Developmental patterns of the hermaphroditic gonad in dicyemid mesozoans (Phylum Dicyemida)

Abstract. The development of the hermaphroditic gonad, the infusorigen, was studied in fixed specimens of 16 species belonging to four genera ( Conocyema, Dicyema, Dicyemennea , and Pseudicyema ) of dicyemid mesozoans. In addition, the literature on the cell lineages of infusorigens of eight other species was reviewed. After an agamete (axoblast) undergoes an unequal first division, excluding a small cell (the paranucleus), the resulting large cell divides to form the infusorigen. Five infursorigen cell lineage patterns can be identified: type I, both the first oogonium and the first spermatogonium are produced at the third division; type II, the first spermatogonium is produced at the second division and the first oogonium is produced at the third division; type III, the first spermatogonium is produced at the second division and the first oogonium is produced at the fourth division; type IV, the first oogonium is produced at the second division and the first spermatogonium is produced at the third division; and type V, the first oogonium is produced at the second division and the first spermatogonium is produced at the fourth division. Later development is similar in members of all genera. Mature infusorigens are similarly organized in all species examined. Sizes of agametes and infusorigen axial cells, the number of infusorigens, and the type of reproductive strategy were not correlated with infusorigen developmental patterns or cell lineage patterns.     

Developmental patterns and cell lineages of vermiform embryos in dicyemid mesozoans.

Patterns of cell division and cell lineages of the vermiform embryos of dicyemid mesozoans were studied in four species belonging to four genera: Conocyema polymorpha, Dicyema apalachiensis, Microcyema vespa, and Pseudicyema nakaoi. During early development, the following common features were apparent: (1) the first cell division produces prospective cells that generate the anterior peripheral region of the embryo; (2) the second cell division produces prospective cells that generate the posterior peripheral region plus the internal cells of the embryo; (3) in the lineage of prospective internal cells, several divisions ultimately result in cell death of one of the daughter cells. Early developmental processes are almost identical in the vermiform embryos of all four dicyemid genera. The cell lineages appear to be invariant among embryos and are highly conserved among species. Species-specific differences appear during later stages of embryogenesis. The number of terminal divisions determines variations in peripheral cell numbers among genera and species. Thus, the numbers of peripheral cells are fixed and hence species-specific.     

Fine structure of dicyemid mesozoans,with special reference to cell junctions

The fine structure of the dicyemid mesozoan, Dicyema acuticephalum, from Octopus vulgaris, was studied with special attention to intercellular junctional complexes between various kinds of cells. Two types of intercellular junction, namely, adherens junctions and gap junctions, were found in both vermiform stages and in infusoriform embryos. Adherens junctions were classified into two types. Zonulae adherentes-like junctions were observed between adjacent peripheral cells at vermiform stages, between adjacent external cells of infusoriform embryos, and between members of groups of internal cells that covered the urn in infusoriform embryos. Maculae adherentes-like junctions were seen between a peripheral cell and an axial cell at vermiform stages. In infusoriform embryos, these junctions were observed between various types of cells, excluding urn cells. Gap junctions were found between adjacent peripheral cells at vermiform stages, whereas in infusoriform embryos these junctions were located between various types of cells excluding urn cells. Dicyemids might be the most primitive multicellular animals to possess these basic types of cell junctions. Ciliary rootlet systems at vermiform stages and in infusoriform embryos were unique in structure compared with those of other primitive multicellular animals. J Morphol 231:297–305, 1997. © 1997 Wiley-Liss, Inc.     

东北地区白桦雌配子体的形成与胚胎发育研究

以20年生白桦(Betula platyphylla)的雌花序为材料,研究了白桦的大孢子发生、雌配子体形成和胚胎发育过程.结果表明:1)大孢子母细胞经过减数分裂形成直线排列的4个大孢子,其中珠孔端的3个大孢子退化,合点端的大孢子进行3次有丝分裂,形成八核胚囊,8个核迅速细胞化,最终发育为成熟胚囊.胚囊发育类型为单孢子蓼型胚囊;2)胚的发育经过原胚、球形胚、心形胚和鱼雷胚等阶段发育为成熟胚;胚柄在球形胚时期最发达,但胚柄短小,只由3个扁平细胞组成,之后逐渐退化.本文同时对雌花的结构和发育特点在桦木科各属中的异同、白桦胚胎发生类型及胚胎发育进程的同步性进行了讨论.     

东北地区白桦雌配子体的形成与胚胎发育研究

以20年生白桦(Betula platyphylla)的雌花序为材料, 研究了白桦的大孢子发生、雌配子体形成和胚胎发育过程。结果表明：1)大孢子母细胞经过减数分裂形成直线排列的4个大孢子, 其中珠孔端的3个大孢子退化, 合点端的大孢子进行3次有丝分裂, 形成八核胚囊, 8个核迅速细胞化, 最终发育为成熟胚囊。胚囊发育类型为单孢子蓼型胚囊; 2)胚的发育经过原胚、球形胚、心形胚和鱼雷胚等阶段发育为成熟胚; 胚柄在球形胚时期最发达, 但胚柄短小, 只由3个扁平细胞组成, 之后逐渐退化。本文同时对雌花的结构和发育特点在桦木科各属中的异同、白桦胚胎发生类型及胚胎发育进程的同步性进行了讨论。     

Biology of dicyemid mesozoans

We reviewed recent advances of some aspects on the biology of dicyemid mesozoans. To date 42 species of dicyemids have been found in 19 species of cephalopod molluscs from Japanese waters. The body of dicyemids consists of 10-40 cells and is organized in a very simple fashion. There are three basic types of cell junction, septate junction, adherens junction, and gap junction. The presence of these junctions suggests not only cell-to-cell attachment, but also cell-to-cell communication. In the development of dicyemids, early stages and cell lineages are identical in vermiform embryos of four genera, Conocyema, Dicyema, Microcyema, and Pseudicyema. Species-specific differences appear during later stages of embryogenesis. In the process of postembryonic growth in some species, the shape of the calotte changes from conical to cap-shaped and discoidal. This calotte morphology appears to result from adaptation to the structure of host renal tissues and help to facilitate niche separation of coexisting species. In most dicyemids distinctly small numbers of sperms are produced in a hermaphroditic gonad (infusorigen). The number of eggs and sperms are roughly equal. An inverse proportional relationship exists between the number of infusorigens and that of gametes, suggesting a trade-off between them. Recent phylogenetic studies suggest dicyemids are a member of the Lophotrochozoa.     

The cleavage program in the 2d cell lineage of Tubifex embryos

Early development in clitellate annelids is characterized by a highly stereotyped sequence of unequal, spiral cleavages. Cell 2d (i.e., the second micromere of the D quadrant) in the oligochaete Tubifex tubifex also undergoes an evolutionarily conserved sequence of cell division to produce four bilateral pairs of ectodermal teloblasts that act as embryonic stem cells. This study was conducted to characterize each of the 15 rounds of cell division that occur in the 2d cell lineage in this clitellate. After its occurrence, cell 2d undergoes three rounds of highly unequal divisions, giving off the first smaller daughter cell toward the posterior right of the larger daughter cell, the second cell toward the posterior left, and the third cell toward the anterior side of the cell; the larger daughter cell that results from the third division (i.e., the great-granddaughter cell of 2d) then divides equally into a bilateral pair of NOPQ proteloblasts. Cell NOPQ on either side of the embryo undergoes 11 rounds of cell division, during which ectoteloblasts N, Q, and O/P are produced in this order. After its appearance, NOPQ undergoes highly unequal divisions twice cutting off the smaller cells toward the anterior end of the embryo and then divides almost equally into ectoteloblast N and proteloblast OPQ. After its appearance, OPQ undergoes highly unequal divisions twice giving off the first smaller cell toward the anterior and the second smaller cell toward the posterior of the embryo and then divides almost equally into ectoteloblast Q and proteloblast OP. Finally, OP undergoes highly unequal division four times after its birth budding off the smaller cells toward the anterior and then cleaves equally into ectoteloblasts O and P. In the unequally dividing cells of the 2d cell lineage, the mitotic apparatus (MA), which forms at the cell's center, moves eccentrically toward the cortical site where the smaller cell will be given off. The moving MA is oriented perpendicular to the surface it approaches, and its peripheral pole becomes closely associated with the cell cortex. In contrast, the MA involved in the equal divisions remains in the cell center throughout mitosis. The key features of the cleavage program in the 2d cell lineage are discussed in light of the present observations. The mechanical aspects of unequal cleavage in the 2d cell lineage and the modes of specification of MA orientation are discussed. A comparison of the cleavage mode in the 2d cell lineage is also performed among six selected clitellate annelid species.     

A new species of Dicyemennea Whitman, 1883 (Phylum Dicyemida) from Sepia latimanus (Mollusca: Cephalopoda: Decapodidae) off Okinawa, Japan

A new species of dicyemid mesozoan is described from a cuttlefish Sepia latimanus Quoy and Gaimard collected off Nago, Naha and Onnason, Okinawa Islands, Ryukyu Islands, Japan. Dicyemennea ryukyuense n. sp. is a large species that reaches about 5 mm in length. The vermiform stage is characterised by 23 peripheral cells, a conical calotte and an axial cell that extends to the base of the propolar cells. The infusoriform embryo consists of 37 cells; two nuclei are present in each urn cell and the refringent bodies are solid. In the type of reproductive strategy, rhombogens of D. ryukyuense form a small number of infusorigens and produce a relatively large number of gametes per infusorigen. In the family Dicyemidae, Dicyemennea Whitman, 1883 is the largest group after Dicyema von Kölliker, 1849. Other dicyemid species, including those belonging to Dicyema, were not detected. Dicyemids have never previously been detected in cephalopods living on corals and rocks off the Ryukyu Islands, even though they are benthonic in habitat. This is the first report of a dicyemid mesozoan from S. latimanus and also from off the Ryukyu Islands. S. latimanus inhabits coral reefs, often swimming over the coral but usually lying on the sandy bottom. Dicyemids cannot possibly infect cephalopods which live exclusively over coral and rock.     

Molecular markers comparing the extremely simple body plan of dicyemids to that of lophotrochozoans: insight from the expression patterns of Hox, Otx, and brachyury

SUMMARY Because of their extremely simple body plan, dicyemids have long been the subject of phylogenetic controversy, regarding whether their body plan reflects their primitiveness or a degeneration from complex metazoans. Several studies have argued that the simple body plan of dicyemids are likely secondarily derived from higher lophotrochozoan animals, as a result of their endoparasitic, or endosymbiotic, lifestyle in the cephalopod kidney. To clarify the evolution of their simple body plan, we investigated the developmental expression patterns of three important regulatory genes, the central type Hox gene ( DoxC ), otx , and brachyury homologs in the dicyemid mesozoa, Dicyema orientale. DoxC was expressed in the trunk and tail of the asexually developing vermiform embryo, with clear anterior boundaries. Do-otx was expressed in the vegetal pole cells of the developing infusoriform embryos, suggesting that the invagination in infusoriform embryo is homologous to the gastrulation of other metazoans. Do-bra is expressed in the presumptive ventral cells, which are ventral to the opening of the urn cavity. The expression of Do-bra suggests that the urn cavity opening of the infusoriform embryo is comparable to the stomodium of trochophore larvae. These gene expression patterns provide molecular clues to trace the evolutionary history of degeneration in the dicyemid embryogenesis and life cycle from those of ancestral lophotrochozoan animals.     

Regulation of the Dorsal Axial Structures in Cell-Deficient Embryos of Xenopus laevis

To study the regulation of the dorsal axial structures, we removed the right animal dorsal and the right vegetal dorsal cells from an 8-cell embryo of Xenopus laevis .
Most of the right dorsal cell-deficient embryos developed to normally proportioned tailbud embryos. No detectable delay was observed in their development. Examinations of serial sections revealed that they had restored bilateral symmetry. The cell numbers of the somite and the notochord had recovered to more than 90% and 70%, respectively, those of controls. Since the right dorsal cell-deficient embryo retained roughly three-quarters of the prospective region for the somites and half of that for the notochord, respectively, the cell number was more than that expected from the remaining prospective regions. Cell lineage analyses showed that progeny of the right ventral cells had formed almost all of the right dorsal axial structures, which are normally formed by the progeny of the right dorsal cells. However, almost all the notochord cells had been derived from the remaining left dorsal cells.
These results indicate that some quantitative aspects of regulation as expressed in terms of the cell number were different between the two tissues examined.     

IAA, ABA, polyamines and free amino acids associated with zygotic embryo development of Ocotea catharinensis

The aim of this work was to study morphological and biochemical aspects during zygotic embryogenesis in O. catharinensis, by measuring changes in the endogenous concentrations of proteins, amino acids, polyamines (PAs), indole-3-acetic acid (IAA) and abscisic acid (ABA). Buffer-soluble and insoluble protein contents were determined by spectrometry, and amino acids, PAs, IAA and ABA concentrations were determined by high performance liquid chromatography. Total amino acid accumulation, predominantly asparagine, occurred when the embryo showed completely developed cotyledons, with posterior reduction in the mature embryo. This decrease in total amino acid concentration in the mature embryo may result from their use in storage␣as well as for LEA protein synthesis. Free putrescine (Put) concentration decreased, while free spermine (Spm) increased during embryo development. This suggest a role for Put in the initial phases of embryogenesis when high rates of cell division occur, while elevated concentration of Spm are essential from the middle to the end of embryo development, when growth is mainly due to cell elongation. An IAA peak in zygotic embryos occurred during initial development, suggesting a link between growth and cellular division as well as with the establishment of bilateral symmetry. ABA concentration declined during initial stages of development then increased at the mature embryo stage, suggesting a possible relationship with dormancy and recalcitrance characteristics. Our results show that changes in the phytohormones (IAA, ABA and PAs) concentrations in combination with amino acids are likely important factors determining the developmental stages of O.␣catharinensis zygotic embryos.     

虎眼万年青的直接体细胞胚胎发生和植株再生

以虎眼万年青(Ornithogalum caudatum Jacq.)子鳞茎的鳞叶为外植体,不经过愈伤组织阶段,直接诱导体细胞胚胎发生并再生植株。组织切片和扫描电镜的观察结果表明:体细胞胚来自鳞叶表皮最外层的单个细胞。这个细胞经第一次平周分裂,成为2-细胞原胚,然后依次经过4-细胞原胚、球形胚、香蕉形胚等阶段发育成小植株,其形态学发育过程与合子胚相似。小植株移入土中,1周后即可成活。此外,还观察到直接体细胞胚只产生于鳞叶的近轴面,而远轴面无体细胞胚产生,这两个部位在细胞结构和形态上有显著差异。     

Redescription of Dicyemennea nouveli (Phylum: Dicyemida) from Enteroctopus dofleini (Mollusca: Cephalopoda: Octopoda)

A species of dicyemid mesozoan is redescribed from the giant Pacific octopus, Enteroctopus dofleini (Wülker, 1910), collected off Iwase in Toyama Bay, Honshu, Japan. Dicyemennea nouveli McConnaughey, 1959, is a large species that reaches about 12,000 microm in length. This species lives in folds of the renal appendages. The vermiform stages are characterized as having 30-41 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the metapolar cells. An anterior abortive axial cell is present in vermiform embryos. Full-grown vermiform embryos have as many as 4 agametes. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid.     

Confocal Microscopy Study of <Emphasis Type="Italic">Arabidopsis</Emphasis> Embryogenesis Using GFP:mTn

Embryogenesis in transgenic Arabidopsis plants with GFP:mTn, a chimeric fusion of soluble shifted green fluorescent protein and a mouse actin binding domain, was studied. Confocal laser scanning microscopy was used to determine patterns of formation and cellular responses during asymmetric cell division. Before such cells divide, the nucleus moves to the position where new cell walls are to be formed. The apical–basal axis of the embryo develops mainly at the zygote to octant stage, and these events are associated with asymmetric divisions of the zygote and hypophyseal cells. Formation of the radial axis is established from the dermatogen to the globular-stage embryo via tangential cell division within the upper tiers. Bilateral symmetry of the embryo primarily happens at the triangular stage through zig-zag cell divisions of initials of the cotyledonary primordia. All stages of embryogenesis are described in detail here.     

Regional specification during embryogenesis in the inarticulate brachiopod Discinisca.

The process of embryogenesis is described for the inarticulate brachiopod Discinisca strigata of the family Discinidae. A fate map has been constructed for the early embryo. The animal half of the egg forms the dorsal ectoderm of the apical and mantle lobes. The vegetal half forms mesoderm and endoderm and is the site of gastrulation; it also forms the ectoderm of the ventral regions of the apical and mantle lobes of the larva. The plane of the first cleavage goes through the animal-vegetal axis of the egg along the future plane of bilateral symmetry of the larva. The timing of regional specification in these embryos was examined by isolating animal, vegetal, or lateral regions at different times from the 2-cell stage through gastrulation. Animal halves isolated at the 8-cell and blastula stages formed an epithelial vesicle and did not gastrulate. When these halves were isolated from blastulae they formed the cell types typical of apical and mantle lobes. Vegetal halves isolated at all stages gastrulated and formed a more or less normal larva; the only defect these larvae had was the lack of an apical tuft, which normally forms from cells at the animal pole of the embryo. When lateral isolates were created at all developmental stages, these halves gastrulated. Cuts which separated presumptive anterior and posterior regions generated isolates at the 4-cell and blastula stages that formed essentially normal larvae; however, at the midgastrula stage these halves formed primarily anterior or posterior structures indicating that regional specification had taken place along the anterior-posterior axis. The plane of the first cleavage, which predicts the plane of bilateral symmetry, can be shifted by either changing the cleavage pattern that generates the bilateral 16-cell blastomere configuration or by isolating embryo halves prior to, or during, the 16-cell stage. These results indicate that while the plane of the first cleavage predicts the axis of bilateral symmetry, the axis is not established until the fourth cleavage. The development of Discinisca is compared to development in the inarticulate brachiopod Glottidia of the family Lingulidae and to Phoronis in the phylum Phoronida.     

Developmental control of cell division in leech embryos

During embryogenesis, cell division must be spatially and temporally regulated with respect to other developmental processes. Leech embryos undergo a series of unequal and asynchronous cleavages to produce individually recognizable cells whose lineages, developmental fates and cell cycle properties have been characterized. Thus, leech embryos provide an opportunity to examine the regulation of cell division at the level of individual well-characterized cells within a community of different types of cells. Isolation of leech homologues of some of the highly conserved regulators of the cell division cycle, and characterization of their patterns of maternal and zygotic expression, indicate that the cell divisions of early leech embryos are regulated by cell type-specific mechanisms. These studies with leech embryos contribute to the emerging appreciation of the diverse mechanisms by which animals regulate cell division during early development.     

生长素在植物胚胎早期发育中的作用

有性生殖是有花植物的一个重要特征, 胚胎则是实现有性生殖和世代交替的重要载体。植物胚胎从双受精开始, 经历了合子极性建立、顶基轴形成、细胞层分化和器官形成等过程, 这些过程都受到生长素的调控。近年来的研究表明, 生长素在生物合成、极性运输和信号转导3个层面上调控胚胎的发育过程。该文以双子叶植物拟南芥(Arabidopsis thaliana)为例, 综述了生长素对胚胎早期发育过程, 包括合子极性和顶基轴建立、表皮原特化和对称模式转变、胚根原特化和根尖分生组织形成及茎尖分生组织形成等发育的调控机制。     

Indeterminate cell lineage of the zebrafish embryo

We have examined the clonal progeny descended from individual blastomeres injected with lineage-tracer dye in the zebrafish embryo. Blastomeres arising by the same cleavages in different embryos generated clones in which the types and positions of cells were highly variable. Several features of early development were correlated with this diversity in cell fate. There was no fixed relationship between the plane of the first cleavage and the eventual plane of bilateral symmetry of the embryo. By blastula stages the cleavages of identified blastomeres were variable in pattern. Moreover, cell fate was not easily related to the longitudinal and dorsoventral position of the clone in the gastrula. These results establish that single blastomeres can potentially generate a highly diverse array of cell types and that the cell lineage is indeterminate.     

Distinct parameters are involved in controlling the number of rounds of cell division in each tissue during ascidian embryogenesis.

We counted cell numbers during embryogenesis of the ascidian, Halocynthia roretzi, every hour. Cell numbers were determined by counting the numbers of nuclei in squashed embryos. The cell number of a larva just after hatching was approximately 3000. Our study addresses the question of what factors control the number of rounds of cell division during development. Three kinds of egg fragments were prepared by cutting unfertilized eggs to alter the volume of cytoplasm and the amount of DNA. After the egg fragments were fertilized, the cell numbers were estimated at the hatching stage. The cell numbers of the resulting larvae differed from those of normal larvae. Precursor blastomeres of various tissues were then isolated from normal and manipulated embryos, and cultured as partial embryos. The cell numbers of the resulting partial embryos were counted to estimate the number of cell divisions in each larval tissue. The results suggested that the number of cell divisions is controlled by a distinct mechanism in each tissue. We propose that the number of rounds of cell division during ascidian embryogenesis is controlled by three mechanisms: the first depending on the volume of cytoplasm; the second on the nucleo-cytoplasmic ratio; and the third depending on neither of these parameters. J. Exp. Zool. 284:379-391, 1999.     

Cleavage pattern,gastrulation, and neurulation in the appendicularian, <Emphasis Type="Italic">Oikopleura dioica</Emphasis>

The appendicularian, Oikopleura dioica is a chordate. Its life cycle is extremely short—approximately 5 days—and its tadpole shape with a beating tail is retained throughout entire life. The tadpole hatches after 3 h of development at 20°C. Here, we describe the cleavage pattern and morphogenetic cell movements during gastrulation and neurulation. Cleavage showed an invariant pattern. It is basically bilateral but also shows various minor left–right asymmetries starting from the four-cell stage. We observed two rounds of unequal cleavage of the posterior-vegetal B-line cells at the posterior pole. The nature of the unequal cleavages is reminiscent of those in ascidian embryos and suggests the presence of a centrosome-attracting body, a special subcellular structure at the posterior pole. The representation of the cell division pattern in this report will aid the identification of each cell, a prerequisite for clarifying the gene expression patterns in early embryos. Gastrulation started as early as the 32-cell stage and progressed in three phases. By the end of the second phase at the 64-cell stage, every vegetal cell had ingressed into the embryo, and animal cells had covered the entire embryo by epiboly. There was no archenteron formation. In the anterior region, eight A-line cells were aligned as a 2 × 4 array along the anterior–posterior axis and become internalized during the 64-cell stage. This process was considered to correspond to neurulation. The simple and accelerated development of Oikopleura, nevertheless giving rise to a conserved chordate body plan, is advantageous for studying developmental mechanisms using molecular and genetic approaches and makes this animal the simplest model organism in the phylum Chordata. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.