优雅蝈螽精子形成期间微丝和微管蛋白的动态变化

【目的】分析微丝和微管蛋白在优雅蝈螽Gampsocleis gratiosa精子形成过程中的作用,为昆虫精子顶体复合体形成和细胞核重建机制研究奠定基础。【方法】应用免疫荧光、PAS-苏木精染色和透射电镜等方法,对优雅蝈螽成虫的精巢、雄性贮精囊和雌性受精囊内精子的发育以及微丝和微管蛋白在精子形成各个时期的分布进行了观察。【结果】精巢中,早期圆形精子细胞中微丝在精子细胞的某区域大量聚集,而微管蛋白随机分布在细胞质中。伸长的精子细胞中,顶体开始形成时,微丝首先在亚顶体区域出现,历经球形、短棒形,然后向细胞核的两侧扩展成倒"Y"形,接着形成箭头形;在顶体的外周即微丝的周围,细胞核周围以及鞭毛中发现微管蛋白。在雄虫贮精囊和雌虫受精囊中,精子和精子束中仅有微管存在,且仅存在于鞭毛中;精子头部的微丝和微管蛋白均消失。【结论】综合分析,我们认为微丝和微管作为"脚手架"结构在优雅蝈螽精子形成期间参与顶体复合体形成和细胞核重建,精子成熟形成精子束过程中"脚手架"结构拆除。     

Dynamics of F-actin during spermiogenesis in Gampsocleis gratiosa ( Orthoptera: Tettigoniidae)

为阐明F-肌动蛋白在优雅蝈螽Gampsocleis gratiosa Brunner von Wattenwyl精子形成过程中的动态变化,本研究利用微分干涉相衬技术和免疫荧光技术首次对优雅蝈螽精子形成过程中的F-肌动蛋白进行了细胞定位,利用透射电镜技术从超微水平观察了优雅蝈螽精子顶体复合体的结构.结果显示:精子形成早期,F-肌动蛋白富集于亚顶体区域,形态由“球状”转变为“棒锥状”;精子形成中期,F-肌动蛋白呈“倒Y型”分布于亚顶体区域和细胞核前端两侧;精子形成后期,亚顶体区域的F-肌动蛋白解聚消失,F-肌动蛋白呈“箭头状”,仅分布于顶体复合体扩张的两翼中.F-肌动蛋白动态变化伴随着细胞核和精子头部的形态改变,F-肌动蛋白的动态装配在精子顶体复合体形态构建和细胞核的形变中起着重要的作用.本研究还发现未成熟的精子尾部有一些富含F-肌动蛋白的细胞质微滴,与精子形成过程中多余细胞质和细胞器的外排有关.F-肌动蛋白的动态变化研究为进一步阐明细胞骨架蛋白在昆虫精子形成过程中的功能和作用机制奠定了基础.     

优雅蝈螽精子形成过程中F-肌动蛋白的动态变化

为阐明F-肌动蛋白在优雅蝈螽Gampsocleis gratiosa Brunner von Wattenwyl精子形成过程中的动态变化, 本研究利用微分干涉相衬技术和免疫荧光技术首次对优雅蝈螽精子形成过程中的F-肌动蛋白进行了细胞定位, 利用透射电镜技术从超微水平观察了优雅蝈螽精子顶体复合体的结构。结果显示： 精子形成早期, F-肌动蛋白富集于亚顶体区域, 形态由“球状”转变为“棒锥状”; 精子形成中期, F-肌动蛋白呈“倒Y型”分布于亚顶体区域和细胞核前端两侧; 精子形成后期, 亚顶体区域的F 肌动蛋白解聚消失, F-肌动蛋白呈“箭头状”, 仅分布于顶体复合体扩张的两翼中。F-肌动蛋白动态变化伴随着细胞核和精子头部的形态改变, F-肌动蛋白的动态装配在精子顶体复合体形态构建和细胞核的形变中起着重要的作用。本研究还发现未成熟的精子尾部有一些富含F-肌动蛋白的细胞质微滴, 与精子形成过程中多余细胞质和细胞器的外排有关。F-肌动蛋白的动态变化研究为进一步阐明细胞骨架蛋白在昆虫精子形成过程中的功能和作用机制奠定了基础。     

暗褐蝈螽与优雅蝈螽精子超微结构的比较研究(直翅目,螽斯科)

研究了暗褐蝈螽Gampsocleis sedakovii(Fischer von Waldheim)和优雅蝈螽G.gratiosa Brunner von Wattenwyl精子的超微结构。这两种蝈螽精子头部的顶体复合体由顶体外层、顶体本体和顶体组成,顶体复合体位于细胞核前端,并包裹部分细胞核;颈部具5纵层细胞器;尾部鞭毛轴丝为典型的9+9+2型,线粒体衍生体部分晶状化。暗褐蝈螽精子较短,顶体复合体夹角较大,精子鞭毛横切面直径稍大;优雅蝈螽精子稍长,顶体复合体夹角较小,精子鞭毛横切面直径较小,两种精子超微结构差异不显著,其生殖隔离机制有待进一步研究。     

中华蝈螽与中华螽斯精子超微结构研究(直翅目,螽斯总科)

研究了中华蝈螽Gampsocleis sinensis与中华螽斯Tettigonia chinensis精子的超微结构.两种精子的顶体复合体侧生于核上且包裹了核的一部分,顶体复合体横切面两侧角分离或两侧角与细胞核远离,或仅见其一侧角;中华蝈螽顶体外层在顶体本体两侧角外多呈晕圈状;轴丝为典型的9 9 2型;轴丝两侧的副纤维结构为2副微管;轴丝与线粒体衍生体之间的连接带3条;中华蝈螽有的横切面中两扁平膜池连接在一起;两种精子的中心粒侧体部位具五纵层细胞器.     

优雅蝈螽与暗褐蝈螽精子束的显微观察

本文应用微分干涉相衬法对优雅蝈螽Ｇampsocleis gratiosa Brunner von Wattenwyl和暗褐蝈螽G. sedakovii （Fischer von Waldheim） 雄性精巢管基部、输精管、贮精囊和精包,及雌性受精囊中精子束的形态变化进行了观察,对探讨螽斯近缘种的生殖隔离机制和生殖生物学具有重要意义.结果表明：这两种蝈螽的精子束通过精包转移到雌性受精囊后,精子束的形态发生了显著变化.精巢管基部的精子为游离的单个精子;输精管、贮精囊和精包中精子成束排列形成较分散的精子束,精子束头部包裹有粘液帽;雌性受精囊中的精子束的精子呈羽状排列,精子的头部汇集在中央轴上.两种蝈螽精子束形态差异不显著.     

优雅蝈螽雄性附腺的超微结构及其腺管提取物对精子束的作用

为阐明优雅蝈螽Gampsocleis gratiosa Brunner von Wattenwyl雄性附腺的结构与功能的关系, 本文利用透射电镜(transmission electron microscope, TEM)技术研究了优雅蝈螽雄性附腺的超微结构, 利用微分干涉相差显微镜(differential interference contrast microscope, DIC)技术并结合雄性附腺匀浆提取物与精子束在体外的短暂培养, 研究了优雅蝈螽雄性附腺对精子束的作用。结果表明： 优雅蝈螽雄性附腺3类腺管组织结构相似, 腺管管壁为单层上皮细胞, 缺少内表皮, 说明其来源于中胚层。上皮细胞富含粗面内质网、 线粒体、 高尔基体, 具有分泌细胞的特点。腺管管腔中分泌物有4种形态, 即电子透明的物质、 电子致密的颗粒物质、 细纤维状物质以及绒球状物质。上皮细胞的分泌方式主要有2种, 即顶质分泌和局部分泌。乳白短腺管的匀浆提取物参与了帽状精子束解聚的过程, 乳白长腺管和透明腺管的匀浆提取物有维持精子束活性的作用。本研究结果为进一步阐明螽斯雄性附腺的生理功能奠定了基础。     

三种螽斯的性选择行为比较研究

本文通过设计Y形迷宫实验,对螽斯属的优雅蝈螽Gampsocleis gratiosa、暗褐蝈螽Gampsocleis sedakovii obscura和鸣螽属的鼓翅鸣螽Uvarovites inflatus雌虫的性选择行为和交配行为进行了观察研究。结果表明:3种螽斯的交配行为谱一致,但交配时间上有差别;3种螽斯雌虫趋于选择体重较大的雄虫交配;体重和精包重之间呈现较复杂的相关性,但均以三次函数的拟合度(R2)最大;雄虫精包重占体重均在10.0%以上,分别为优雅蝈螽11.1%、暗褐蝈螽12.5%、鼓翅鸣螽14.9%。     

刘氏蝎蛉的精子发生和精子超微结构

【目的】精子超微结构在不同昆虫类群中变异较大,在昆虫种类鉴别和系统发育分析中具有重要意义。但截止目前,长翅目(Mecoptera)昆虫的精子发生和精子超微结构研究还很不充分。【方法】采用光学显微镜和透射电子显微镜技术,观察了刘氏蝎蛉Panorpa liui Hua的精子发生和精子超微结构。【结果】刘氏蝎蛉精原细胞在精囊内共分裂7次,产生128个精子细胞。这些精子细胞同步发育,成熟后结合形成数目略低于128的精子束。精子形成期,精子细胞内高尔基复合体产生的原顶体颗粒物质形成精子顶体;球状细胞核伸长、核内染色质凝集,形成致密的线形细胞核;分散的线粒体聚集、融合产生的副核转化形成2条线粒体衍生物。成熟精子由头部、颈区和长鞭毛组成。精子头部包括双层顶体和具有2条侧沟的细胞核两部分;颈区主要由中心粒和致密的鞘状中心粒侧体组成。鞭毛螺旋状,主要由1条9+2型轴丝、2条大小不等的线粒体衍生物和2条副体组成。【结论】刘氏蝎蛉精子束内精子数目略低于128,可能与精子复杂的形成过程及细胞的吞噬作用有关。线粒体衍生物在不同类群间差异显著,可为长翅目系统发育分析提供有用的特征。     

精子形成中的表观遗传调控

精子形成是指单倍体球形精子细胞分化发育成为精子的过程。这一连续进程包含一系列复杂的生化事件和剧烈的形态变化,涉及顶体和尾部形成、组蛋白-鱼精蛋白转换、细胞核压缩和细胞质丢弃等。近期研究发现,表观遗传调控在精子形成过程中发挥重要作用,对确保精子细胞正常发育和精子生成至关重要。现总结近期的相关研究进展,从DNA甲基化和组蛋白修饰两个方面简介精子形成过程中的表观遗传调控功能和机制。     

Spermiogenesis in the Australian cockatiel Nymphicus hollandicus

Information on the ultrastructure of parrot spermatids and spermatozoa is limited to only four species with no comprehensive study of spermiogenesis conducted within the order Psittaciformes. The present study was undertaken to describe the development of the cockatiel spermatid using electron microscopy. Four phases of spermatid maturation were documented on the basis of nuclear morphology, development of the acrosome, perforatorium, and axial filament. These phases included 1) round nuclei, 2) irregular nuclei, 3) elongated nuclei with granular chromatin, and 4) elongated nuclei with homogenous chromatin. While development of the cockatiel spermatid was comparable to that of other domestic avian species, we have noted the hollow nature of some chromatin granules, an abnormal formation of the axoneme, the absence of the fibrous sheath around the axoneme of the principal piece, and the absence of an annulus. J. Morphol. , 2012. © 2012 Wiley Periodicals, Inc.     

Ultrastructural examination of spermiogenesis within the testis of the ground skink, Scincella laterale (Squamata, Sauria, Scincidae)

Although the events of spermiogenesis are commonly studied in amniotes, the amount of research available for lizards (Sauria) is lacking. Many studies have described the morphological characteristics of mature spermatozoa in lizards, but few detail the ultrastructural changes that occur during spermiogenesis. The purpose of this study was to gain a better understanding of the subcellular events of spermiogenesis within the temperate ground skink (Scincella laterale). The morphological data presented here represent the first complete ultrastructural study of spermiogenesis within the Scincidae clade. Samples of testes from 20 specimens were prepared using standard techniques for transmission electron microscopy. Many of the ultrastructural changes occurring during spermiogenesis within the ground skink are similar to that of other saurians. However, there were a few unique characteristics that to date have not been described during spermiogenesis in other lizards. For example, during early round spermatid development within the ground skink testis, proacrosomal granules begin to form within the acrosomal vesicle before making contact with the apex of the nucleus. Also, a prominent microtubular manchette develops during spermiogenesis; however, the circular component of the manchete is absent in this species of skink. This developmental difference in manchette formation may lead to the more robust and straight mature spermatozoa that are common within the Scincidae family. These anatomical character differences may be valuable nontraditional sources that along with more traditional sources (i.e., mitochondrial DNA) may help elucidate phylogenetic relationships, which are historically considered controversial at best, among species within Scincidae and Sauria.     

Spermiogenesis in the bullfrog (Rana catesbeiana): a study of cytoplasmic events including cell volume changes and cytoplasmic elimination

The process by which spermatid cytoplasmic volume is reduced and cytoplasm eliminated during spermiogenesis was investigated in the bullfrog Rana catesbeiana. At early phases of spermiogenesis, newly formed, rounded spermatids were found within spermatocysts. As acrosomal development, nuclear elongation, and chromatin condensation occurred, spermatid nuclei became eccentric within the cell. A cytoplasmic lobe formed from the caudal spermatid head and flagellum and extended toward the seminiferous tubule lumen. The cytoplasmic lobe underwent progressive condensation whereby most of its cytoplasm became extremely electron dense and contrasted sharply with numerous electron-translucent vesicles contained therein. At the completion of spermiogenesis, many spermatids with their highly condensed cytoplasm still attached were released from their Sertoli cell into the lumen of the seminiferous tubule. There was no evidence of the phagocytosis of residual bodies by Sertoli cells. Because spermatozoa are normally retained in the testis in winter and are not released until the following breeding season, sperm were induced to traverse the duct system with a single injection of hCG. Some spermatids remained attached to their cytoplasm during the sojourn through the testicular and kidney ducts; however, by the time the sperm reached the Wolffian duct, separation had occurred. The discarded cytoplasmic lobe (residual body) appeared to be degraded with the epithelium of the Wolffian duct. It was determined that the volume of the spermatid was reduced by 87% during spermiogenesis through a nuclear volume decrease of 76% and cytoplasmic volume decrease of 95.3%.     

Defect of sperm assembly in a neurological mutant of the mouse, wobbler (WR)

In the wobbler (WR) mouse, a neuromuscular mutant characterized by a motoneuron degeneration and male infertility, the cellular basis of the defect in spermiogenesis was studied by light and electron microscopy as well as by lectin binding. Spermatozoa of the wobbler mutant had rounded heads, and their motility was reduced. In histological sections of WR testes, spermatogenesis appeared normal up to the stage of round spermatids, but the elongation and flattening of the nucleus during late spermiogenesis did not occur. Numbers of spermatid nuclei in WR testes were reduced to 70%-80% of controls. The acrosomal marker glycoprotein, peanut agglutinin receptor, was synthesized, but the acrosomal membrane did not attach to the nucleus. The disturbance in spermiogenesis of the wobbler mouse is not due to impaired descent of the testis, nor to a lack of testosterone, and is distinct from that observed in other mouse mutants (quaking, QK; Purkinje cell degeneration, PCD) with combined neurological and spermiogenesis defects.     

Ordered arrangement and rearrangement of chromosomes during spermatogenesis in two species of planarians (Plathelminthes)

The pattern of distribution of telomeric DNA (TTAGGG), 28S rDNA, and 5S rDNA has been studied using fluorescence in situ hybridization (FISH) and primed in situ labelling during spermatogenesis and sperm formation in the filiform spermatozoa of two species of planarians, Dendrocoelum lacteum and Polycelis tenuis (Turbellaria, Plathelminthes). In both species, the positions of FISH signals found with each probe sequence are constant from cell to cell in the nuclei of mature sperm. Chromosome regions containing 5S and 28S rDNA genes are gathered in distinct bundles of spiral form. In early spermatids with roundish nuclei, the sites of a given sequence on different chromosomes remain separate. Centromeres (marked by 5S rDNA) gather into a single cluster in the central region of the slightly elongated sperm nucleus. During spermatid maturation, this cluster migrates to the distal pole of the nucleus. In Polycelis, telomeric sites gather into three distinct clusters at both ends and in the middle of the moderately elongated nucleus. These clusters retain their relative positions as the spermatid matures. All the chromosome ends bearing 28S rDNA gather only into the proximal cluster. Our data suggest that structures in the nucleus selectively recognise chromosome regions containing specific DNA sequences, which helps these regions to find their regular places in the mature sperm nucleus and causes clustering of the sites of these sequences located on different chromosomes. This hypothesis is supported by observations on elongated sperm of other animals in which a correlation exists between ordered arrangement of chromosomes in the mature sperm nucleus and clustering of sites of the same sequence from different chromosomes during spermiogenesis. Received: 15 December 1997; in revised form: 24 March 1998 / Accepted: 14 April 1998     

Components and development of the centriolar complex during and beyond spermiogenesis in a passeridan bird, the Masked weaver (Ploceus velatus)

The fate of the proximal centriole in passeridan birds is an area of controversy and relative lack of knowledge in avian spermatogenesis and spermatology. This study examines, for the first time, spatiotemporal changes in the centriolar complex in various phases of spermiogenesis in a passerine bird, the Masked weaver (Ploceus velatus). It also describes the configuration of the centriolar complex and the relationship between it and the granular body in both intra- and extra-testicular spermatozoa. It is shown that the proximal centriole is retained and attaches, at its free end, to the granular body of spermatids in every step of spermiogenesis, as well as in mature intra-testicular and post-testicular spermatozoa, including those in the lumen of the seminal glomus. As the centriolar complex, along with its attached granular body, approaches the nucleus in the early spermatid, the proximal centriole articulates with the distal centriole at an acute angle of about 45°, and thereafter, both centrioles, still maintaining this conformation, implant, by means of their articulating proximal ends, at the implantation fossa of the nucleus. In the mature spermatid and spermatozoon, the granular body winds itself helically around the centriolar complex in the neck/midpiece region of the cell, and, thus, becomes the granular helix. The significance of this observation must await future studies, including possible phylogenetic re-evaluation and classification of birds.     

Ultrastructure of spermatozoa correlated with phylogenetic relationship between Heteropneustes fossilis and Rana tigrina

A study using transmission electron microscopy (TEM) of the spermatozoa of Rana tigrina and Heteropneustes fossilis in all phases of the annual reproductive cycle revealed that there was a phylogenetic relationship between them. The spermatozoa of H. fossilis appeared horseshoe-shaped, somewhat oval or wedge-shaped at the anterior end and broader at the posterior end. The horseshoe-shaped spermatozoan nucleus was observed during spermiogenesis of R. tigrina but later changed into a finger shape at maturity. The posterior end of the nuclei of mature spermatozoa of R. tigrina was blunt. The extremely dense homogenized nucleus was capped with an acrosomal vesicle in both species suggesting a definite phylogenetic inter-relationship between them.     

Ultrastructure of Spermatogenesis in the Testis of Paragonimus heterotremus

Lung fluke, Paragonimus heterotremus, is a flatworm causing pulmonary paragonimiasis in cats, dogs, and humans in Southeast Asia. We examined the ultrastructure of the testis of adult P. heterotremus with special attention to spermatogenesis and spermiogenesis using scanning and transmission electron microscopy. The full sequence of spermatogenesis and spermiogenesis, from the capsular basal lamina to the luminal surface, was demonstrated. The sequence comprises spermatogonia, spermatocytes with obvious nuclear synaptonemal complexes, spermatids, and eventual spermatozoa. Moreover, full steps of spermatid differentiation were shown which consisted of 1) early stage, 2) differentiation stage representing the flagella, intercentriolar body, basal body, striated rootlets, and electron dense nucleus of thread-like lamellar configuration, and 3) growing spermatid flagella. Detailed ultrastructure of 2 different types of spermatozoa was also shown in this study.     

Differentiation of binuclear spermatozoa in mice

In an electron microscopy study of abnormal spermatogenesis in mice, we have found that two discrete haploid nuclei may be located in a single spermatid cytoplasm after the second meiotic division. The spermatid continues to differentiate and forms a binucleate spermatozoon with both nuclei separately packaged within the sperm head. The Golgi apparatus of the double spermatid forms a single proacrosome that attaches to both nuclei. Apparently, one acrosomal structure differentiates to cover and compartmentalize the two haploid nuclei within the sperm head. Chromatin condensation appears normal. The head morphology and number of flagella vary in mature spermatozoa produced by this process. This work demonstrates one pathway by which polyploid spermatids continue to differentiate to spermatozoa after failure of cytoplasmic division or possibly cellular fusion.