蓝尾石龙子精子的超微结构

蓝尾石龙子(Eumeces elegans)附睾以2．5％戊二醛和1％锇酸双重固定,按常规制作超薄切片,用H-600透射电镜研究观察精子的超微结构。精子由头部和尾组成,头部由顶体复合体和核组成,尾由颈段、中段、主段和末段组成。头部的顶体囊前部扁平,分为皮质和髓质,顶体下锥由类结晶状的顶体下物质组成,穿孔器顶端尖,、穿孔器基板塞子状,细胞核延长,核内小管缺,核伸展部前端具一电子透明区,核肩圆,核陷窝锥形。颈段具片层结构,近端中心粒和远端中心粒的长轴呈直角,9束外周致密纤维与远端中心粒相应的9束三联微管相联,向后与轴丝相应的9束双联微管相联,中央纤维与2个中央单微管相联。中段短,含有线状嵴的柱状线粒体,由连续的规则小卵状或小梯形致密体组成线粒体间的环状结构,纤维鞘伸入中段,终环紧贴于细胞膜的内表面。线粒体与环状结构的模式为：rs1／mi1,rs2／mi2,rs3／mi3,rs4／mi4,横切面上每圈线粒体数目为10个。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。轴丝复合体呈“9 2”型。蓝尾石龙子精子超微结构与已描述的石龙子科种类比较发现,与蜓蜥群和胎生群的石龙子相似;但没有发现石龙子科精子的独征。     

Ultrastructure of the spermatozoon of the American Alligator,Alligator mississippiensis (Reptilia: Alligatoridae)

This study details the ultrastructure of the spermatozoa of the American Alligator, Alligator mississippiensis. American Alligator spermatozoa are filiform and slightly curved. The acrosome is tapered at its anterior end and surrounded by the acrosome vesicle and an underlying subacrosomal cone, which rests just cephalic to the nuclear rostrum. One endonuclear canal extends from the subacrosomal cone through the rostral nucleus and deep into the nuclear body. The neck region separates the nucleus and midpiece and houses the proximal centriole and pericentriolar material. The distal centriole extends through the midpiece and has 9 × 3 sets of peripheral microtubules with a central doublet pair within the axoneme that is surrounded by a dense sheath. The midpiece is composed of seven to nine rings of mitochondria, which have combinations of concentrically and septate cristae. The principal piece has a dense fibrous sheath that surrounds an axoneme with a 9 + 2 microtubule arrangement. The sheath becomes significantly reduced in size caudally within the principal piece and is completely missing from the endpiece. Dense peripheral fibers, especially those associated with microtubule doublets 3 and 8, penetrate into the anterior portion of the principal piece axoneme. The data reported here hypothesize that sperm morphology is highly conserved in Crocodylia; however, specific morphological differences can exist between species. J. Morphol. 2011. © 2011 Wiley‐Liss, Inc.     

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.     

The ultrastructure of amberjack (Seriola dumerilii) sperm

Scanning and transmission electron microscopy were used to investigate the fine structure of sperm of the Mediterranean amberjack Seriola dumerilii. Each spermatozoon has an ovoid head which lacks an acrosome, a short, irregularly-shaped midpiece and a long flagellar tail. The midpiece houses eight spherical mitochondria, which are separated from the axoneme by the cytoplasmic canal. The centrioles are arranged approximately at right angles to each other. The proximal centriole lies inside, and the distal centriole outside, the nuclear fossa. The flagellum is inserted eccentrically into the head and is tangential to the nucleus, so that the spermatozoon is asymmetrical. It contains the conventional 9 + 2 axoneme, shows intratubular differentiations in the A microtubules of doublets 1, 2, 5 and 6, and possesses one pair of lateral fins. On the basis of its ultrastructural organization, the amberjack sperm resembles type II sperm as defined previously, except for the presence of the proximal centriole inside the nuclear fossa. This could result from a partial rotation of the nucleus during spermiogenesis.     

黄姑鱼（ Nibea albiflora） 与大黄鱼（ Pseudosciaena crocea） 精子超微结构的观察与比较

应用扫描电镜（SEM）与透射电镜（TEM）观察了黄姑鱼和大黄鱼精子的超微结构。结果显示,黄姑鱼和大黄鱼精子无论在形态、大小还是超微结构上都十分相似。黄姑鱼和大黄鱼精子均由头部、中段和尾部（鞭毛）3部分组成。精子头部形状近似椭圆形,无顶体,细胞核呈肾形。中心粒复合体位于细胞核背侧,近、远端中心粒相互垂直,远端中心粒分化成基体并形成轴丝。中段的袖套呈筒状,4～5个圆形的线粒体围绕轴丝呈环形排列。精子尾部为单鞭毛,轴丝为典型“9＋2”结构,鞭毛表面质膜形成不规则侧鳍。     

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.     

中国石龙子成熟精子的超微结构

利用透射电镜观察中国石龙子附睾成熟精子的超微结构。顶体囊前部扁平、由皮质和髓质组成 ,穿孔器中度倾斜、顶端尖 ,穿孔器基板塞子状 ,细胞核长形 ,核内小管缺 ,核前电子透亮区小 ,核肩圆 ,核陷窝锥形。颈段具片层结构 ,近端中心粒和远端中心粒的长轴呈直角 ,9束外周致密纤维与远端中心粒相应的 9束三联微管相联 ,向后与轴丝相应的 9束双联微管相联 ,中央纤维与 2个中央单微管相联。中段短 ,多层膜结构缺 ,含有线状嵴的柱状线粒体 ,不规则卵状致密体组成不连续的环状结构 ,纤维鞘伸入中段 ,具终环。线粒体与环状结构的模式为 :rs1 /mi1 ,rs2 /mi2 ,rs3/mi3,rs4 /mi4。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。中国石龙子精子超微结构具有塞子状的穿孔器基板、致密体形成不连续的环状结构和纤维鞘始于ms2等特征与巨石龙子群和蜓蜥 -胎生群不同。没有发现石龙子科精子的独征     

Ultrastructure of the spermatid of Caprimulgus europaeus Linnaeus 1758, the European nightjar (Aves; Caprimulgidae), with phylogenetic implications

The sperm of Caprimulgus europaeus is typical of other nonpasserines in many respects. Features shared with Paleognathae and Galloanserae are the conical acrosome, shorter than the nucleus; the presence of a perforatorium and endonuclear canal; the presence of a proximal as well as distal centriole; the elongate midpiece with mitochondria grouped around a central axis (here maximally six mitochondria in approximately 10 tiers); and the presence of a fibrous or amorphous sheath around the principal piece of the axoneme. A major (apomorphic) difference from paleognaths and galloanserans is the short distal centriole, the midpiece being penetrated for most of its length by the axoneme and for only a very short proximal portion by the centriole. Nonpasserines differ from paleognaths in that the latter have a transversely ribbed fibrous sheath, whereas in nonpasserines it is amorphous, as in Caprimulgus, or absent. The absence of an annulus is an apomorphic feature of Caprimulgus, apodiform, psittaciform, gruiform, and passerine sperm, homoplastic in at least some of these. In contrast to passerines, in Caprimulgus the cytoplasmic microtubules in the spermatid are restricted to a transient longitudinal manchette. The structure of the spermatid and spermatozoon is consistent with placement of the Caprimulgidae near the Psittacidae, but is less supportive of close proximity to the Apodidae, from DNA-DNA hybridization and some other analyses.     

Ultrastructure of spermiogenesis in a freshwater stingray, Himantura signifer

Ultrastructural changes of spermatids during spermiogenesis in a freshwater stingray, Himantura signifer, are described. Differentiation of spermatids begins with modification of the nuclear envelope adjacent to the Golgi apparatus, before the attachment of the acrosomal vesicle. A fibrous nuclear sheath extends over the nuclear surface from the site of acrosomal adherence. The conical apical acrosome is formed during nuclear elongation. At the same time, chromatin fibers shift from an initially random arrangement, assume a longitudinal orientation, and become helical before final nuclear condensation. An axial midpiece rod is formed at the posterior end of nucleus and connects to the base of the sperm tail. Numerous spherical mitochondria surround the midpiece axis. The tail originating from the posterior end of the midpiece is composed of the usual 9 + 2 axoneme accompanied by two longitudinal columns, which are equal in size and round in cross section. The two longitudinal columns are absent at the end piece. A distinctive feature of freshwater stingray sperm is its spiral configuration.     

A novel apical midpiece defect in the spermatozoa of a bull without an apparent decrease in motility and fertility. A case study

Despite some limitations as predictors of fertility, evaluation of sperm morphology and progressive motility is the commonest method to assess viability of frozen/thawed semen. In this article we describe by light and transmission electron microscopy a novel midpiece structural defect observed in 24-36% of frozen/thawed sperm cells from a Charolais bull, used in artificial insemination programs without any apparent ill effect to the fertility. After thawing, the sperm progressive motility ranged from 65 to 80% and the pregnancy rate for all artificial inseminations performed (43%) did not differ (p>0.05) from results obtained with insemination with semen of other bulls (40%). The defect consisted in mitochondrial aplasia at the neck region, mitochondrial segmental elongation and gaps and thickening of the outer dense fibers at the apical region of the midpiece, and loss of the cementing substance and development of plasma membrane extensions in the entire midpiece. No structural abnormalities were found in the capitulum, proximal centriole, striated columns, axoneme, annulus and fibrous sheath. The thickness of the outer fibers returned to normal at the distal region of the midpiece. Based on the examination it is suggested that the alterations might be originally caused by loss of the cementing substance that links mitochondria to the plasma membrane in association with mitochondrial aplasia at the neck region of the midpiece. The abnormality appeared not related to other described sperm defect syndromes, although it shared particular characteristics with the dag defect, segmental aplasia of the mitochondrial sheath, corkscrew defect and pseudodroplet defect.     

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.     

Ultrastructure of Thunnus thynnus and Euthynnus allettevatus spermatozoa

The spermatozoa of Thunnus thynnus and Euthynnus alletteratus consist of an acrosome-less head (comprising the ovoid nucleus and the short midpiece) and a long flagellar tail that contains the conventional 9 + 2 axoneme and lacks lateral fins. The centrioles are arranged at approximately right angles and lie outside of a shallow nuclear groove. The flagellum inserts laterally on the nucleus, therefore the spermatozoon is asymmetrical. The midpiece contains a few mitochondria which are separated from the axoneme by the cytoplasmic canal; they are spherical in T. thynnus and elongate, somewhat irregular in E. alletteratus . Although the main ultrastructural characteristics of the spermatozoa appear to indicate a great homogeneity in the sperm morphology within the family Scombridae, small species-specific divergences may be of use in systematics.     

Ultrastructure of spermiogenesis in the gastropod Calliotropis glyptus Watson (Prosobranchia: Trochidae)with special reference to the embedded acrosome

Testicular spermatozoa and sperm development in the archaeogastropod Calliotropis glyptus Watson (Trochoidae: Trochidae) are examined using transmission electron microscopy and formalin-fixed tissues. During spermiogenesis, the acrosome, formed evidently through fusion of Golgi-derived proacrosomal vesicles, becomes deeply embedded in the condensing spermatid nucleus. Two centrioles (proximal and distal), both showing triplet microtubular substructure, are present in spermatids—the distal centriole giving rise to the sperm tail and its associated rootlet. During formation of the basal invagination in the spermatid nucleus, centrioles, and rootlet move towards the nucleus and come to lie totally within the basal invagination. Mitochondria are initially positioned near the base of the nucleus but subsequently become laterally displaced. Morphology of the mature spermatozoon is modified from that of the classic primitive or ect-aquasperm type by having 1) the acrosome embedded in the nucleus (the only known example within the Mollusca), 2) a deep basai invagination in the nucleus containing proximal and distal centrioles and an enveloping matrix (derived from the rootlet), 3) laterally displaced periaxonemal mitochondria, and 4) a tail extending from the basal invagination of the nucleus. Implantation of the acrosomal complex and centrioles within imaginations of the nucleus and lateral displacement of mitochondria effectively minimize the length of the sperm head and midpiece. Such modifications may be associated with motility demands, but this remains to be established. The unusual features of C. glyptus spermatozoa, though easily derivable from ‘typical’ trochoid sperm architecture, may prove useful in delineating the genus Calliotropis or tracing its relationship to other genera within the trochid subfamily Margaritinae.     

Immunogold localization of the regulatory subunit of a type II cAMP- dependent protein kinase tightly associated with mammalian sperm flagella

We have shown previously that the regulatory subunit (RII) of a type II cAMP-dependent protein kinase is an integral component of the mammalian sperm flagellum (Horowitz, J.A., H. Toeg, and G.A. Orr. 1984. J. Biol. Chem. 259:832-838; Horowitz, J.A., W. Wasco, M. Leiser, and G.A. Orr. 1988. J. Biol. Chem. 263:2098-2104). The subcellular localization of this flagellum-associated RII in bovine caudal epididymal sperm was analyzed at electron microscope resolution with gold-conjugated secondary antibody labeling techniques using anti-RII monoclonal antibodies. By immunoblot analysis, the flagellum-associated RII was shown to interact with mAb 622 which cross reacts with both neural and nonneural isoforms of RII. In contrast, a neural specific monoclonal antibody (mAb 526) failed to interact with flagellar RII. In the midpiece of the demembranated sperm tail, gold label after mAb 622 incubation was primarily associated with the outer mitochondrial membrane. Although almost all specific labeling in the midpiece can be assigned to the mitochondria, in the principal piece, there is some labeling of the fibrous sheath. Labeling of the outer dense fibers and the axoneme was sparse. Specific labeling was virtually absent in the sperm head. Sections of sperm tails incubated in the absence of primary antisera or with mAb 526 showed little labeling. A beta-tubulin monoclonal antibody localized only to the 9 + 2 axoneme. These results raise the possibility that a type II cAMP-dependent protein kinase located at the outer mitochondrial membrane plays a role in the direct cAMP stimulation of mitochondrial respiration during sperm activation.     

Ultrastructure of spermiogenesis in <Emphasis Type="Italic">Cristatella mucedo</Emphasis> Cuvier (Bryozoa: Phylactolaemata: Cristatellidae)

In Cristatella mucedo spermiogenesis occurs in a morula consisting of a large number of spermatids connected with a central cytophore. The mature sperm cell is filiform and consists of a head, a midpiece and a tail region, the latter two separated by a deep circular constriction. The comparatively short head contains a drop-shaped, bilaterally symmetrical and pointed nucleus capped by a minute acrosome. The single centriole is placed in a deep posterior invagination of the nucleus followed by the axoneme with the typical 9 + 2 pattern. The elongated midpiece is 0.9–1.1 μm thick and contains several helices of mitochondria surrounding the axoneme. The tail is thicker (1.3 μm) and richer in cytoplasm with many compact accumulations of an electron-dense substance lying peripherally and another less dense material wrapped around the axoneme. The course of the spermiogenesis and the fine structure of the sperm are very similar to that of Plumatella fungosa. Comparison with other species shows that the same sperm type is recognizable in four of the five families of Phylactolaemata and, provided it occurs also in the fifth family, the Stephanellidae, is a synapomorphy of the entire class.     

Fate of sperm tail components after incorporation into the hamster egg

The changes in the cytoplasmic organelles of sperm tail in golden hamster eggs fertilized in vivo were observed by electron microscopy. Eggs were obtained from oviducts of hamsters that had been superovulated and inseminated by injection of cauda epididymal spermatozoa into the uteri. In the egg cytoplasm 10 hours after insemination, some of the mitochondria of the spermatozoon midpiece had begun to swell, and a number of multivesicular bodies were observed surrounding the midpiece. The fibrous sheath of the principal piece quickly disappeared prior to the first cleavage, whereas the axoneme and outer dense fibers were unaltered. During the two-cell stage, numerous multivesicular bodies gathered around the midpiece and fused with the mitochondria. The heterophagic vacuoles thus formed then gradually separated from the axial fibers. The outer dense fibers were disarranged and partially torn into small segments; then they seemed to dissociate into substructural granular components. The axonemal microtubules had begun to swell but were still present in the two blastomeres. It is indicated from these observations that at least the mitochondria of the tail constituents carried into the oocyte are digested into small molecular elements by the multivesicular bodies and are possibly distributed as nutrients for the blastomeres during the early stage of development.     

Ultrastructure of Sperm in <Emphasis Type="Italic">Mercenaria stimpsoni</Emphasis> and <Emphasis Type="Italic">Mactra chinensis</Emphasis> (Mollusca: Bivalvia) from the Sea of Japan

The ultrastructure of the sperm of the common bivalve species Mercenaria stimpsoni and Mactra chinensis from Peter the Great Bay is described. The sperm structure is typical for animals with external insemination. The sperm consists of a head, middle part, and flagellum. The sperm head of M. stimpsoni has a curved crescent form and includes the nucleus and acrosome; the head length is 9.8 μm. The acrosome is subdivided to the acrosome granule and the periacrosomal material. There are 4 mitochondria of about 0.8 μm in size in the middle part of the spermatozoon. The mitochondria surround the centriolar apparatus, which consists of proximal and distal centrioles located at a right angle. The axoneme originates from the distal centriole. The sperm of M. chinensis is barrel-shaped, with a head length of 3.2 μm. The acrosome is relatively larger, and its height is 1–1.2 μm. There are also 4 mitochondria 0.6–0.8 μm in the middle part of the spermatozoon. The sperm structure of the described species is typical of the families to which the mollusks belong, with insignificant variations.     

Ultrastructural differentiation of spermiogenesis in Scincus scincus (Scincidae,Reptilia)

BackgroundKnowledge of spermiogenesis in reptiles, especially in lizards, is very limited. Lizards found in Arabian deserts have not been considered for detailed studies due to many reasons and the paucity of these animals. Therefore, we designed a study on the differentiation and morphogenesis of spermiogenesis, at an ultrastructural level, in a rare lizard species, Scincus scincus.ResultsThe spermiogenesis process includes the development of an acrosomal vesicle, aggregation of acrosomal granules, formation of subacrosomal nuclear space, and nuclear elongation. A role for manchette microtubules was described in nuclear shaping and organelle movement. During head differentiation, the fine granular chromatin of the early spermatid is gradually replaced by highly condensed contents in a process called chromatin condensation. Furthermore, ultrastructural features of sperm tail differentiation in S. scincus were described in detail. The commencement was with caudal migration toward centrioles, insertion of the proximal centriole in the nuclear fossa, and extension of the distal centrioles to form the microtubular axoneme. Subsequently, tail differentiation consists of the enlargement of neck portion, middle piece, the main and end pieces.ConclusionsThis study aids in the understanding of different aspects of spermiogenesis in the lizard family and unfurls evolutionary links within and outside reptiles.