Spermatogenesis in animals as revealed by electron microscopy

Summary The first indication of differentiation of the Jensen's ring has been detected in an early stage of spermiogenesis of Felis catus Linné when the pair of centrioles takes up a position immediately beneath the plasma membrane. The chromatoid bodies appear in the early spermatid cytoplasm through the nuclear pore complex. In a more advanced stage, such bodies have been found in association with the striated columns, the distal centriole or the proximal part of flagellum and the Jensen's ring. As the spermiogenesis proceeds, the bodies have decreased their size and density, and finally disappear in mature spermatozoa. The chromatoid bodies seem, therefore, to share with the centriole the capacity to form the connecting piece. As a consequence of disorganization of triplet microtubules of the centriole, a noticeable material appears in the center of lumen of the centriole to be identifiable as a distinct precursor of the central pair of axonemal complex. Microtubules are first developed as the sheath of principal piece of the sperm flagellum, originating from the plasma membrane surrounding the axonemal complex.     

The Spermatozoa of the Thysanuran Insects Petrobius brevistylis Carp. and Lepisma saccharina L.

The spermatozoa of Petrobius and Lepisma share a few general insect features (filamentous shape, two mitochondria, compact acrosome vesicle, bilateral symmetry) but differ fundamentally with regard to specializations. In Petrobius, a long coiled acrosome, a coiled nucleus, and a “body” with axonema, two mitochondria, and a pair of lateral bodies follow each other in normal sequence. In Lepisma the acrosome is a small vestige in the spoon-shaped anterior end, the centriole is dislocated anteriorly, and nucleus, two mitochondria and axonema run like parallel filaments through most of the spermatozoon. The centriole adjunct develops into a postnuclear body in Lepisma but forms a pair of complicated “lateral bodies” in Petrobius. It is concluded that ancestral forms must have had fairly primitive spermatozoa and that specialization has proceeded independently within each evolutionary line.     

The spermatogenesis and the sperm structure of Terebrantia (Thysanoptera, Insecta)

Spermatogenesis and the sperm structure of the terebrantian Aeolothrips intermedius Bagnall are described. Spermatogenesis consists of two mitotic divisions; the second is characterized by the loss of half of the spermatids, which have pyknotic nuclei. Early spermatids have two centrioles, but when spermiogenesis starts, a third centriole is produced. The three basal bodies give rise to three flagella; later these fuse into a single flagellum which contains three 9 + 0 axonemes. The basal bodies are surrounded by a large amount of centriole adjunct material. During spermiogenesis this material contributes to the shifting of the three axonemes towards the anterior sperm region parallel to the elongating nucleus, and it is transformed into a dense cylinder. In the mature spermatids the three axonemes amalgamate to create a bundle of 27 doublet microtubules. Near the end of spermiogenesis the dense cylinder of the centriole adjunct lies parallel to the nucleus and the axonemes. It ends where the mitochondrion appears at half-sperm length. We confirm that Terebrantia testes have a single sperm cyst; their sperm are characterized by a cylindrical nucleus, three axonemes fused into one, a small mitochondrion and a short cylindrical centriole adjunct which corresponds to the dense body described in a previous work. The acrosome is lacking. At the midpoint of the anterior half of the sperm the outline of the cross-section is bilobed, with the nucleus contained in a pocket evagination of the plasma membrane. These characters are discussed in light of a comparison between Tubulifera and Terebrantia.     

Annulate lamellae and chromatoid bodies in the testes of a cyprinid fish (Pimephales notatus)

Summary Observations on annulate lamellae and chromatoid bodies in spermatogonia of the cyprinid fish Pimephales notatus have revealed several commonly occurring features heretofore unreported: These include (a) the presence of annulate lamellae in close association with chromatoid bodies; (b) the existence of a chromatoid band or shell between the nuclear envelope and some chromatoid bodies with connections among them; (c) the presence of annulate pore complexes in the absence of well developed membrane envelopes as well as in association with such envelopes; (d) the presence of material just outside the nucleus and contiguous with nuclear pores which is of a similar density and texture to that of the chromatoid bands and chromatoid bodies; (e) filamentous material between the cytoplasmic sides of nuclear pores and the chromatoid band, bridging a distance of approximately 1000 Å and similar threads extending a like distance between chromatoid bodies (and bands) and annulate lamellae associated with them; and (f) mitochondria closely arranged about some chromatoid bodies.     

Ultrastructural labeling of the chromatoid body and the centriole-associated body using the DNase-gold colloidal complex in monkey spermatids

In Monkey spermatids at different steps of spermiogenesis, the use of DNase-gold complex showed, at the ultrastructural level, a labeling over the chromatin and concomitantly over the chromatoid body, centriole associated body and annular chromatoid body. The results obtained with the DNase-gold complex containing either DNA or actin led to discuss the nature of the substances revealed by the labeling in the cytoplasmic structures.     

Spermatogenesis and sperm structure of Acerella muscorum, (Ionescu, 1930) (Hexapoda, Protura)

The general organization of the male genital system, the spermatogenesis and the sperm structure of the proturan Acerella muscorum have been described. At the apex of testis apical huge cells are present; their cytoplasm contains a conventional centriole, a large amount of dense material and several less electron-dense masses surrounded by mitochondria. Spermatocytes have normal centrioles and are interconnected by cytoplasmic bridges. Such bridges seem to be absent between spermatid cells and justify the lack of synchronization of cell maturation. Spermatids are almost globular cells with a spheroidal nucleus and a large mass of dense material corresponding to the centriole adjunct. Within this mass a centriole is preserved. Mitochondria of normal structure are located between the nucleus and the plasma membrane. The spermatids are surrounded by a thick membrane. No flagellar structure is formed. Sperm have a compact spheroidal nucleus, a large cap of centriole adjunct material within which a centriole is still visible. A layer of mitochondria is located over the nucleus. The cytoplasm is reduced in comparison to spermatids; many dense bodies are interspersed with sperm in the testicular lumen. The sperm are small, immotile cells of about 2.5-3 μm in diameter.     

Spatiotemporal changes of levels of a moonlighting protein,phospholipid hydroperoxide glutathione peroxidase,in subcellular compartments during spermatogenesis in the rat testis

We studied temporal changes in the subcellular localization and levels of a moonlighting protein, phospholipid hydroperoxide glutathione peroxidase (PHGPx), in spermatogenic cells and mature sperm of the rat by immunofluorescence and immunoelectron microscopy. The PHGPx signals were detected in chromatoid bodies, clear nucleoplasm, mitochondria-associated material, mitochondrial aggregates, granulated bodies, and vesicles in residual bodies in addition to mitochondria, nuclei, and acrosomes as previously reported. Within mitochondria, PHGPx moved from the matrix to the outermost membrane region in step 19 spermatid, suggesting that this spatiotemporal change is synchronized with the functional change of PHGPx in mitochondria. In the nucleus, PHGPx was associated with electron-lucent spots and with the nuclear envelope, and PHGPx in the latter region increased after step 16. In early pachytene spermatids, PHGPx signals were noted in the nuclear material exhibiting a very similar density to chromatoid bodies and in the intermitochondrial cement, supporting the previous proposal that chromatoid bodies originate from the nucleus and intermitochondrial cement. The presence of PHGPx in such various compartments suggested versatile roles for this protein in spermatogenesis. Quantitative immunoelectron microscopic analysis also revealed dynamic changes in the labeling density of PHGPx in different subcellular compartments as follows: 1). Total cellular PHGPx rapidly increased after step 5 and reached a maximum at step 18; 2). mitochondrial labeling density increased after step 1 and achieved a maximum in steps 15-17; 3). nuclear labeling density suddenly increased in steps 12-14 to a maximum; 4). in cytoplasmic matrix, the density remained low in all steps; and 5). the labeling density in chromatoid bodies gradually decreased from pachytene spermatocytes to spermatids at step 18. These spatiotemporal changes in the level of PHGPx during the differentiation of spermatogenic cells to sperm infer that PHGPx plays a diverse and important biological role in spermatogenesis.     

Localization of mitochondrial ribosomal RNA on the chromatoid bodies of marine planarian polyclad embryos

Electron-dense cytoplasmic structures, referred to as chromatoid bodies, are observed in the somatic stem cells, called neoblasts, and germline cells in adult planarians. Although it has been revealed that the chromatoid bodies morphologically resemble germline granules in Drosophila and Xenopus embryos, what essential role it plays in the planarian has remained unclear. In the present study, to examine whether chromatoid bodies in planarian embryos are responsible for germline formation, the presence and behavior of chromatoid bodies during embryogenesis were examined. Mitochondrial large ribosomal RNA and mitochondrial small ribosomal RNA were used as candidate markers for components of the chromatoid body. Starting from the fertilized egg, extramitochondrial signals of both RNA (mtrRNA) were observed. At the ultrastructural level, mtrRNA were localized on the surface of the chromatoid bodies. At subsequent stages, the signals of mtrRNA were observed in certain restricted blastomeres that contribute to the formation of larval structures. The signals gradually decreased from the gastrula stage. These results suggest that the chromatoid bodies associated with mtrRNA in embryogenesis are not germline granules. The chromatoid bodies of blastomeres may be concerned with the toti- or pluripotency and cell differentiation as proposed in adult planarian neoblasts.     

Fine structure of the chromatoid body during spermatogenesis in the water-strider,Gerris remigis (SAY)

Summary During spermatogenesis in Gerris remigis, chromatoid bodies appear in the spermatocytes and persist to the-mid-spermatid stage. These structures consist of numerous, parallel tubules, which measure approximately 500 Å in diameter. The tubules are arranged in hexagonal array, and contain dense granules that resemble ribosomes. The chromatoid body may be secretory in function, or may be involved in intracellular transport.The technical assistance of Mr. Roy R. Keppie and Mrs. Mona Brandreth is gratefully acknowledged.     

Ultrastructural localization of RNA in the chromatoid bodies of undifferentiated cells (neoblasts) in planarians by the RNase–gold complex technique

Undifferentiated cells of planarians (Platyhelminthes, Turbellaria), also called neoblasts, are totipotent stem cells, which give rise to all differentiated cell types, while maintaining their own density by cell proliferation. Neoblasts are the only somatic cells of planarians bearing chromatoid bodies in their cytoplasm; these organelles disappear as differentiation takes place. Studies on germinal cells of several groups of organisms have shown that chromatoid bodies contain substantial amounts of RNA. To test its presence in neoblasts, we have used an RNase–gold technique. We found chromatoid bodies labeled with RNase–gold particles. Heterogeneity in the density of the label, may be correlated with the functionality and complexity of these organelles. The gold marker was also present over the nucleus and rough endoplasmic reticulum, but mitochondria, secretory granules, and the extracellular space were devoid of label. This specific localization of RNA in planarian chromatoid bodies supports earlier findings on germ cells and embryonic cells in a variety of organisms, indicating that chromatoid bodies are information-storage structures, essential during the process of cell differentiation. © 1993 Wiley-Liss, Inc.     

线粒体可能参与鲫鱼精子发生中拟染色体的形成

运用电子显微镜观察了鲫鱼生精细胞发育过程中拟染色体的形成和解体,以及拟染色体和线粒体的关系。在精子细胞阶段之前的各期生精细胞中都存在拟染色体。仅在精原细胞中观察到拟染色体的形成过程。拟染色体的形成方式与其它鱼类中拟染色体的形成方式相似。在生精细胞的发育过程中,线粒体的形态和数量发生变化。在初级精原细胞阶段,线粒体较大,多为球形,嵴少,基质电子密度低。随着生精细胞的发育,线粒体逐渐变小,多为长条状,嵴多,基质的电子密度升高。拟染色体形成后往往与线粒体结合。与拟染色体结合的线粒体往往解体,部分或全部的外膜和内膜破裂以至消失。线粒体解体后,其中的物质可能会转移到拟染色体中[动物学报52(2):328-334,2006]。     

Polysome-like structures in the chromatoid body of rat spermatids

A procedure for isolating the chromatoid body from the testis of 40-day-old rats was developed. Electron-microscopical analysis indicated that about 70% of the isolated organelles were chromatoid bodies, while the remaining structures corresponded to dense bodies and probably to satellites. Negative staining of the isolated organelles revealed the presence of polysome-like structures in about 20% of the chromatoid bodies suggesting that the polysomes were not due to contamination with cytoplasmic polysomes. Moreover, the presence of RNA in the stroma of the chromatoid body was confirmed by RNAse-gold staining. Preliminary electrophoretic analysis of the RNA extracted from the organelles revealed the presence of a complex population of RNAs including 5.8 and 5 S ribosomal RNAs but no tRNA.     

The sperm structure of Cryptocercus punctulatus Scudder (Blattodea) and sperm evolution in Dictyoptera

Sperm of the dictyopteran key taxon Cryptocercus punctulatus was examined. It has largely maintained a blattodean groundplan condition, with a three‐layered acrosome, an elongate nucleus, a single centriole, a conspicuous centriole adjunct material, two connecting bands (=accessory bodies), and a long functional flagellum with a 9+9+2 axoneme provided with accessory tubules with 16 protofilaments and intertubular material. These sperm characters are shared with several other polyneopterans. The sperm of C. punctulatus is very similar to what is found in Periplaneta americana and species of other groups of roaches, including the sperm of Loboptera decipiens described here for the first time. The general sperm organization here described can be assumed for the groundplan of Insecta and Pterygota. The following evolutionary path can be suggested: after the split between Cryptocercidae and the common ancestor of Isoptera, the typical pattern of sperm formation was altered very distinctly, resulting in a duplication or multiplication (Mastotermitidae) of the centrioles. Mastotermes has maintained a certain sperm motility, but with a very unusual apparatus of multiple flagella with a 9+0 axoneme pattern. After the split into Mastotermitidae and the remaining Isoptera, sperm motility was completely abandoned, and different modifications of sperm components occurred, and even the loss of the sperm flagellum. J. Morphol. 276:361–369, 2015. © 2014 Wiley Periodicals, Inc.     

Spermiogenesis in three species of cicadas (Hemiptera: Cicadidae)

Spermiogenesis in three species of cicadas representing one cicadettine (Monomatapa matoposa Boulard) and two cicadines (Diceroprocta biconica [Walker] and Kongota punctigera [Walker]) was investigated by light and electron microscopy. Although spermiogenesis was occurring in the testis of adult males of all species, earlier spermiogenic stages were observed in D. biconica only. While spermiogenesis was similar to that described for other insects, some differences were noted. For example granular material did not assemble around the centriole to form a centriolar adjunct but did accumulate in the cytoplasm of early spermatids adjacent to a region of the nuclear membrane where nuclear pores were aggregated. In late spermatids this material accumulated anterior to the mitochondrial derivatives in a developing postero‐lateral nuclear groove. While this material has been named the ‘centriolar adjunct’ by previous authors, its formation away from the centriole raises questions about its true identity. Second, during acrosome maturation an ante‐acrosomal region of cytoplasm develops. Although present in later spermatids, this region is lost in spermatozoa. Interspecific variations in chromatin condensation patterns and the number of microtubule layers encircling the spermatid nucleus during spermiogenesis were noted.     

A ribonucleoprotein inclusion body in Entamoeba invadens

Summary Cytochemical tests have shown that the chromatoid bodies contain mainly ribonucleic acid and protein. There is no evidence to suggest the presence of any desoxyribonucleic acid, lipid, glycogen, neutral fat, or metaphosphate. The cyclic changes in the chromatoid body have been studied by means of light and electron microscopy and estimations of the total RNA present. The results indicate that the chromatoid bodies arise by a process of aggregation of small groups of 250–300 A units form polycrystalline masses in precystic and early cysts. At the same time there is a steady rise in the amount of RNA present. In the maturing cysts, the crystalline masses fragment into separate particles, but there is no corresponding fall in the amount of RNA. The significance of these results in relation to the previous literature on chromatoid bodies and similarly named cellular inclusions in spermatogenic and plant cells, as well as in the protozoa, is discussed. Attention is drawn to the striking similarity of composition, formation, development and ultimate fate of these various inclusion bodies but as the terminology is still confused it is proposed that the term chromatoid body be retained solely for particulate or lamellar ribonucleo-protein accumulations. Acknowledgements. Most of this study was presented in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the University of Edinburgh. I should like to express my deep appreciation to Prof. M. M. Swann F. R. S. for his interest and helpful guidance in this work. I am grateful to the Melville Trust for Cancer Research who provided the electron microscope.     

Inhibitor induced alterations of chromatoid bodies in male germ line cells of Xenopus laevis

The action of inhibitors of protein synthesis on the structure of cytoplasmic inclusions found in the male germ cell line of the anuran, Xenopus laevis, has been studied by light and electron microscopy. Results indicate that one such inclusion, the chromatoid body, is sensitive to treatment with either chloramphenicol or puromycin. These drugs administered in vivo or in vitro cause up to a thirty-fold increase in the volume of the chromatoid body in all stages where it is normally present. Maximum size increase obtainable is the same for either drug, but is different and characteristic for each germ cell stage. Drug action is dose dependent, with "chromatoid body syndrome" occurring over a relatively narrow concentration range. Cyclohexamide, in contrast to chloramphenicol or puromycin, does not produce a clear increase in the size of chromatoid bodies, and is capable of blocking the action of the other drugs at normally effective concentrations. Results obtained in this investigation suggest that primary spermatogonia contain enough chromatoid body material to account for the total amount present in all subsequent germ cell stages. This fact, coupled with other studies where chromatoid-like bodies have been observed, suggests the hypothesis that the chromatoid body represents at least in part an aggregation stage of materials associated with the microtubule population of the germ cell line. Alternately, or in addition, ribonucleoprotein may contribute to the structure of the chromatoid body.     

Sperm structure of Mecoptera and Siphonaptera (Insecta) and the phylogenetic position of<Emphasis Type="Italic"> Boreus hyemalis</Emphasis>

Sperm ultrastructure has been studied in three species of the taxa Mecoptera and Siphonaptera. The spermatozoon of the scorpion fly Panorpa germanica shows an apical bilayered acrosome, a helicoidal nucleus, a centriolar region and a 9+2 flagellar axoneme helicoidally arranged around a long mitochondrial derivative. A second mitochondrial derivative is very short and present only in the centriolar region. A single accessory body is present and it is clearly formed as a prolongation of the centriole adjunct material. Two lateral lamellae run parallel to the nucleus. The snow fly Boreus hyemalis has a conventional sperm structure and shows a bilayered acrosome, a long nucleus, a centriolar region, two mitochondrial derivatives and two accessory bodies. The axoneme is of the 9+2 type and is flattened at the tail tip. Both P. germanica and B. hyemalis have two longitudinal extra-axonemal rods and have a glycocalyx consisting of longitudinal parallel ridges or filaments. The spermatozoon of the flea Ctenocephalides canis has a long apical bilayered acrosome, a nucleus, a centriolar region, a 9+2 axoneme wound around two unequally sized mitochondrial derivatives, and two triangular accessory bodies. In the posterior tail end the flagellar axoneme disorganises and a few microtubular doublets run helicoidally around the remnant mitochondrial derivative. The glycocalyx consists of fine transverse striations. In all three species, the posterior tail tip is characterised by a dense matrix embedding the disorganised axoneme. From this comparative analysis of the sperm structure it is concluded that Mecoptera, as traditionally defined, is monophyletic and that B. hyemalis is a member of Mecoptera rather than of Siphonaptera.     

褐菖■精细胞晚期的变化及精子结构研究

本文研究卵胎生硬骨鱼褐菖（Ｓｅｂａｓｔｉｓｃｕｓｍａｒｍｏｒａｔｕｓ）精细胞的成熟变化和精子结构。褐菖精细胞发育晚期已具有硬骨鱼类精子的结构雏形：细胞核的背面较平坦,腹面稍外鼓,呈弧面;染色质浓缩成团块状,核的腹侧和后端的染色质较致密;中心粒复合体由近端中心粒和基体组成,近端中心粒和基体排成“Ｌ”形;近端中心粒向细胞核的背侧伸出中心粒附属物,中心粒附属物由９条微管组成,９条微管围成一筒状结构,类似轴丝。在晚期精细胞形成精子的过程中,中心粒附属物和近端中心粒相继退缩以至消失不见,同时细胞核后端的形状也随着发生变化。中心粒附属物和近端中心粒的相继消失可以看作是成熟的最后标志。精子的中心粒复合体由基体及其上方的基体帽组成,袖套接于核的后端,其中约有３０～４０个线粒体;鞭毛从袖套腔中伸出,鞭毛的中心结构是轴丝;轴丝外方为细胞质形成的侧鳍,在鞭毛的近核段,轴丝两侧的侧鳍较宽且不对称。     

Sperm ultrastructure and spermiogenesis in the relic species Tricholepidion gertschi Wygodzinsky (Insecta, Zygentoma)

The silverfish Tricholepidion gertschi is of interest in that it is the most basal representative of Zygentoma. An ultrastructural study of its spermiogenesis was performed to find out whether there are traits which resemble those of other, more advanced insects. This was found to be the case; spermiogenesis can be considered to be of a common insectan type, leading to the formation of elongated sperm cells with acrosome, nucleus, neck region and a tail with axoneme and two mitochondrial derivatives. Total cell length, 50 microm, is short for an insect. There are some specializations, which probably represent autapomorphies. The acrosome has a posterior canal or cleft that makes a U-turn. The centriole adjunct forms a prominent post-nuclear ring surrounding the centriole and have a posterior extension, and further originates nine intertubular fibers with a longitudinal periodicity and two accessory bodies. The mitochondrial derivatives have five rows of regularly spaced cristae within a crystalline matrix. The axoneme has accessory tubules consisting of 16 protofilaments, formed at the B-tubules of the doublets and placed at some distance from them in the posterior part of the sperm tail.