峨眉髭蟾精子形态结构及分类学意义

应用透射电镜、扫描电镜和光学显微镜对峨眉髭蟾(Vibrissaphoraboringii)精子的形态和超微结构研究的结果表明:峨眉髭蟾的精子具角蟾科物种精子基本的形态和结构特征,即精子头部呈螺旋状,尾部呈弯曲状;精子具锥形的顶体、纤维束构成的穿孔器、平行排列的中心粒和双轴丝;线粒体位于尾部;精子核窝不明显、无轴纤维和波动膜等特征。此外,对已有报道的角蟾科和无尾类物种精子的特征进行分析比较表明:(1)角蟾科精子细胞核呈螺旋状,中心粒平行排列,尾部具双轴丝等结构不同于无尾类其他科精子的结构,具有明显的科间差别;(2)角蟾科精子各部的量度,尾部线粒体的分布和数量,以及轴丝的排列等特征在属间和种间表现出明显的差异;(3)峨眉髭蟾和东南亚拟髭蟾指名亚种精子的形态和超微结构存在明显的差异。     

Ultrastructure of the spermatozoa of the flatworms Phaenocelis peleca and Boninia divae (Platyhelminthes, Polycladida)

The ultrastructure of spermatozoa of the acotylean Phaenocelis peleca and the cotylean Boninia divae is described. All spermatozoa are filiform and biflagellate with a 9+"1" microtubular pattern in the axoneme. Sperm characters in P. peleca follow the morphologies described for other acotyleans, with axonemes exiting the sperm shaft at the distal end and remaining in close contact with the sperm membrane. The nucleus occupies the proximal region of the shaft, and two types of dense bodies and mitochondria are located at the distal end. Unlike other members of the Cotylea, the axonemes of B. divae spermatozoa are incorporated into the sperm shaft, leaving the shaft at some distance from the distal end and then remaining free. This type of morphology is characteristic for acotyleans. Additionally, the spermatozoa of B. divae contain only one type of dense bodies plus a unique structure, which we call a central core. The nucleus in this species is unique as well; it shows periodic constrictions and rings of electron-dense granules, characters that further contribute to the distinct status of Boniniidae.     

Spermiogenesis in Paratomella rubra(Platyhelminthes, Acoela): Ultrastructural, Immunocytochemical, Cytochemical Studies and Phylogenetic Implications

Spermiogenesis and sperm structure of the primitive acoel Paratomella rubra from the Ligurian Sea, Italy, were investigated by several methods. During spermiogenesis, after flagellar incorporation by formation of two longitudinal lateral grooves, spermatid elongation is characterized in Paratomella by the presence of four membranes encircling each axoneme plus two membranes encircling both the axonemes and the nucleus. These structures were interpreted as being three cytoplasmic canals situated one inside the other. The filiform spermatozoon has two incorporated axonemes of 9+2 type, a nucleus almost as long as the sperm cell itself, a single elongate mitochondrion, and two types of membrane-bound granules, respectively, small and gastrula-shaped, and large. Organelles are highly ordered, the sperm is bilaterally symmetrical with a single long mitochondrion on the ventral side and a regular row of large granules, for some length embedded in the nucleus, on the dorsal side. Immunocytochemical studies and the use of fluorescent nuclear dyes reveal the spatial relationships of the axonemes with the nucleus. The granules were shown by Thiéry, PTA and enzyme digestion tests to contain glycoproteins and/or polysaccharides and very little protein. Glycogen particles were detected in the cytoplasm. Cells containing coiled spermatozoa undergoing resorption were found in the parenchyma. New apomorphies of the taxon Paratomella based on sperm structure are proposed: a very long nucleus, a highly bilaterally symmetrical pattern of organelles, a single long mitochondrion. These characters are not found in other Acoela, and particularly in Hesiolicium , sometimes considered closely related to Paratomella . © 1997 Published by Elsevier Science Ltd on behalf of The Royal Swedish Academy of Sciences.     

Sperm structure of Trichoptera. III. Hydropsychidae,polycentropodidae and philopotamidae (Annulipalpia)

Spermatozoa from 9 species belonging to 3 families of trichopteran suborder Annulipalpia (Philopotamidae : Philopotamus ludificatus, P. montanus, Wormaldia occipitalis, W. copiosa; Polycentropodidae: Plectrocnemia geniculata, Polycentropus mortoni, P. irroratus, Cyrnus trimaculatus; Hydropsychidae: Hydropsyche pellucidula) were studied by light, scanning, and transmission electron microscopy. The absence of axonemal dynein arms and a consequent sperm immotility are characteristic of the species studied. The presence of 7, rather than 2, central microtubules is shared by Polycentropodidae and some Philopotamidae. A greater variability in the sperm axoneme was found within Philopotamidae than in the other families. The species of the genus Wormaldia have an axial vesicle rather than microtubules and their axonemes have a 9 + 9 + 0 or 13 + 13 + 0 pattern. Hydropsychidae have spermatozoa provided with numerous finger-like appendages containing microtubular doublets. The progressive loss of sperm flagellum and motility within the group is discussed.     

Electron microscope study of spermatogenesis in two cestodes acanthobothrium filicolle benedenii Loennberg, 1889 and Onchobothrium uncinatum (Rud., 1819) (Tetraphyllidea, Onchobothriidae) (author's transl)]

The spermatogenesis, spermatozoon differentiation and fine structure of Acanthobothrium filicolle benedenii Loennberg, 1889 and Onchobothrium uncinatum (Rud., 1819) were studied by means of light and electron microscopy. Spermatogenesis in both species is of the rosette type. During sperm differentiation, the five following stages have been distinguished: (1) formation of arching membranes and differentiation zone; (2) nuclear elongation; (3) formation of two flagella with a median cytoplasmic process; (4) flagellar rotation; (5) fusion of two flagella with the median cytoplasmic process induced by the migration of nucleus into the latter. The mature spermatozoa of both species are threadlike structures about 250 mum long. They consist of two axonemes of the platyhelminth type (9+1 pattern), elongated nucleus and cortical microtubules embedded in a cytoplasmic matrix containing numerus beta-glycogen particles. The body which appears on cross-sections as crest, lateral with respect to the axoneme, is present in both species. Such a body is reported for the first time in Cestode spermatozoa. There is no mitochondrion in the two Onchobothriidae sperms studied.     

滇蛙多盘吸虫精子超微结构

利用透射电镜研究多盘吸虫精子的形态和结构。它属于单殖类中Ｐｏｌｙｏｐｉｓｔｈｏｃｏｔｙｌｅａ类型。精子细线形,含两根融入精子内的９＋１型轴丝,细长的细胞核,纤细的线粒体和微管。在第二根轴丝出现前的区段无微管的分布,具两根轴丝的主干段轴丝外无外侧管;细胞核消失后微管呈整圈排列;精子后部含糖原颗粒,背腹面中央向髓布内凹呈哑铃状并两侧向外突出形成波动膜;由精细胞则来的区段具外饰物。     

Implications phylogénétiques de I'ultrastructure de la spermatogenèse, du spermatozoïde et de I'ovogenèse du TurbellariéUrastoma cyprinae ("Prolecithophora", Urastomidae)

Spermiogenesis in Urastoma cyprinae (Graff. 1882) involvcs a progressive lengthening of the spermatid. Free flagella are only transitory. The mature spermatozoon is fusibrm. 45μm in length and 2.5 μm in width. It contains two incorporated axonermes of the flatworm 9+"1" pattern, two elongated mitochondria. an elongated nucleus and a row or cortical longitudinal microtubules. Observations on oogenesis concerncd only the immuturc ovary. lmmature oocytes contain few dense granules and accssory cells were not ohserved. Phylogenetic implications of a biflagellate spermatozoon in a Prolecithophora are important. The presence of two 9 +"1" axonemes confirms that Urastoma (and the Prolecithophora) belongs to the taxon Trepaxonemata Ehlers. Previous electron microscope studies on spermitozoa of Prolecithophora (four genera) only dealt with aflagellate spermatozoa. On this basis, Ehlers (1985) proposed two autapomorphics for the taxon Prolecithophora: aflagellate spermatozoon and spermatozoal mitochondrial derivatives with abundant membranes. The present observations on Urastoma contradict these two autapomorphics. The taxon Prolecithophora cannot be defined by autapomorphies of the spermatozoon.     

Sperm motility in fishes. (II) Effects of ions and osmolality: a review

The spermatozoa of most fish species are immotile in the testis and seminal plasma. Therefore, motility is induced after the spermatozoa are released into the aqueous environment during natural reproduction or into the diluent during artificial reproduction. There are clear relationships between seminal plasma composition and osmolality and the duration of fish sperm motility. Various parameters such as ion concentrations (K+, Na+, and Ca2+), osmotic pressure, pH, temperature and dilution rate affect motility. In the present paper, we review the roles of these ions on sperm motility in Salmonidae, Cyprinidae, Acipenseridae and marine fishes, and their relationship with seminal plasma composition. Results in the literature show that: 1. K+ is a key ion controlling sperm motility in Salmonidae and Acipenseridae in combination with osmotic pressure; this control is more simple in other fish species: sperm motility is prevented when the osmotic pressure is high (Cyprinidae) or low (marine fishes) compared to that of the seminal fluid. 2. Cations (mostly divalent, such as Ca2+) are antagonistic with the inhibitory effect of K+ on sperm motility. 3. In many species, Ca2+ influx and K+ or Na+ efflux through specific ionic channels change the membrane potential and eventually lead to an increase in cAMP concentration in the cell, which constitutes the initiation signal for sperm motility in Salmonidae. 4. Media that are hyper- and hypo-osmotic relative to seminal fluid trigger sperm motility in marine and freshwater fishes, respectively. 5. The motility of fish spermatozoa is controlled through their sensitivity to osmolality and ion concentrations. This phenomenon is related to ionic channel activities in the membrane and governs the motility mechanisms of axonemes.     

Initiation of Sperm Motility Induced by Cyclic AMP in Hamster and Boar

When the plasma membrane of hamster and boar spermatozoa was extraced by treatment with Triton X-100 and the demembranated spermatozoa were transferred to a reactivating medium containing only ATP, axonemes were initially immotile, and then gradually became motile. Under these experimental conditions, the cAMP content in the reactivating medium increased soon. This suggests that cAMP is synthesized from ATP by adenylate cyclase involved in incompletely removed or solubilized residual sperm membrane and that the autosynthesized cAMP causes the delay in motility initiation. This delayed initiation of motility did not occur when phosphodiesterase was added to the reactivating medium and the phosphodiesterase-dependent quiescent sperm became motile instantaneously at any time when excess cAMP was supplemented. Furthermore, demembranated sperm which were diluted in the reactivating medium containing ATP and cAMP, immediately became motile. cAMP levels in the cell increased during the initiation of sperm motility in both species. These results suggest that cAMP is the real factor indispensable for the initiation of sperm motility at ejaculation in mammals.     

A new viviparous acoel Childia vivipara sp. nov. with observations on the developing embryos, sperm ultrastructure, body wall and stylet musculatures

A free‐living viviparous acoel, Childia vivipara sp. nov., from the Gullmar fjord of the Swedish coast is described. The new species is assigned to the taxon Childia based on histological, ultrastructural and molecular sequence similarities. All available molecular markers (18S rRNA, 28S rRNA and histone H3) and several morphological characters, obtained using transmission electron microscopy and confocal scanning laser microscopy of whole mount specimen stained with TRITC‐labelled phalloidin, support the placement of C. vivipara in the taxon Childia. Childia vivipara and other Childia species share the following morphological synapomorphies: well‐developed copulatory organs built of tightly packed stylet needles, proximal part of the stylet inserted into the seminal vesicle, reversed body‐wall musculature, absence of ventral diagonal muscles, presence of dorsal diagonal muscles, and presence of ventral straight longitudinal muscles between frontal pore and mouth, 9 + 1 sperm axoneme structure, six distal sperm cytoplasmic microtubules, and extensive overlap of axonemes and nucleus. The new species can be easily distinguished from other Childia species by its viviparous mode of reproduction and single curved stylet. Observations on late embryonic development based on the oldest developing embryos are discussed.     

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

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

A monoclonal antibody against the dynein IC1 peptide of sea urchin spermatozoa inhibits the motility of sea urchin, dinoflagellate, and human flagellar axonemes.

To investigate the role of axonemal components in the mechanics and regulation of flagellar movement, we have generated a series of monoclonal antibodies (mAb) against sea urchin (Lytechinus pictus) sperm axonemal proteins, selected for their ability to inhibit the motility of demembranated sperm models. One of these antibodies, mAb D1, recognizes an antigen of 142 kDa on blots of sea urchin axonemal proteins and of purified outer arm dynein, suggesting that it acts by binding to the heaviest intermediate chain (IC1) of the dynein arm. mAb D1 blocks the motility of demembranated sea urchin spermatozoa by modifying the beating amplitude and shear angle without affecting the ATPase activity of purified dynein or of demembranated immotile spermatozoa. Furthermore, mAb D1 had only a marginal effect on the velocity of sliding microtubules in trypsin-treated axonemes. This antibody was also capable of inhibiting the motility of flagella of Oxyrrhis marina, a primitive dinoflagellate, and those of demembranated human spermatozoa. Localization of the antigen recognized by mAb D1 by immunofluorescence reveals its presence on the axonemes of flagella from sea urchin spermatozoa and O. marina but not on the cortical microtubule network of the dinoflagellate. These results are consistent with a dynamic role for the dynein intermediate chain IC1 in the bending and/or wave propagation of flagellar axonemes.     

Comparative spermatology of selected polyclad flatworms (platyhelminthes)

Sperm ultrastructure of four acotylean (Idioplana atlantica, Armatoplana leptalea, Styloplanocera fasciata, Melloplana ferruginea) and three cotylean polyclads (Pseudoceros bicolor, Phrikoceros mopsus, Enchiridium evelinae) was investigated. All spermatozoa are biflagellate, exhibiting a 9+"1" axoneme pattern. All acotylean axonemes originate and extend within the sperm shaft, and once exiting the shaft, remain attached to it. The flagella of all cotylean spermatozoa exit the shaft immediately and remain free. Structures shared by all species include: an elongated nucleus, in acotyleans located only in the posterior part of the shaft, whereas in cotyleans it extends along the entire sperm body; mitochondria along with small and large dense bodies arranged in a specific pattern; and a ring of microtubules that extends along the entire sperm shaft just beneath the cell membrane. A unique spermatozoon has been found in E. evelinae, where round vesicle-like structures fill the anterior part of the nucleus, and a different type of large dense bodies is present. The spermatozoa of all studied species exhibit numerous characters (axoneme/flagella position, distribution and position of large and small dense bodies, of mitochondria, presence of nuclear vesicles) that may be of phylogenetic value at the family and higher taxonomic levels.     

Study of demembranated, reactivated human spermatozoa with decondensed nuclei

Reactivated movement of the axonemes in demembranated spermatozoa with decondensed nuclei allows decondensation to be monitored in vitro with minimal disruption, and provides access to the nucleus for ultrastructural investigation and experimental manipulation. In the present study, fresh liquefied semen samples with sperm concentrations > or = 13 x 10(6)/ml were diluted 1:10 with a demembranating solution containing 0.01-0.022% Triton X-100. Inter-sample variation in the concentration of Triton X-100 required to permeabilize the sperm membrane was observed as judged by the ability of the spermatozoa to be reactivated by ATP but not by an ATP-free control solution, with the extent of demembranation being checked by transmission electron microscopy. After exposure to DTT and heparin, coordinated and sometimes progressive movement of partially decondensed spermatozoa occurred in a reactivating solution. Unlike ram, human sperm heads required decondensation with heparin. An unusual ultrastructural feature of the decondensing human sperm nuclei, not previously reported, was the appearance of dense globular material extruding from the nucleus. Enzymatic treatment of the sections with protease but not with deoxyribonuclease removed this material, which was presumably protamine.     

Double spermatogenesis in Chelicerata

Sperm dimorphism is a rare phenomenon in Chelicerata. Until now, it was known only from three species of the opilionid genus Siro (Sironidae, Cyphophthalmi). Fertilizing (eusperm) and nonfertilizing spermatozoa (parasperm) develop in the same cyst and are thus sister cells. The fine structure of the spermatozoa of two species has been examined and is compared here. In contrast to Siro rubens, S. duricorius spermatozoa lack an acrosomal complex. Both sperm types produce a transitional process, a more or less modified flagellum, which is later retracted. Hence, the spermatozoa are aflagellate. Eusperm and parasperm of all three species form highly ordered sperm balls that are stored in the deferent duct. Reviewing and adding new results about the sperm dimorphism in this arachnid taxon provides the basis for some considerations of another enigmatic morphological character found in Uropygi and Amblypygi, i.e., the tubular accessory genital glands that show holocrine extrusion. These glands are suggested to represent modified, infertile derivatives of the testis anlage. Their secretion is produced in a way reminiscent of a strongly degenerated spermatogenesis. Consequently, these products may be regarded as strongly degenerated germ cells representing a line of germ cell development, which has been separated very early in spermatogenesis from the usual line leading to fertilizing sperm cells. This further, although less evident, case of probable dichotomous germ cell development is discussed with respect to the controversial phylogenetic-systematic relationships between Uropygi (Thelyphonida and Schizomida), Amblypygi, and Araneae.     

Sperm of Doradidae (Teleostei: Siluriformes)

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

Spermatozoon structure and motility in the anuran Lepidobatrachus laevis

Synthetic human gonadotropin releasing hormone (GnRH) injections were used for induction of spermatozoon release followed by cloacal lavage or mechanical stimulation of sperm release in Lepidobatrachus laevis . Light microscopic observations of Lepidobatrachus laevis spermatozoa indicated an acrosomal segment with a length of 4.1 μm delineated by an indentation, a nuclear region of 12.6 μm in length and a midpiece of 0.87 μm in length. The tail was 54.9 μm long by 1.35 μm wide with two lateral axial fibers and a central undulating membrane. At the electron microscopic level, the unusual tail had two complete axonemes that emanated from the distal centriole. The tail also contained two axial fibers 77 nm in diameter medial to the axonemes and was connected by an undulating membrane. An unusual accessory cell adherent to the head of the spermatozoon was noted in freshly obtained suspensions of spermatozoa. Spermatozoa with the accessory cell were motile and a subsequent loss of motility was correlated with the shedding of the accessory cell.     

Ultrastructure of paraspermatozoa,euspermatozoa and eusperm-like spermatozoa ofObtortio cf.fulva (Prosobranchia: Cerithiacea)

The cerithiaceanObtortio cf.fulva produces three distinct types of spermatozoa: (1) paraspermatozoa, (2) euspermatozoa and (3) eusperm-like spermatozoa. Like most mesogastropods, euspermatozoa ofObtortio are composed of a conical acrosome, short posteriorly invaginated nucleus, elongate midpiece and glycogen piece, and short terminal region. The midpiece, however, is distinctly cerithiacean in structure and is composed of four non-helical midpiece elements. Eusperm-like spermatozoa closely resemble euspermatozoa, but have a very short nucleus only one half to one third the length of the euspermatozoon nucleus. Paraspermatozoa of this species are composed of (1) head (mosaic sheath of dense blocks enveloping multiple axonemes which attach anteriorly to a long apical structure), (2) midpiece (multiple axonemes interspersed with elongate mitochondria), and (3) multiple tail tuft (axonemes each ensheathed by glycogen granules). The possible role of eusperm-like spermatozoa is briefly discussed together with the taxonomic implications of the structure of the three sperm types.