulldog bat,Noctilio leporinus

Sperm of the greater bulldog bat Noctilio leporinus display an architecture that is totally unique among mammalian spermatozoa. The sperm head of Noctilio is extraordinarily large and flat and lies eccentrically with respect to the sperm tail. The major portion of the atypically large acrosome lies anterior to the nucleus and is shaped into a dozen accordionlike folds that run parallel to the long axis of the sperm. The ridge of each fold is shaped into ∼60 minute, evenly spaced rises that extend along the entire length of the fold. We speculate that acrosome ridges may serve to strengthen the sperm head during transport. Mol. Reprod. Dev. 48:90–94, 1997 © 1997 Wiley-Liss, Inc.     

瘤背石磺的精子结构

用光学显微镜和透射电子显微镜技术研究了瘤背石磺精子的结构特点,分析了其生理生态适应性以及在肺螺亚纲系统演化中的意义。瘤背石磺的精子由头部、中段和末段组成。头部由奶嘴形的顶体和长圆筒状的细胞核构成。顶体包括顶体囊和顶体构架体两部分;两者的内含物都分布均匀,电子密度稍低于细胞核;顶体基部平整,与核前端之间有一空隙,内含物电子密度极低。细胞核由电子密度高的均匀颗粒物质组成,并出现核泡;核的后端有一"杯形"的凹陷,称为核后窝。中段结构复杂,主要包括一对位于核后窝内的中心粒、轴丝、质膜、线粒体及由线粒体衍生的糖原质螺旋体、基质层和类晶体层等。末段由"9 2"结构的轴丝及外包的质膜组成,无糖原质螺旋体和其它线粒体衍生物。比较瘤背石磺精子与肺螺亚纲其它物种的精子结构,我们认为该物种的精子属于"进化型",是一类在进化地位中比基眼目高等的动物。     

An ultrastructural analysis of the mature spermatozoon of Anguispira alternata (say) (pulmonata,stylommatophora)

Mature spermatozoa from the hermaphroditic duct of adult snails were examined using various techniques of light microscopy as well as scanning and transmission electron microscopy. The sperm are approximately 557 μm in length including a dextrally spiral head approximately 13 μm long. The head consists of an electron-dense nucleus sculptured into a double-ridged spiral and an acrosome projecting approximately 0.45 μm beyond the apex of the nucleus. The acrosome consists of a membrane-bound vesicle approximately 0.1 μm in diameter and a column of homogeneous material which extends along one side of the terminal spiral of the nucleus. This material is separated from the nucleus by the nuclear envelope. The neck region, though similar to that found in other pulmonates, possesses a unique coiled structure surrounding the central doublet of the axoneme. The midpiece axoneme possesses a 9+9+2 configuration anteriorly grading into a 9+2 pattern for the majority of its length. There are three mitochondrial helices – one primary and two secondary – in the midpiece. Only the primary helix persists throughout the midpiece.     

The spermatozoon of the Echiniscidae (Tardigrada,Heterotardigrada) and its phylogenetic significance

The spermatozoon ultrastructure of four species of moss-dwelling Heterotardigrada belonging to four genera of Echiniscidae, namely Pseudechiniscus juanitae, Echiniscus duboisi, Novechiniscus armadilloides and Antechiniscus parvisentus, was investigated. In all species, the testicular male gamete is similar in morphology and in length. The spermatozoon is made up of a long head, consisting of a cylindrical acrosome and an oval or rod-shaped nuclear region which contains a nucleus with osmiophilic and electron-dense chromatin, and a tapering tail, with a "9+2" axoneme. An elongated sack-like structure originates from the posterior part of the head, extending beyond the main axis of the cell and running parallel to the tail. It consists of two parallel tubular regions which sometimes form a strict double helix and contain two voluminous, "free" mitochondria with unmodified cristae. In addition, a voluminous vesicle is present laterally to the centriole or between the end of the nucleus and the beginning of the mitochondria, limited by two cytomembranes and filled with electron-lucent and granular material. The male gametes representative of these moss-dwelling Echiniscidae are very similar to the spermatozoa of the marine Echiniscoididae Echiniscoides sigismundi. This close similarity emphasises that habitat changes have had little influence on the organisation of the sperm cell representative of Echiniscoidea. Spermatozoon characters which could be useful for phylogenetic studies on Tardigrada are discussed.     

Ultrastructure of the biflagellate spermatozoon of tomopteris helgolandica greef, 1879 (annelida,polychaeta)

The spermatozoon of the polychaete Tomopteris helgolandica is of an aberrant type with two flagella, each measuring about 40μm. The nucleus is roughly conical and weakly bent. At the anterior end it is rounded and covered only by the nuclear and plasma membranes. Membraneous, electron-dense structures are applied laterally to the nucleus. These structures may have a helical arrangement. The middle piece contains about ten mitochondria, two centrioles, and two centriolar satellite complexes. The centriolar regions are connected with the posterior part of the nucleus. The axonemes of the two tail flagella lack the usual central complex with central tubules, radial spokes, or related structures. No arms seem to be present on the A tubules of the doublets. In the middle piece the tail flagella are surrounded by invaginations of the plasma membrane forming flagellar canals. The sperm has a bilateral symmetry whereas the primitive sperm has a radial symmetry. The occurrence of two tail flagella in this spermatozoon has no phylogenetical connection with biflagellate spermatozoa in other animal groups. A series of mutations has resulted in the development of two flagella emerging from the two centrioles, the lack of a central complex in the axoneme, and the lack of a typical acrosome. In the Polychaeta, sperm structure is generally more related to function that to phylogenetics. During swimming the spermatozoon of Tomopteris rotates around its longitudinal axis.     

NUCLEAR CHANGES DURING SPERMIOGENESIS IN A PULMONATE SNAIL

Changes in both external form and internal arrangement of nuclear material have been investigated in the differentiation of the sperm of the pulmonate snail, Otala lactea. Sperm head differentiation begins with a flattening of the previously spherical nucleus and a thickening of the nuclear envelope covering the anterior and posterior surfaces of that nucleus. Tail filaments can be seen in the cytoplasm at this time. At a slightly later period the mitochondria begin to form the tail filament sheath and at this time the nucleus begins to elongate in a direction parallel to the future axis of the sperm. At the same time the nuclear material begins to orient itself at right angles to the nuclear surface which lines the pit occupied by the centriole. As nuclear elongation proceeds, this orientation of nuclear substance takes on the appearance of 60 A thick sheets bent in a direction parallel to the sperm axis. Soon the sheets fill the entire nucleus. The nucleus then begins to twist along its axis so that it starts to take on the mature, flame-shaped form. At this time the flat sheets begin to disappear and in the mature sperm head they are no longer visible (see Text-fig. 2 B).     

Sperm morphology in four species of African platypleurine cicadas (Hemiptera: Cicadomorpha: Cicadidae)

Mature spermatozoa from four species of platypleurine cicadas (Albanycada albigera, Azanicada zuluensis, Platypleura capensis and P. hirtipennis) were examined by light and electron microscopy. The filiform sperm have a similar ultrastructure in all species but notable variations were found in sperm dimensions. All species produce more than one discrete length of nucleated, motile sperm, a form of polymorphism termed polymegaly. Polymegaly is expressed in two ways: sperm have bi- or trimodal head and tail lengths. The anterior parts of sperm heads are embedded in an elongate homogenous matrix forming a spermatodesm. The conical acrosome is deeply invaginated posteriorly, and sits on top of the nucleus. The acrosomal contents are differentiated internally with an electron-lucent central medulla and a denser cortex. The homogenously electron-dense nucleus is pointed anteriorly and is generally cylindrical, although posteriorly there is a lateral invagination that extends part-way along the nucleus. This invagination houses fine granular material of the centriolar adjunct. Vesicle-like elements that are associated with both the posterior nucleus and the centriolar adjunct are also found within the invagination. Immediately posterior of and adjoining the centriolar adjunct is a pair of mitochondrial derivatives that are elongated and extend for almost the entire length of the tail. The absence of accessory bodies in cicada sperm suggests that within the Cicadomorpha, the families Cicadidae and Cercopidae are closely affiliated.     

Fine structure of spermatophores of some Oribatid mites (Acari,Arachnida)

The ultrastructure of spermatophores was studied in seven species of Oribatei. Each spermatophore is composed of a head and a stalk which is attached to the substratum. The spherical head consists of two distinct portions: (1) the sperm package; and (2) the head matrix. Position and structure of the sperm package are described in the different species, as well as the various structures in the head matrix. The sperm package contains sperm embedded in secretory products. The most characteristic feature of the sperm is an electron-dense chromatin body. Mitochondria lie adjacent to it or are partly or completely incorporated into the chromatin body. The cytoplasmic surface of the plasma membrane regularly bears an electron-dense layer; the plasma membrane is covered by a secretory sheath. The spermatophores of the Oribatei are structurally complicated and not uniform. The possible role of the spermatophore's structural elements is discussed and an attempt is made to evaluate characteristic features taxonomically.     

Organization of the Acrosome and Helical Structures in Sperm of the Aplysiid, Aplysia kurodai (Gastropoda, Opisthobranchia)

Spermiogenesis in the aplysiid, Aplysia kurodai (Gastropoda, Opisthobranchia) was studied by transmission electron microscopy, with special attention to acrosome formation and the helical organization of the nucleus and the other sperm components. In the early spermatid, the periphery of the nucleus differentiates into three characteristics parts. The first part is that electron-dense deposits accumulate on the outer nuclear envelope. This part is destined to be the anterior side of the sperm because a tiny acrosome is organized on its mid-region at the succeeding stage of spermiogenesis. The second part, in which electron-dense material attaches closely to the inner side of the nuclear envelope, is the presumptive posterior side. A centriolar fossa is formed in this part and the axoneme of the flagellum extends from the fossa. A number of lamellar vesicles derived from mitochondria assemble around the axoneme and form the flagellum complex. The third part is recognized by the chromatin which condenses locally along the inner nuclear envelope. During development of the spermatid, this part extends to form a spiral nucleus accompanied by chromatin condensation and formation of microtubular lamellae outside the extending nucleus.
Finally, in the mature sperm, a tiny, spherical acrosomal vesicle is detected at the apex. The slender nucleus, overlapping both the primary and secondary helices which are composed of different structural elements, winds around the flagellum axoneme.     

Structure and formation of the unusual sperm of Patelloida latistrigata (Mollusca : Patellogastropoda): implications for fertilization biology

The structure of the spermatozoa and spermatogenesis of the lottiid limpet Patelloida latistrigata is described by transmission electron microscopy. Although the lengths of the spermatozoa (about 60 μm) and their head region (about 12 μm) are similar to those of other patellogastropods, the structure of the sperm head and midpiece are very different. The head consists of an unusually large acrosome (about 11-μm long) with a broad posterior invagination that houses the relatively small nucleus. The midpiece mitochondria, which are rather elongate with large folded tubular cristae, are housed in a cytoplasmic sheath posterior to the nucleus. The proximal centriole is unusually elongate (about 2-μm long). The axoneme that emerges from the distal centriole is surrounded anteriorly by the cytoplasmic sheath in which the cytoplasmic side of the plasma membrane has electron-dense material. The flagellum is enlarged at its terminal end. Spermatogenesis is similar to that described for other patellogastropods. Patelloida latistrigata, therefore, has spermatozoa that seem to meet the morphological criteria of ent-aquasperm, which raises the question of whether fertilization is truly external in this limpet. However, it is also possible that the modifications to the sperm are linked to unknown specializations of the egg or egg envelope.     

Identification of a sperm nuclear annulus: a sperm DNA anchor

We report the isolation and characterization of a new nuclear structure from spermatozoa of the golden Syrian hamster which we term the nuclear annulus. The nuclear annulus was located at the implantation fossa, the point at which the tail is joined to the sperm head, inside the nucleus adjacent to the inner nuclear envelope. Extraction of sperm nuclei with 2 M NaCl and 10 mM dithiothreitol caused the solubilization of the protamines and DNA decondensation. This released the DNA from its structural constraints except that the DNA remained anchored to the nuclear annulus, which survived the extraction procedure. Nuclear annuli were isolated and examined by scanning electron microscopy. The nuclear annulus was a double-crescent, ring-shaped structure, 2.10 +/- 0.16 micron long and 1.36 +/- 0.13 micron wide. We believe that the nuclear annulus may play an important role in the organization of sperm DNA.     

Interspecific Variation in Structural Organisation of the Spermatozoon in the Asian Bandicoot Rats,Bandicota Species (family Muridae)

The structural organisation of the spermatozoon from two species of bandicoot rats Bandicota bengalensis and Bandicota indica was investigated by light and electron microscopy together with the effect of incubation in Triton-X 100 and sodium dodecyl sulphate. The sperm head of B. bengalensis is invariably falciform, has a uniform electron-dense nucleus capped by an acrosome with a posteriolateral equatorial segment, a subacrosomal cytoskeleton with a large rostral perforatorium, and a sperm tail, attached to the lower concave surface of the sperm head, with typical coarse fibres and fibrous sheath. By contrast, the sperm head shapes of B. indica are generally conical or bulbous, the nucleus contains a few large vacuoles, the acrosome lacks an equatorial segment, no recognisable perforatorium occurs, and the sperm tail, which is attached basally, is very short with only modest development of coarse fibres and fibrous sheath. These results indicate that, within the genus Bandicota, huge interspecific differences in morphology of the spermatozoon have evolved. The spermatozoa of B. bengalensis are similar to those of Rattus and many other murids and thus presumably represent the ancestral condition, whereas those of B. indica (and B. savilei) are unlike spermatozoa from any other eutherian mammal so far described. © 1998 The Royal Swedish Academy of Sciences. Published by Elsevier Science Ltd. All rights reserved     

Role of the cytoskeleton in sperm entry during fertilization in the freshwater bivalve Dreissena polymorpha

The present study examined the role of the cytoskeleton in sperm entry and migration through the egg cytoplasm during fertilization in the zebra mussel, Dreissena polymorpha (Bivalvia: Veneroida: Dreissenidae). Fertilization in this freshwater bivalve occurs outside the mantle cavity, permitting detailed observations of fertilization. After its initial binding to the egg surface, the sperm is incorporated in two stages: (1) a gradual incorporation of the sperm nucleus into the egg cortex, followed by (2) a more rapid incorporation of the sperm axoneme, and translocation of the sperm head through the egg cytoplasm. Initial incorporation into the egg cortex was shown to be microfilament dependent. Microfilaments were found in the sperm's preformed acrosomal filament, the microvilli on the egg surface, and in an actin-filled insemination cone surrounding the incorporating sperm. Treatment of eggs with cytochalasin B inhibited sperm entry in a dose- and time-dependent manner. Microtubule polymerization was not necessary for initial sperm entry. Following incorporation of the sperm head, the flagellar axoneme entered the egg cytoplasm and remained active for several minutes. Associated with the incorporated axoneme was a flow of cytoplasmic particles originating near the proximal end of the flagella. Inhibition of microtubule polymerization prevented entry of the sperm axoneme, and the subsequent cytoplasmic current was not observed. After sperm incorporation into the egg cortex, no appreciable microfilaments were associated with the sperm nucleus. A diminutive sperm aster was associated with the sperm nucleus during its decondensation, but no obvious extension toward the female pronucleus was observed. The sperm aster was significantly smaller than the spindle associated with the female pronucleus, suggesting a reduced role for the sperm aster in amphimixis.     

Sperm morphology in the black coral Cirrhipathes sp. (Anthozoa, Antipatharia)

Abstract. Male polyps of the antipatharian Cirrhipathes sp., collected along the coral reef of Siladen Island (Sulawesi, Indonesia), were studied in order to gain an insight into the reproductive biology. Spermatocysts (maximum size 120 μm) are located within the primary gametogenic mesenteries and are separated by mesenteric cell cytoplasmic extensions. Sperm, maturing along radial rows, have a fairly round shape and contain a series of electron-dense vesicles in the apical nuclear region. A single mitochondrion flanks the nucleus. A peculiar cup-like electron-dense body, edged with regularly spaced electron-dense granules, is interposed between the nucleus and the tail, and delimits a central region that includes two centrioles. Cross-sections of the cup-like body reveal that the distal centriole has a pericentriolar system, consisting of nine arms arranged in a radial pattern. Each arm branches into three processes that are connected to the electron-dense granules. Indirect evidence of spawning is derived from the accumulation of sperm in the gastric cavity. This process takes place through the lysis of the cells bordering the mesenteries. Intact cells of this bordering layer appear to be involved in the phagocytosis of non-expelled gametes.     

Morphological aspects of fertilization in Phallusia (Ascidia) nigra (Ascidiacea,Tunicata)

The spermatozoa of Phallusia (Ascidia) nigra have an elongated head (approximately 5 m in length) in which a nucleus and a single mitochondrion are located side by side. There is no midpiece. The apex of the head is wedge-shaped. Acrosomal vesicles (approximately 55–65 nm in diameter) and moderately electron-dense material (MEDM) are present between the plasmalemma and the nuclear membranes in the anterior tip of the head. The MEDM occupies a central position and three or four acrosomal vesicles are seen in a line alongside it. The acrosomal vesicles disappear as the sperm makes contact with the surface of the chorion. Gamete fusion most likely occurs between a small process extending from the peripheral margin of the sperm apex and the egg surface, resulting in incorporation of the sperm into the egg from the anterior region of its head.