Nuclear morphogenesis during spermiogenesis in the nudibranch molluscHypselodoris tricolor (Gastropoda,opisthobranchia)

An electron microscope study was carried out on Hypselodoris tricolor spermatids to describe the development of the nuclear morphogenesis and investigate the possible cause(s) of the change in the shape of the spermatid nucleus during spermiogenesis. Three different stages may be distinguished in the course of the nuclear morphogenesis on the basis of the morphology and inner organization of the nucleus. Stage 1 spermatid nuclei are spherical or ovoid in shape and the nucleoplasm finely granular in appearance. Stage 2 nuclei exhibit a disc- or cup-shaped morphology, and the chromatin forms short, thin filaments. During stage 3, a progressive nuclear elongation takes place, accompanied by chromatin rearrangement, first into fibers and then into lamellae, both formations helically oriented. A row of microtubules attached to the nuclear envelope completely surrounds the nucleus. Interestingly, the microtubules always lie parallel to the chromatin fibers adjacent to them. Late stage 3 spermatids show the highest degree of chromatin condensation and lack the manchette at the end of spermiogenesis. Our findings indicate the existence of a clear influence exerted on the chromatin by the manchette microtubules, which appear to be involved in determining the specific pattern of chromatin condensation in Hypselodoris tricolor.     

四种癞蝗精子的比较研究（直翅目：蝗总科）

本文通过对丽突鼻蝗ＲｈｉｎｏｔｍｅｔｈｉｓｐｕｌｃｈｒｉｓＸｉｅｔＺｈｅｎｇ、景泰突颜蝗ＥｏｔｍｅｔｈｉｓｊｉｎｔａｉｅｎｓｉｓＸｉｅｔＺｈｅｎｇ、友谊华癞蝗ＳｉｎｏｔｍｅｔｈｉｓａｍｉｃｕｓＢ．－Ｂｉｅｎｋｏｔ和短翅华癞蝗ＳｉｎｏｔｍｅｔｈｉｓｂｒａｃｈｙｐｔｅｒｕｓＺｈｅｎｇｅｔＸｉ精子的比较研究,发现它们的精子的线粒体衍生体超微形态结构及精子发生过程中第７、８两期精细胞线粒体衍生体周围微管排数十分相似,而在精子的外形长度、中心粒侧体超微形态结构及线粒体衍生体的大小等项特征方面存在着一定差异,这些差异只是在种间精子的鉴别方面有一定的意义,而不具有属级水平的鉴别意义。     

两种网翅蝗精子的比较研究：直翅目：蝗总科

通过对青脊竹蝗Ｃｅｒａｃｒｉｓ ｎｉｇｇｒｉｃｏｒｎｉｓ ｎｉｇｒｉｃｏｒｎｉｓ Ｗａｌｋ及宽翅曲背蚯 Ｐａｒａｒｃｙｐｔｅｒａ ｍｉｃｒｏｐｔｅｒａ ｍｅｒｉｄｉｏｎａｌｉｓ 精子的比较。发它们的精子的线粒体衍生体和中心粒 本的超微形态结构以及精子发生过程中七,八两期精 细胞线粒体衍生体周围微的排数十分相似。     

斑腿蝗科三个种类精子的比较研究：直翅目：蝗总科

本文通过对日本黄脊蝗Ｐａｔａｎｇａｊａｐｏｉｎｃａ（Ｉ．Ｂｄ）,短角直斑腿蝗ＳｔｅｎｏｃａｔａｎｔｏｐｓｍｉｓｔｓｈｅｎｋｏｉＷｉｌｌ和霍山蹦蝗ＳｉｎｏｐｏｄｉｓｍａｈｏｕｓｈａｎａＨｕａｎｇ精子的比较,发现了它们的精子的线粒体衍生体和中心粒侧体的超微形态结构十分相似,在精子发生过程中７、８两期精细胞线粒体衍生体周围微管的排数相一致,这３个种精子间的差异主要反映在外形头和尾的长度,线粒体衍生体的大     

Sperm morphology in five species of cicadettine cicadas (Hemiptera: Cicadomorpha: Cicadidae)

Mature spermatozoa from five species of cicadas of the subfamily Cicadettinae (Quintilia wealei, Melampsalta leucoptera, Stagira simplex, Xosopsaltria thunbergi and Monomatapa matoposa) were examined by light and electron microscopy. In each species sperm are elongate, aggregated into organized bundles with their heads embedded in a homogenous matrix to form spermatodesmata, and exhibit polymegaly. The head of the sperm consist of an anteriorly positioned conical acrosome that has a tubular substructure and a deep, posterior invagination that forms the subacrosomal space (eccentrically positioned anteriorly). The acrosome is flattened anteriorly; posteriorly it extends along either side of the nucleus as two tubular processes that gradually decrease in diameter. The filiform nucleus tapers anteriorly and intrudes into the subscrosomal space. Posteriorly the nucleus has a lateral invagination that houses material of the so-called centriolar adjunct. Posterior to the centriolar adjuct and the nucleus are two crystalline mitochondrial derivatives and a centriole, respectively, the latter giving rise to the axoneme, which has a 9 + 9 + 2 arrangement of microtubules. In these respects the sperm are similar to those of platypleurine cicadas. However, some features seem unique to cicadettines, including the structural organization of an enlarged centriolar adjunct and the dimensions of the tails. The enlarged centriolar adjunct has a lamella-like substructure and can be considered a synapomorphic character in the Cicadettinae. It is, therefore, potentially useful in the separation of this subfamily from the Cicadinae. In addition, the great length of the sperm nucleus of long-headed sperm in M. matoposa could be a synapomorphy of this genus and related taphurine and cicadettine species.     

Shoreward orientation involving geomagnetic cues in the nudibranch mollusc Tritonia diomedea

The marine nudibranch mollusc Tritonia diotnedea orients to the geomagnetic field in the laboratory, and has identifiable brain cells (Pd5, 6) which respond electrically when the ambient magnetic field is rotated artificially. Field studies reported here seek to determine if and why Tritonia diomedea uses geomagnetic cues to orient in the field. Animals were collected in their natural habitat using SCUBA, and placed on lines with magnetic headings parallel to the shore, at different locations with respect to their site of origin. Observations made at two or more tidal cycles later indicate that most animals move from the line in a direction corresponding to the original shoreward direction, regardless of the actual shoreward direction at the site of release, suggesting guidance by geomagnetic cues. Tritonia diomedea are close to neutrally buoyant, and subject to transport over great distances when dislodged by tidal currents or during escape swimming behavior. Since the natural distribution of food and mates is along the shoreline, shoreward orientation using geomagnetic cues, particularly when other cues are weak or ambiguous, may have adaptive value.     

Ultrastructure of the renopericardial complex in Hypselodoris tricolor (Gastropoda, Nudibranchia)

The histology and ultrastructure of the renopericardial complex of Hypselodoris tricolor (Gastropoda, Nudibranchia, Doridoidea) have been investigated by means of semithin serial sections and transmission electron microscopy (TEM). The examinations revealed a functional metanephridial system comprising a monotocardian heart with ventricle and auricle in a spacious pericardium that is linked with the single, large kidney by a renopericardial duct with prominent ciliation towards its opening. Podocytes as the site of ultrafiltration were not only detected in the auricular epicardium, but also line the entire outer pericardial epithelium. The cuboidal, highly vacuolated excretory cells of the kidney epithelium with extensive basal infoldings and an apical microvillous border indicate secretory and reabsorptive activity. Solitary rhogocytes (pore cells) of the connective tissue and haemocoel represent additional loci of ultrafiltration with a fine structure identical to that of the podocytes (slits between cytoplasmic processes, bridged by fine diaphragms and covered by extracellular matrix). The presence of podocytes situated in the epicardial wall of the auricle is regarded as plesiomorphic for the Mollusca and is confirmed for the Nudibranchia. An additional, extensive and separate ultrafiltration site in the outer pericardial wall is not known from any other taxon of the Mollusca and strongly suggests a significantly increased ultrafiltration activity in H. tricolor.     

Polymorphism of spermatozoa in Largus rufipennis Laporte 1832 (Heteroptera: Pyrrhocoroidea: Largidae)

The production of polymorphic spermatozoa has been registered in various insect orders such as Diptera, Lepidoptera, and Hemiptera. In this work, morphology of two types of spermatozoa produced by Largus rufipennis was reported for the first time in the Largidae family. For this, techniques including optical and transmission electron microscopy were used. Spermatozoa measured, on the average, 260 and 200 μm, and both types possessed a nucleus measuring on the average 65 μm. No ultrastructural differences were observed between the two spermatozoa types from L. rufipennis. The head region is composed of an acrosome, a nucleus, and part of the centriolar adjunct. The centriolar adjunct is in parallel with the nucleus and followed by mitochondrial derivates. The flagellum consists of an axoneme (9 + 9 + 2 microtubules) and two mitochondrial derivatives; no other accessory bodies were observed. The mitochondrial derivatives are symmetric in size and diameter. A similar quantity of the two spermatozoa types was observed in the seminal vesicle (57% of the large type and 43% of the small type), while in the spermatheca of the female, the larger spermatozoa were preferentially stored (87%). These results permit discussions concerning of the species biology reproduction, most specifically sperm competition strategies.     

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.     

Sperm morphology in four species of African platypleurine cicadas (Hemiptera: Cicadomorpha: Cicadidae)

Mature spermatozoa from four species of platypleurine cicadas (Albanycada albigera, Azanicada zuluensis, Platypleura capensis and P. hirtipennis) were examined by light and electron microscopy. The filiform sperm have a similar ultrastructure in all species but notable variations were found in sperm dimensions. All species produce more than one discrete length of nucleated, motile sperm, a form of polymorphism termed polymegaly. Polymegaly is expressed in two ways: sperm have bi- or trimodal head and tail lengths. The anterior parts of sperm heads are embedded in an elongate homogenous matrix forming a spermatodesm. The conical acrosome is deeply invaginated posteriorly, and sits on top of the nucleus. The acrosomal contents are differentiated internally with an electron-lucent central medulla and a denser cortex. The homogenously electron-dense nucleus is pointed anteriorly and is generally cylindrical, although posteriorly there is a lateral invagination that extends part-way along the nucleus. This invagination houses fine granular material of the centriolar adjunct. Vesicle-like elements that are associated with both the posterior nucleus and the centriolar adjunct are also found within the invagination. Immediately posterior of and adjoining the centriolar adjunct is a pair of mitochondrial derivatives that are elongated and extend for almost the entire length of the tail. The absence of accessory bodies in cicada sperm suggests that within the Cicadomorpha, the families Cicadidae and Cercopidae are closely affiliated.     

Giant spiral shaped spermatozoa of Diasemopsis comoroensis (Diptera,Diopsidae) with a unique ultrastructural component

In this study we describe a new kind of sperm gigantism in the stalk-eyed fly, Diasemopsis comoroensis (Diptera, Diopsidae). The sperm cells have a length of up to 1.7 mm and can be coiled into a compact ‘slinky’ spiral. Their ultrastructure involves a prominent electron dense central band, which runs the entire length of the sperm tail and in some regions constitutes its largest element in cross section. We propose that this organelle is either a giant centriole adjunct or a kind of accessory body derived from it, and that it takes part in coiling the sperm tail. To our knowledge, no comparable structure has been described before.     

Sperm ultrastructure of five species of the Neotropical ant genus Pseudomyrmex (Hymenoptera: Formicidae)

The seminal vesicles of adult males of five species of Pseudomyrmex were prepared for light and transmission electron microscopy. The Pseudomyrmex spermatozoa are long and slender with similar morphology. The head region has an acrosome and a nucleus. In all the studied species, two morphologically distinct types of acrosomal vesicles were observed, a long structure, as observed in all known ants, and a pear‐shaped one, never before observed in ants. The nucleus is elongated and both condensed and loose chromatin are present. The flagellum has an axoneme, a centriolar adjunct, two mitochondrial derivatives and two accessory bodies. The centriolar, the mitochondrial derivatives and the accessory bodies are similar to observations in most ant species that have been studied. The axoneme presents an uncommon 9 + 9 + 1 microtubule arrangement and the central microtubule has 13 protofilaments. The acrosomal dimorphism and the different levels of chromatin organization are exclusive characteristics of Pseudomyrmex. Furthermore, the 9 + 9 + 1 microtubule arrangement is different from all Hymenoptera, as well as from most insects, which present a 9 + 9 + 2 arrangement. These new morphological characters that are specific to Pseudomyrmex, are valuable synapomorphies of the genus and can be used in taxonomic characterization of the Pseudomyrmecinae subfamily and in phylogenetic analyses in Formicidae family.     

Behavior of centrioles during meiosis in the male silkworm, Bombyx mori (Lepidoptera)

The behavior of centrioles during eupyrene and apyrene meiosis was examined in the silkworm, Bombyx mori , by transmission electron microscopy and indirect immunofluorescence for tubulin. In eupyrene spermatocytes the centrioles, accompanied by axonemes, attached temporarily to the nucleus at diplotene, then detached from the nucleus in diakinesis. After the separation, a beret-shaped structure consisting of a double membrane covered the proximal region of the pair of centrioles. The structure disappeared after breakdown of the nuclear membrane. The centriole, with the axoneme, reattached to the nucleus at telophase I. The process was repeated during meiosis II until the centrioles maintained their nuclear attachment in newly developed spermatids. In stark contrast to their eupyrene counterparts, apyrene spermatocytes were conspicuously devoid of any attachment of the centrioles to the nucleus. These eupyrene-specific and apyrene-specific relationships were consistently and repeatedly found between the nuclear membrane and centrioles, giving rise to suspicion that the behavioral phenomena may be related to differentiation of the dimorphic sperm types.     

Comparative studies on the histology of the ovotestis in Hypselodoris tricolor and Godiva banyulensis (Gastropoda,Opisthobranchia), with special reference to yolk formation

The histology of the ovotestis was studied by light and electron microscopy in two nudibranch gastropod species. While in Hypselodoris tricolor the ovotestis is intimately associated with the digestive gland tissue, the large gonadal mass of Godiva banyulensis is placed freely in the haemocoele. This fact results in great histological differences between both species. As is common among Mollusca, the immature yolk granule in Hypselodoris and Godiva presumably originates from membrane-rich cytoplasmic inclusions, which we have termed dense multivesicular bodies. Such inclusions consist of an outer membrane enclosing membrane remnants and a granular, electron-dense material. These elements are accumulated and mixed in the center of the dense multivesicular body and could be actually transformed into the paracrystalline core of the immature yolk granule, the cortex of which is made up of part of the central accumulation materials that have not spread into the crystal. During vitellogenesis, some mitochondria are subjected to a process of transformation affecting mainly their inner membrane (including mitochondrial cristae) and matrix. However, the conversion of modified mitochondria into yolk precursors, as reported for other gastropod species, could not be determined with absolute certainty on the basis of our observations on static material. The mature yolk granule consists of a central paracrystalline core, similar in structure to that of the immature yolk granule, and a peripheral membranous cortex, which seems to spread centripetally, thus permitting the crystal to grow. The cortical material consumed in synthesizing the central core appears to be restored by addition of degenerative mitochondria to the yolk granule surface.     

Ultrastructure of the biflagellate spermatozoon of tomopteris helgolandica greef, 1879 (annelida,polychaeta)

The spermatozoon of the polychaete Tomopteris helgolandica is of an aberrant type with two flagella, each measuring about 40μm. The nucleus is roughly conical and weakly bent. At the anterior end it is rounded and covered only by the nuclear and plasma membranes. Membraneous, electron-dense structures are applied laterally to the nucleus. These structures may have a helical arrangement. The middle piece contains about ten mitochondria, two centrioles, and two centriolar satellite complexes. The centriolar regions are connected with the posterior part of the nucleus. The axonemes of the two tail flagella lack the usual central complex with central tubules, radial spokes, or related structures. No arms seem to be present on the A tubules of the doublets. In the middle piece the tail flagella are surrounded by invaginations of the plasma membrane forming flagellar canals. The sperm has a bilateral symmetry whereas the primitive sperm has a radial symmetry. The occurrence of two tail flagella in this spermatozoon has no phylogenetical connection with biflagellate spermatozoa in other animal groups. A series of mutations has resulted in the development of two flagella emerging from the two centrioles, the lack of a central complex in the axoneme, and the lack of a typical acrosome. In the Polychaeta, sperm structure is generally more related to function that to phylogenetics. During swimming the spermatozoon of Tomopteris rotates around its longitudinal axis.     

Ultrastructure of the spermatozoid of Lycopodium obscurum (Lycopodiaceae)

Ultrastructural observations reveal that the spermatozoid of Lycopodium obscurum is crescent shaped and contains two posteriorly directed flagella that are inserted at the front of the cell. The nucleus is broad and elongated with a narrow posterior projection or nuclear diverticulum. Spline microtubules (MTs) number 180 at their maximum and provide the framework for the cell. These MTs extend from the anterior of the locomotory apparatus and along the outermost surface of the nucleus, with a central shank of 14–17 MTs encircling the cell for at least one-third gyre beyond the nucleus. The two basal bodies are slightly staggered and positioned at the front of the cell over a highly elongated multilayered structure (MLS). The MLS extends laterally around the cell anterior and curves posteriorly over the nucleus. One large anterior mitochondrion is situated subjacent to the MLS, while numerous small mitochondria are scattered near or among the lobes of the single plastid. The plastid rests on the inner nuclear surface and contains numerous large starch grains. This cell differs from that of L. cernuum, the only other species of Lycopodium examined to date, in that it is more elongated and has an anterior-posterior orientation of the nucleus, basal bodies, MLS, and spline. Comparisons with coiled gametes of bryophytes and Selaginella suggest that some degree of coiling and cell streamlining may be ancestral in archegoniate spermatozoids.     

Sperm ultrastructure in the nudibranch genus Halgerda with reference to other Discodorididae and to Chromodorididae (Mollusca: Opisthobranchia)

Comparative sperm ultrastructure within the molluscan nudibranch genus Halgerda (Discodorididae) was examined for the first time using transmission electron microscopy (TEM), based on 17 of the 35 known species. In addition, observations on two other discodorids are made to facilitate outgroup comparison with Halgerda, including one species of Discodoris (D. boholiensis) and Asteronotus cespitosus (currently accepted as the closest sister taxon to Halgerda). Comparison was also made with some genera of the Chromodorididae in view of sperm similarities. Spermatozoa of all species examined were of the complex, helical, elongate ( approximately 300-400 micro m) type characteristic of most heterobranch gastropods. These cells exhibit the following discrete regions (in anteroposterior sequence) : an acrosomal complex (composed of a rounded, membrane-bound vesicle and a column-like pedestal); a solid, helical nucleus; an elongate, helical midpiece (composed of an axoneme and associated nine coarse fibers, an enveloping mitochondrial derivative of matrix, and paracrystalline materials and glycogen helix); an annular complex; and a short glycogen piece. Of these regions, the midpiece is by far the longest, occupying over 90% of the total sperm length. Comparison with other members of the radula-bearing cryptobranch dorids reveals several sperm similarities to other genera in the clade, particularly those of other Discodorididae and also with the Chromodorididae. Comparison with previously studied genera reveals noteworthy sperm differences within the Discodorididae. The most notable differences are the internal structure of the acrosomal pedestal (long and homogeneous in Halgerda, Discodoris; short and homogeneous in Asteronotus; long and finely striated in Rostanga; oblong with angular electron-lucent striations in Jorunna) and the internal structure of the glycogen piece. The pronounced helical keels of most Halgerda and Discodoris nuclei contrast with the weakly helical nucleus of Asteronotus. Sperm features alone do not provide a means of defining the genus Halgerda or the family Discodorididae nor do they support the monophyletic status of the caryophyllidia-bearing dorids. Important sperm characters such as the acrosome, nucleus, and midpiece can often still be determined from specimens that have been initially fixed in formalin, then stored in ethanol for extended periods of time (i.e., museum material). Of all sperm features, the mitochondrial derivative of the midpiece is the most resistant to long-term fixation : the survival of acrosomal, nuclear, and axonemal components is variable, presumably a factor of prefixation autolysis, varied primary fixation times and temperatures, formalin quality, and duration of alcohol storage.     

The spatial and temporal expression of Tekt1, a mouse tektin C homologue, during spermatogenesis suggest that it is involved in the development of the sperm tail basal body and axoneme

Tektins comprise a family of filament-forming proteins that are known to be coassembled with tubulins to form ciliary and flagellar microtubules. Recently we described the sequence of the first mammalian tektin protein, Tekt1 (from mouse testis), which is most homologous with sea urchin tektin C. We have now investigated the temporal and spatial expression of Tekt1 during mouse male germ cell development. By in situ hybridization analysis TEKT1 RNA expression is detected in spermatocytes and in round spermatids in the mouse testis. Immunofluorescence microscopy analysis with anti-Tekt1 antibodies showed no distinct labeling of any subcellular structure in spermatocytes, whereas in round spermatids anti-Tekt1 antibodies co-localize with anti-ANA antibodies to the centrosome. At a later stage, elongating spermatids display a larger area of anti-Tektl staining at their caudal ends; as spermiogenesis proceeds, the anti-Tekt1 staining disappears. Together with other evidence, these results provide the first intraspecies evidence that Tekt1 is transiently associated with the centrosome, and indicates that Tekt1 is one of several tektins to participate in the nucleation of the flagellar axoneme of mature spermatozoa, perhaps being required to assemble the basal body.     

Phylogeographic differentiation in the mitochondrial control region in the koala, Phascolarctos cinereus (Goldfuss 1817)

The koala, Phascolarctos cinereus, is a geographically widespread species endemic to Australia, with three currently recognized subspecies: P.c. adustus, P.c. cinereus, and P.c. victor. Intraspecific variation in the mitochondrial DNA (mtDNA) control region was examined in over 200 animals from 16 representative populations throughout the species’ range. Eighteen different haplotypes were defined in the ≈ 860 bp mtDNA control region, as determined by heteroduplex analysis/temperature gradient gel electrophoresis (HDA/TGGE). Any single population typically possessed only one or two haplotypes yielding an average within-population haplotypic diversity of 0.180 ± 0.003, and nucleotide diversity of 0.16%. Overall, mtDNA control region sequence diversity between populations averaged 0.67%, and ranged from 0% to 1.56%. Nucleotide divergence between populations averaged 0.51%, and ranged from 0% to 1.53%. Neighbour-joining methods revealed limited phylogenetic distinction between geographically distant populations of koalas, and tentative support for a single evolutionarily significant unit (ESU). This is consistent with previous suggestions that the morphological differences formalized by subspecific taxonomy may be interpreted as clinal variation. Significant differentiation in mtDNA-haplotype frequencies between localities suggested that little gene flow currently exists among populations. When combined with microsatellite analysis, which has revealed substantial differentiation among koala populations, we conclude that the appropriate short-term management unit (MU) for koalas is the local population.