无蹼壁虎精子头部形成的研究

本文用透射电镜观察了无蹼壁虎精子头形成的过程。早期精细胞具有显著的高尔基复合体、线粒体集合及细胞质桥、接着高尔基体成熟面分泌出前顶体囊泡,并逐渐向核移动。以后精子形成可分四个时间：时间Ⅰ,当前顶体囊泡移至核膜时,核膜凹陷形成封闭的顶体囊泡,囊泡底部靠近核膜有一电子致密的顶体颗粒;时间Ⅱ,细胞核延长,顶体囊泡变扁平;时期Ⅲ,细胞核进一步延长,核内染色质纤维变粗并沿核纵轴方向排列有序;时间Ⅳ,精子发育     

中国石龙子精子形成的超微结构研究

采用透射电镜观察中国石龙子精子的形成过程。结果表明:早期精细胞中有高尔基复合体和线粒体集合,由高尔基复合体所分泌的前顶体囊泡,逐渐向核移动,以后的过程可分为四个时期。时期Ⅰ:前顶体囊泡移至核膜时,核膜凹陷形成封闭的顶体囊泡,囊泡底部靠近核膜处有一电子致密的顶体颗粒,近端中心粒及鞭毛开始出现。时期Ⅱ:顶体囊泡变扁平,细胞核延长,染色质浓缩成短丝状的染色质纤维。时期Ⅲ:核进一步延长,染色质纤维变粗变长,按核纵向排列有序。时期Ⅳ:染色质纤维浓缩至最大限度,电子透明的核质消失,核呈高电子致密,顶体复合体发育完全。     

中华鳖精子头部的形成

应用透射电镜技术详细研究了中华鳖精子头部形成过程的超微结构变化。结果显示,中华鳖精子头部的形成过程可分成5个连续时期：第1期前顶体泡形成并移向细胞核一侧,同侧核膜凹陷成浅窝。前顶体泡底部中央出现小的顶体颗粒,纤维物质层位于核前端与前顶体泡底壁之间,其核膜一侧的中央形成更小的顶体下颗粒,将与核内小管的形成有关。细胞核开始端移和变形。第Ⅱ期核浅窝逐渐外推,前顶体泡变成扁囊状覆盖于隆突的核顶端,顶体颗粒弥散成中等电子致密物分布于顶体帽中,纤维物质层发育为顶体下锥。环形核套微管在顶体后端的核周围逐渐形成,核内染色质开始浓缩成圆形颗粒,核膜下出现明显问隙,细胞核体积变小。顶体下颗粒消失,但其下端的核质中可见2-4条核内小管开始发生。第Ⅲ期拉长的细胞核前端突出于精子细胞外,表面有顶体复合体覆盖,核后端最宽并出现植入窝。染色质进一步浓缩,颗粒间隙变小,细胞核更细长。第Ⅳ期染色质浓缩成致密均质物,核肩之前的细胞核变细,成为核前突。环形核套微管先后改建为斜行和纵行核套微管,支持细胞突起形成“袖领”包绕顶体。第Ⅴ期核套微管解聚而消失,顶体周围的“袖领”也消失,顶体下间隙出现。结果显示中华鳖精子头部的形成过程,即核质浓缩的形态变化过程、顶体的形成和核内小管的发育与变化方面,存在许多与其他爬行动物不同之处。     

东方扁虾精子的超微结构

利用电镜研究了东方扁虾（Ｔｈｅｎｕｓ ｏｒｉｅｎｔａｌｉｓ）精子的形态和结构。精子由核、膜复合物区和顶体区３部分组成。核内含非浓缩的染色质、微管及细纤维丝,外被核膜;５～６条辐射臂自核部位伸出,臂内充满微管。膜复合物区位于核与顶体之间,由许多膜片层结构及其衍生的囊泡共同组成。顶体区由顶体囊和围顶体物质组成,顶体结构复杂,由顶体帽、内顶体物质和外顶体物质等构成;围顶体物质呈细颗粒状,主要分布于顶体囊     

三疣梭子蟹精子顶体反应过程中的形态和结构变化

用离子载体A2 3187和卵水人工诱导三疣梭子蟹精子的顶体反应 ,分别获得 75 33%和 84 83%的顶体反应率。应用光镜和电镜技术观察了顶体反应前后精子形态和结构的变化。未处理精子呈陀螺形 ,由顶体、核杯和 5 - 10条核辐射臂组成。顶体包括顶体囊和顶体管。顶体囊的伞形头帽拥有约 70条辐射肋。连续发生的精子顶体反应过程被人为地分为四个阶段 :(1)头帽鼓起 ;(2 )顶体囊外翻 ;(3)穿孔器前伸 ,顶体囊膜翻转 ;(4 )顶体囊膜脱落 ,顶体丝形成。直到第四阶段才观察到钉状精子的辐射臂开始收缩。探讨了辐射臂和穿孔器前冲在精子入卵中的功能     

长江华溪蟹精子形成的研究

1994年9-11月,对采自安徽省宁国县的长江华溪蟹(Sinopotamon yangtsekiense),利用透射电镜技术,并结合细胞化学方法,研究了其精子形成过程。结果显示:早期精细胞圆形,胞质丰富,内含大量内质同小泡及线粒体。核也为圆形,较小。精细胞开始分化,细胞膨胀为长椭圆形,核质重新分布,分别移向细胞的两端。精子的顶体由高尔基体产生,其过程为:高尔基体分泌产生囊泡,继而形成原顶体囊,进一步发育成顶体囊,最后形成顶体。在顶体囊与核之间有膜复合体。中心粒位于核内面凹陷处。细胞化学反应显示,核杯为Feulgen阳性,顶体为PAS阳性。       

OEP及卵黄浓度对蓝狐冻融精子质量的影响

人工采取 6只优质芬兰雄性蓝狐精液 ,利用不同OEP及卵黄含量的Tris 果糖 -柠檬酸钠稀释液稀释 ,制成细管冻精 ,透射电镜下观察精子冷冻前后质膜和顶体超微结构 ,荧光免疫方法检测不同培养时间冻融精子的质量。结果表明 ,蓝狐精子顶体外膜双层膜的厚度为 0 0 2 0 μm ,冷冻 -解冻过程中易发生质膜膨胀、顶体外膜融合现象。顶体产生的囊泡分两种类型 ,一种是体积较大的中空囊泡 ,平均直径为 1 2 5 μm。另一种是体积较小的实体囊泡 ,内充满顶体内容物 ,平均直径为 0 83μm ,两种囊泡的数量不定。OEP能有效抑制顶体囊泡形成 ,影响顶体囊泡类型、体积大小及囊泡数量 ,添加适宜剂量OEP能使顶体囊泡的体积明显缩小 ,囊泡的总数及中空囊泡的数量显著降低。蓝狐冻融精子质量与OEP及卵黄剂量有关 ,在卵黄存在的前提下 ,OEP有利于维持冻融过程中质膜 (5 6 3% )、顶体的完整性 (5 7 8% ) ,显著提高冻融精子活力 (5 4 7% )。在蓝狐精液稀释液中 ,OEP、卵黄的适宜含量分别为 1 %、 2 0 %     

三疣梭子蟹精子顶体反应前后胞内Ca~(2+)的变化

应用激光扫描共聚焦显微镜(LSCM)和Fluo-3/AM荧染技术对三疣梭子蟹精子顶体反应前后的胞内Ca2 变化进行了观察和检测.结果显示,在精子顶体反应过程中,胞内Ca2 主要分布在细胞核、穿孔器和胞质膜残存处,胞内Ca2 浓度([Ca2 ]I)总体上呈现先上升后下降的趋势.顶体反应前精子的平均荧光强度为35.95±5.71;穿孔器前伸、顶体囊膜翻转阶段精子的平均荧光强度为66.80±7.35;顶体囊膜脱落、顶体丝形成阶段精子的平均荧光强度为3.87±2.82;上述各阶段间精子荧光强度有极显著差异(P<0.01).顶体反应穿孔器前伸、顶体囊膜翻转阶段的精子相比顶体反应前精子,[Ca2 ]I显著提高;而在顶体囊膜脱落、顶体丝形成阶段,[Ca2 ]I则急剧下降,只在顶体丝基部胞质膜残存处有微量Ca2 存在.初步探讨了三疣梭子蟹精子顶体反应前后胞内Ca2 变化的功能.     

菲律宾蛤仔的精子发生和精子超微结构

用透射电镜研究了菲律宾蛤仔(Ruditapes philippinarum)精子结构和精子发生过程中细胞形态结构的变化及细胞器的演变规律。菲律宾蛤仔雄性生殖细胞的形态由椭圆形渐变为辣椒状,细胞核的形态由椭圆形逐渐拉长,渐变为锥形。染色质的凝集经历：小颗粒团块状一较大颗粒均匀状一粗颗粒均匀状的过程。线粒体在演化过程中数量先增多后逐渐减少,嵴数逐渐增多,电子密度和体积逐渐增大。高尔基体在初级精母细胞期已经发育,随后的各期中发育良好,分泌旺盛。精细胞Ⅱ期,高尔基体分泌的潴泡开始融合,形成前顶体囊。精细胞Ⅲ期,高尔基体的分泌物仍不断融合。精细胞分化的后期,前顶体囊逐渐发育形成顶体。菲律宾蛤仔成熟精子呈长辣椒状,为原生型,由头部、中段和尾部构成。头部的顶体为细长柱形,末端渐细,电子密度较小;细胞核为锥形;中段线粒体4个,尾部鞭毛为典型的“9 2”型结构。此外在成熟精子线粒体环横切面有一特殊“风车状”结构。     

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。     

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.     

MORPHOLOGICAL OBSERVATIONS ON THE SPERMATOZOA OF ECHINOTHURID SEA URCHINS*

The morphology of the spermatozoa of three species of echinothurid sea urchins, Asthenosoma ijimai, Araeosoma owstoni, Hapalosoma gemmiferum, was investigated by means of transmission and scanning electron microscopy. The spermatozoa of these three species of echinothurid sea urchins have similar fine structure, but they differ in several features from the more familiar regular sea urchins. 1) The external anatomy of the head region of the echinothurid spermatozoon is diagnostic in that it has a highly elongated head. 2) The spermatozoon of echinothurid sea urchins has a very long slender nucleus, protruding on its proximal end, so that the shape of the nucleus resembles a sperhead. 3) The acrosomal granule in the acrosomal vesicle of the echinothurid spermatozoon is not a mass of homogenous particulate material but an electron opaque rod condensed in the central part of the acrosomal vesicle. Scanning electron microscopic examination revealed that echinothurid spermatozoa form acrosomal processes similar to those of other regular sea urchins. 4) The basal body is situated just beneath the middle of the posterior protrusion of the nucleus. The distal centriole is located beside the basal body almost in contact with it. The axis of the distal centriole is almost but not quite parallel to that of the basal body. A satellite complex can be recognized around the posterior part of the proximal centriole.     

Spermiogenesis in caecilians Ichthyophis tricolor and Uraeotyphlus cf. narayani (Amphibia: Gymnophiona): analysis by light and transmission electron microscopy

Spermiogenesis, known as spermateleosis in lower vertebrates, is the transformation of the round spermatid into a highly specialized spermatozoon with a species-specific structure. Spermateleosis and sperm morphology of two species of caecilians, Ichthyophis tricolor and Uraeotyphlus cf. narayani, from the Western Ghats of Kerala, India, were studied using light and transmission electron microscopy. Spermateleosis is described in early, mid-, and late phases. During the early phase, the spermatid nucleus does not elongate, but the acrosome vesicle is Golgi-derived and its material is produced as a homogeneous substance rather than as discrete granules. In development of the acrosome, the centrioles shift in position to the lower half of the cell. The acrosomal vesicles take the full shape of the acrosome with the establishment of the perforatorium in midphase. An endonuclear canal develops and accommodates the perforatorium. The incipient flagellum is laid down when the proximal centriole attaches to the posterior side of the nucleus and the distal centriole connects to the proximal centriole, which forms the basal granule of the acrosome. The axial fiber also appears during midphase. The mitochondria shift in position to the posterior pole of the cell to commence establishment of the midphase. Late phase is characterized by nuclear condensation and elongation. Consequently, the final organization of the sperm is established with the head containing the nucleus and the acrosome. The undulating membrane separates the axoneme and axial fiber. Most of the cytoplasm is lost as residual bodies.     

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.     

Fine structure of the spermatozoon of Marthasterias glacialis (Linnaeus) (Echinodermata; Asteroidea), with special reference to acrosomal morphology

The sperm of Marthasterias glacialis (Linnaeus) was studied by light and electron microscopy. It is a long uniflagellated cell of the “primitive” type. The head has a spherical shape and contains a nucleus with a spheroid acrosome lying in a cup-shaped anterior fossa. The acrosome is formed by an acrosomal vesicle surrounded by the periacrosomal material. The basal specializations of the acrosomal vesicle show a clear differentiation of its constituents resembling the structure of membrane. The midpiece contains a very large annular mitochondrion which encircles two perpendicular centrioles. The distal centriole is in close association with a pericentriolar radial complex. The tail, containing a common microtubular axoneme, is projected to a variable position.     

ELECTRON MICROSCOPE STUDIES ON SPERM DIFFERENTIATION IN MARINE ANNELID WORMS. I. SPERM FORMATION IN IKEDOSOMA GOGOSHIMENSE

The ultrastructur of spermatozoa and the changes through which they are differentiated during sperm formation in an echiuroid were observed under the electron microscope. Many spermatids are connected to one central cytoplasmic mass and the sperm differentiation proceeds synchronously in one sperm-ball. Dense plate-like structures appear in the cytoplasm of early spermatids and disappear soon. In the process of nuclear condensation, many electron-dense aggregates appear in homogeneously textured chromonema and the aggregates are packed together to form a uniformly dense nucleus. Near the centriole at the opposite side from the central mass, the mitochondria fuse together to form one large middle-piece mitochondrion and the acrosomal vesicle is formed from the Golgi-complex. The differentiating acrosome in the late spermatid moves to the anterior tip of the head. In the completed acrosome, a flocculent substance accumulates in the conspicuously expanded invaginated pocket of the acrosomal vesicle and two kinds of material of different electron density fill the inside of the acrosomal vesicle. The spermatozoa remain connected to the central mass at the lateral side of the head until they become fully mature and are packed into the nephridia before spawning.     

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.     

Microtubule configurations and post-translational alpha-tubulin modifications during mammalian spermatogenesis

The mechanisms underlying cell cycle progression and differentiation are tightly entwined with changes associated in the structure and composition of the cytoskeleton. Mammalian spermatogenesis is a highly intricate process that involves differentiation and polarization of the round spermatid. We found that pachytene spermatocytes and round spermatids have most of the microtubules randomly distributed in a cortical network without any apparent centrosome. The Golgi apparatus faces the acrosomal vesicle and some microtubules contact its surface. In round spermatids, at step 7, there is an increase in short microtubules around and over the nucleus. These microtubules are located between the rims of the acrosome and may be the very first sign in the formation of the manchette. This new microtubular configuration is correlated with the beginning of the migration of the Golgi apparatus from the acrosomal region towards the opposite pole of the cell. Next, the cortical microtubules form a bundle running around the nucleus perpendicular to the main axis of the cell. At later stages, the nuclear microtubules increase in size and a fully formed manchette appears at stage 9. On the other hand, acetylated tubulin is present in a few microtubules in pachytene spermatocytes and in the axial filament (precursor of the sperm tail) in round spermatids. Our results suggest that at step 7, the spermatid undergoes a major microtubular reordering that induces or allows organelle movement and prepares the cell for the formation of the manchette and further nuclear shaping. This new microtubular configuration is associated with an increase in short microtubules over the nucleus that may correspond to the initial step of the manchette formation. The new structure of the cytoskeleton may be associated with major migratory events occurring at this step of differentiation.     

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

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