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]。     

Transport of material between the nucleus,the chromatoid body and the golgi complex in the early spermatids of the rat

Summary The movement and transport of material between intranuclear dense particles, the chromatoid body and the Golgi complex have been studied in early spermatids of the rat. The analyses involved observation of living accurately identified cells, time-lapse cinemicrography and electron microscopy.The chromatoid body establishes transient contacts with intranuclear material during early spermiogenesis. The chromatoid body also makes contacts with the Golgi complex. It is suggested that the chromatoid body receives material from the nucleus during the postmeiotic period and participates in the early formation of the acrosomic system.This work was supported by the Finnish National Research Council for Medical Sciences. The authors are grateful to Mrs. Marita Aaltonen and Mrs. Raija Andersen for their skilful technical assistance     

Spoltud-1 is a chromatoid body component required for planarian long-term stem cell self-renewal

Freshwater planarians exhibit a striking power of regeneration, based on a population of undifferentiated totipotent stem cells, called neoblasts. These somatic stem cells have several characteristics resembling those of germ line stem cells in other animals, such as the presence of perinuclear RNA granules (chromatoid bodies). We have isolated a Tudor domain-containing gene in the planarian species Schmidtea polychroa, Spoltud-1, and show that it is expressed in neoblast cells, germ line cells and central nervous system, and during embryonic development. Within the neoblasts, Spoltud-1 protein is enriched in chromatoid bodies. Spoltud-1 RNAi eliminates protein expression after 3 weeks, and abolishes the power of regeneration of planarians after 7 weeks. Neoblast cells are eliminated by the RNAi treatment, disappearing at the end rather than gradually during the process. Neoblasts with no detectable Spoltud-1 protein are able to proliferate and differentiate. These results suggest that Spoltud-1 is required for long term stem cell self renewal.     

Relationship between the chromatoid body and the acrosomal system in early spermatids of Myxine glutinosa L.

Summary A transient close relationship between the chromatoid body and the developing acrosome is demonstrated in early spermatids of Myxine glutinosa.This work was supported by the Norwegian Research Council for Science and Humanities (NAVF, Grant Nr. D 61.44) and the Austrian Fonds zur Förderung der wissenschaftlichen Forschung, Projekt 2183     

Localization of mouse vasa homolog protein in chromatoid body and related nuage structures of mammalian spermatogenic cells during spermatogenesis

The localization of vasa homolog protein in the spermatogenic cells of mice, rats, and guinea pigs was studied by immunofluorescence and electron microscopies with the antibody against mouse vasa homolog (MVH) protein. By immunofluorescence microscopy, four types of granular staining patterns were identified: (1) fine particles observed in diplotene and meiotic cells, (2) small granules associated with a mitochondrial marker and appearing in pachytene spermatocytes after stage V, (3) strands lacking the mitochondrial marker in late spermatocytes, and (4) large irregularly shaped granules in round spermatids. Immunoelectron microscopy defined the ultrastructural profiles of these MVH protein-positive granules: the first type consisted of small dense particles, the second had intermitochondrial cement (IMC), the third type, consisting of strands, had loose aggregates of either material dissociated from IMC or 70–90-nm particles, and the fourth had typical chromatoid bodies (CBs). The results suggest that MVH proteins function in these components of nuage. MVH protein-positive structures other than CBs disappeared during meiosis and CB appeared first in early spermatids. The results suggest that the formation of nuage is discontinued between spermatocytes and spermatids. The formation of nuage in spermatocytes and of CB in spermatids is discussed.     

Tudor-related proteins TDRD1/MTR-1, TDRD6 and TDRD7/TRAP: domain composition, intracellular localization, and function in male germ cells in mice

The germ-line cells of many animals possess a characteristic cytoplasmic structure termed nuage or germinal granules. In mice, nuage that is prominent in postnatal male germ cells is also called intermitochondrial cement or chromatoid bodies. TDRD1/MTR-1, which contains Tudor domain repeats, is a specific component of the mouse nuage, analogously to Drosophila Tudor, a constituent of polar granules/nuage in oocytes and embryos. We show that TDRD6 and TDRD7/TRAP, which also contain multiple Tudor domains, specifically localize to nuage and form a ribonucleoprotein complex together with TDRD1/MTR-1. The characteristic co-localization of TDRD1, 6 and 7 was disrupted in a mutant of mouse vasa homologue/DEAD box polypeptide 4 (Mvh/Ddx4), which encodes another evolutionarily conserved component of nuage. In vivo over-expression experiments of the TDRD proteins and truncated forms during male germ cell differentiation showed that a single Tudor domain is a structural unit that localizes or accumulates to nuage, but the expression of the truncated, putative dominant negative forms is detrimental to meiotic spermatocytes. These results indicate that the Tudor-related proteins, which contain multiple repeats of the Tudor domain, constitute an evolutionarily conserved class of nuage components in the germ-line, and their localization or accumulation to nuage is likely conferred by a Tudor domain structure and downstream of Mvh, while the characteristic repeated architecture of the domain is functionally essential for the differentiation of germ cells.     

Ultrastructural observations on the germ line of Xenopus laevis

Summary The development of the male germ line in Xenopus laevis has been examined by electron microscopy. Findings have been compared to the parallel process in the female. Three structures unique to the germ line were found in both male and female cells: a fibrillar nuclear region free of DNA; largely proteinaceous masses of nuage material; and a chromatoid body. Germ plasm bodies of the egg and early embryo appear to represent a form of nuage material. The finding of a structure which can be identified as a chromatoid body in the female germ line is unique, as is its presence in sexually undifferentiated primordial germ cells. The chromatoid body in Xenopus, unlike that in mammals, does not persist in the spermatozoon. Instead, it dissociates into a series of coated vesicles during spermatogenesis. The chromosomal ultrastructure of meiotic prophase stages in Xenopus is similar in both sexes until diplotene, when male bivalents condense and enter meiotic metaphase instead of entering the extended lampbrush stage characteristic of the female. The multiple nucleoli present in gonia are lost at the onset of meiotic prophase, but no obvious mechanism for this process was observed.The author would like to thank Drs. Joseph Gall and Bernard Tandler for their helpful suggestions during the course of this investigation. The author is a postdoctoral fellow of the National Institutes of Health, U.S.A. This research was supported by N.I.H. Grants 51823 and 12427.     

Sm proteins, the constituents of the spliceosome, are components of nuage and mitochondrial cement in Xenopus oocytes

A conserved feature of germ cells in many animal species is the presence of perinuclear electron-dense material called the "nuage" that is believed to be a precursor of germinal (or polar or P) granules. In Xenopus oogenesis the nuage is first observed near the nuclear envelope and subsequently in close contact with mitochondria, at which stage it is called the mitochondrial cement. In this study, we found that, in Xenopus pre-stage I and stage I oocytes, nuage and mitochondrial cement contain the spliceosomal Sm proteins, Xcat2 mRNA, and DEAD-box RNA helicase XVLG1. Other components of Cajal bodies or splicing machinery such as coilin, SMN protein, and snRNAs are absent from the nuage and mitochondrial cement. We suggest that Xenopus Sm proteins have adapted to a role independent of pre-mRNA splicing and that instead of binding to their traditional spliceosomal partner such as snRNA, they bind mRNAs that are the components of germinal granules (i.e., Xcat2 mRNA) and facilitate the transport of these mRNAs from the nucleus to the nuage that is a precursor of germinal granules. In addition, the presence of Vasa-like DEAD-box helicase in Xenopus nuage suggests involvement of nuage in the microRNA and/or RNAi pathway, similar to the role of nuage in Drosophila.     

Ultrastructure of germ cells, the Leydig cells, and Sertoli cells during spermatogenesis in Boleophthalmus pectinirostris (Teleostei, Perciformes, Gobiidae)

The ultrastructures of germ cells, Leydig cells, and Sertoli cells during spermatogenesis in male Boleophthalmus pectinirostris were investigated by electron microscopic observations. During the period of maturation divisions, well-developed Leydig cells have three major morphological characteristics: a vesicular nucleus, mitochondria with tubular cristae, and a number of smooth endoplasmic reticulum. Based on cytoplasmic features, it appears that Leydig cells are responsible for the synthesis of male sex steroids. Although no clear evidence of steroidogenesis was found in the Sertoli cells, they were found to perform a phagocytic function in the seminiferous lobules. Most Sertoli cells contain granules thought to represent deposited glycogen or lipid but there is no indication of a transfer of nutrients to the spermatids. During the period of germ cell degeneration, several characteristics of phagocytosis appear in the cytoplasm of the Sertoli cells. In particular, it is assumed that the Sertoli cells are involved in the degeneration and resorption of undischarged spermatids after spermiation. No acrosome of the sperm is formed. The structure of the spermatozoon in B. pectinirostris is very similar and closely resembles to those of suborder Gobioidei (perciform type teleosts). The flagellum or sperm tail shows the typical 9+2 array of microtubules.     

黑脊倒刺鲃卵子发生中生殖质的产生

采用光学和电子显微镜对黑脊倒刺的卵原细胞和各期卵母细胞中生殖质的形成过程进行观察。仅在卵原细胞和Ⅱ时相卵中观察到了生殖质的产生过程 ,在Ⅲ、Ⅳ时相卵中未观察到生殖质的产生过程。生殖质来源于核仁。它从细胞核中释放出来 ,进入细胞质。卵原细胞和卵母细胞产生生殖质的方式不同。在卵原细胞的生殖质形成过程中 ,生殖质的前体物质先移到核膜内侧 ,核膜解体 ,形成囊泡 ,随后 ,在生殖质前体物质所在处的内侧重新形成核膜 ,这样 ,细胞核表面就形成一个凹陷 ,生殖质前体物质位于核表面的凹陷之中 ,就这样被隔离于细胞核之外而进入细胞质中 ,成为生殖质。生殖质形成后 ,在细胞质中与线粒体相结合。生殖质的这种形成方式与精原细胞中拟染色体的形成方式相似。而在卵母细胞的生殖质形成过程中 ,生殖质的前体物质移到核膜内侧之后 ,核膜并无囊泡化 ,核孔明显 ,生殖质的前体物质通过核孔离开细胞核 ,位于细胞核表面 ,形成生殖质。生殖质产生之后也与线粒体结合。以后 ,生殖质连同线粒体离开细胞核。生殖质最终与线粒体分离 ,分散于细胞质中。在Ⅲ时相卵中 ,生殖质细小 ;在Ⅳ时相卵中 ,几乎见不到生殖质     

Meiosis and retrotransposon silencing during germ cell development in mice

In mammals, germ cells derive from the pluripotent cells that are present early in embryogenesis, and then differentiate into male sperm or female eggs as development proceeds. Fusion between an egg and a sperm at fertilization allows genetic information from both parents to be transmitted to the next generation, and produces a pluripotent zygote to initiate the next round of embryogenesis. Meiosis is a central event in this self-perpetuating cycle that creates genetic diversity by generating new combinations of existing genetic alleles, and halves the number of chromosomes in the developing male and female germ cells to allow chromosome number to be maintained through successive generations. The developing germ cells also help to maintain genetic and chromosomal stability through the generations by protecting the genome from excessive de novo mutation. Several mouse mutants have recently been characterised whose germ cells exhibit defects in silencing the potentially mutagenic endogenous retroviruses and other retrotransposons that are prevalent in mammalian genomes, and these germ cells also exhibit defects in progression through meiosis. Here we review how mouse germ cells develop and proceed through meiosis, how mouse germ cells silence endogenous retroviruses and other retrotransposons, and discuss why silencing of endogenous retroviruses and other retrotransposons may be required for meiotic progression in mice.     

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.     

The immunolocalization of small nuclear ribonucleoprotein particles in testicular cells during the cycle of the seminiferous epithelium of the adult rat

The objective of this study was to determine the cellular and subcellular distribution of small nuclear ribonucleoprotein particles (snRNPs) in the adult rat testis in relation to the different cell types at the various stages of the cycle of the seminiferous epithelium. The distribution of snRNPs in the nucleus and cytoplasm of germ cells was quantitated in an attempt to correlate RNA processing with morphological and functional changes occurring during the development of these cells. Light-microscopic immunoperoxidase staining of rat testes with polyclonal anti-Sm and monoclonal anti-Y12 antibodies localized spliceosome snRNPs in the nuclei and cytoplasm of germ cells up to step 10 spermatids. Nuclear staining was intense in Sertoli cells, spermatogonia, spermatocytes, and in the early steps of round spermatid development. Although comparatively weaker, cytoplasmic staining for snRNPs was strongest in mid and late pachytene spermatocytes and early round spermatids. Quantitative electron-microscopic immunogold labeling of Lowicryl embedded testicular sections confirmed the light-microscopic observations but additionally showed that the snRNP content peaked in the cytoplasm of midpachytene spermatocytes and in the nuclei of late pachytene spermatocytes. The immunogold label tended to aggregate into distinct loci over the nuclear chromatin. The chromatoid body of spermatids and spermatocytes and the finely granular material in the interstices of mitochondrial aggregates of spermatocytes were found to be additional sites of snRNP localization and were intensely labeled. This colocalization suggests that these dense cytoplasmic structures may be functionally related. Anti-U1 snRNP antibodies applied to frozen sections showed the same LM localization pattern as spliceosome snRNPs. Anti-U3 snRNP antibodies applied to frozen sections stained nucleoli of germ cells where pre-rRNA is spliced.     

The identification and characterization of a testis-specific cDNA during spermatogenesis

Using bioinformatics and experimental validation, we obtained a cDNA (named srsf) which was exclusively expressed in the mouse testes. RT-PCR analysis showed that srsf mRNA was not expressed in the gonad during the sex determination period or during embryogenesis. In developing mouse tests, srsf expression was first detected on post-natal day 10, reached its highest level on day 23, and then reduced to and remained at a moderate level throughout adulthood. In situ hybridization analysis demonstrated that srsf mRNA was expressed in pachytene spermatocytes and round spermatids in the testes. The predicted protein contains one RNA-binding domain (RBD) and a serine-arginine rich domain (RS), which are characterized by some splicing factors of SR family members. These findings indicate that srsf may play a role during spermatogenesis.     

不同壳色福寿螺形态性状与体质量的关系

【背景】福寿螺因其食性杂、抗逆性和繁殖力强以及自然天敌少等不断扩散,侵害农作物,被列为我国首批外来入侵物种。国内外学者一直致力于研究对其的防治与监控。自然界中福寿螺存在2种壳色——黄色和黑色,壳色受遗传因素和环境因素的双重影响。广东省福寿螺多以黑色为主,福寿螺倾向于与不同壳色的螺交配。壳色在一定程度上影响其交配的选择性,但2种壳色的福寿螺繁殖力指标差异不显著。而关于这2种壳色的螺在形态学上的差异鲜有报道。【方法】利用生物统计软件和分析方法进行相关性分析、通径分析及多元回归分析,计算相关系数、通径系数和决定系数,研究2种壳色福寿螺形态性状与体质量的关系。【结果】2种壳色福寿螺的体质量、层高的变异系数较大,且黄色比黑色变异系数大。对黄色福寿螺体质量影响较大的依次为壳高、口宽;对黑色福寿螺体质量影响较大的依次为口宽、层高。【结论与意义】2种壳色福寿螺在形态性状方面差异显著,可以将壳色作为特征标记,为福寿螺的监测与灾害评估提供参考。     

Effects of sub-chronic aluminum chloride on spermatogenesis and testicular enzymatic activity in male rats

Aims

The aim of this study was to determine the effects of sub-chronic aluminum chloride (AlCl₃) on spermatogenesis and testicular enzymatic activity in male rats.

Main methods

Forty Wistar male rats were randomly divided into four groups: control group (CG, 0), low-dose group (LG, 64.18 mg/kg BW AlCl₃), mid-dose group (MG, 128.36 mg/kg BW AlCl₃) and high-dose group (HG, 256.72 mg/kg BW AlCl₃). The rats were orally administered with AlCl₃ for 120 days. At the end of the experiment, the contents of Al, Fe, Cu and Zn, the enzyme activities of testicular acid phosphatase (ACP), succinate dehydrogenase (SDH), lactate dehydrogenase (LDH), lactate dehydrogenase isoenzyme (LDH-x), the sperm count and the sperm malformation rate were examined.

Key findings

The results showed that the Al and Cu contents, sperm count and the enzyme activities of testicular ACP, SDH, LDH and LDH-x decreased, while the Zn and Fe contents and sperm malformation rate increased in AlCl₃-treated rats.

Significance

It suggests that sub-chronic AlCl₃ disorders the balance of trace element and decreases the spermatogenesis and the activities of testicular enzymes, indicating that AlCl₃ has adverse effect on the testicular function in male rats.     

Effect of short- and medium-term toxicity of doxorubicin on spermatogenesis in adult Wistar rats

Doxorubicin (DXR) is a widely used chemotherapeutic anticancer agent that has potent activity against several solid and non-solid human malignant tumors, including childhood malignancies. However, DXR has serious toxic effects on tissues with rapid cell cycles, such as myeloid and lymphatic tissues, intestinal mucosa, testes and ovaries. In the present study, the short- and medium-term toxic effects of DXR on the reproductive system of male Wistar rats were evaluated using morphometric and stereological tools to quantify damage to the seminiferous epithelium. Adult male Wistar rats were treated with dose of 7.5?mg/kg of DXR and were sacrificed at seven, 14, 21 and 28?days after treatment. The testes were fixed in glutaraldehyde solution, routinely processed and embedded in plastic for evaluation under a light microscope. A significant reduction in testis weight was found as a result of massive germ cell apoptosis. Differences in comparison to the control group were found in the relative frequency of all stages of the seminiferous epithelium cycle, with significant differences for stages VIII–XI. Apoptosis significantly decreased the number of pachytene spermatocytes in the stages evaluated (I, II–III and VIII) at seven and 14?days. At 21 and 28?days after treatment, the testes exhibited the massive loss of germ cells that resulted in a missing cell layer. Moreover, reductions in the height of seminiferous tubules, tubular diameter and tubular compartment as well as an increase in the intertubular compartment were found in the period studied.